USP32 Phan Tich Thuoc PDF

USP 32 MONOGRAPH LIST / 1
Acarbose Acetaminophen and Pseudoephedrine Hydrochloride Tablets

Acebutolol Hydrochloride Acetazolamide
Acebutolol Hydrochloride Capsules Acetazolamide for Injection
Acepromazine Maleate Acetazolamide Oral Suspension
Acepromazine Maleate Injection Acetazolamide Tablets
Acepromazine Maleate Tablets Glacial Acetic Acid
Acetaminophen Acetic Acid Irrigation
Acetaminophen Capsules Acetic Acid Otic Solution
Acetaminophen Oral Solution Acetohexamide
Acetaminophen for Effervescent Oral Solution Acetohexamide Tablets
Acetaminophen Suppositories Acetohydroxamic Acid
Acetaminophen Oral Suspension Acetohydroxamic Acid Tablets
Acetaminophen Tablets Acetylcholine Chloride
Acetaminophen Extended-Release Tablets Acetylcholine Chloride for Ophthalmic Solution
Acetaminophen and Aspirin Tablets Acetylcysteine
Acetaminophen, Aspirin, and Caffeine Tablets Acetylcysteine Solution
Acetaminophen and Caffeine Tablets Acetylcysteine and Isoproterenol Hydrochloride Inhalation
Solution
Capsules Containing at Least Three of the FollowingAcet-
aminophen and Salts of Chlorpheniramine, Dex- Acitretin
tromethorphan, and Phenylpropanolamine
Acitretin Capsules
Oral Solution Containing at Least Three of the Following
Acetaminophen and Salts of Chlorpheniramine, Dex- Acyclovir
tromethorphan, and Phenylpropanolamine Acyclovir Capsules
Tablets Containing at Least Three of the FollowingAcet- Acyclovir for Injection
aminophen and Salts of Chlorpheniramine, Dex-
tromethorphan, and Phenylpropanolamine Acyclovir Ointment
Capsules Containing at Least Three of the FollowingAcet- Acyclovir Oral Suspension
aminophen and Salts of Chlorpheniramine, Dex-
tromethorphan, and Pseudoephedrine Acyclovir Tablets
Oral Powder Containing at Least Three of the Following Adenine

Acetaminophen and Salts of Chlorpheniramine, Dex- Adenosine
tromethorphan, and Pseudoephedrine
Adenosine Injection
Oral Solution Containing at Least Three of the Following
Acetaminophen and Salts of Chlorpheniramine, Dex- Medical Air
tromethorphan, and Pseudoephedrine
Alanine
Tablets Containing at Least Three of the FollowingAcet-
aminophen and Salts of Chlorpheniramine, Dex- Albendazole
tromethorphan, and Pseudoephedrine Albendazole Oral Suspension
Acetaminophen, Chlorpheniramine Maleate, and Dex- Albendazole Tablets
tromethorphan Hydrobromide Tablets
Albumin Human
Acetaminophen and Codeine Phosphate Capsules
Albuterol
Acetaminophen and Codeine Phosphate Oral Solution
Albuterol Sulfate
Acetaminophen and Codeine Phosphate Oral Suspension
Albuterol Tablets
Acetaminophen and Codeine Phosphate Tablets
Alclometasone Dipropionate
Acetaminophen, Dextromethorphan Hydrobromide, Doxyl-
amine Succinate, and Pseudoephedrine Hydrochloride Oral Alclometasone Dipropionate Cream
Solution
Alclometasone Dipropionate Ointment
Acetaminophen and Diphenhydramine Citrate Tablets
Alcohol
Acetaminophen, Diphenhydramine Hydrochloride, and
Pseudoephedrine Hydrochloride Tablets Dehydrated Alcohol
Copyright 2008 The United States Pharmacopeial Convention. All Rights Reserved.
For Discussion Purposes Only Not for Dissemination
2 / MONOGRAPH LIST USP 32
Dehydrated Alcohol Injection Aluminum Chloride

Rubbing Alcohol Aluminum Chlorohydrate
Alcohol in Dextrose Injection Aluminum Chlorohydrate Solution
Alendronate Sodium Aluminum Chlorohydrex Polyethylene Glycol
Alendronate Sodium Tablets Aluminum Chlorohydrex Propylene Glycol
Alfentanil Hydrochloride Aluminum Dichlorohydrate
Alfentanil Injection Aluminum Dichlorohydrate Solution
Alfuzosin Hydrochloride Aluminum Dichlorohydrex Polyethylene Glycol
Allantoin Aluminum Dichlorohydrex Propylene Glycol
Allopurinol Aluminum Hydroxide Gel
Allopurinol Oral Suspension Dried Aluminum Hydroxide Gel
Allopurinol Tablets Dried Aluminum Hydroxide Gel Capsules
Allyl Isothiocyanate Dried Aluminum Hydroxide Gel Tablets
Aloe Aluminum Phosphate Gel
Alprazolam Aluminum Sesquichlorohydrate
Alprazolam Oral Suspension Aluminum Sesquichlorohydrate Solution
Alprazolam Tablets Aluminum Sesquichlorohydrex Polyethylene Glycol
Alprostadil Aluminum Sesquichlorohydrex Propylene Glycol
Alprostadil Injection Aluminum Subacetate Topical Solution
Alteplase Aluminum Sulfate
Alteplase for Injection Aluminum Sulfate and Calcium Acetate for Topical Solution
Altretamine Aluminum Sulfate and Calcium Acetate Tablets for Topical
Solution
Altretamine Capsules
Aluminum Zirconium Octachlorohydrate
Ammonium Alum
Aluminum Zirconium Octachlorohydrate Solution
Potassium Alum
Aluminum Zirconium Octachlorohydrex Gly
Alumina and Magnesia Oral Suspension
Aluminum Zirconium Octachlorohydrex Gly Solution
Alumina and Magnesia Tablets
Aluminum Zirconium Pentachlorohydrate
Alumina, Magnesia, and Calcium Carbonate Oral Suspension
Aluminum Zirconium Pentachlorohydrate Solution
Alumina, Magnesia, and Calcium Carbonate Tablets
Aluminum Zirconium Pentachlorohydrex Gly
Alumina, Magnesia, and Calcium Carbonate Chewable
Tablets Aluminum Zirconium Pentachlorohydrex Gly Solution
Alumina, Magnesia, Calcium Carbonate, and Simethicone Aluminum Zirconium Tetrachlorohydrate
Tablets
Aluminum Zirconium Tetrachlorohydrate Solution
Alumina, Magnesia, Calcium Carbonate, and Simethicone
Chewable Tablets Aluminum Zirconium Tetrachlorohydrex Gly
Alumina, Magnesia, and Simethicone Oral Suspension Aluminum Zirconium Tetrachlorohydrex Gly Solution
Alumina, Magnesia, and Simethicone Tablets Aluminum Zirconium Trichlorohydrate
Alumina, Magnesia, and Simethicone Chewable Tablets Aluminum Zirconium Trichlorohydrate Solution
Alumina and Magnesium Carbonate Oral Suspension Aluminum Zirconium Trichlorohydrex Gly
Alumina and Magnesium Carbonate Tablets Aluminum Zirconium Trichlorohydrex Gly Solution
Alumina, Magnesium Carbonate, and Magnesium Oxide Amantadine Hydrochloride

Tablets Amantadine Hydrochloride Capsules
Alumina and Magnesium Trisilicate Oral Suspension Amantadine Hydrochloride Oral Solution
Alumina and Magnesium Trisilicate Tablets Amcinonide
Aluminum Acetate Topical Solution
Amcinonide Cream Amitriptyline Hydrochloride

Amcinonide Ointment Amitriptyline Hydrochloride Injection
Amifostine Amitriptyline Hydrochloride Tablets
Amifostine for Injection Amlodipine Besylate
Amikacin Aromatic Ammonia Spirit
Amikacin Sulfate Ammonium Chloride
Amikacin Sulfate Injection Ammonium Chloride Injection
Amiloride Hydrochloride Ammonium Chloride Delayed-Release Tablets
Amiloride Hydrochloride Tablets Ferric Ammonium Citrate
Amiloride Hydrochloride and Hydrochlorothiazide Tablets Ferric Ammonium Citrate for Oral Solution
Amiloxate Ammonium Molybdate
Aminobenzoate Potassium Ammonium Molybdate Injection
Aminobenzoate Potassium Capsules Amobarbital Sodium
Aminobenzoate Potassium for Oral Solution Amobarbital Sodium for Injection
Aminobenzoate Potassium Tablets Amodiaquine
Aminobenzoate Sodium Amodiaquine Hydrochloride
Aminobenzoic Acid Amodiaquine Hydrochloride Tablets
Aminobenzoic Acid Gel Amoxapine
Aminobenzoic Acid Topical Solution Amoxapine Tablets
Aminocaproic Acid Amoxicillin
Aminocaproic Acid Injection Amoxicillin Boluses
Aminocaproic Acid Oral Solution Amoxicillin Capsules
Aminocaproic Acid Tablets Amoxicillin Intramammary Infusion
Aminoglutethimide Amoxicillin for Injectable Suspension
Aminoglutethimide Tablets Amoxicillin Oral Suspension
Aminohippurate Sodium Injection Amoxicillin for Oral Suspension
Aminohippuric Acid Amoxicillin Tablets
Aminopentamide Sulfate Amoxicillin Tablets for Oral Suspension
Aminopentamide Sulfate Injection Amoxicillin and Clavulanate Potassium for Oral Suspension
Aminopentamide Sulfate Tablets Amoxicillin and Clavulanate Potassium Tablets
Aminophylline Amphetamine Sulfate
Aminophylline Injection Amphetamine Sulfate Tablets
Aminophylline Oral Solution Amphotericin B
Aminophylline Rectal Solution Amphotericin B Cream
Aminophylline Suppositories Amphotericin B for Injection
Aminophylline Tablets Amphotericin B Lotion
Aminophylline Delayed-Release Tablets Amphotericin B Ointment
Aminosalicylate Sodium Ampicillin
Aminosalicylate Sodium Tablets Ampicillin Boluses
Aminosalicylic Acid Ampicillin Capsules
Aminosalicylic Acid Tablets Ampicillin for Injection
Amitraz Ampicillin Soluble Powder
Amitraz Concentrate for Dip Ampicillin for Injectable Suspension
Ampicillin for Oral Suspension Arginine Hydrochloride

Ampicillin Tablets Arginine Hydrochloride Injection
Ampicillin and Probenecid for Oral Suspension Arsanilic Acid
Ampicillin Sodium Ascorbic Acid
Ampicillin and Sulbactam for Injection Ascorbic Acid Injection
Amprolium Ascorbic Acid Oral Solution
Amprolium Soluble Powder Ascorbic Acid Tablets
Amprolium Oral Solution Aspartic Acid
Amyl Nitrite Aspirin
Amyl Nitrite Inhalant Aspirin Boluses
Anileridine Aspirin Capsules
Anileridine Injection Aspirin Delayed-Release Capsules
Anileridine Hydrochloride Aspirin Suppositories
Anileridine Hydrochloride Tablets Aspirin Tablets
Antazoline Phosphate Buffered Aspirin Tablets
Anthralin Aspirin Delayed-Release Tablets
Anthralin Cream Aspirin Effervescent Tablets for Oral Solution
Anthralin Ointment Aspirin Extended-Release Tablets
Anthrax Vaccine Adsorbed Aspirin, Alumina, and Magnesia Tablets
Anticoagulant Citrate Dextrose Solution Aspirin, Alumina, and Magnesium Oxide Tablets
Anticoagulant Citrate Phosphate Dextrose Solution Aspirin, Caffeine, and Dihydrocodeine Bitartrate Capsules
Anticoagulant Citrate Phosphate Dextrose Adenine Solution Aspirin and Codeine Phosphate Tablets
Anticoagulant Heparin Solution Aspirin, Codeine Phosphate, Alumina, and Magnesia Tablets
Anticoagulant Sodium Citrate Solution Astemizole
Antihemophilic Factor Astemizole Tablets
Cryoprecipitated Antihemophilic Factor Atenolol
Antimony Potassium Tartrate Atenolol Injection
Antimony Sodium Tartrate Atenolol Oral Solution
Antipyrine Atenolol Tablets
Antipyrine and Benzocaine Otic Solution Atenolol and Chlorthalidone Tablets
Antipyrine, Benzocaine, and Phenylephrine Hydrochloride Atovaquone
Otic Solution
Atovaquone Oral Suspension
Antithrombin III Human
Atracurium Besylate
Antivenin (Crotalidae) Polyvalent
Atracurium Besylate Injection
Antivenin (Latrodectus mactans)
Atropine
Antivenin (Micrurus fulvius)
Atropine Sulfate
Apomorphine Hydrochloride
Atropine Sulfate Injection
Apomorphine Hydrochloride Tablets
Atropine Sulfate Ophthalmic Ointment
Apraclonidine Hydrochloride
Atropine Sulfate Ophthalmic Solution
Apraclonidine Ophthalmic Solution
Atropine Sulfate Tablets
Aprotinin
Activated Attapulgite
Aprotinin Injection
Colloidal Activated Attapulgite
Arginine
Aurothioglucose
Aurothioglucose Injectable Suspension Azathioprine Tablets

Avobenzone Azathioprine Sodium for Injection
Azaperone Azithromycin
Azaperone Injection Azithromycin Capsules
Azatadine Maleate Azithromycin for Oral Suspension
Azatadine Maleate Tablets Aztreonam
Azathioprine Aztreonam Injection
Azathioprine Oral Suspension Aztreonam for Injection
USP 32 Official Monographs / Acarbose 1
Acarbose
132270
Analysis
(Comment on this Monograph)id=m115=Acarbose=A- Samples: Standard solution and Sample solution
Monos.pdf) Calculate the percentage of C25H43NO18 in the portion of
Acarbose taken:
Result = (rU/rS) (CS/CU) 100
rU = peak response from the Sample solution

rS = peak response from the Standard solution
CS = concentration of USP Acarbose RS in the
Standard solution (mg/mL)
C25H43NO18 645.60 CU = concentration of the Sample solution (mg/mL)
D-Glucose, O-4,6-dideoxy-4-[[[1S-(1,4,5,6)]-4,5,6- Acceptance criteria: 95.0%102.0%
trihydroxy-3-(hydroxymethyl)-2-cyclohexen-1-yl]amino]--D- IMPURITIES
glucopyranosyl-(14)-O--D-glucopyranosyl-(14)-; Inorganic Impurities
O-4,6-Dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3- RESIDUE ON IGNITION 281: NMT 0.2%, determined on 1.0 g
(hydroxymethyl)-2-cyclohexen-1-yl]amino]--D- HEAVY METALS, Method II 231: NMT 20 ppm
glucopyranosyl-(14)-O--D-glucopyranosyl-(14)-D- Organic Impurities
glucose [56180-94-0]. PROCEDURE
DEFINITION Mobile phase, System suitability solution, and
Acarbose is produced by certain strains of Actinoplanes Chromatographic system: Proceed as directed in the
utahensis. It contains NLT 95.0% and NMT 102.0% of Assay.
C25H43NO18, calculated on the anhydrous basis. Sample stock solution: Use the Sample solution, as
directed in the Assay.
IDENTIFICATION Sample solution: Sample stock solution and water (1:99)
A. INFRARED ABSORPTION 197K Analysis
B. The retention time of the Sample solution corresponds to Samples: Sample stock solution and Sample solution
that of the Standard solution, as obtained in the Assay. Calculate the percentage of each impurity in the portion
of Acarbose taken:
ASSAY
PROCEDURE Result = (ri/ra) RRF 100
Solution A: 0.6 mg/mL of monobasic potassium phosphate
and 0.35 mg/mL of dibasic sodium phosphate ri = peak response for each impurity
Mobile phase: Acetonitrile and Solution A (3:1) ra = response of the main acarbose peak in the
System suitability solution: Reconstitute a vial of USP chromatogram of the Sample solution
Acarbose System Suitability Mixture RS in 1 mL of water. RRF = relative response factor for each impurity, as
Standard solution: Reconstitute a vial of USP Acarbose RS listed in the Impurity Table
in 5.0 mL of water. Acceptance criteria
Sample solution: 20 mg/mL of Acarbose in water Individual impurities: See Impurity Table.
Chromatographic system Total impurities: NMT 3.0%
(See Chromatography 621, System Suitability.)
Mode: LC Impurity Table
Detector: UV 210 nm
Column: 4-mm 25-cm; packing L8 Relative Relative Acceptance
Temperature: 35 Retention Response Criteria,
Flow rate: 2 mL/min Name Time Factor NMT (%)
Injection size: 10 L Impurity Aa 0.9 1 0.6
System suitability Impurity Bb 0.8 1.6 0.5
Sample: System suitability solution
[NOTEIdentify the acarbose peak and the peaks due to Impurity Cc 1.2 1 1.5
the impurities listed in the Impurity Table.] Impurity Dd 0.5 1.33 1.0
Suitability requirements Impurity Ee 1.7 0.8 0.2
Height to valley ratio: NLT 1.2 for the impurity A peak,
and the valley between the impurity A peak and the Impurity Ff 1.9 0.8 0.3
acarbose peak Impurity Gg 2.2 0.8 0.3
[NOTEThe chromatogram obtained is similar to the Impurity Hh 0.6 1 0.2
chromatogram supplied with USP Acarbose System
Suitability Mixture RS.]
2 Acarbose / Official Monographs USP 32
Impurity Table (continued) USP Acarbose System Suitability Mixture RS

Relative Relative Acceptance
Retention Response Criteria,
Name Time Factor NMT (%) Acebutolol Hydrochloride
Any individual 0.2 (Comment on this Monograph)id=m125=Acebutolol
unknown impurity Hydrochloride=A-Monos.pdf)
aO-4,6-Dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-
(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl-(14)-O--D-
glucopyranosyl-(14)-D-arabino-hex-2-ulopyranose.
b(1R,4R,5S,6R)-4,5,6-Trihydroxy-2-(hydroxymethyl)cyclohex-2-enyl 4-O-
[4,6-dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-
(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl]--D-
glucopyranoside.
c-D-Glucopyranosyl 4-O-[4,6-dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-
trihydroxy-3-(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl]- C18H28N2O4 HCl 372.89

-D-glucopyranoside. Butanamide, N-[3-acetyl-4-[2-hydroxy-3-[(1-
d4-O-[4,6-Dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3- methylethyl)amino]propoxy]phenyl]-, monohydrochloride,
(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl]-D- ()-;
glucopyranose. ()-3-Acetyl-4-[2-hydroxy-3-(isopropylamino)propoxy]-
eO-4,6-Dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3- butyranilide monohydrochloride [34381-68-5].
(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl-(14)-O--D-
DEFINITION
glucopyranosyl-(14)-O--D-glucopyranosyl-(14)-D-arabino-hex-2-
Acebutolol Hydrochloride contains NLT 98.0% and NMT
ulopyranose (4-O--acarbosyl-D-fructopyranose).
fO-4,6-Dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-
102.0% of C18H28N2O4 HCl, calculated on the dried basis.
(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl-(14)-O--D- IDENTIFICATION
glucopyranosyl-(14)-O--D-glucopyranosyl-(14)-D-glucopyranose (4-O- A. INFRARED ABSORPTION 197K
-acarbosyl-D-glucopyranose)O-4,6-Dideoxy-4-[[(1S,4R,5S,6S)-4,5,6- B. PROCEDURE
trihydroxy-3-(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl- Sample solution: Prepare a mixture of the Standard solution
(14)-O--D-glucopyranosyl-(14)-O--D-glucopyranosyl-(14)-D- and the Sample solution (1:1) from the Assay.
glucopyranose (4-O--acarbosyl-D-glucopyranose). Analysis: Chromatograph the Sample solution as directed in
g-D-Glucopyranosyl O-4,6-dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-
the Assay.
(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl-(14)-O--D- Acceptance criteria: The chromatogram thus obtained
glucopyranosyl-(14)-O--D-glucopyranoside (-D-glucopyranosyl - exhibits a single major peak due to acebutolol.
acarboside)-D-Glucopyranosyl O-4,6-dideoxy-4-[[(1S,4R,5S,6S)-4,5,6- C. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the
trihydroxy-3-(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl- requirements, when tested as directed for alkaloidal
(14)-O--D-glucopyranosyl-(14)-O--D-glucopyranoside (-D- hydrochlorides
glucopyranosyl -acarboside).
hO-4,6-Dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3- ASSAY
(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl-(14)-O-6- PROCEDURE
deoxy--D-glucopyranosyl-(14)-D-glucopyranoseO-4,6-Dideoxy-4-[ Mobile phase: Methanol, glacial acetic acid, and 0.3%
[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydroxymethyl)cyclohex-2-enyl]amino]- aqueous solution of sodium dodecyl sulfate (675:20:325)
-D-glucopyranosyl-(14)-O-6-deoxy--D-glucopyranosyl-(14)-D- [NOTEMake adjustments if necessary to achieve a
glucopyranose. retention time for acebutolol of between 4 and 7 min.]
Standard solution: 0.14 mg/mL of USP Acebutolol
Hydrochloride RS
SPECIFIC TESTS Sample solution: 0.14 mg/mL of Acebutolol Hydrochloride
OPTICAL ROTATION, Specific Rotation 781S: +168 to +183 Chromatographic system
Sample solution: 10 mg/mL (See Chromatography 621, System Suitability.)
PH 791: 5.57.5, in a solution containing 50 mg/mL
WATER DETERMINATION, Method Ic 921: NMT 4.0%
ADDITIONAL REQUIREMENTS
PACKAGING AND STORAGE: Preserve in tight containers.
USP REFERENCE STANDARDS 11
USP Acarbose RS
USP 32 Official Monographs / Acebutolol 3
Mode: LC ADDITIONAL REQUIREMENTS

Detector: UV 254 nm PACKAGING AND STORAGE: Preserve in tight containers, and
Column: 3.9-mm 30-cm; packing L1 store at controlled room temperature.
Flow rate: 2 mL/min USP REFERENCE STANDARDS 11
Injection size: 10 L USP Acebutolol Hydrochloride RS
System suitability
Sample: Standard solution
Suitability requirements
Column efficiency: NLT 1500 theoretical plates Acebutolol Hydrochloride Capsules
Tailing factor: NMT 2.5 (Comment on this Monograph)id=m127=Acebutolol
Relative standard deviation: NMT 2.0% Hydrochloride Capsules=A-Monos.pdf)
Analysis
Samples: Standard solution and Sample solution DEFINITION
Calculate the percentage of C18H28N2O4 HCl in the Acebutolol Hydrochloride Capsules contain the equivalent of
Acebutolol Hydrochloride taken: NLT 90.0% and NMT 110.0% of the labeled amount of
acebutolol (C18H28N2O4).
IDENTIFICATION
rU = peak response from the Sample solution The retention time of the Sample solution corresponds to that
rS = peak response from the Standard solution of the Standard solution, as obtained in the Assay.
CS = concentration of USP Acebutolol Hydrochloride
RS in the Standard solution (mg/mL) ASSAY
CU = concentration of the Sample solution (mg/mL) PROCEDURE
Acceptance criteria: 98.0%102.0% Solution A: Dissolve 2.4 g of sodium 1-decanesulfonate in
1000 mL of water. Adjust with glacial acetic acid to a pH of
IMPURITIES 3.5.
Inorganic Impurities Mobile phase: Methanol and Solution A (3:2)
RESIDUE ON IGNITION 281: NMT 0.1% Standard solution: 0.22 mg/mL of USP Acebutolol
HEAVY METALS, Method II 231: NMT 20 ppm Hydrochloride RS in methanol
Organic Impurities [NOTEThis is equivalent to 0.2 mg/mL of acebutolol.]
PROCEDURE Sample stock solution: Weigh and mix, as completely as
Standard solution: 1.0 mg/mL of USP Acebutolol possible, the contents of NLT 20 Capsules. Transfer a portion
Hydrochloride RS in methanol of the powder, equivalent to 200 mg of acebutolol, to a
Reference solution A: Dilute 3.0 mL of the Standard 200-mL volumetric flask. Add 180 mL of methanol, and stir
solution with methanol to 100 mL. by mechanical means for 30 min. Dilute with methanol to
Reference solution B: Dilute 5.0 mL of the Standard volume.
solution with methanol to 100 mL. Sample solution: Dilute 5.0 mL of the Sample stock solution
Sample solution A: 10 mg/mL of Acebutolol with methanol to 25 mL.
Hydrochloride in methanol Chromatographic system
Sample solution B: Sample solution A and methanol (1:9) (See Chromatography 621, System Suitability.)
Developing solvent system: Butyl alcohol, glacial acetic Mode: LC
acid, and water (4:1:5) Detector: UV 254 nm
Chromatographic system Column: 3.9-mm 15-cm; 5-m packing L1
(See Chromatography 621, Thin-Layer Chromatography.) Flow rate: 1 mL/min
Mode: TLC Injection size: 20 L
Adsorbent: 0.25-mm layer of chromatographic silica gel System suitability
mixture Sample: Standard solution
Application volume: 20 L Suitability requirements
Analysis Tailing factor: NMT 1.5
Samples: Standard solution, Reference solution A, Reference Relative standard deviation: NMT 2.0%
solution B, Sample solution A, and Sample solution B Analysis
Proceed as directed in the chapter. Develop the Samples: Standard solution and Sample solution
chromatogram in the Developing solvent system until the Calculate the percentage of C18H28N2O4 in the Capsules
solvent front has moved three-fourths the length of the taken:
plate. Examine the plate under short-wavelength UV
light. Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
Acceptance criteria: The chromatograms from Sample
solution B and the Standard solution show principal spots at rU = peak response from the Sample solution
the same RF value. No secondary spot from Sample solution rS = peak response from the Standard solution
A, excluding the area at the point of application, is more CS = concentration of USP Acebutolol Hydrochloride
intense than the principal spot of Reference solution A RS in the Standard solution (mg/mL)
(0.3%); NMT two secondary spots from Sample solution A CU = nominal concentration of the Sample solution
are more intense than the principal spot of Reference (mg/mL)
solution B (0.1%); and the total of all impurities detected in Mr1 = molecular weight of acebutolol, 336.44
the chromatogram of Sample solution A is NMT 0.5%. Mr2 = molecular weight of acebutolol hydrochloride,
372.89
SPECIFIC TESTS
MELTING RANGE OR TEMPERATURE 741: 140144
PH 791: 4.57.0, in a solution (1 in 100)
LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT
1.0% of its weight
4 Acebutolol / Official Monographs USP 32
Acceptance criteria: 90.0%110.0% rS = peak response from the Standard solution

CS = concentration of USP Acebutolol Hydrochloride
PERFORMANCE TESTS RS in the Standard solution (g/mL)
DISSOLUTION 711 CU = nominal concentration of the Sample solution
Medium: Water; 900 mL (g/mL)
Apparatus 2: 50 rpm Mr1 = molecular weight of acebutolol, 336.44
Time: 30 min Mr2 = molecular weight of acebutolol hydrochloride,
Sample solution: Sample per Dissolution 711. 372.89
Standard solution: USP Acebutolol Hydrochloride RS in Test 2
Medium Solution A: Prepare as directed in the Assay.
Spectrometric conditions Mobile phase: Methanol and Solution A (1:1)
Mode: UV Standard solution: Transfer 30 mg of USP Acebutolol
Analytical wavelength: 232 nm Hydrochloride RS to a 50-mL volumetric flask. Add 12 mL
Analysis: Determine the amount of C18H28N2O4 dissolved of methanol, swirl to dissolve, and dilute with Mobile
by employing the UV absorption on filtered portions of the phase to volume. Dilute a volume of this stock solution
Sample solution in comparison with a Standard solution in with Mobile phase to obtain a concentration of 1.4 g/mL
the same Medium. of USP Acebutolol Hydrochloride RS.
Tolerances: NLT 80% (Q) of the labeled amount of Sample solution: Transfer a portion of the contents of 20
C18H28N2O4 is dissolved. opened Capsules, equivalent to 250 mg of acebutolol, to
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements a 100-mL volumetric flask, add 25 mL of methanol, and
shake by mechanical means for 15 min. Dilute with Mobile
IMPURITIES phase to volume. Centrifuge a portion of this solution,
Organic Impurities and transfer 10.0 mL of the clear supernatant to a 100-mL
PROCEDURE volumetric flask. Dilute with Mobile phase to volume.
Test 1 Chromatographic system
Solution A: Prepare as directed in the Assay. (See Chromatography 621, System Suitability.)
Mobile phase: Methanol and Solution A (44:56) Mode: LC
Diluent: Methanol and Solution A (1:1) Detector: UV 240 nm
Standard solution: Transfer 30 mg of USP Acebutolol Column: 3.9 mm 15 cm; 4-m packing L1
Hydrochloride RS to a 50-mL volumetric flask. Add 12 mL Flow rate: 1 mL/min
of methanol, swirl to dissolve, and dilute with Diluent to Injection size: 70 L
volume. Dilute a volume of this solution with Diluent to System suitability
obtain a concentration of 1.4 g/mL of USP Acebutolol Sample: Standard solution
Hydrochloride RS. Suitability requirements
Sample solution: Transfer a portion of the contents of 20 Relative standard deviation: NMT 6.0%
opened Capsules, equivalent to 250 mg of acebutolol, to Analysis
a 100-mL volumetric flask, add 25 mL of methanol, and Samples: Mobile phase, Standard solution, and Sample
shake by mechanical means for 15 min. Dilute with solution
Diluent to volume. Centrifuge a portion of this solution, Calculate the percentage of each impurity eluting after
and transfer 10.0 mL of the clear supernatant to a 100-mL the acebutolol peak in the portion of Capsules taken:
volumetric flask. Dilute with Diluent to volume.
Chromatographic system Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
Mode: LC rU = peak response for any individual impurity from
Detector: UV 240 nm the Sample solution
Column: 3.9-mm 15-cm; 4-m packing L1 rS = peak response from the Standard solution
Flow rate: 1 mL/min CS = concentration of USP Acebutolol Hydrochloride
Injection size: 35 L RS in the Standard solution (g/mL)
System suitability CU = nominal concentration of the Sample solution
Sample: Standard solution (g/mL)
Suitability requirements Mr1 = molecular weight of acebutolol, 336.44
Relative standard deviation: NMT 6.0% Mr2 = molecular weight of acebutolol hydrochloride,
Analysis 372.89
Samples: Diluent, Standard solution, and Sample solution Acceptance criteria
[NOTERecord the chromatograms for two times the Test 1: NMT 0.5% of any individual impurity
retention time of acebutolol, and measure the Test 2: NMT 0.5% of any individual impurity
responses for all the peaks, disregarding any peaks Sum of impurities found in Test 1 and Test 2: NMT
corresponding to those obtained from the Diluent.] 1.0%
Calculate the percentage of each impurity eluting prior
to the acebutolol peak in the portion of Capsules taken: ADDITIONAL REQUIREMENTS
PACKAGING AND STORAGE: Preserve in tight containers, and
Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 store at controlled room temperature.
rU = peak response for any individual impurity from USP Acebutolol Hydrochloride RS
the Sample solution
USP 32 Official Monographs / Acepromazine 5
Acepromazine Maleate Acceptance criteria: 98.0%101.0%

(Comment on this Monograph)id=m134=Acepromazine IMPURITIES
Maleate=A-Monos.pdf) Inorganic Impurities
RESIDUE ON IGNITION 281: NMT 0.2%
Organic Impurities
PROCEDURE
[NOTEConduct this test without exposure to daylight, and
with the minimum necessary exposure to artificial light.]
Diluent: Methanol and diethylamine (19:1)
Standard solution: Dilute 1 volume of the Sample solution
C19H22N2OS C4H4O4 442.53 with 200 volumes of Diluent (0.1 mg/mL).
Ethanone, 1-[10-[3-(dimethylamino)propyl]-10H-phenothiazin-2- Sample solution: 20.0 mg/mL in Diluent
yl]-, (Z)-2-butenedioate (1:1); Developing solvent system: n-Heptane, isobutyl alcohol,
10-[3-(Dimethylamino)propyl]phenothiazin-2-yl methyl ketone and diethylamine (75:17:8)
maleate (1:1) [3598-37-6]. Chromatographic system
(See Chromatography 621, Thin-Layer Chromatography.)
DEFINITION Mode: TLC
Acepromazine Maleate contains NLT 98.0% and NMT 101.0% Adsorbent: 0.25-mm layer of chromatographic silica gel
of C19H22N2OS C4H4O4, calculated on the anhydrous basis. mixture
[NOTEThroughout the following procedures, protect samples, Application volume: 10 L
the USP Reference Standard, and solutions containing them, Analysis
by conducting the procedures without delay, under subdued Samples: Standard solution and Sample solution
light, or using low-actinic glassware.] Develop the chromatogram in the Developing solvent
system until the solvent front has moved three-fourths
IDENTIFICATION the length of the plate. Examine the plate under short-
A. INFRARED ABSORPTION 197K wavelength UV light.
B. The retention time of the peak from the Sample solution Acceptance criteria: No spot, other than the principal
corresponds to that of the Standard solution, as obtained in acepromazine spot and any at the origin, observed in the
the Assay. chromatogram of the Sample solution is more intense than
the principal spot observed in the chromatogram of the
ASSAY Standard solution (0.5%).
PROCEDURE
Solution A: Add 6 mL of triethylamine to 700 mL of water, SPECIFIC TESTS
and adjust with phosphoric acid to a pH of 2.5. MELTING RANGE OR TEMPERATURE 741: 136139
Mobile phase: Acetonitrile and Solution A (300:700) PH 791: 4.05.5 in a solution (1 in 100)
Standard stock solution: 1 mg/mL of USP Acepromazine WATER DETERMINATION, Method I 921: NMT 1.0%
Maleate RS in 0.05 N hydrochloric acid
Standard solution: 0.1 mg/mL of USP Acepromazine ADDITIONAL REQUIREMENTS
Maleate RS in water, from Standard stock solution PACKAGING AND STORAGE: Preserve in well-closed containers,
Sample stock solution: 1 mg/mL of Acepromazine Maleate protected from light. Store at room temperature.
in 0.05 N hydrochloric acid LABELING: Label it to indicate that it is for veterinary use
Sample solution: 0.1 mg/mL of Acepromazine Maleate in only.
water, from Sample stock solution USP REFERENCE STANDARDS 11
Chromatographic system USP Acepromazine Maleate RS
Mode: LC
Detector: UV 280 nm
Column: 4 mm 15 cm; 5-m packing L7 Acepromazine Maleate Injection
Flow rate: 1 mL/min (Comment on this Monograph)id=m137=Acepromazine
Injection size: 10 L Maleate Injection=A-Monos.pdf)
System suitability
Sample: Standard solution DEFINITION
Suitability requirements Acepromazine Maleate Injection is a sterile solution of
Column efficiency: NLT 1500 theoretical plates Acepromazine Maleate in Water for Injection. It contains NLT
Tailing factor: NMT 2.5 90.0% and NMT 110.0% of the labeled amount of
Relative standard deviation: NMT 2.0% acepromazine maleate (C19H22N2OS C4H4O4).
Analysis [NOTEThroughout the following procedures, protect samples,
Samples: Standard solution and Sample solution the Reference Standard, and solutions containing them, by
Calculate the percentage of C19H22N2OS C4H4O4 in the conducting the procedures without delay, under subdued
Acepromazine Maleate taken: light, or using low-actinic glassware.]
Result = (rU/rS) (CS/CU) 100 IDENTIFICATION

A. INFRARED ABSORPTION 197K
rU = peak response from the Sample solution Sample: To a volume of Injection equivalent to 20 mg of
rS = peak response from the Standard solution acepromazine maleate, add 2 mL of water and 3 mL of 2 N
CS = concentration of USP Acepromazine Maleate RS sodium hydroxide, and extract with two 5-mL portions of
in the Standard solution (mg/mL) cyclohexane. Combine the cyclohexane extracts, and
CU = concentration of the Sample solution (mg/mL)
6 Acepromazine / Official Monographs USP 32
evaporate to dryness under vacuum, using gentle heat, if Acepromazine Maleate Tablets
necessary. (Comment on this Monograph)id=m140=Acepromazine
B. The retention time of the Sample solution corresponds to Maleate Tablets=A-Monos.pdf)
that of the Standard solution, as obtained in the Assay.
DEFINITION
ASSAY Acepromazine Maleate Tablets contain NLT 90.0% and NMT
PROCEDURE 110.0% of the labeled amount of acepromazine maleate
Solution A: Add 6 mL of triethylamine to 700 mL of water (C19H22N2OS C4H4O4).
and adjust with phosphoric acid to a pH of 2.5. [NOTEThroughout the following procedures, protect samples,
Mobile phase: Acetonitrile and Solution A (300:700) the Reference Standard, and solutions containing them, by
Standard stock solution: 1 mg/mL of USP Acepromazine conducting the procedures without delay, under subdued
Maleate RS in 0.05 N hydrochloric acid light, or using low-actinic glassware.]
Standard solution: 0.1 mg/mL of USP Acepromazine
Maleate RS in water, from Standard stock solution IDENTIFICATION
Sample stock solution: 1 mg/mL of acepromazine maleate A. INFRARED ABSORPTION 197K
in 0.05 N hydrochloric acid, from an appropriately diluted Sample: To a quantity of powdered Tablets equivalent to 20
volume of Injection mg of acepromazine maleate, add 2 mL of water and 3 mL
Sample solution: 0.1 mg/mL of acepromazine maleate in of 2 N sodium hydroxide, and extract with two 5-mL
water, from Sample stock solution portions of cyclohexane. Combine the cyclohexane extracts,
Chromatographic system and evaporate to dryness under vacuum, using gentle heat
(See Chromatography 621, System Suitability.) if necessary.
Mode: LC B. The retention time of the Sample solution corresponds to
Detector: UV 280 nm that of the Standard solution, as obtained in the Assay.
Column: 4 mm 15 cm; 5-m packing L7
Flow rate: 1 mL/min ASSAY
Injection size: 10 L PROCEDURE
System suitability Solution A: Add 6 mL of triethylamine to 700 mL of water,
Sample: Standard solution and adjust with phosphoric acid to a pH of 2.5.
Suitability requirements Mobile phase: Acetonitrile and Solution A (300:700)
Column efficiency: NLT 1500 theoretical plates Standard stock solution: 1 mg/mL of USP Acepromazine
Tailing factor: NMT 2.5 Maleate RS in 0.05 N hydrochloric acid
Relative standard deviation: NMT 2.0% Standard solution: 0.1 mg/mL of USP Acepromazine
Analysis Maleate RS in water, from Standard stock solution
Samples: Standard solution and Sample solution Sample stock solution: Transfer NLT 10 Tablets to a 200-
Calculate the percentage of C19H22N2OS C4H4O4 in the mL volumetric flask, add 100 mL of 0.05 N hydrochloric
volume of Injection taken: acid, and sonicate for 10 min. Shake by mechanical means
for 30 min, and dilute with 0.05 N hydrochloric acid to
Result = (rU/rS) (CS/CU) 100 volume.
Sample solution: 0.1 mg/mL of acepromazine maleate in
rU = peak area from the Sample solution water, from Sample stock solution. Pass a portion of this
rS = peak area from the Standard solution solution through a filter having a 0.5-m or finer porosity.
CS = concentration of USP Acepromazine Maleate RS Chromatographic system
in the Standard solution (mg/mL) (See Chromatography 621, System Suitability.)
CU = nominal concentration of the Sample solution Mode: LC
(mg/mL) Detector: UV 280 nm
Acceptance criteria: 90.0%110.0% Column: 4 mm 15 cm; 5-m packing L7
Flow rate: 1 mL/min
SPECIFIC TESTS Injection size: 10 L
PH 791: 4.55.8 System suitability
OTHER REQUIREMENTS: It meets the requirements under Sample: Standard solution
Injections 1. Suitability requirements
BACTERIAL ENDOTOXINS TEST 85: NMT 4.5 USP Endotoxin Column efficiency: NLT 1500 theoretical plates
Units/mg of acepromazine maleate Tailing factor: NMT 2.5
STERILITY TESTS 71: It meets the requirements when tested Relative standard deviation: NMT 2.0%
as directed for Test for Sterility of the Product to be Examined, Analysis
Membrane Filtration. Samples: Standard solution and Sample solution
ADDITIONAL REQUIREMENTS Calculate the percentage of C19H22N2OS C4H4O4 in the
PACKAGING AND STORAGE: Preserve in tight, light-resistant, Tablets taken:
single-dose or multiple-dose Containers for Injections as Result = (rU/rS) (CS/CU) 100
described under Injections 1, preferably of Type I glass, and
store at controlled room temperature. rU = peak area from the Sample solution
LABELING: Label it to indicate that it is for veterinary use rS = peak area from the Standard solution
only. CS = concentration of USP Acepromazine Maleate RS
USP REFERENCE STANDARDS 11 in the Standard solution (mg/mL)
USP Acepromazine Maleate RS
USP Endotoxin RS
USP 32 Official Monographs / Acetaminophen 7
CU = nominal concentration of the Sample solution Acceptance criteria: 98.0%101.0%

(mg/mL)
Acceptance criteria: 90.0%110.0% IMPURITIES
Inorganic Impurities
ADDITIONAL REQUIREMENTS RESIDUE ON IGNITION 281: NMT 0.1%
PACKAGING AND STORAGE: Preserve in tight, light-resistant CHLORIDE AND SULFATE, Chloride 221
containers, and store at controlled room temperature. Sample solution: Shake 1.0 g of Acetaminophen with 25
LABELING: Label the Tablets to indicate that they are for mL of water, filter, and add 1 mL of 2 N nitric acid and 1
veterinary use only. mL of silver nitrate TS.
USP REFERENCE STANDARDS 11 Acceptance criteria: The filtrate shows no more chloride
USP Acepromazine Maleate RS than corresponds to 0.20 mL of 0.020 N hydrochloric acid
(0.014%).
CHLORIDE AND SULFATE, Sulfate 221
Sample solution: Shake 1.0 g of Acetaminophen with 25
Acetaminophen mL of water, filter, and add 2 mL of 1 N acetic acid, then
(Comment on this Monograph)id=m150=Acetaminophen=A- add 2 mL of barium chloride TS.
Monos.pdf) Acceptance criteria: The mixture shows no more sulfate
than corresponds to 0.20 mL of 0.020 N sulfuric acid
(0.02%).
SULFIDE
Sample: 2.5 g
Analysis: Place the Sample in a 50-mL beaker. Add 5 mL of
alcohol and 1 mL of 3 N hydrochloric acid. Moisten a piece
of lead acetate test paper with water, and fix to the
underside of a watch glass. Cover the beaker with the
C8H9NO2 151.16 watch glass so that part of the lead acetate paper hangs
Acetamide, N-(4-hydroxyphenyl)-; down near the pouring spout of the beaker. Heat the
4-Hydroxyacetanilide [103-90-2]. contents of the beaker on a hot plate just to boiling.
Acceptance criteria: No coloration or spotting of the test
DEFINITION paper occurs.
Acetaminophen contains NLT 98.0% and NMT 101.0% of HEAVY METALS, Method II 231: NMT 10 ppm
C8H9NO2, calculated on the anhydrous basis. Organic Impurities
IDENTIFICATION PROCEDURE 1
A. INFRARED ABSORPTION 197K Alkaline nitroferricyanide solution: Dissolve1 g of sodium
B. ULTRAVIOLET ABSORPTION 197U nitroferricyanide and 1 g of anhydrous sodium carbonate in
Sample solution: 5 g/mL in a mixture of 0.1 N 100 mL of water.
hydrochloric acid in methanol (1 in 100) Sample solution: Transfer 5.0 g of Acetaminophen to a
C. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 100-mL volumetric flask, and dissolve in 75 mL of a
Sample solution: 1 mg/mL in methanol mixture of equal volumes of methanol and water. Add 5.0
Developing solvent system: Methylene chloride and mL of Alkaline nitroferricyanide solution, dilute with a
methanol (4:1) mixture of equal volumes of methanol and water to
Acceptance criteria: Meets the requirements of the test volume, and allow to stand for 30 min.
Standard solution: 2.5 g/mL of p-aminophenol, similarly
ASSAY prepared as Sample solution, using the same quantities of
PROCEDURE the same reagents
Sample solution: Dissolve 120 mg of Acetaminophen in 10 Blank: 5.0 mL of Alkaline nitroferricyanide solution, diluted
mL of methanol in a 500-mL volumetric flask, and dilute in a mixture of equal volumes of methanol and water to
with water to volume. Dilute 5.0 mL of the resulting 100 mL
solution with water to 100 mL. Spectrometric conditions
Standard solution: 12 g/mL of USP Acetaminophen RS in Mode: UVVis
the same medium as Sample solution Analytical wavelength: 710 nm
Spectrometric conditions Cell: 1 cm
Mode: UV Analysis
Analytical wavelength: 244 nm Samples: Sample solution, Standard solution, and Blank
Cell: 1 cm Acceptance criteria: The absorbance of the Sample
Blank: Water solution does not exceed that of the Standard solution,
Analysis corresponding to NMT 50 ppm of p-aminophenol.
Samples: Standard solution and Sample solution PROCEDURE 2
Calculate the percentage of C8H9NO2 in the portion of Standard solution: 10 g/mL of p-chloroacetanilide in
Acetaminophen taken: ether
Sample solution: Transfer 1.0 g of Acetaminophen to a
Result = (AU/AS) (CS/CU) 100 glass-stoppered, 15-mL centrifuge tube, add 5.0 mL of
ether, shake by mechanical means for 30 min, and
AU = absorbance of the Sample solution centrifuge at 1000 rpm for 15 min or until a clean
AS = absorbance of the Standard solution separation is obtained. The supernatant is used as the
CS = concentration of USP Acetaminophen RS in the Sample solution.
Standard solution (g/mL)
CU = concentration of the Sample solution (g/mL)
8 Acetaminophen / Official Monographs USP 32
Chromatographic system having a 0.5-m or finer porosity, discarding the first 10 mL

(see Chromatography 621, Thin-Layer Chromatography.) of the filtrate.
Mode: TLC Chromatographic system
Adsorbent: 0.25-mm layer of chromatographic silica gel (See Chromatography 621, System Suitability.)
mixture Mode: LC
Application volume: Apply 200 L of the Sample solution, Detector: UV 243 nm
in 40-L portions, to obtain a single spot NMT 10 mm in Column: 3.9-mm 30-cm; packing L1
diameter. Flow rate: 1.5 mL/min
Developing solvent system: Solvent hexane and acetone Injection size: 10 L
(3:1) System suitability
Analysis Sample: Standard solution
Samples: Standard solution and Sample solution Suitability requirements
Develop the chromatogram in an unsaturated chamber Column efficiency: NLT 1000 theoretical plates
with the Developing solvent system until the solvent front Tailing factor: NMT 2
has moved three-fourths of the length of the plate. Relative standard deviation: NMT 2.0%
Examine the plate under short-wavelength UV light. Analysis
Acceptance criteria: Any spot in the chromatogram of Samples: Standard solution and Sample solution
the Sample solution, at an RF value corresponding to the Calculate the percentage of C8H9NO2 in the portion of
principal spot from the Standard solution, is not greater in Capsules taken:
size or intensity than the principal spot of the Standard
solution (NMT 10 ppm). Result = (rU/rS) (CS/CU) 100
SPECIFIC TESTS rU = peak response from the Sample solution
MELTING RANGE OR TEMPERATURE 741: 168172 rS = peak response from the Standard solution
WATER DETERMINATION, Method I 921: NMT 0.5% CS = concentration of USP Acetaminophen RS in the
READILY CARBONIZABLE SUBSTANCES TEST 271: Dissolve 0.50 Standard solution (mg/mL)
g in 5 mL of sulfuric acid TS: the solution has no more color CU = nominal concentration of acetaminophen in the
than Matching Fluid A. Sample solution (mg/mL)
Acceptance criteria: 90.0%110.0%
PACKAGING AND STORAGE: Preserve in tight, light-resistant PERFORMANCE TESTS
containers, and store at room temperature. Protect from DISSOLUTION 711
moisture and heat. Medium: Water; 900 mL
USP REFERENCE STANDARDS 11 Apparatus 2: 50 rpm
USP Acetaminophen RS Time: 45 min
Detector: UV 249 nm
Sample solution: Sample per Dissolution 711.
Standard solution: USP Acetaminophen RS in Medium
Acetaminophen Capsules Analysis: Determine the amount of C8H9NO2 dissolved by
(Comment on this Monograph)id=m160=Acetaminophen using UV absorption on filtered portions of the Sample
Capsules=A-Monos.pdf) solution suitably diluted with Medium, if necessary, in
comparison with the Standard solution having a known
DEFINITION concentration of USP Acetaminophen RS.
Acetaminophen Capsules contain NLT 90.0% and NMT 110.0% Tolerances: NLT 75% (Q) of the labeled amount of
of the labeled amount of acetaminophen (C8H9NO2). C8H9NO2 is dissolved.
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
IDENTIFICATION
A. The retention time of the Sample solution corresponds to ADDITIONAL REQUIREMENTS
that of the Standard solution, as obtained in the Assay. PACKAGING AND STORAGE: Preserve in tight containers, and
B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 store at a controlled room temperature.
Sample solution: Equivalent to 1 mg/mL of acetaminophen USP REFERENCE STANDARDS 11
from contents of the Capsules in methanol. Filter and use USP Acetaminophen RS
filtrate.
Developing solvent system: Methylene chloride and
methanol (4:1)
Acceptance criteria: Meet the requirements Acetaminophen Oral Solution
(Comment on this Monograph)id=m180=Acetaminophen Oral
ASSAY Solution=A-Monos.pdf)
PROCEDURE
Mobile phase: Methanol and water (1:3) DEFINITION
Standard solution: 0.01 mg/mL of USP Acetaminophen RS Acetaminophen Oral Solution contains NLT 90.0% and NMT
in Mobile phase 110.0% of the labeled amount of acetaminophen (C8H9NO2).
Sample solution: Weigh the contents of NLT 20 Capsules,
and calculate the average weight of the contents of each IDENTIFICATION
Capsule. Mix the combined contents of the Capsules, and A. The retention time of the Sample solution corresponds to
transfer a portion, equivalent to 100 mg of acetaminophen, that of the Standard solution, as obtained in the Assay.
to a 200-mL volumetric flask. Add 100 mL of Mobile phase, B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
shake by mechanical means for 10 min, and dilute with Sample solution: Equivalent to 1 mg/mL of acetaminophen
Mobile phase to volume. Transfer 5.0 mL of this solution to a in methanol
250-mL volumetric flask and dilute with Mobile phase to Developing solvent system: Methylene chloride and
volume. Pass a portion of this solution through a filter methanol (4:1)
Acceptance criteria: Meets the requirements Acetaminophen for Effervescent Oral

ASSAY
Solution
PROCEDURE (Comment on this Monograph)id=m183=Acetaminophen for
Mobile phase: Methanol and water (1:3) Effervescent Oral Solution=A-Monos.pdf)
Standard solution: 0.01 mg/mL of USP Acetaminophen RS
in Mobile phase DEFINITION
Sample solution: Transfer a measured volume of Oral Acetaminophen for Effervescent Oral Solution contains, in each
Solution, equivalent to 500 mg of acetaminophen, to a 250- 100 g, NLT 5.63 g and NMT 6.88 g of acetaminophen
mL volumetric flask, and dilute with Mobile phase to volume. (C8H9NO2).
Transfer 5.0 mL of this solution to a second 250-mL IDENTIFICATION
volumetric flask, and dilute with Mobile phase to volume. A. A 10-g portion dissolves, with effervescence, in water
Transfer 25.0 mL of this solution to a 100-mL volumetric when prepared as directed for the Sample solution in the
flask, and dilute with Mobile phase to volume. Pass a portion Assay.
of this solution through a filter having a 0.5-m or finer B. The retention time of the Sample solution corresponds to
porosity, discarding the first 10 mL of the filtrate. that of the Standard solution, as obtained in the Assay.
Chromatographic system C. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
(See Chromatography 621, System Suitability.) Sample solution: Triturate 0.4 g of the powder with 25 mL
Mode: LC of methanol, and filter.
Detector: UV 243 nm Developing solvent system: Methylene chloride and
Column: 3.9-mm 30-cm; packing L1 methanol (4:1)
Flow rate: 1.5 mL/min Acceptance criteria: Meets the requirements of the test
Injection size: 10 L
System suitability ASSAY
Sample: Standard solution PROCEDURE
Suitability requirements Mobile phase: Methanol and water (1:3)
Column efficiency: NLT 1000 theoretical plates Standard solution: 0.01 mg/mL of USP Acetaminophen RS
Tailing factor: NMT 2.0 in Mobile phase
Relative standard deviation: NMT 2.0% Sample solution: Dissolve 10 g of Acetaminophen for
Analysis Effervescent Oral Solution in 200 mL of water in a 1000-mL
Samples: Standard solution and Sample solution volumetric flask, using gentle heat if necessary, until
Calculate the percentage of C8H9NO2 in each mL of the effervescence subsides, then dilute with water to volume.
Oral Solution taken: Transfer 10.0 mL of this solution to a 100-mL volumetric
flask, and dilute with water to volume. Transfer 8.0 mL of
Result = (rU/rS) (CS/CU) 100 this solution to a 50-mL volumetric flask, and dilute with
Mobile phase to volume. Pass a portion of this solution
rU = peak response from the Sample solution through a filter having a 0.5-m or finer porosity, discarding
rS = peak response from the Standard solution the first 10 mL of the filtrate.
CS = concentration of USP Acetaminophen RS in the Chromatographic system
Standard solution (mg/mL) (See Chromatography 621, System Suitability.)
CU = nominal concentration of acetaminophen in the Mode: LC
Sample solution (mg/mL) Detector: UV 243 nm
Acceptance criteria: 90.0%110.0% Column: 3.9-mm 30-cm; packing L1
PERFORMANCE TESTS Flow rate: 1.5 mL/min
DELIVERABLE VOLUME 698: Meets the requirements for oral Injection size: 10 L
solution packaged in multiple-unit containers System suitability
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements Sample: Standard solution
for oral solution packaged in single-unit containers Suitability requirements
Column efficiency: NLT 1000 theoretical plates
SPECIFIC TESTS Tailing factor: NMT 2.0
PH 791: 3.86.1 Relative standard deviation: NMT 2.0%
ALCOHOL DETERMINATION, Method II 611: Between 90.0% Analysis
and 115.0% of the labeled amount of C2H5OH, determined Samples: Standard solution and Sample solution
by the gasliquid chromatographic procedure, with acetone Calculate the quantity, in g/100 g, of C8H9NO2 in the
being used as the internal standard portion of Acetaminophen for Effervescent Oral Solution
taken:
PACKAGING AND STORAGE: Preserve in tight containers, and Result = (rU/rS) (CS/CU) F
store at a controlled room temperature.
USP REFERENCE STANDARDS 11 rU = peak response from the Sample solution
USP Acetaminophen RS rS = peak response from the Standard solution
CS = concentration of USP Acetaminophen RS in the Mode: LC

Standard solution (mg/mL) Detector: UV 243 nm
CU = concentration of the Sample solution (mg Column: 3.9-mm 30-cm; packing L1
Acetaminophen for Effervescent Oral Flow rate: 1.5 mL/min
Solution/mL) Injection size: 10 L
F = conversion factor (100) System suitability
Acceptance criteria: 5.63 g6.88 g/100 g of C8H9NO2 Sample: Standard solution
PERFORMANCE TESTS Column efficiency: NLT 1000 theoretical plates
MINIMUM FILL 755 Tailing factor: NMT 2.0
For solid packaged in multiple-unit containers: Meets the Relative standard deviation: NMT 2.0%
requirements Analysis
UNIFORMITY OF DOSAGE UNITS 905 Samples: Standard solution and Sample solution
For solid packaged in single-unit containers: Meets the Calculate the percentage of C8H9NO2 in each Suppository
requirements taken:
ADDITIONAL REQUIREMENTS Result = (rU/rS) (CS/CU) 100
PACKAGING AND STORAGE: Preserve in air tight containers,
and store at controlled room temperature. rU = peak response from the Sample solution
USP REFERENCE STANDARDS 11 rS = peak response from the Standard solution
USP Acetaminophen RS CS = concentration of USP Acetaminophen RS in the
CU = nominal concentration of the Sample solution
Acetaminophen Suppositories (mg/mL)
(Comment on this Monograph)id=m197=Acetaminophen
Suppositories=A-Monos.pdf) ADDITIONAL REQUIREMENTS
DEFINITION store at controlled room temperature or in a cool place.
Acetaminophen Suppositories contain NLT 90.0% and NMT USP REFERENCE STANDARDS 11
110.0% of the labeled amount of acetaminophen (C8H9NO2). USP Acetaminophen RS
IDENTIFICATION
A. The retention time of the chromatogram of the Sample
solution corresponds to that of the Standard solution, as Acetaminophen Oral Suspension
obtained in the Assay. (Comment on this Monograph)id=m190=Acetaminophen Oral
B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Suspension=A-Monos.pdf)
Sample solution: Transfer a portion of Suppositories,
equivalent to 20 mg of acetaminophen, to a beaker, add 20 DEFINITION
mL of methanol, and heat on a steam bath until melted. Acetaminophen Oral Suspension is a suspension of
Remove the beaker from the steam bath, allow to cool with Acetaminophen in a suitable aqueous vehicle. It contains NLT
occasional stirring, and filter. 90.0% and NMT 110.0% of the labeled amount of C8H9NO2.
Developing solvent system: Methylene chloride and
methanol (4:1) IDENTIFICATION
Acceptance criteria: Meet the requirements INFRARED ABSORPTION 197K
Sample: Transfer a volume of Oral Suspension, equivalent
ASSAY to 240 mg of acetaminophen, to a separator, add 50 mL of
PROCEDURE ethyl acetate, and shake. Filter the ethyl acetate extract
Mobile phase: Methanol and water (1:3) through a funnel containing glass wool and 10 g of
Standard solution: 0.01 mg/mL of USP Acetaminophen RS anhydrous sodium sulfate. Collect the filtrate in a beaker,
in Mobile phase and evaporate on a steam bath to dryness. Dry the residue
Sample stock solution: Tare a small dish and a glass rod, in a vacuum over silica gel.
place in the dish NLT 5 Suppositories, heat gently on a
steam bath until melted, then stir, cool while stirring, and ASSAY
weigh. Transfer a weighed portion of the mass, equivalent to PROCEDURE
100 mg of acetaminophen, to a separator, add 30 mL of Mobile phase: Methanol and water (1:3)
solvent hexane, and dissolve. Add 30 mL of water, shake Standard solution: 0.01 mg/mL of USP Acetaminophen RS
gently, and allow the phases to separate. [NOTEIf an in Mobile phase
emulsion forms, allow sufficient time for it to separate.] Sample solution: Transfer a volume of Oral Suspension,
Transfer the aqueous layer to a 200-mL volumetric flask, previously well-shaken, equivalent to 100 mg of
wash the solvent hexane in the separator with three 30-mL acetaminophen, to a 200-mL volumetric flask. Add 100 mL
portions of water, adding the washings to the volumetric of Mobile phase, and shake by mechanical means for 10
flask, and dilute with Mobile phase to volume. min. Dilute with Mobile phase to volume. Transfer 5.0 mL of
Sample solution: Transfer 5.0 mL of this Sample stock this solution to a 250-mL volumetric flask, and dilute with
solution to a 250-mL volumetric flask, and dilute with Mobile Mobile phase to volume. Pass a portion of this solution
phase to volume. Pass a portion of this solution through a through a filter having a 0.5-m or finer porosity, discarding
filter having a 0.5-m or finer porosity, discarding the first the first 10 mL of the filtrate.
10 mL of the filtrate. Chromatographic system
Chromatographic system (See Chromatography 621, System Suitability.)
Mode: LC IDENTIFICATION
Detector: UV 243 nm A. The retention time of the Sample solution corresponds to
Column: 3.9-mm 30-cm; packing L1 that of the Standard solution, as obtained in the Assay.
Flow rate: 1.5 mL/min B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
Injection size: 10 L Sample solution: Equivalent to 1 mg/mL of acetaminophen
System suitability from powdered Tablets in methanol; filtered
Sample: Standard solution Developing solvent system: Methylene chloride and
Suitability requirements methanol (4:1)
Column efficiency: NLT 1000 theoretical plates Acceptance criteria: Meet the requirements
Tailing factor: NMT 2.0
Relative standard deviation: NMT 2.0% ASSAY
Analysis PROCEDURE
Samples: Standard solution and Sample solution Mobile phase: Methanol and water (1:3)
Calculate the percentage of C8H9NO2 in each mL of the Standard solution: 0.01 mg/mL of USP Acetaminophen RS
Oral Suspension taken: in Mobile phase
Sample solution: Weigh and finely powder NLT 20 Tablets.
Result = (rU/rS) (CS/CU) 100 Dissolve a portion of the powder in Mobile phase to prepare
a solution containing 0.01 mg/mL of acetaminophen. To
rU = peak response from the Sample solution facilitate dissolution, shake powder in Mobile phase by
rS = peak response from the Standard solution mechanical means for 10 min, and sonicate for 5 min before
CS = concentration of USP Acetaminophen RS in the makeup with Mobile phase to volume. Pass a portion of this
Standard solution (mg/mL) solution through a filter having a 0.5-m or finer porosity,
CU = nominal concentration of acetaminophen in the discarding the first 10 mL of the filtrate. Use the clear
Sample solution (mg/mL) filtrate.
Acceptance criteria: 90.0%110.0% Chromatographic system
PERFORMANCE TESTS Mode: LC
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements Detector: UV 243 nm
for oral suspension packaged in single-unit containers Column: 3.9-mm 30-cm; packing L1
DELIVERABLE VOLUME 698 Meets the requirements for oral Flow rate: 1.5 mL/min
suspension packaged in multiple-unit containers Injection size: 10 L
System suitability
IMPURITIES Sample: Standard solution
Organic Impurities Suitability requirements
PROCEDURE: LIMIT OF 4-AMINOPHENOL Column efficiency: NLT 1000 theoretical plates
Diluent: Methanol, formic acid, and water (75:2:425) Tailing factor: NMT 2
Mobile phase: 0.01 M sodium butanesulfonate in Diluent Relative standard deviation: NMT 2.0%
Standard solution: 24 g/mL of USP 4-Aminophenol RS in Analysis
Mobile phase Samples: Standard solution and Sample solution
Sample solution: Equivalent to 4.8 mg/mL of Calculate the percentage of C8H9NO2 in the portion of
acetaminophen in Mobile phase Tablets taken:
Chromatographic system
(See Chromatography 621, System Suitability.) Result = (rU/rS) (CS/CU) 100
Mode: LC
Detector: UV 272 nm rU = peak response from the Sample solution
Column: 4.6-mm 20-cm; 10-m packing L1 rS = peak response from the Standard solution
Flow rate: 2 mL/min CS = concentration of USP Acetaminophen RS in the
Injection size: 20 L Standard solution (mg/mL)
Analysis CU = nominal concentration of the Sample solution
Samples: Standard solution and Sample solution (mg/mL)
Acceptance criteria: The peak area for 4-aminophenol Acceptance criteria: 90.0%110.0%
from the Sample solution is not greater than the
corresponding peak area from the Standard solution. PERFORMANCE TESTS
DISSOLUTION 711
SPECIFIC TESTS Medium: pH 5.8 phosphate buffer (see Reagents, Indicators,
PH 791: 4.06.9 and SolutionsBuffer Solutions); 900 mL
Apparatus 2: 50 rpm
ADDITIONAL REQUIREMENTS Time: 30 min
PACKAGING AND STORAGE: Preserve in tight containers, and Detector: UV 243 nm
store at a controlled room temperature. Standard solution: USP Acetaminophen RS in Medium
USP REFERENCE STANDARDS 11 Sample solution: Sample per Dissolution 711.
USP Acetaminophen RS Analysis: Determine the amount of C8H9NO2 dissolved by
USP 4-Aminophenol RS using UV absorption on filtered portions of the Sample
solution suitably diluted with Medium, if necessary, in
comparison with a Standard solution having a known
Acetaminophen Tablets concentration of USP Acetaminophen RS.
Acceptance criteria: NLT 80% (Q) of the labeled amount
(Comment on this Monograph)id=m200=Acetaminophen of C8H9NO2
Tablets=A-Monos.pdf) For Tablets labeled as chewable
DEFINITION Medium: pH 5.8 phosphate buffer (see Reagents,
Acetaminophen Tablets contain NLT 90.0% and NMT 110.0% Indicators, and SolutionsBuffer Solutions); 900 mL
of the labeled amount of acetaminophen (C8H9NO2).
Apparatus 2: 75 rpm rS = peak response from the Standard solution

Time: 45 min CS = concentration of USP Acetaminophen RS in the
Detecor, Sample solution, Standard solution, and Standard solution (mg/mL)
Analysis: Proceed as directed in Dissolution. CU = nominal concentration of the Sample solution
Tolerances: NLT 75% (Q) of the labeled amount of (mg/mL)
C8H9NO2 is dissolved. Acceptance criteria: 90.0%110.0%
PERFORMANCE TESTS
ADDITIONAL REQUIREMENTS DISSOLUTION 711
PACKAGING AND STORAGE: Preserve in tight containers, and Test 1
store at controlled room temperature. Medium: Simulated gastric fluid TS (without enzyme); 900
LABELING: Label Tablets that must be chewed to indicate mL
that they are to be chewed before swallowing. Apparatus 2: 50 rpm
USP REFERENCE STANDARDS 11 Time: 15 min, 1 h, and 3 h
USP Acetaminophen RS Sample solution: Sample per Dissolution 711.
Standard solution: USP Acetaminophen RS in Medium
Spectrometric conditions
Mode: UV
Acetaminophen Extended-Release Analytical wavelength: 280 nm
Tablets Analysis: Determine the amount of C8H9NO2 dissolved
(Comment on this Monograph)id=m205=Acetaminophen from UV absorbances, using a filtered portion of the Sample
Extended-Release Tablets=A-Monos.pdf) solution in comparison with a Standard solution.
Tolerances: The percentages of the labeled amount of
DEFINITION C8H9NO2 dissolved at the times specified conform to
Acetaminophen Extended-Release Tablets contain NLT 90.0% Acceptance Table 2.
and NMT 110.0% of the labeled amount of acetaminophen
(C8H9NO2). Time Amount Dissolved
IDENTIFICATION 15 min 45%65%
A. INFRARED ABSORPTION 197K 1h 60%85%
Sample: Use a portion of powdered Tablets.
3h NLT 85%
B. The retention time of the Sample solution corresponds to
For gelatin-coated Tablets
ASSAY Medium, Apparatus, Sample solution, Standard solution,
PROCEDURE Spectrometric conditions, and Analysis: Proceed as
Solution A: Phosphoric acid and water (1:9) directed in Test 1.
Mobile phase: Methanol, Solution A, and water (300:1:700) Times: 30 min, 90 min, and 4 h
Standard solution: Dissolve a quantity of USP Tolerances: The percentage of the labeled amount of
Acetaminophen RS in methanol, and dilute with Mobile C8H9NO2 dissolved at the times specified conform to
phase to 0.65 mg/mL. Acceptance Table 2.
Sample stock solution: Transfer 10 Tablets into a 250-mL
volumetric flask containing 50 mL of water and a magnetic Time Amount Dissolved
stir bar. Stir at least 30 min or until the coating has 30 min 40%60%
dissolved. Add 150 mL of methanol, and stir for 45 min.
Tablet cores should be disintegrated at least 15 min prior to 90 min 55%85%
ending the stirring. Remove the magnetic stir bar and rinse 4h NLT 80%
into the flask with methanol. Dilute with methanol to
volume, and centrifuge, using the clear supernatant as the Test 2: If the product complies with this test, the labeling
Sample stock solution. indicates that it meets USP Dissolution Test 2.
Sample solution: Dilute 5 mL of the Sample stock solution Medium, Apparatus, Sample solution, Standard solution,
with Mobile phase to 200 mL. Spectrometric conditions, and Analysis: Proceed as
Chromatographic system directed in Test 1.
(See Chromatography 621, System Suitability.) Times: 15 min, 1 h, and 3 h
Mode: LC Tolerances: The percentages of the labeled amount of
Detector: UV 295 nm C8H9NO2 dissolved at the times specified conform to
Column: 3.9-mm 15-cm; packing L1 Acceptance Table 2.
Flow rate: 2 mL/min
Injection size: 20 L Time Amount Dissolved
System suitability
Sample: Standard solution 15 min 40%60%
Suitability requirements 1h 55%75%
Tailing factor: NMT 3.0 3h NLT 80%
Relative standard deviation: NMT 2.0%
Analysis UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Samples: Standard solution and Sample solution
Calculate the percentage of C8H9NO2 in the portion of ADDITIONAL REQUIREMENTS
Tablets taken: PACKAGING AND STORAGE: Preserve in tight containers.
LABELING: Where the Tablets are gelatin-coated, the label so
Result = (rU/rS) (CS/CU) 100 states. When more than one Dissolution Test is given, the
labeling states the Dissolution Test used only if Test 1 is not CS = concentration of the corresponding USP
used. Reference Standard in the Standard solution
USP REFERENCE STANDARDS 11 (mg/mL)
USP Acetaminophen RS CU = nominal concentration of the corresponding
analyte in the Sample solution (mg/mL)
Acceptance criteria: 90.0%110.0% of the labeled
amounts of C8H9NO2 and C9H8O4
Acetaminophen and Aspirin Tablets
(Comment on this Monograph)id=m230=Acetaminophen and PERFORMANCE TESTS
Aspirin Tablets=A-Monos.pdf) DISSOLUTION, Procedure for a Pooled Sample 711
Medium: Water; 900 mL
DEFINITION Apparatus 2: 50 rpm
Acetaminophen and Aspirin Tablets contain NLT 90.0% and Time: 45 min
NMT 110.0% of the labeled amounts of acetaminophen Mobile phase, Solution A, and Chromatographic system:
(C8H9NO2) and aspirin (C9H8O4). Prepare as directed in the Assay.
Internal standard solution: 1 mg/mL of benzoic acid in
IDENTIFICATION methanol
The retention times of the Sample solution correspond to Sample stock solution: Sample per Dissolution 711.
those of the Standard solution, as obtained in the Assay. Sample solution: Combine 4.0 mL of the Sample stock
solution and 1.0 mL of the Internal standard solution.
ASSAY Standard stock solution A: 70 g/mL USP Salicylic Acid RS
PROCEDURE in Solution A
[NOTEInject the Standard solution and the Sample solution Standard solution A: Combine 4.0 mL of the Standard
promptly after preparation.] stock solution A and 1.0 mL of the Internal standard solution.
Solution A: Chloroform, methanol, and glacial acetic acid Standard stock solution B: 360 g/mL each of USP
(78:20:2) Acetaminophen RS and USP Aspirin RS in Solution A
Mobile phase: Transfer 225 mg of tetramethylammonium Standard solution B: Combine 4.0 mL of the Standard
hydroxide pentahydrate to a 1000-mL flask, and add 750 stock solution B and 1.0 mL of the Internal standard solution.
mL of water, 125 mL of methanol, 125 mL of acetonitrile, Analysis
and 1.0 mL of glacial acetic acid. Stir for 3 min, and pass Samples: Sample solution, Standard solution A, and
through a membrane filter having a 0.5-m or finer Standard solution B
porosity. [NOTEThe relative retention times for acetaminophen,
Internal standard solution: 20 mg/mL of benzoic acid in salicylic acid, aspirin, and benzoic acid are about 0.3, 0.4,
Solution A 0.6, and 1.0, respectively.]
Standard solution: Transfer 325 mg of USP Acetaminophen Analyze using a 20-L injection size.
RS and 325 mg of USP Aspirin RS to a 100-mL volumetric Determine the amount of C8H9NO2 dissolved:
flask, add 10.0 mL of Internal standard solution, and dilute
with Solution A to volume. Result = 90 (C/W) (RU/RS)
Sample solution: Transfer an equivalent to 325 mg of
acetaminophen, from finely powdered Tablets (NLT 20), to a C = concentration of USP Acetaminophen RS in
100-mL volumetric flask, add 10.0 mL of Internal standard Standard solution B (g/mL)
solution and 50 mL of Solution A, and sonicate for 3 min. W = labeled amount of acetaminophen (mg)
Dilute with Solution A to volume. Pass a portion of this RU = relative peak response ratio from the Sample
solution through a filter having a 2.5-m or finer porosity solution
and use the filtrate. RS = relative peak response ratio from the Standard
Chromatographic system solution B
(See Chromatography 621, System Suitability.) Determine the amount of C9H8O4 dissolved:
Mode: LC
Detector: UV 280 nm Result = {[90C1(RU1/RS1)] + [90C2(RU2/RS2)(1.3044)]}/W
Column: 3.9-mm 30-cm; packing L1
Flow rate: 2 mL/min C1 = concentration of USP Aspirin RS in Standard
Injection size: 5 L solution B (g/mL)
System suitability RU1 = ratio of the relative peak response for the aspirin
Sample: Standard solution peak and the internal standard peak from the
[NOTEThe retention times for acetaminophen, salicylic Sample solution
acid (if present), aspirin, and benzoic acid are about 2, 3, RS1 = ratio of the relative peak response for the aspirin
5, and 8 min, respectively.] peak and the internal standard peak from
Suitability requirements Standard solution B
Relative standard deviation: NMT 3.0% for either C2 = concentration of USP Salicylic Acid RS in
analyte Standard solution A (g/mL)
Analysis RU2 = ratio of the relative peak response for the
Samples: Standard solution and Sample solution salicylic acid peak and the internal standard
Calculate, individually, the percentages of C8H9NO2 and peak from the Sample solution
C9H8O4 in the portion of Tablets taken: RS2 = ratio of the relative peak response for the
salicylic acid peak and the internal standard
Result = (RU/RS) (CS/CU) 100 peak from Standard solution A
W = labeled amount of aspirin (mg)
RU = ratio of the peak responses of the analyte and Tolerances: NLT 75% (Q) of the labeled amounts of
benzoic acid from the Sample solution C8H9NO2 and C9H8O4 is dissolved.
RS = ratios of the peak responses of the analyte and
benzoic acid from the Standard solution
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements the labeled amount, in mg, of acetaminophen per Tablet;
for Content Uniformity with respect to acetaminophen and to and J being the ratio of the labeled amount, in mg, of
aspirin caffeine to the labeled amount, in mg, of acetaminophen
per Tablet.
IMPURITIES Standard solution: Transfer 20.0 mL of the Standard stock
Organic Impurities solution and 3.0 mL of the Internal standard solution to a 50-
PROCEDURE: LIMIT OF SALICYLIC ACID mL volumetric flask, and dilute with Solution A to volume.
Solution A, Mobile phase, Internal standard solution, This solution contains 0.1 mg/mL of USP Acetaminophen
Sample solution, and Chromatographic system: RS, 0.1J mg/mL of USP Aspirin RS, and 0.1J mg/mL of USP
Proceed as directed in the Assay. Caffeine RS.
Standard stock solution: 1.0 mg/mL of USP Salicylic Acid Sample stock solution: Transfer an equivalent to 250 mg,
RS in Solution A from finely powdered Tablets (NLT 20), of acetaminophen to
Standard solutions: Transfer 1.0-mL, 5.0-mL, and 10.0-mL a 100-mL volumetric flask. Add 75 mL of Solution A and
portions of Standard stock solution to separate 100-mL shake by mechanical means for 30 min. Dilute with Solution
volumetric flasks, add 10.0 mL of Internal standard solution A to volume.
to each flask, and dilute with Solution A to volume. Sample solution: Transfer 2.0 mL of the Sample stock
Analysis solution to a 50-mL volumetric flask. Add 3.0 mL of Internal
Samples: Sample solution and Standard solutions standard solution, and dilute with Solution A to volume.
Plot the ratios of the peak responses for salicylic acid and Chromatographic system
benzoic acid for each of the Standard solutions versus (See Chromatography 621, System Suitability.)
concentrations, in mg/mL, of salicylic acid, and draw the Mode: LC
straight line best fitting the three plotted points. From the Detector: UV 275 nm
graph so obtained, and from the ratio of the peak Column: 4.6-mm 10-cm; 5-m packing L1
responses for salicylic acid and benzoic acid in the Column temperature: 45 1
chromatogram of the Sample solution as obtained in the Flow rate: 2 mL/min
Assay, determine the concentration, in mg/mL, of salicylic Injection size: 10 L
acid (C7H6O3) in the Sample solution, and calculate the System suitability
percentage of salicylic acid in relation to the concentration Sample: Standard solution
of aspirin in the Sample solution, as determined in the [NOTEThe relative retention times for acetaminophen,
Assay. caffeine, aspirin, benzoic acid, and salicylic acid are about
Acceptance criteria: NMT 3.0% 0.3, 0.5, 0.8, 1.0, and 1.2, respectively.]
ADDITIONAL REQUIREMENTS Tailing factor: NMT 1.2 for each analyte peak
PACKAGING AND STORAGE: Preserve in tight containers, and Resolution: NLT 1.4 between any of the analyte and
store at controlled room temperature. internal standard peaks
USP REFERENCE STANDARDS 11 Relative standard deviation: NMT 2.0%
USP Acetaminophen RS Analysis
USP Aspirin RS Samples: Standard solution and Sample solution
USP Salicylic Acid RS Calculate, individually, the percentages of C8H9NO2,
C9H8O4, and C8H10N4O2 in the portion of Tablets taken:
Acetaminophen, Aspirin, and Caffeine Result = (RU/RS) (CS/CU) 100

Tablets RU = ratio of the peak responses of the analyte and
(Comment on this Monograph)id=m240=Acetaminophen, internal standard peak from the Sample solution
Aspirin, and Caffeine Tablets=A-Monos.pdf) RS = ratio of the peak responses of the analyte and
internal standard peak from the Standard
DEFINITION solution
Acetaminophen, Aspirin, and Caffeine Tablets contain NLT CS = concentration of the corresponding USP
90.0% and NMT 110.0% of the labeled amounts of Reference Standard in the Standard solution
acetaminophen (C8H9NO2), aspirin (C9H8O4), and caffeine (mg/mL)
(C8H10N4O2). CU = nominal concentration of the analyte in the
Sample solution (mg/mL)
IDENTIFICATION Acceptance criteria: 90.0%110.0%
The retention times of the Sample solution correspond to
those of the Standard solution, as obtained in the Assay. PERFORMANCE TESTS
DISSOLUTION 711
ASSAY Medium: Water; 900 mL
PROCEDURE Apparatus 2: 100 rpm
[NOTEInject the Standard solution and the Sample solution Time: 60 min
promptly after preparation.] Mobile phase, Internal standard solution, Solution A,
Solution A: Methanol and glacial acetic acid (95:5) Standard stock solution, and Chromatographic system:
Mobile phase: Methanol, glacial acetic acid, and water Proceed as directed in the Assay.
(28:3:69) Sample stock solution: Sample per Dissolution 711.
Internal standard solution: 6 mg/mL of benzoic acid in Sample solution: Transfer 20.0 mL of a filtered portion of
methanol the Sample stock solution to a 50-mL volumetric flask. Add
Standard stock solution: Dissolve quantities of USP 3.0 mL of Internal standard solution and 20 mL of Solution A.
Acetaminophen RS, USP Aspirin RS, and USP Caffeine RS in Mix, and allow to stand for 30 s. Dilute with Solution A to
Solution A to obtain a solution having known concentrations volume.
of 0.25 mg/mL of USP Acetaminophen RS, 0.25J mg/mL of Standard solution: Transfer 20.0 mL of Standard stock
USP Aspirin RS, and 0.25J mg/mL of USP Caffeine RS, J solution, 3.0 mL of Internal standard solution, and 20 mL of
being the ratio of the labeled amount, in mg, of aspirin to
water to a 50-mL volumetric flask. Mix, and allow to stand USP Salicylic Acid RS
for about 30 s. Dilute with Solution A to volume. Use within
8 h.
Analysis: Proceed as directed for Analysis in the Assay.
Calculate, individually, the percentages of C8H9NO2, C9H8O4, Acetaminophen and Caffeine Tablets
and C8H10N4O2 dissolved: (Comment on this Monograph)id=m246=Acetaminophen and
Caffeine Tablets=A-Monos.pdf)
Result = (RU/RS) (CS/CU) 100
DEFINITION
RU = ratio of the peak responses of the analyte and Acetaminophen and Caffeine Tablets contain NLT 90.0% and
internal standard peak from the Sample solution NMT 110.0% of the labeled quantities of acetaminophen
RS = ratio of the peak responses of the analyte and (C8H9NO2) and caffeine (C8H10N4O2).
internal standard peak from the Standard
solution IDENTIFICATION
CS = concentration of the corresponding USP The retention times of the Sample solution correspond to
Reference Standard in the Standard solution those of the Standard solution, relative to the internal
(mg/mL) standard, obtained in the Assay.
CU = nominal concentration of the analyte in the
Sample solution (mg/mL) ASSAY
Tolerances: NLT 75% (Q) of the labeled amounts of PROCEDURE
C8H9NO2, C9H8O4, and C8H10N4O2 is dissolved. Mobile phase: Methanol, glacial acetic acid, and water
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements (28:3:69)
for Content Uniformity with respect to acetaminophen, Internal standard solution: 6 mg/mL of benzoic acid in
aspirin, and caffeine methanol
Solution A: Methanol and glacial acetic acid (95:5)
IMPURITIES Standard stock solution: 0.25 mg/mL of USP
Organic Impurities Acetaminophen RS and 0.25J mg/mL USP Caffeine RS in
PROCEDURE: LIMIT OF SALICYLIC ACID Solution A; J being the ratio of the labeled amount, in mg, of
Mobile phase and Solution A: Prepare as directed in the caffeine to the labeled amount, in mg, of acetaminophen
Assay. per Tablet.
Standard solution: 0.02 mg/mL of USP Salicylic Acid RS in Standard solution: Transfer 20.0 mL of Standard stock
Solution A solution and 3.0 mL of Internal standard solution to a 50-mL
Sample solution: Transfer an equivalent to 250 mg of volumetric flask, and dilute to volume with Solution A (0.1
aspirin, from finely powdered Tablets (NLT 20), to a 100- mg/mL of USP Acetaminophen RS and 0.1J mg/mL of USP
mL volumetric flask. Add 75 mL of Solution A, and shake by Caffeine RS).
mechanical means for 30 min. Dilute with Solution A to Sample stock solution: Finely powder NLT 20 Tablets.
volume. Transfer a portion of the powder, equivalent to 250 mg of
Chromatographic system acetaminophen, to a 100-mL volumetric flask. Add 75 mL of
(See Chromatography 621, System Suitability.) Solution A, and shake by mechanical means for 30 min.
Mode: LC Dilute with Solution A to volume.
Detector: UV 302 nm Sample solution: Transfer 2.0 mL of the Sample stock
Column: 4.6-mm 10-cm; 5-m packing L1 solution and 3.0 mL of Internal standard solution to a 50-mL
Temperature: 45 1 volumetric flask, and dilute with Solution A to volume.
Flow rate: 2 mL/min Chromatographic system
Injection size: 10 L (See Chromatography 621, System Suitability.)
System suitability Mode: LC
Sample: Standard solution Detector: UV 275 nm
Suitability requirements Column: 4.6-mm 10-cm; 5-m packing L1
Tailing factor: NMT 1.6 Column temperature: 45 1
Relative standard deviation: NMT 3.0% Flow rate: 2 mL/min
Analysis Injection size: 10 L
Samples: Standard solution and Sample solution System suitability
Calculate the percentage of salicylic acid in the portion of Sample: Standard solution
Tablets taken: [NOTEThe relative retention times for acetaminophen,
caffeine, and benzoic acid are about 0.3, 0.5, and 1.0,
Result = (rU/rS) (CS/CU) 100 respectively.]
rU = peak response from the Sample solution Resolution: NLT 1.4 between any of the analyte and
rS = peak response from the Standard solution internal standard peaks
CS = concentration of USP Salicylic Acid RS in the Tailing factor: NMT 1.2 for each analyte peak
Standard solution (mg/mL) Relative standard deviation: NMT 2.0%
CU = nominal concentration of aspirin in the Sample Analysis
solution (mg/mL) Samples: Standard solution and Sample solution
Acceptance criteria: NMT 3.0% Calculate, individually, the percentages of C8H9NO2 and
C8H10N4O2 in the Tablets:
PACKAGING AND STORAGE: Preserve in tight containers, and Result = (RU/RS) (CS/CU) 100
USP REFERENCE STANDARDS 11 RU = ratio of the peak responses of the analyte and
USP Acetaminophen RS internal standard peaks from the Sample
USP Aspirin RS solution
USP Caffeine RS
RS = ratio of the peak responses of the analyte and Chlorpheniramine, Dextromethorphan, and
internal standard peaks from the Standard Phenylpropanolamine=A-Monos.pdf)
solution
CS = concentration of the corresponding USP DEFINITION
Reference Standard in the Standard solution Capsules Containing at Least Three of the Following
(mg/mL) Acetaminophen and Salts of Chlorpheniramine,
CU = nominal concentration of the corresponding Dextromethorphan, and Phenylpropanolamine contain NLT
analyte in the Sample solution (mg/mL) 90.0% and NMT 110.0% of the labeled amounts of
Acceptance criteria: 90.0%110.0% of C8H9NO2 and acetaminophen (C8H9NO2), chlorpheniramine maleate
C8H10N4O2 (C16H19ClN2 C4H4O4), dextromethorphan hydrobromide
(C18H25NO HBr H2O), and phenylpropanolamine
PERFORMANCE TESTS hydrochloride (C9H13NO HCl).
DISSOLUTION 711 [NOTEThe heading of this monograph does not constitute the
Medium: Water; 900 mL official title. It is not intended that the name described herein
Apparatus 2: 100 rpm be recognized as the official title or the common or usual
Time: 60 min name. The name for each article encompassed by this
Mobile phase, Internal standard solution, Solution A, monograph shall be composed of the names of the active
Standard stock solution, and Chromatographic system: ingredients contained therein, as well as the quantitative
Proceed as directed in the Assay. amount of each active ingredient, and a statement of the
Sample stock solution: Sample per Dissolution 711. function (or purpose) of the ingredient in the article.]
Sample solution: Transfer an aliquot of a filtered portion of
Sample stock solution to a 50-mL volumetric flask in order to IDENTIFICATION
obtain an expected concentration of 0.1 mg/mL of A. If phenylpropanolamine hydrochloride is claimed in the
acetaminophen and 0.1J mg/mL of caffeine. Add 3.0 mL of labeling to be present, the chromatogram of the Sample
Internal standard solution and 20 mL of Solution A, and allow solution, obtained as directed in the Assay for
to stand for 30 s. Dilute with Solution A to volume. Phenylpropanolamine hydrochloride, exhibits a major peak for
[NOTEJ is defined for the Standard stock solution.] phenylpropanolamine, the retention time of which
Standard solution: Transfer 20.0 mL of the Standard stock corresponds to that exhibited by the Standard solution.
solution, 3.0 mL of Internal standard solution, and 20 mL of B. If acetaminophen is claimed in the labeling to be
water to a 50-mL volumetric flask and allow to stand for 30 present, the chromatogram of the Sample solution, obtained
s. Dilute with Solution A to volume. Use within 8 h. as directed in the Assay for Acetaminophen, exhibits a major
Analysis: Proceed as directed for Analysis in the Assay, using peak for acetaminophen, the retention time of which
the Sample solution and Standard solution prepared within corresponds to that exhibited by the Standard solution.
Dissolution. C. If chlorpheniramine maleate is claimed in the labeling to
Calculate the percentages of C8H9NO2 and C8H10N4O2 be present, the chromatogram of the Sample solution,
dissolved: obtained as directed in the Assay for Chlorpheniramine
maleate, exhibits a major peak for chlorpheniramine, the
Result = (RU/RS) (CS/CU) 100 retention time of which corresponds to that exhibited by the
Standard solution.
RU = ratio of the peak responses of the analyte and D. If dextromethorphan hydrobromide is claimed in the
internal standard peaks from the Sample labeling to be present, the chromatogram of the Sample
solution solution, obtained as directed in the Assay for
RS = ratio of the peak responses of the analyte and Dextromethorphan hydrobromide, exhibits a major peak for
internal standard peaks from the Standard dextromethorphan, the retention time of which corresponds
solution to that exhibited by the Standard solution.
CS = concentration of the corresponding USP
Reference Standard in the Standard solution ASSAY
(mg/mL) PHENYLPROPANOLAMINE HYDROCHLORIDE (if present)
CU = nominal concentration of the corresponding Mobile phase: Methanol and water (60:40) containing
analyte in the Sample solution (mg/mL) 0.34 g of monobasic potassium phosphate, 0.15 g of
Tolerances: NLT 75% (Q) of the labeled amounts of triethylamine hydrochloride, 0.25 g of sodium lauryl sulfate,
C8H9NO2 and C8H10N4O2 is dissolved. and 0.1 mL of phosphoric acid in each 100 mL of solution
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Standard stock solution: 0.5 mg/mL USP
Phenylpropanolamine Hydrochloride RS in Mobile phase
ADDITIONAL REQUIREMENTS Standard solution: 1 mL of Standard stock solution and 8
PACKAGING AND STORAGE: Preserve in tight containers, and mL of Mobile phase. Dilute with water to 10 mL.
store at controlled room temperature. Chlorpheniramine standard solution: Prepare as directed
USP REFERENCE STANDARDS 11 for Standard solution in the Assay for Chlorpheniramine
USP Acetaminophen RS maleate.
USP Caffeine RS Dextromethorphan standard solution: Prepare as directed
for Standard solution in the Assay for Dextromethorphan
hydrobromide.
System suitability solution 1 (for Capsules that contain
Capsules Containing at Least Three of chlorpheniramine salt): Standard solution and
the FollowingAcetaminophen and Chlorpheniramine standard solution (1:1)
Salts of Chlorpheniramine, System suitability solution 2 (for Capsules that contain no
Dextromethorphan, and chlorpheniramine salt): Standard solution and
Dextromethorphan standard solution (1:1)
Phenylpropanolamine Sample stock solution: Transfer NLT 10 Capsules to a 500-
(Comment on this Monograph)id=m247=Capsules Containing mL volumetric flask. Add 100 mL of water and 10 mL of 5%
at Least Three of the Following-Acetaminophen and Salts of
phosphoric acid, and gently heat until the Capsules are fully Mode: LC
dispersed. Cool the solution to room temperature, dilute Detector: UV 280 nm
with water to volume, and filter. Column: 4.6-mm 15-cm; packing L7
Sample solution: Equivalent of 0.05 mg/mL of Flow rate: 1 mL/min
phenylpropanolamine hydrochloride from Sample stock Injection size: 5 L
solution diluted with water System suitability
Chromatographic system Sample: Standard solution
(See Chromatography 621, System Suitability.) Suitability requirements
Mode: LC Tailing factor: NMT 2.0
Detector: UV 214 nm Relative standard deviation: NMT 2.0%
Column: 4.6-mm 15-cm; packing L11 Analysis
Flow rate: 1.5 mL/min Samples: Standard solution and Sample solution
Injection size: 10 L Calculate the percentage of C8H9NO2:
System suitability
Sample: Standard solution and System suitability solution 1 Result = (rU/rS) (CS/CU) 100
or System suitability solution 2
Suitability requirements rU = acetaminophen peak response from the Sample
Resolution: NLT 2.0 between phenylpropanolamine and solution
chlorpheniramine or between phenylpropanolamine and rS = peak response from the Standard solution
dextromethorphan for the appropriate System suitability CS = concentration of USP Acetaminophen RS in the
solution Standard solution (mg/mL)
Tailing factor: NMT 2.0 for the phenylpropanolamine CU = nominal concentration of acetaminophen in the
peak, Standard solution Sample solution (mg/mL)
Relative standard deviation: NMT 2.0%, Standard Acceptance criteria: 90.0%110.0%
solution CHLORPHENIRAMINE MALEATE (if present)
Analysis Mobile phase, System suitability solutions,
Samples: Standard solution and Sample solution Chromatographic system, and System suitability:
Calculate the percentage of C9H13NO HCl: Proceed as directed in the Assay for Phenylpropanolamine
hydrochloride.
Result = (rU/rS) (CS/CU) 100 Standard stock solution: 0.8 mg/mL of USP
Chlorpheniramine Maleate RS
rU = phenylpropanolamine peak response from the Standard solution: 8 g/mL of USP Chlorpheniramine
Sample solution Maleate RS from the Standard stock solution diluted with
rS = peak response from the Standard solution 0.1% phosphoric acid
CS = concentration of USP Phenylpropanolamine Sample stock solution: Transfer NLT 10 Capsules to a 500-
Hydrochloride RS in the Standard solution mL volumetric flask. Add 100 mL of water and 10 mL of 5%
(mg/mL) phosphoric acid, and gently heat until the Capsules are fully
CU = nominal concentration of the dispersed. Cool the solution to room temperature, dilute
phenylpropanolamine hydrochloride in the with water to volume, and filter.
Sample solution (mg/mL) Sample solution: Equivalent of 8 g/mL of
Acceptance criteria: 90.0%110.0% chlorpheniramine maleate from Sample stock solution diluted
ACETAMINOPHEN (if present) with 0.1% phosphoric solution
Mobile phase: Methanol, water, and glacial acetic acid Analysis
(20:79:1) Samples: Standard solution and Sample solution
Standard solution: Transfer 25 mg of USP Acetaminophen Calculate the percentage of C16H19ClN2 C4H4O4:
RS to a 100-mL volumetric flask. Dissolve in 4 mL of
methanol. Add 0.2 mL of phosphoric acid, and dilute with Result = (rU/rs) (CS/CU) 100
water to volume.
Sample stock solution: Transfer NLT 10 Capsules to a 500- rU = chlorpheniramine maleate peak response from
mL volumetric flask. Add 100 mL of water and 10 mL of 5% the Sample solution
phosphoric acid, and gently heat until the Capsules are fully rS = peak response from the Standard solution
dispersed. Cool the solution to room temperature, and CS = concentration of USP Chlorpheniramine Maleate
dilute with water to volume. RS in the Standard solution (mg/mL)
Sample solution: Equivalent of 0.25 mg/mL CU = nominal concentration of chlorpheniramine
acetaminophen from Sample stock solution diluted with 0.1% maleate in the Sample solution (mg/mL)
phosphoric acid
Acceptance criteria: 90.0%110.0% Oral Solution Containing at Least Three

DEXTROMETHORPHAN HYDROBROMIDE (if present) of the FollowingAcetaminophen and
Mobile phase, System suitability solutions, Salts of Chlorpheniramine,
Chromatographic system, and System suitability:
Proceed as directed in the Assay for Phenylpropanolamine Dextromethorphan, and
hydrochloride. Phenylpropanolamine
Standard stock solution: 0.6 mg/mL of USP (Comment on this Monograph)id=m248=Oral Solution
Dextromethorphan Hydrobromide RS Containing at Least Three of the Following-Acetaminophen and
Standard solution: 0.06 mg/mL of USP Dextromethorphan Salts of Chlorpheniramine, Dextromethorphan, and
Hydrobromide RS from Standard stock solution diluted with Phenylpropanolamine=A-Monos.pdf)
0.1% phosphoric acid
Sample stock solution: Transfer NLT 10 Capsules to a 500- DEFINITION
mL volumetric flask. Add 100 mL of water and 10 mL of 5% Oral Solution Containing at Least Three of the Following
phosphoric acid, and gently heat until the Capsules are fully Acetaminophen and Salts of Chlorpheniramine,
dispersed. Cool the solution to room temperature, dilute Dextromethorphan, and Phenylpropanolamine contains NLT
with water to volume, and filter. 90.0% and NMT 110.0% of the labeled amounts of
Sample solution: Equivalent of 0.06 mg/mL of acetaminophen (C8 H9NO2), chlorpheniramine maleate (C16H19
dextromethorphan hydrobromide from Sample stock solution ClN2 C4H4O4), dextromethorphan hydrobromide (C18H25NO
diluted with 0.1% phosphoric acid HBr H2O), and phenylpropanolamine hydrochloride (C9H13NO
Analysis HCl).
Samples: Standard solution and Sample solution [NOTEThe heading of this monograph does not constitute the
Calculate the percentage of C18H25NO HBr H2O: official title. It is not intended that the name described herein
be recognized as the official title or the common or usual
Result = (rU/rS) (CS/Cu) (Mr1/Mr2) 100 name. The name for each article encompassed by this
monograph shall be composed of the names of the active
rU = dextromethorphan hydrobromide peak response ingredients contained therein, as well as the quantitative
from the Sample solution amount of each active ingredient, and a statement of the
rS = peak response from the Standard solution function (or purpose) of the ingredient in the article.]
CS = concentration of USP Dextromethorphan
Hydrobromide RS in the Standard solution IDENTIFICATION
(mg/mL) A. If phenylpropanolamine hydrochloride is claimed in the
CU = nominal concentration of dextromethorphan labeling to be present, the retention time of the peak of the
hydrobromide in the Sample solution (mg/mL) Sample solution corresponds to that of the Standard solution,
Mr1 = molecular weight of dextromethorphan as obtained in the Assay for Phenylpropanolamine
hydrobromide monohydrate, 370.33 hydrochloride.
Mr2 = molecular weight of dextromethorphan B. If acetaminophen is claimed in the labeling to be
hydrobromide anhydrous, 352.32 present, the retention time of the peak of the Sample
Acceptance criteria: 90.0%110.0% solution corresponds to that of the Standard solution, as
obtained in the Assay for Acetaminophen.
PERFORMANCE TESTS C. If chlorpheniramine maleate is claimed in the labeling to
DISSOLUTION, Procedure for a Pooled Sample 711 be present, the retention time of the peak of the Sample
Medium: Water; 900 mL solution corresponds to that of the Standard solution, as
Apparatus 1: 50 rpm obtained in the Assay for Chlorpheniramine maleate.
Time: 45 min D. If dextromethorphan hydrobromide is claimed in the
Sample solution: Mix 9.0 mL of a filtered portion of the labeling to be present, the retention time of the peak of the
solution under test with 1.0 mL of 1% phosphoric acid Sample solution corresponds to that of the Standard solution,
solution. as obtained in the Assay for Dextromethorphan hydrobromide.
Analysis: Determine the amounts of phenylpropanolamine
hydrochloride, acetaminophen, chlorpheniramine maleate, ASSAY
and dextromethorphan hydrobromide dissolved, employing PHENYLPROPANOLAMINE HYDROCHLORIDE (if present)
the procedures set forth in the Assay for Mobile phase: 3.4 mg/mL of monobasic potassium
Phenylpropanolamine hydrochloride, Assay for Acetaminophen, phosphate, 1.5 mg/mL of triethylamine hydrochloride, 2.5
Assay for Chlorpheniramine maleate, and Assay for mg/mL of sodium lauryl sulfate, and 1L/mL of phosphoric
Dextromethorphan hydrobromide, respectively, making any acid in methanol and water (60:40)
necessary volumetric adjustments. Standard stock solution: 0.5 mg/mL of USP
Tolerances: NLT 75% (Q) of the labeled amounts of Phenylpropanolamine Hydrochloride RS in water
C9H13NO HCl, C8H9NO2, C16H19ClN2 C4H4O4, and Standard solution: Standard stock solution, Mobile phase,
C18H25NO HBr H2O and water (1:8:1)
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Chlorpheniramine standard solution: Use Standard
solution from the Assay for Chlorpheniramine maleate.
ADDITIONAL REQUIREMENTS Dextromethorphan standard solution: Use Standard
PACKAGING AND STORAGE: Preserve in tight containers. solution from the Assay for Dextromethorphan hydrobromide.
LABELING: The label for each article encompassed by this System suitability solution A (for Oral Solution that contains
monograph bears a name composed of the active either all the four ingredients or a combination of three
ingredients. The label states the name and quantity of each containing chlorpheniramine salt): Standard solution and
active ingredient and indicates its function (or purpose) in the Chlorpheniramine standard solution (1:1)
the article. System suitability solution B (for Oral Solution that contains
USP REFERENCE STANDARDS 11 no chlorpheniramine salt): Standard solution and
USP Acetaminophen RS Dextromethorphan standard solution (1:1)
USP Chlorpheniramine Maleate RS
USP Dextromethorphan Hydrobromide RS
USP Phenylpropanolamine Hydrochloride RS
Sample solution: Equivalent to 0.05 mg/mL of Acceptance criteria: 90.0%110.0%

phenylpropanolamine hydrochloride in Mobile phase and CHLORPHENIRAMINE MALEATE (if present)
water (4:1) Mobile phase, System suitability solutions,
Chromatographic system Chromatographic system, and System suitability:
(See Chromatography 621, System Suitability.) Proceed as directed in the Assay for Phenylpropanolamine
Mode: LC hydrochloride.
Detector: UV 214 nm Standard stock solution: 0.08 mg/mL of USP
Column: 4.6-mm 15-cm; packing L11 Chlorpheniramine Maleate RS in water
Flow rate: 1.5 mL/min Standard solution: Standard stock solution, Mobile phase,
Injection size: 20 L and water (1:8:1)
System suitability Sample solution: Equivalent to 8 g/mL of acetaminophen
Samples: Standard solution, System suitability solution A (10 maleate in Mobile phase and water (4:1)
L), or System suitability solution B (10 L) Analysis
Suitability requirements Samples: Standard solution and Sample solution
Resolution: NLT 2.0 between phenylpropanolamine and Calculate the percentage of C16H19ClN2 C4H4O4 in the
chlorpheniramine or between phenylpropanolamine and volume of Oral Solution taken:
dextromethorphan, System suitability solution A or B
Tailing factor: NMT 2.0 for the phenylpropanolamine Result = (rU/rS) (CS/CU) 100
peak, Standard solution
Relative standard deviation: NMT 2.0%, Standard rU = chlorpheniramine peak response from the
solution Sample solution
Analysis rS = peak response from the Standard solution
Samples: Standard solution and Sample solution CS = concentration of USP Chlorpheniramine Maleate
Calculate the percentage of C9H13NO HCl in the volume of RS in the Standard solution (mg/mL)
Oral Solution taken: CU = nominal concentration of chlorpheniramine
maleate in the Sample solution (mg/mL)
Result = (rU/rS) (CS/CU) 100 Acceptance criteria: 90.0%110.0%
DEXTROMETHORPHAN HYDROBROMIDE (if present)
rU = phenylpropanolamine peak response from the Mobile phase, System suitability solutions,
Sample solution Chromatographic system, and System suitability:
rS = peak response from the Standard solution Proceed as directed in the Assay for Phenylpropanolamine
CS = concentration of USP Phenylpropanolamine hydrochloride.
Hydrochloride RS in the Standard solution Standard stock solution: 0.4 mg/mL of USP
(mg/mL) Dextromethorphan Hydrobromide RS in water
CU = nominal concentration of phenylpropanolamine Standard solution: Standard stock solution, Mobile phase,
hydrochloride in the Sample solution (mg/mL) and water (1:8:1)
Acceptance criteria: 90.0%110.0% Sample solution: Equivalent to 0.04 mg/mL of
ACETAMINOPHEN (if present) dextromethorphan hydrobromide in Mobile phase and water
Mobile phase: Methanol, glacial acetic acid, and water (4:1)
(20:1:79). Analysis
Standard solution: Dissolve 16.5 mg of USP Samples: Standard solution and Sample solution
Acetaminophen RS in 2.5 mL of methanol, then dilute to Calculate the percentage of C18H25NO HBr H2O in the
100 mL with water (0.165 mg/L). volume of Oral Solution taken:
Sample solution: Equivalent to 0.165 mg/mL of
acetaminophen in methanol and water (1:39) Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
(See Chromatography 621, System Suitability.) rU = dextromethorphan peak response from the
Mode: LC Sample solution
Detector: UV 280 nm rS = peak response from the Standard solution
Column: 4.6-mm 15-cm; packing L7 CS = concentration of USP Dextromethorphan
Flow rate: 1 mL/min Hydrobromide RS in the Standard solution
Injection size: 5 L (mg/mL)
System suitability CU = nominal concentration of dextromethorphan
Sample: Standard solution hydrobromide in the Sample solution (mg/mL)
Suitability requirements Mr1 = molecular weight of dextromethorphan
Tailing factor: NMT 2.0 hydrobromide monohydrate, 370.33
Relative standard deviation: NMT 2.0% Mr2 = molecular weight of anhydrous
Analysis dextromethorphan hydrobromide, 352.32
Samples: Standard solution and Sample solution Acceptance criteria: 90.0%110.0%
Calculate the percentage of C8H9NO2 in the volume of Oral
Solution taken: OTHER COMPONENTS
ALCOHOL DETERMINATION (if present), Method II 611:
Result = (rU/rS) (CS/CU) 100 90.0%110.0% of the labeled amount of C2 H5 OH
rU = acetaminophen peak response from the Sample SPECIFIC TESTS

solution MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED
rS = peak response from the Standard solution MICROORGANISMS 62: It meets the requirements of the
CS = concentration of USP Acetaminophen RS in the tests for absence of Salmonella species and Escherichia coli.
Standard solution (mg/mL) The total aerobic microbial count does not exceed 100
CU = nominal concentration of acetaminophen in the cfu/g, and the total combined molds and yeast count does
Sample solution (mg/mL) not exceed 10 cfu/g.
PH 791: 2.67.5 Standard stock solution: 2.5 mg/mL of USP

Phenylpropanolamine Hydrochloride RS in water
ADDITIONAL REQUIREMENTS Standard solution: Standard stock solution, methanol, and
PACKAGING AND STORAGE: Preserve in tight containers. 0.1% phosphoric acid (1:5:44)
LABELING: The label for each article encompassed by this Chlorpheniramine standard solution: Use Standard
monograph bears a name composed of the active solution from the Assay for Chlorpheniramine maleate.
ingredients. The label states the name and quantity of each Dextromethorphan standard solution: Use Standard
active ingredient and indicates its function (or purpose) in solution from the Assay for Dextromethorphan hydrobromide.
the article. System suitability solution A (for Tablets that contain either
USP REFERENCE STANDARDS 11 all the four ingredients or a combination of three containing
USP Acetaminophen RS chlorpheniramine salt): Standard solution and the
USP Chlorpheniramine Maleate RS Chlorpheniramine standard solution (1:1)
USP Dextromethorphan Hydrobromide RS System suitability solution B (for Tablets that contain no
USP Phenylpropanolamine Hydrochloride RS chlorpheniramine salt): Standard solution and
Sample solution: Transfer an equivalent to 2.5 mg of
Tablets Containing at Least Three of phenylpropanolamine hydrochloride, from finely powdered
Tablets (NLT 20), to a 50-mL volumetric flask. Add 5 mL of
the FollowingAcetaminophen and methanol, and sonicate to disperse the powder. Dilute with
Salts of Chlorpheniramine, 0.1% phosphoric acid to volume and filter.
Dextromethorphan, and Chromatographic system
Phenylpropanolamine (See Chromatography 621, System Suitability.)
Mode: LC
(Comment on this Monograph)id=m255=Tablets Containing at Detector: UV 214 nm
Least Three of the Following-Acetaminophen and Salts of Column: 4.6-mm 15-cm; packing L11
Chlorpheniramine, Dextromethorphan, and Flow rate: 2 mL/min
Phenylpropanolamine=A-Monos.pdf) Injection size: 20 L
DEFINITION System suitability
Tablets Containing at Least Three of the Following Samples: Standard solution, System suitability solution A (10
Acetaminophen and Salts of Chlorpheniramine, L), or System suitability solution B (10 L)
Dextromethorphan, and Phenylpropanolamine contain NLT Suitability requirements
90.0% and NMT 110.0% of the labeled amounts of Resolution: NLT 2.0 between phenylpropanolamine and
acetaminophen (C8H9NO2), chlorpheniramine maleate chlorpheniramine or between phenylpropanolamine and
(C16H19ClN2 C4H4O4), dextromethorphan hydrobromide dextromethorphan, System suitability solution A or B
(C18H25NO HBr H2O), and phenylpropanolamine Tailing factor: NMT 2.0 for the phenylpropanolamine
hydrochloride (C9H13NO HCl). peak, Standard solution
[NOTEThe heading of this monograph does not constitute the Relative standard deviation: NMT 2.0%, Standard
official title. It is not intended that the name described herein solution
be recognized as the official title or the common or usual Analysis
name. The name for each article encompassed by this Samples: Standard solution and Sample solution
monograph shall be composed of the names of the active Calculate the percentage of C9H13NO HCl in portion of
ingredients contained therein as well as the quantitative Tablets taken:
amount of each active ingredient, and a statement of the Result = (rU/rS) (CS/CU) 100
function (or purpose) of the ingredient in the article.]
IDENTIFICATION rU = phenylpropanolamine peak response from the
A. If phenylpropanolamine hydrochloride is claimed in the Sample solution
labeling to be present, the retention time of the peak of the rS = peak response from the Standard solution
Sample solution corresponds to that of the Standard solution, CS = concentration of USP Phenylpropanolamine
as obtained in the Assay for Phenylpropanolamine Hydrochloride RS in the Standard solution
hydrochloride. (mg/mL)
B. If acetaminophen is claimed in the labeling to be CU = nominal concentration of phenylpropanolamine
present, the retention time of the peak of the Sample hydrochloride in the Sample solution (mg/mL)
solution corresponds to that of the Standard solution, as Acceptance criteria: 90.0%110.0%
obtained in the Assay for Acetaminophen. ACETAMINOPHEN (if present)
C. If chlorpheniramine maleate is claimed in the labeling to Mobile phase: Methanol, glacial acetic acid, and water
be present, the retention time of the peak of the Sample (20:1:79).
solution corresponds to that of the Standard solution, as Standard solution: Dissolve 50 mg of USP Acetaminophen
obtained in the Assay for Chlorpheniramine maleate. RS in 4 mL of methanol, then dilute with 0.1% phosphoric
D. If dextromethorphan hydrobromide is claimed in the acid to 100 mL.
labeling to be present, the retention time of the peak of the Sample solution: Transfer an equivalent to 100 mg of
Sample solution corresponds to that of the Standard solution, acetaminophen, from finely powdered Tablets (NLT 20), to a
as obtained in the Assay for Dextromethorphan hydrobromide. 50-mL volumetric flask. Add 7.5 mL of methanol, and
sonicate to disperse the powder. Add 0.5 mL of phosphoric
ASSAY acid, dilute with water to volume, and filter. Transfer 25.0
PHENYLPROPANOLAMINE HYDROCHLORIDE mL of the filtered solution to a 100-mL volumetric flask, and
Mobile phase: 3.4 mg/mL of monobasic potassium dilute with water to volume.
phosphate, 1.5 mg/mL of triethylamine hydrochloride, 2.5 Chromatographic system
mg/mL of sodium lauryl sulfate, and 1L/mL of phosphoric (See Chromatography 621, System Suitability.)
acid in methanol and water (60:40)
Mode: LC Injection size: 20 L

Detector: UV 280 nm Analysis
Column: 4.6-mm 15-cm; packing L7 Samples: Standard solution and Sample solution
Flow rate: 1 mL/min Calculate the percentage of C18H25NO HBr H2O in the
Injection size: 5 L portion of Tablets taken:
System suitability
Sample: Standard solution Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
Tailing factor: NMT 2.0 rU = dextromethorphan peak response from the
Relative standard deviation: NMT 2.0% Sample solution
Samples: Standard solution and Sample solution CS = concentration of USP Dextromethorphan
Calculate the percentage of C8H9NO2 in the portion of Hydrobromide RS in the Standard solution
Tablets taken: (mg/mL)
CU = nominal concentration of dextromethorphan
Result = (rU/rS) (CS/CU) 100 hydrobromide in the Sample solution (mg/mg)
Mr1 = molecular weight of dextromethorphan
rU = acetaminophen peak response from the Sample hydrobromide monohydrate, 370.33
solution Mr2 = molecular weight of anhydrous
rS = peak response from the Standard solution dextromethorphan hydrobromide, 352.32
CS = concentration of USP Acetaminophen RS in the Acceptance criteria: 90.0%110.0%
CU = nominal concentration of acetaminophen in the PERFORMANCE TESTS
Sample solution (mg/mL) DISSOLUTION, Procedure for a Pooled Sample 711
Acceptance criteria: 90.0%110.0% Medium: 0.1 M hydrochloric acid; 900 mL
CHLORPHENIRAMINE MALEATE (if present) Apparatus 2: 50 rpm
Mobile phase, System suitability solutions, Time: 45 min
Chromatographic system, and System suitability: Sample solution: Mix 9.0 mL of a filtered portion of the
Proceed as directed in the Assay for Pseudoephedrine solution under test with 1.0 mL of 1% phosphoric acid
hydrochloride. solution.
Standard stock solution: 0.8 mg/mL of USP Analysis: Determine the amounts of phenylpropanolamine
Chlorpheniramine Maleate RS in water hydrochloride, acetaminophen, chlorpheniramine maleate,
Standard solution: 8 g/mL of USP Chlorpheniramine and dextromethorphan hydrobromide dissolved, employing
Maleate RS from Standard stock solution in 0.1% phosphoric the Analyses set forth in the Assays for Phenylpropanolamine
acid hydrochloride, Acetaminophen, Chlorpheniramine maleate, and
Sample solution: Transfer an equivalent to 0.4 mg of Dextromethorphan hydrobromide, respectively, making any
chlorpheniramine maleate, from finely powdered Tablets necessary volumetric adjustments.
(NLT 20), to a 50-mL volumetric flask. Add 5 mL of Tolerances: NLT 75% (Q) of the labeled amount of
methanol, and sonicate to disperse the powder. Add 0.2 mL C9H13NO HCl, C8H9NO2, C16H19ClN2 C4H4O4, and
of phosphoric acid, dilute with water to volume, and filter. C18H25NO HBr H2O
Injection size: 20 L UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Analysis
Samples: Standard solution and Sample solution ADDITIONAL REQUIREMENTS
Calculate the percentage of C16H19ClN2 C4H4O4 in the PACKAGING AND STORAGE: Preserve in tight containers.
portion of Tablets taken: LABELING: The label for each article encompassed by this
monograph bears a name composed of the active
Result = (rU/rS) (CS/CU) 100 ingredients. The label states the name and quantity of each
active ingredient and indicates its function (or purpose) in
rU = chlorpheniramine peak response from the the article.
Sample solution USP REFERENCE STANDARDS 11
rS = peak response from the Standard solution USP Acetaminophen RS
CS = concentration of USP Chlorpheniramine Maleate USP Chlorpheniramine Maleate RS
RS in the Standard solution (mg/mL) USP Dextromethorphan Hydrobromide RS
CU = nominal concentration of the Sample solution USP Phenylpropanolamine Hydrochloride RS
(mg/mL)
DEXTROMETHORPHAN HYDROBROMIDE (if present) Capsules Containing at Least Three of
Mobile phase, System suitability solutions,
Chromatographic system, and System suitability: the FollowingAcetaminophen and
Proceed as directed in the Assay for Phenylpropanolamine Salts of Chlorpheniramine,
hydrochloride. Dextromethorphan, and
Standard stock solution: 0.6 mg/mL of USP
Dextromethorphan Hydrobromide RS in water
Pseudoephedrine
Standard solution: 60 g/mL of USP Dextromethorphan (Comment on this Monograph)id=m249=Capsules Containing
Hydrobromide RS from Standard stock solution in 0.1% at Least Three of the Following-Acetaminophen and Salts of
phosphoric acid Chlorpheniramine, Dextromethorphan, and
Sample solution: Transfer an equivalent to 3 mg of Pseudoephedrine=A-Monos.pdf)
dextromethorphan hydrobromide, from finely powdered DEFINITION
Tablets (NLT 20), to a 50-mL volumetric flask. Add 5 mL of Capsules Containing at Least Three of the Following
methanol, and sonicate to disperse the powder. Add 0.2 mL Acetaminophen and Salts of Chlorpheniramine,
of phosphoric acid, dilute with water to volume, and filter.
Dextromethorphan, and Pseudoephedrine contain NLT 90.0% Mode: LC

and NMT 110.0% of the labeled amounts of acetaminophen Detector: UV 214 nm
(C8H9NO2), chlorpheniramine maleate (C16H19ClN2 C4H4O4), Column: 4.6-mm 15-cm; packing L11
dextromethorphan hydrobromide (C18H25NO HBr H2O), and Flow rate: 2 mL/min
pseudoephedrine hydrochloride (C10H15NO HCl) or Injection size: 10 L
pseudoephedrine sulfate [(C10H15NO)2 H2SO4]. System suitability
[NOTEThe heading of this monograph does not constitute the Samples: Standard solution and System suitability solution A
official title. It is not intended that the name described herein or System suitability solution B
be recognized as the official title or the common or usual Suitability requirements
name. The name for each article encompassed by this Resolution: NLT 2.0 between pseudoephedrine and
monograph shall be composed of the names of the active chlorpheniramine or between pseudoephedrine and
ingredients contained therein, as well as the quantitative dextromethorphan, System suitability solution A or B
amount of each active ingredient, and a statement of the Tailing factor: NMT 2.5 for pseudoephedrine peak,
function (or purpose) of the ingredient in the article.] Standard solution
Relative standard deviation: NMT 2.0%, Standard
IDENTIFICATION solution
A. If pseudoephedrine hydrochloride or pseudoephedrine Analysis
sulfate is claimed in the labeling to be present, the retention Samples: Standard solution and Sample solution
time of the peak of the Sample solution corresponds to that Calculate the percentage of C10H15NO HCl in the Capsules
of the Standard solution, as obtained in the Assay for taken:
Pseudoephedrine sulfate.
B. If acetaminophen is claimed in the labeling to be Result = (rU/rS) (CS/CU) 100
present, the retention time of the peak of the Sample
solution corresponds to that of the Standard solution, as rU = pseudoephedrine peak response from the Sample
obtained in the Assay for Acetaminophen. solution
C. If chlorpheniramine maleate is claimed in the labeling to rS = peak response from the Standard solution
be present, the retention time of the peak of the Sample CS = concentration of USP Pseudoephedrine
solution corresponds to that of the Standard solution, as Hydrochloride RS in the Standard solution
obtained in the Assay for Chlorpheniramine maleate. (mg/mL)
D. If dextromethorphan hydrobromide is claimed in the CU = nominal concentration of the Sample solution
labeling to be present, the retention time of the peak of the (mg/mL)
Sample solution corresponds to that of the Standard solution, Acceptance criteria: 90.0%110.0%
as obtained in the Assay for Dextromethorphan hydrobromide. PSEUDOEPHEDRINE SULFATE (where pseudoephedrine sulfate is
the salt form used, if present in the formulation)
ASSAY Mobile phase, System suitability solutions A and B,
PSEUDOEPHEDRINE HYDROCHLORIDE (where pseudoephedrine Chromatographic system, and System suitability:
hydrochloride is the salt form used, if present in the Proceed as directed in the Assay for Pseudoephedrine
formulation) hydrochloride.
Mobile phase: 3.4 mg/mL of monobasic potassium Chlorpheniramine standard solution: Use Standard
phosphate, 1.5 mg/mL of triethylamine hydrochloride, 2.5 solution from the Assay for Chlorpheniramine maleate.
mg/mL of sodium lauryl sulfate, and 1L/mL of phosphoric Dextromethorphan standard solution: Use Standard
acid in methanol and water (60:40). solution from the Assay for Dextromethorphan hydrobromide.
Standard stock solution: 1.5 mg/mL of USP Standard stock solution: 3.0 mg/mL of USP
Pseudoephedrine Hydrochloride RS in water Pseudoephedrine Sulfate RS in water
Standard solution: Standard stock solution, Mobile phase, Standard solution: Standard stock solution, Mobile phase,
and water (1:8:1) and water (1:4:5)
Chlorpheniramine standard solution: Use Standard Sample solution: Proceed as directed for the Sample
solution from the Assay for Chlorpheniramine maleate. solution in the Assay for Pseudoephedrine hydrochloride to
Dextromethorphan standard solution: Use Standard obtain a solution having a concentration of 0.24 mg/mL of
solution from the Assay for Dextromethorphan hydrobromide. pseudoephedrine sulfate.
System suitability solution A (for Oral Solution that contains Analysis
either all the four ingredients or a combination of three Samples: Standard solution and Sample solution
containing chlorpheniramine salt): Standard solution and Calculate the percentage of (C10H15NO)2 H2SO4 in the
Chlorpheniramine standard solution (1:1) Capsules taken:
System suitability solution B (for Oral Solution that contains
no chlorpheniramine salt): Standard solution and Result = (rU/rS) (CS/CU) 100
Sample stock solution: Transfer NLT 10 Capsules to a 500- rU = pseudoephedrine peak response from the Sample
mL volumetric flask. Add 100 mL of water and 10 mL of 5% solution
phosphoric acid, and gently heat until the Capsules are fully rS = peak response from the Standard solution
dispersed. Cool the solution to room temperature, dilute CS = concentration of USP Pseudoephedrine Sulfate
with water to volume, and filter. RS in the Standard solution (mg/mL)
Sample solution: 0.12 mg/mL of pseudoephedrine CU = nominal concentration of the Sample solution
hydrochloride, from Sample stock solution in water (mg/mL)
Acceptance criteria: 90.0%110.0% CU = nominal concentration of the Sample solution

ACETAMINOPHEN (if present) (mg/mL)
Mobile phase: Methanol, glacial acetic acid, and water Acceptance criteria: 90.0%110.0%
(20:1:79) DEXTROMETHORPHAN HYDROBROMIDE (if present)
Standard solution: Dissolve 25 mg of USP Acetaminophen Mobile phase, System suitability solutions,
RS in 4 ml of methanol, then add 0.2 mL of phosphoric acid Chromatographic system, and System suitability:
and dilute to 100 mL with water (0.25 mg/mL). Proceed as directed in the Assay for Pseudoephedrine
Sample solution: Transfer NLT 10 Capsules to a 500-mL hydrochloride.
volumetric flask. Add 100 mL of water and 10 mL of 5% Standard stock solution: 1.5 mg/mL of USP
phosphoric acid, and gently heat until the Capsules are fully Dextromethorphan Hydrobromide RS in water
dispersed. Cool the solution to room temperature, and Standard solution: Standard stock solution, Mobile phase,
dilute with water to volume. Quantitatively dilute a portion and water (1:16:3)
of this solution, if necessary, with 0.1% phosphoric acid to Sample solution: Transfer NLT 10 Capsules to a 500-mL
obtain a solution having a concentration of 0.25 mg/mL of volumetric flask. Add 100 mL of water and 10 mL of 5%
acetaminophen. phosphoric acid, and gently heat until the Capsules are fully
Chromatographic system dispersed. Cool the solution to room temperature, dilute
(See Chromatography 621, System Suitability.) with water to volume, and filter. Quantitatively dilute a
Mode: LC portion of this solution, if necessary, with 0.1% phosphoric
Detector: UV 280 nm acid to obtain a solution having a concentration of 0.04
Column: 4.6-mm 15-cm; packing L7 mg/mL of dextromethorphan hydrobromide.
Flow rate: 1 mL/min Injection size: 10 L
Injection size: 10 L Analysis
System suitability Samples: Standard solution and Sample solution
Sample: Standard solution Calculate the percentage of C18H25NO HBr H2O in the
Suitability requirements Capsules taken:
Relative standard deviation: NMT 2.0% Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
Analysis
Samples: Standard solution and Sample solution rU = dextromethorphan peak response from the
Calculate the percentage of C8H9NO2 in the Capsules Sample solution
taken: rS = peak response from the Standard solution
CS = concentration of USP Dextromethorphan
Result = (rU/rS) (CS/CU) 100 Hydrobromide RS in the Standard solution
(mg/mL)
rU = acetaminophen peak response from the Sample CU = nominal concentration of dextromethorphan
solution hydrobromide in the Sample solution (mg/mL)
rS = peak response from the Standard solution Mr1 = molecular weight of dextromethorphan
CS = concentration of USP Acetaminophen RS in the hydrobromide monohydrate, 370.33
Standard solution (mg/mL) Mr2 = molecular weight of anhydrous
CU = nominal concentration of acetaminophen in the dextromethorphan hydrobromide, 352.32
Sample solution (mg/mL) Acceptance criteria: 90.0%110.0%
CHLORPHENIRAMINE MALEATE (if present) PERFORMANCE TESTS
Mobile phase, System suitability solutions, DISSOLUTION, Procedure for a Pooled Sample 711
Chromatographic system, and System suitability: Medium: Water; 900 mL
Proceed as directed in the Assay for Pseudoephedrine Apparatus 1: 100 rpm
hydrochloride. Time: 45 min
Standard stock solution: 1 mg/mL of USP Sample solution: Mix 9.0 mL of a filtered portion of the
Chlorpheniramine Maleate RS in water solution under test with 1.0 mL of 1% phosphoric acid
Standard solution: Standard stock solution, Mobile phase, solution.
and water (1:80:19) Analysis: Determine the amounts of pseudoephedrine
Sample solution: Transfer NLT 10 Capsules to a 500-mL hydrochloride or pseudoephedrine sulfate (as appropriate),
volumetric flask. Add 100 mL of water and 10 mL of 5% acetaminophen, chlorpheniramine maleate, and
phosphoric acid, and gently heat until the Capsules are fully dextromethorphan hydrobromide dissolved, employing the
dispersed. Cool the solution to room temperature, dilute Analyses set forth in the Assays for Pseudoephedrine
with water to volume, and filter. Quantitatively dilute a hydrochloride or Pseudoephedrine sulfate, Acetaminophen,
portion of this solution, if necessary, with 0.1% phosphoric Chlorpheniramine maleate, and Dextromethorphan
acid to obtain a solution having a concentration of 8 g/mL hydrobromide, respectively, making any necessary volumetric
of chlorpheniramine maleate. adjustments.
Injection size: 10 L Tolerances: NLT 75% (Q) of the labeled amounts of
Analysis C10H15NO HCl or (C10H15 NO)2 H2SO4, C8H9NO2,
Samples: Standard solution and Sample solution C16H19ClN2 C4H4O4, and C18H25NO HBr H2O
Calculate the percentage of C16H19ClN2 C4H4O4 in the UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Capsules taken:
Result = (rU/rS) (CS/CU) 100 PACKAGING AND STORAGE: Preserve in tight containers, and
store at controlled room temperature.
rU = chlorpheniramine peak response from the LABELING: The label for each article encompassed by this
Sample solution monograph bears a name composed of the active
rS = peak response from the Standard solution ingredients contained in the article. The label states the
CS = concentration of USP Chlorpheniramine Maleate name and quantity of each active ingredient and indicates its
RS in the Standard solution (mg/mL) function (or purpose) in the article.
USP REFERENCE STANDARDS 11 Standard solution: 0.24 mg/mL by diluting Standard stock
USP Acetaminophen RS solution with 0.1% phosphoric acid
USP Chlorpheniramine Maleate RS Chlorpheniramine standard solution: Prepare as directed
USP Dextromethorphan Hydrobromide RS for Standard solution in the Assay for Chlorpheniramine
USP Pseudoephedrine Hydrochloride RS maleate.
USP Pseudoephedrine Sulfate RS Dextromethorphan standard solution: Prepare as directed
for Standard solution in the Assay for Dextromethorphan
hydrobromide.
System suitability solution A (for Oral Powder that contains
Oral Powder Containing at Least Three either all of the four ingredients or a combination of three
of the FollowingAcetaminophen and that includes chlorpheniramine salt): Standard solution and
Salts of Chlorpheniramine, the Chlorpheniramine standard solution (1:1)
Dextromethorphan, and System suitability solution B (for Oral Powder that contains
no chlorpheniramine salt): Standard solution and the
Pseudoephedrine Dextromethorphan standard solution (1:1)
(Comment on this Monograph)id=m251=Oral Powder Sample stock solution: Transfer the contents of 10 unit-
Containing at Least Three of the Following-Acetaminophen and dose containers of the Oral Powder to a 2000-mL volumetric
Salts of Chlorpheniramine, Dextromethorphan, and flask. Add 1000 mL of water and 2.0 mL of phosphoric acid.
Pseudoephedrine=A-Monos.pdf) Gently heat to 60 until the powder is fully dispersed. Cool
the flask to room temperature, add 40 mL of methanol, and
DEFINITION dilute with water to volume.
Oral Powder Containing at Least Three of the Following Sample solution: Equivalent of 0.24 mg/mL of
Acetaminophen and Salts of Chlorpheniramine, pseudoephedrine hydrochloride from Sample stock solution in
Dextromethorphan, and Pseudoephedrine contains NLT 90.0% 0.1% phosphoric acid
and NMT 110.0% of the labeled amounts of acetaminophen Chromatographic system
(C8H9NO2), chlorpheniramine maleate (C16H19ClN2 C4H4O4), (See Chromatography 621, System Suitability.)
dextromethorphan hydrobromide (C18H25NO HBr H2O), and Mode: LC
pseudoephedrine hydrochloride (C10H15NO HCl) or Detector: UV 214 nm
pseudoephedrine sulfate [(C10H15NO)2 H2SO4]. Column: 4.6-mm 15-cm; packing L11
[NOTEThe heading of this monograph does not constitute the Flow rate: 2 mL/min
official title. It is not intended that the name described herein Injection volume: 10 L
be recognized as the official title or the common or usual System suitability
name. The name for each article encompassed by this Samples: Standard solution and System suitability solution A
monograph shall be composed of the names of the active or System suitability solution B
ingredients contained therein, as well as the quantitative Suitability requirements
amount of each active ingredient, and a statement of the Resolution: NLT 2.0 between pseudoephedrine and
function (or purpose) of the ingredient in the article.] chlorpheniramine or between pseudoephedrine and
IDENTIFICATION dextromethorphan, System suitability solution A or System
A. If pseudoephedrine hydrochloride or pseudoephedrine suitability solution B
sulfate is claimed in the labeling to be present, the retention Tailing factor: NMT 2.5 for the pseudoephedrine peak,
time of the major peak for pseudoephedrine of the Sample Standard solution
solution corresponds to that of the Standard solution, as Relative standard deviation: NMT 2.0%, Standard
obtained in the Assay for Pseudoephedrine hydrochloride or solution
the Assay for Pseudoephedrine sulfate. Analysis
B. If acetaminophen is claimed in the labeling to be Samples: Standard solution and Sample solution
present, the retention time of the major peak for Calculate the percentage of C10H15NO HCl in each unit-
acetaminophen of the Sample solution corresponds to that of dose container of Oral Powder taken:
the Standard solution, as obtained in the Assay for Result = (rU/rS) (CS/CU) 100
Acetaminophen.
C. If chlorpheniramine maleate is claimed in the labeling to rU = peak response for pseudoephedrine
be present, the retention time of the major peak for hydrochlorde from the Sample solution
chlorpheniramine of the Sample solution corresponds to that rS = peak response from the Standard solution
of the Standard solution, as obtained in the Assay for CS = concentration of USP Pseudoephedrine
Chlorpheniramine maleate. Hydrochloride RS in the Standard solution
D. If dextromethorphan hydrobromide is claimed in the (mg/mL)
labeling to be present, the retention time of the major peak CU = nominal concentration of pseudoephedrine
for dextromethorphan of the Sample solution corresponds to hydrochloride in the Sample solution (mg/mL)
that of the Standard solution, as obtained in the Assay for Acceptance criteria: 90.0%110.0%
Dextromethorphan hydrobromide. PSEUDOEPHEDRINE SULFATE (where pseudoephedrine sulfate is
ASSAY the salt form used, if present in the formulation)
PSEUDOEPHEDRINE HYDROCHLORIDE (where pseudoephedrine Mobile phase, System suitability solutions,
hydrochloride is the salt form used, if present in the Chromatographic system, and System suitability:
formulation) Proceed as directed in the Assay for Pseudoephedrine
Mobile phase: Mixture of methanol and water (60:40) hydrochloride.
containing 0.34 g of monobasic potassium phosphate, 0.3 g Chlorpheniramine standard solution: Prepare as directed
of triethylamine hydrochloride, 0.15 g of sodium lauryl for Standard solution in the Assay for Chlorpheniramine
sulfate, and 0.1 mL of phosphoric acid in each 100 mL of maleate.
solution Dextromethorphan standard solution: Prepare as directed
Standard stock solution: 3.0 mg/mL of USP for Standard solution in the Assay for Dextromethorphan
Pseudoephedrine Hydrochloride RS hydrobromide.
Standard stock solution: 6.0 mg/mL of USP rU = peak response from the Sample solution
Pseudoephedrine Sulfate RS rS = peak response from the Standard solution
Standard solution: 0.48 mg/mL by diluting Standard stock CS = concentration of USP Chlorpheniramine Maleate
solution with 0.1% phosphoric acid RS in the Standard solution (g/mL)
Sample solution: Proceed as directed for the Sample CU = nominal concentration of chlorpheniramine
solution in the Assay for Pseudoephedrine hydrochloride to maleate in the Sample solution (g/mL)
obtain a solution having a nominal concentration of 0.48 Acceptance criteria: 90.0%110.0%
mg/mL of pseudoephedrine sulfate. DEXTROMETHORPHAN HYDROBROMIDE
Analysis: Proceed as directed for Analysis in the Assay for (if present)
Pseudoephedrine hydrochloride. Mobile phase, System suitability solutions,
Calculate the percentage of (C10H15NO)2 H2SO4 in each Chromatographic system, and System suitability:
unit-dose container of Oral Powder taken: Proceed as directed in the Assay for Pseudoephedrine
Hydrochloride.
Dextromethorphan Hydrobromide RS
rU = peak response for pseudoephedrine Standard solution: 0.08 mg/mL solution from Standard
hydrochloride from the Sample solution stock solution and 0.1% phosphoric acid
rS = peak response from the Standard solution Sample stock solution: Transfer the contents of 10 unit-
CS = concentration of USP Pseudoephedrine Sulfate RS dose containers of Oral Powder to a 2000-mL volumetric
in the Standard solution (mg/mL) flask. Add 1000 mL of water and 2 mL of phosphoric acid.
CU = nominal concentration of pseudoephedrine Gently heat to 60 until the powder is fully dispersed. Cool
sulfate in the Sample solution (mg/mL) the flask to room temperature, add 40 mL of methanol, and
Acceptance criteria: 90.0%110.0% dilute with water to volume.
ACETAMINOPHEN (if present) Sample solution: Nominally 0.08 mg/mL of
Mobile phase, Standard solution, and Chromatographic dextromethorphan hydrobromide from Sample stock solution
system: Proceed as directed in the Assay for and 0.1% phosphoric acid
Pseudoephedrine hydrochloride. Analysis
Sample stock solution: Transfer the contents of 10 unit- Samples: Standard solution and Sample solution
dose containers of the Oral Powder to a 2000-mL volumetric Calculate the percentage of C18H25NO HBr H2O in each
flask. Add 1000 mL of water and 2 mL of phosphoric acid. unit-dose container of Oral Powder taken:
Gently heat to 60 until the powder is fully dispersed. Cool
the flask to room temperature, add 40 mL of methanol, and Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
dilute with water to volume.
Sample solution: Nominally 0.50 mg/mL of acetaminophen rU = peak response for dextromethorphan
from Sample stock solution and 0.1% phosphoric acid hydrobromide from the Sample solution
Samples: Standard solution and Sample solution CS = concentration of USP Dextromethorphan
Calculate the percentage of C8H9NO2 in each unit-dose Hydrobromide RS in the Standard solution
container of Oral Powder taken: (mg/mL)
CU = nominal concentration of dextromethorphan
Result = (rU/rS) (CS/CU) x100 hydrobromide in the Sample solution (mg/mL)
Mr1 = molecular weight of dextromethorphan
rU = peak response from the Sample solution hydrobromide monohydrate, 370.33
rS = peak response from the Standard solution Mr2 = molecular weight of anhydrous
CS = concentration of USP Acetaminophen RS in the dextromethorphan hydrobromide, 352.32
CU = nominal concentration of acetaminophen in the Acceptance criteria: 90.0%110.0%
Sample solution (mg/mL) PERFORMANCE TESTS
Acceptance criteria: 90.0%110.0% UNIFORMITY OF DOSAGE UNITS 905
CHLORPHENIRAMINE MALEATE (if present) For oral powder packaged in single-unit containers:
Mobile phase and Chromatographic system: Proceed as Meets the requirements
directed in the Assay for Pseudoephedrine hydrochloride. MINIMUM FILL 755: Meets the requirements
Chlorpheniramine Maleate RS ADDITIONAL REQUIREMENTS
Standard solution: 8 g/mL of chlorpheniramine maleate PACKAGING AND STORAGE: Preserve in tight containers, and
from Standard stock solution and 0.1% phosphoric acid store at controlled room temperature.
Sample stock solution: Transfer the contents of 10 unit- LABELING: The label for each article encompassed by this
dose containers of Oral Powder to a 2000-mL volumetric monograph bears a name composed of the active
flask. Add 1000 mL of water and 2 mL of phosphoric acid. ingredients. The label states the name and quantity of each
Gently heat to 60 until the powder is fully dispersed. Cool active ingredient and indicates its function (or purpose) in
the flask to room temperature, add 40 mL of methanol, the article.
dilute with water to volume, and mix. USP REFERENCE STANDARDS 11
Sample solution: Nominally 8 g/mL solution of USP Acetaminophen RS
chlorpheniramine maleate from Sample stock solution and USP Chlorpheniramine Maleate RS
0.1% phosphoric acid USP Dextromethorphan Hydrobromide RS
Analysis USP Pseudoephedrine Hydrochloride RS
Samples: Standard solution and Sample solution USP Pseudoephedrine Sulfate RS
Calculate the percentage of C16H19ClN2 C4H4O4 in each
unit-dose container of Oral Powder taken:
Oral Solution Containing at Least Three Sample solution: Equivalent to 0.15 mg/mL of
of the FollowingAcetaminophen and pseudoephedrine hydrochloride from Oral solution in Mobile
Salts of Chlorpheniramine, phase and water (4:1)
Dextromethorphan, and (See Chromatography 621, System Suitability.)
Pseudoephedrine Mode: LC
(Comment on this Monograph)id=m254=Oral Solution Detector: UV 214 nm
Containing at Least Three of the Following-Acetaminophen and Column: 4.6-mm 15-cm; packing L11
Salts of Chlorpheniramine, Dextromethorphan, and Flow rate: 2 mL/min
Pseudoephedrine=A-Monos.pdf) Injection size: 10 L
System suitability
DEFINITION Samples: Standard solution and System suitability solution A
Oral Solution Containing at Least Three of the Following or System suitability solution B
Acetaminophen and Salts of Chlorpheniramine, Suitability requirements
Dextromethorphan, and Pseudoephedrine contains NLT 90.0% Resolution: NLT 2.0 between pseudoephedrine and
and NMT 110.0% of the labeled amounts of acetaminophen chlorpheniramine or between pseudoephedrine and
(C8H9NO2), chlorpheniramine maleate (C16H19ClN2 C4H4O4), dextromethorphan, System suitability solution A or B
dextromethorphan hydrobromide (C18H25NO HBr H2O), and Tailing factor: NMT 2.5 for pseudoephedrine peak,
pseudoephedrine hydrochloride (C10H15NO HCl) or Standard solution
pseudoephedrine sulfate [(C10H15NO)2 H2SO4]. Relative standard deviation: NMT 2.0%, Standard
[NOTEThe heading of this monograph does not constitute the solution
official title. It is not intended that the name described herein Analysis
be recognized as the official title or the common or usual Samples: Standard solution and Sample solution
name. The name for each article encompassed by this Calculate the percentage of C10H15NO HCl in each mL of
monograph shall be composed of the names of the active the Oral Solution taken:
ingredients contained therein, as well as the quantitative
amount of each active ingredient, and a statement of the Result = (rU/rS) (CS/CU) 100
IDENTIFICATION rS = pseudoephedrine peak response from the
A. If pseudoephedrine hydrochloride or pseudoephedrine Standard solution
sulfate is claimed in the labeling to be present, the retention CS = concentration of USP Pseudoephedrine
time of the peak of the Sample solution corresponds to that Hydrochloride RS in the Standard solution
of the Standard solution, as obtained in the Assay for (mg/mL)
Pseudoephedrine sulfate. CU = nominal concentration of pseudoephedrine
B. If acetaminophen is claimed in the labeling to be hydrochloride in the Sample solution (mg/mL)
present, the retention time of the peak of the Sample Acceptance criteria: 90.0%110.0%
solution corresponds to that of the Standard solution, as PSEUDOEPHEDRINE SULFATE (where pseudoephedrine sulfate is
obtained in the Assay for Acetaminophen. the salt form used, if present in the formulation)
C. If chlorpheniramine maleate is claimed in the labeling to Mobile phase, System suitability solutions A and B,
be present, the retention time of the peak of the Sample Chromatographic system, and System suitability:
solution corresponds to that of the Standard solution, as Proceed as directed in the Assay for Pseudoephedrine
obtained in the Assay for Chlorpheniramine maleate. hydrochloride.
D. If dextromethorphan hydrobromide is claimed in the Chlorpheniramine standard solution: Use Standard
labeling to be present, the retention time of the peak of the solution from the Assay for Chlorpheniramine maleate.
Sample solution corresponds to that of the Standard solution, Dextromethorphan standard solution: Use Standard
as obtained in the Assay for Dextromethorphan hydrobromide. solution from the Assay for Dextromethorphan hydrobromide.
ASSAY Pseudoephedrine Sulfate RS in water
PSEUDOEPHEDRINE HYDROCHLORIDE (where pseudoephedrine Standard solution: Standard stock solution, Mobile phase,
hydrochloride is the salt form used, if present in the and water (1:4:5)
formulation) Sample solution: Equivalent to 0.3 mg/mL of
Mobile phase: 3.4 mg/mL of monobasic potassium pseudoephedrine sulfate in Mobile phase and water (4:1)
phosphate, 1.5 mg/mL of triethylamine hydrochloride, 2.5 Analysis
mg/mL of sodium lauryl sulfate, and 1L/mL of phosphoric Samples: Standard solution and Sample solution
acid in methanol and water (60:40) Calculate the percentage of (C10H15NO)2 H2SO4 in each mL
Standard stock solution: 1.5 mg/mL of USP of Oral Solution taken:
Pseudoephedrine Hydrochloride RS in water
Standard solution: Standard stock solution, Mobile phase, Result = (rU/rS) (CS/CU) 100
and water (1:8:1)
Chlorpheniramine standard solution: Use Standard rU = pseudoephedrine peak response from the Sample
solution from the Assay for Chlorpheniramine maleate. solution
Dextromethorphan standard solution: Use Standard rS = peak response from the Standard solution
solution from the Assay for Dextromethorphan hydrobromide. CS = concentration of USP Pseudoephedrine Sulfate
System suitability solution A (for Oral Solution that contains RS in the Standard solution (mg/mL)
either all the four ingredients or a combination of three CU = nominal concentration of the Sample solution
containing chlorpheniramine salt): Standard solution and (mg/mL)
Chlorpheniramine standard solution (1:1) Acceptance criteria: 90.0%110.0%
System suitability solution B (for Oral Solution that contains ACETAMINOPHEN (if present)
no chlorpheniramine salt): Standard solution and Mobile phase: Methanol, glacial acetic acid, and water
Dextromethorphan standard solution (1:1) (20:1:79)
Standard solution: 0.165 mg/mL of USP Acetaminophen Sample solution: Equivalent to 0.075 mg/mL of
RS, in methanol and water (1:39) [NOTEFirst dissolve in dextromethorphan hydrobromide, from Oral Solution in
methanol, and mix until dissolved and then dilute with Mobile phase and water (4:1)
water to volume.] Injection size: 10 L
Sample solution: Equivalent to 0.165 mg/mL of Analysis
acetaminophen, from Oral Solution in methanol and water Samples: Standard solution and Sample solution
(1:39) Calculate the percentage of C18H25NO HBr H2O in each
Chromatographic system mL of Oral Solution taken:
Mode: LC Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
Detector: UV 280 nm
Column: 4.6-mm 15-cm; packing L7 rU = dextromethorphan peak response from the
Flow rate: 1 mL/min Sample solution
Injection size: 10 L rS = peak response from the Standard solution
System suitability CS = concentration of USP Dextromethorphan
Sample: Standard solution Hydrobromide RS in the Standard solution
Suitability requirements (mg/mL)
Tailing factor: NMT 2.0 CU = nominal concentration of dextromethorphan
Relative standard deviation: NMT 2.0% hydrobromide in the Sample solution (mg/mL)
Analysis Mr1 = molecular weight of dextromethorphan
Samples: Standard solution and Sample solution hydrobromide monohydrate, 370.33
Calculate the percentage of C8H9NO2 in each mL of Oral Mr2 = molecular weight of anhydrous
Solution taken: dextromethorphan hydrobromide, 352.32
OTHER COMPONENTS
rU = acetaminophen peak response from the Sample ALCOHOL DETERMINATION (if present), Method II 611:
solution 90.0%110.0% of the labeled amount of C2H5OH
CS = concentration of USP Acetaminophen RS in the PERFORMANCE TESTS
Standard solution (mg/mL) UNIFORMITY OF DOSAGE UNITS 905
CU = nominal concentration of acetaminophen in the For Oral Solution packaged in single-unit containers:
Sample solution (mg/mL) Meets the requirements
Acceptance criteria: 90.0%110.0% DELIVERABLE VOLUME 698
CHLORPHENIRAMINE MALEATE (if present) For Oral Solution packaged in multiple-unit containers:
Mobile phase, System suitability solutions, Meets the requirements
Chromatographic system, and System suitability: SPECIFIC TESTS
Proceed as directed in the Assay for Pseudoephedrine MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED
hydrochloride. MICROORGANISMS 62: The total bacterial count does not
Standard stock solution: 1 mg/mL of USP exceed 100 cfu/g, the total combined molds and yeasts
Chlorpheniramine Maleate RS in water count does not exceed 10 cfu/g, and it meets the
Standard solution: Standard stock solution, Mobile phase, requirements of the tests for absence of Salmonella species
and water (1:80:19) and Escherichia coli.
Sample solution: Equivalent to 0.01 mg/mL of PH 791: 3.77.5
chlorpheniramine maleate from Oral Solution in Mobile
phase and water (4:1) ADDITIONAL REQUIREMENTS
Injection size: 10 L PACKAGING AND STORAGE: Preserve in tight containers, and
Analysis store at controlled room temperature.
Samples: Standard solution and Sample solution LABELING: The label for each article encompassed by this
Calculate the percentage of C16H19ClN2 C4H4O4 in each mL monograph bears a name composed of the active
of the Oral Solution taken: ingredients. The label states the name and quantity of each
active ingredient and indicates its function (or purpose) in
Result = (rU/rS) (CS/CU) 100 the article.
rU = chlorpheniramine peak response from the USP Acetaminophen RS
Sample solution USP Chlorpheniramine Maleate RS
rS = peak response from the Standard solution USP Dextromethorphan Hydrobromide RS
CS = concentration of USP Chlorpheniramine Maleate USP Pseudoephedrine Hydrochloride RS
RS in the Standard solution (mg/mL) USP Pseudoephedrine Sulfate RS
(mg/mL)
DEXTROMETHORPHAN HYDROBROMIDE (if present) Tablets Containing at Least Three of
Mobile phase, System suitability solutions, the FollowingAcetaminophen and
Chromatographic system, and System suitability: Salts of Chlorpheniramine,
Proceed as directed in the Assay for Pseudoephedrine
hydrochloride. Dextromethorphan, and
Standard stock solution: 1.5 mg/mL of USP Pseudoephedrine
Dextromethorphan Hydrobromide RS in water (Comment on this Monograph)id=m256=Tablets Containing at
Standard solution: Standard stock solution, Mobile phase, Least Three of the Following-Acetaminophen and Salts of
and water (1:16:3)
Chlorpheniramine, Dextromethorphan, and Mode: LC

Pseudoephedrine=A-Monos.pdf) Detector: UV 214 nm
DEFINITION Flow rate: 2 mL/min
Tablets Containing at Least Three of the Following Injection size: 10 l
Acetaminophen and Salts of Chlorpheniramine, System suitability
Dextromethorphan, and Pseudoephedrine contain NLT 90.0% Samples: Standard solution and System suitability solution A
and NMT 110.0% of the labeled amounts of acetaminophen or System suitability solution B
(C8H9NO2), chlorpheniramine maleate (C16H19ClN2 C4H4O4), Suitability requirements
dextromethorphan hydrobromide (C18H25NO HBr H2O), and Resolution: NLT 2.0 between pseudoephedrine and
pseudoephedrine hydrochloride (C10H15NO HCl) or chlorpheniramine or between pseudoephedrine and
pseudoephedrine sulfate [(C10H15NO)2 H2SO4]. dextromethorphan
[NOTEThe heading of this monograph does not constitute the Tailing factor: NMT 2.5 for pseudoephedrine peak,
official title. It is not intended that the name described herein Standard solution
be recognized as the official title or the common or usual Relative standard deviation: NMT 2.0%
name. The name for each article encompassed by this Analysis
monograph shall be composed of the names of the active Samples: Standard solution and Sample solution
ingredients contained therein, as well as the quantitative Calculate the percentage of C10H15NO HCl in each mL of
amount of each active ingredient, and a statement of the the Oral Solution taken:
IDENTIFICATION
A. If pseudoephedrine hydrochloride or pseudoephedrine rU = pseudoephedrine peak response from the Sample
sulfate is claimed in the labeling to be present, the retention solution
time of the peak of the Sample solution corresponds to that rS = peak response from the Standard solution
of the Standard solution, as obtained in the Assay for CS = concentration of USP Pseudoephedrine
Pseudoephedrine sulfate. Hydrochloride RS in the Standard solution
B. If acetaminophen is claimed in the labeling to be (mg/mL)
present, the retention time of the peak of the Sample CU = nominal concentration of the Sample solution
solution corresponds to that of the Standard solution, as (mg/mL)
obtained in the Assay for Acetaminophen. Acceptance criteria: 90.0%110.0%
C. If chlorpheniramine maleate is claimed in the labeling to PSEUDOEPHEDRINE SULFATE (where pseudoephedrine sulfate is
be present, the retention time of the peak of the Sample the salt form used, if present in the formulation)
solution corresponds to that of the Standard solution, as Mobile phase, System suitability solutions,
obtained in the Assay for Chlorpheniramine maleate. Chromatographic system, and System suitability:
D. If dextromethorphan hydrobromide is claimed in the Proceed as directed in the Assay for Pseudoephedrine
labeling to be present, the retention time of the peak of the hydrochloride.
Sample solution corresponds to that of the Standard solution, Chlorpheniramine standard solution: Use the Standard
as obtained in the Assay for Dextromethorphan hydrobromide. solution from the Assay for Chlorpheniramine maleate.
Dextromethorphan standard solution: Use the Standard
ASSAY solution from the Assay for Dextromethorphan hydrobromide.
PSEUDOEPHEDRINE HYDROCHLORIDE (where pseudoephedrine Standard stock solution: 3.0 mg/mL of USP
hydrochloride is the salt form used, if present in the Pseudoephedrine Sulfate RS in water
formulation) Standard solution: Standard stock solution, Mobile phase,
Mobile phase: 3.4 mg/mL of monobasic potassium and water (1:4:5)
phosphate, 1.5 mg/mL of triethylamine hydrochloride, 2.5 Sample solution: Equivalent to 0.24 mg/mL of
mg/mL of sodium lauryl sulfate, and 1L/mL of phosphoric pseudoephedrine sulfate, from powdered Tablets, in
acid in methanol and water (60:40) methanol and 0.1% phosphoric acid (1:9) [NOTEFinely
Standard stock solution: 1.5 mg/mL of USP powder NLT 20 Tablets. First add methanol, and sonicate to
Pseudoephedrine Hydrochloride RS in water disperse the powder, and then make up to volume with
Standard solution: Mobile phase, Standard stock solution, 0.1% phosphoric acid.]
and water (8:1:1) Analysis: Calculate the percentage of (C10H15NO)2 H2SO4 in
Chlorpheniramine standard solution: Use the Standard each mL of the Oral Solution taken:
solution from the Assay for Chlorpheniramine maleate.
Dextromethorphan standard solution: Use the Standard Result = (rU/rS) (CS/CU) 100
solution from the Assay for Dextromethorphan hydrobromide.
System suitability solution A (for Oral Solution that contains rU = pseudoephedrine peak response from the Sample
either all the four ingredients or a combination of three solution
containing chlorpheniramine salt): Standard solution and rS = peak response from the Standard solution
Chlorpheniramine standard solution (1:1) CS = concentration of USP Pseudoephedrine Sulfate RS
System suitability solution B (for Oral Solution that contains in the Standard solution (mg/mL)
no chlorpheniramine salt): Standard solution and CU = nominal concentration of the Sample solution
Dextromethorphan standard solution (1:1) (mg/mL)
Sample solution: Equivalent to 0.12 mg/mL of Acceptance criteria: 90.0%110.0%
pseudoephedrine hydrochloride, from powdered Tablets, in ACETAMINOPHEN (if present)
methanol and 0.1% phosphoric acid (1:9) [NOTEFinely Mobile phase: Methanol, glacial acetic acid, and water
powder NLT 20 Tablets. First add methanol, and sonicate to (20:1:79)
disperse the powder, and then make up to volume with
0.1% phosphoric acid.]
Standard solution: 0.165 mg/mL of USP Acetaminophen Sample solution: Equivalent to 0.075 mg/mL of
RS in methanol and water (1:39) [NOTEFirst dissolve in dextromethorphan hydrobromide, from Oral solution in
methanol, and mix until solution is complete, and then Mobile phase and water (4:1)
dilute with water to volume.] Injection size: 10 L
Sample solution: Equivalent to 0.165 mg/mL of Analysis
acetaminophen, from Oral Solution in methanol and water Samples: Standard solution and Sample solution
(1:39) Calculate the percentage of C18H25NO HBr H2O in each
Chromatographic system mL of the Oral Solution taken:
Mode: LC Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
Detector: UV 280 nm
Column: 4.6-mm 15-cm; packing L7 rU = dextromethorphan peak response from the
Flow rate: 1 mL/min Sample solution
Injection size: 10 L rS = dextromethorphan peak response from the
System suitability Standard solution
Sample: Standard solution CS = concentration of USP Dextromethorphan
Suitability requirements Hydrobromide RS in the Standard solution
Tailing factor: NMT 2.0 (mg/mL)
Relative standard deviation: NMT 2.0% CU = nominal concentration of the Sample solution
Analysis (mg/mL)
Samples: Standard solution and Sample solution Mr1 = molecular weight of dextromethorphan
Calculate the percentage of C8H9NO2 in each mL of the hydrobromide monohydrate, 370.33
Oral Solution taken: Mr2 = molecular weight of anhydrous
dextromethorphan hydrobromide, 352.32
rU = acetaminophen peak response from the Sample PERFORMANCE TESTS
solution DISSOLUTION 711
rS = peak response from the Standard solution Test 1
CS = concentration of USP Acetaminophen RS in the Medium: pH 5.8 phosphate buffer (see Reagents,
Standard solution (mg/mL) Indicators, and SolutionsBuffer Solutions ); 900 mL
CU = nominal concentration of the Sample solution Apparatus 2: 50 rpm
(mg/mL) Time: 45 min
Acceptance criteria: 90.0%110.0% Sample solution: Mix 9.0 mL of a filtered portion of the
CHLORPHENIRAMINE MALEATE (if present) solution under test with 1.0 mL of 1% phosphoric acid
Mobile phase, System suitability solutions, solution.
Chromatographic system, and System suitability: Analysis: Determine the amounts of pseudoephedrine
Proceed as directed in the Assay for Pseudoephedrine hydrochloride or pseudoephedrine sulfate (as appropriate),
hydrochloride. acetaminophen, chlorpheniramine maleate, and
Standard stock solution: 1 mg/mL of USP dextromethorphan hydrobromide dissolved, using the
Chlorpheniramine Maleate RS in water Analyses set forth in the Assay for Pseudoephedrine
Standard solution: Standard stock solution, Mobile phase, hydrochloride or Assay for Pseudoephedrine sulfate, Assay for
and water (1:80:19) Acetaminophen, Assay for Chlorpheniramine maleate, and
Sample solution: Equivalent to 0.01 mg/mL of Assay for Dextromethorphan hydrobromide, respectively,
chlorpheniramine maleate from Oral Solution in Mobile making any necessary volumetric adjustments.
phase and water (4:1) Tolerances: NLT 75% (Q) of the labeled amounts of
Injection size: 10 L pseudoephedrine hydrochloride or pseudoephedrine
Analysis sulfate, acetaminophen, chlorpheniramine maleate, and
Samples: Standard solution and Sample solution dextromethorphan hydrobromide
Calculate the percentage of C16H19ClN2 C4H4O4 in each mL Test 2: If the product complies with this test, the labeling
of the Oral Solution: indicates that it meets USP Dissolution Test 2.
Result = (rU/rS) (CS/CU) 100 Apparatus, Time, Sample solution, Analysis, and
Tolerances: Proceed as directed for Test 1.
rU = chlorpheniramine peak response from the UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Sample solution
rS = peak response from the Standard solution ADDITIONAL REQUIREMENTS
CS = concentration of USP Chlorpheniramine Maleate PACKAGING AND STORAGE: Preserve in tight containers, and
RS in the Standard solution (mg/mL) store at controlled room temperature.
CU = nominal concentration of the Sample solution LABELING: The label for each article encompassed by this
(mg/mL) monograph bears a name composed of the active
Acceptance criteria: 90.0%110.0% ingredients. The label states the name and quantity of each
DEXTROMETHORPHAN HYDROBROMIDE (if present) active ingredient and indicates its function (or purpose) in
Mobile phase, Chromatographic system, and System the article. When more than one Dissolution Test is given, the
suitability: Proceed as directed in the Assay for labeling states the Dissolution Test used only if Test 1 is not
Pseudoephedrine hydrochloride. used.
Standard stock solution: 1.5 mg/mL of USP USP REFERENCE STANDARDS 11
Dextromethorphan Hydrobromide RS in water USP Acetaminophen RS
Standard solution: Standard stock solution, Mobile phase, USP Chlorpheniramine Maleate RS
and water (1:16:3)
USP Dextromethorphan Hydrobromide RS CS = concentration of USP Acetaminophen RS in the

USP Pseudoephedrine Hydrochloride RS Standard solution (mg/mL)
USP Pseudoephedrine Sulfate RS CU = nominal concentration of the Sample solution
(mg/mL)
CHLORPHENIRAMINE MALEATE
Acetaminophen, Chlorpheniramine Mobile phase: Methanol and water (3:2) containing 0.34 g
Maleate, and Dextromethorphan of monobasic potassium phosphate, 0.3 g of triethylamine
Hydrobromide Tablets hydrochloride, 0.15 g of sodium lauryl sulfate, and 0.1 mL
(Comment on this Monograph)id=m2310=Acetaminophen, of phosphoric acid in each 100 mL of solution
Chlorpheniramine Maleate, and Dextromethorphan Pseudoephedrine hydrochloride standard stock solution:
Hydrobromide Tablets=A-Monos.pdf) 3.0 mg/mL of USP Pseudoephedrine Hydrochloride RS
Pseudoephedrine hydrochloride standard solution:
DEFINITION Transfer 1.0 mL of Pseudoephedrine hydrochloride standard
Acetaminophen, Chlorpheniramine Maleate, and stock solution to a 25-mL volumetric flask. Add 2.5 mL of
Detromethorphan Hydrobromide Tablets contain NLT 90.0% methanol, and dilute with 0.1% phosphoric acid to volume.
and NMT 110.0% of the labeled amounts of acetaminophen Standard stock solution: 0.8 mg/mL of USP
(C8H9NO2), chlorpheniramine maleate (C16H19ClN2 C4H4O4), Chlorpheniramine Maleate RS
and dextromethorphan hydrobromide (C18H25NO HBr H2O). Standard solution: 8 g/mL of USP Chlorpheniramine
Maleate RS in 0.1% phosphoric acid, from Standard stock
A. The retention time of the major peak for acetaminophen System suitability solution: Pseudoephedrine hydrochloride
from the Sample solution corresponds to that of the Standard stock solution and Standard solution (1:1)
solution, as obtained in the Assay for Acetaminophen. Sample solution: Transfer an equivalent to 2 mg of
B. The retention time of the major peak for chlorpheniramine maleate, from powdered Tablets (NLT 20),
chlorpheniramine from the Sample solution corresponds to to a 250-mL volumetric flask. Add 25 mL of methanol, and
that of the Standard solution, as obtained in the Assay for sonicate to disperse the powder. Add 1 mL of phosphoric
Chlorpheniramine maleate. acid, dilute with water to volume, and filter.
C. The retention time of the major peak for Chromatographic system
dextromethorphan from the Sample solution corresponds to (See Chromatography 621, System Suitability.)
that of the Standard solution, as obtained in the Assay for Mode: LC
Dextromethorphan hydrobromide. Detector: UV 214 nm
ASSAY Flow rate: 2 mL/min
ACETAMINOPHEN Injection size: 10 L
Mobile phase: Methanol, water, and glacial acetic acid System suitability
(20:79:1) Samples: Standard solution and System suitability solution
Standard solution: Transfer 50 mg of USP Acetaminophen Suitability requirements
RS to a 100-mL volumetric flask. Add 4 mL of methanol, Resolution: NLT 2.0 between pseudoephedrine and
and mix until solution is complete. Dilute with 0.1% chlorpheniramine, System suitability solution
phosphoric acid to volume. Tailing factor: NMT 2.5 for the pseudoephedrine peak,
Sample stock solution: Transfer an equivalent to 100 mg of System suitability solution
acetaminophen, from powdered Tablets (NLT 20), to a 50- Relative standard deviation: NMT 2.0%, Standard
mL volumetric flask. Add 7.5 mL of methanol, and sonicate solution
to disperse the powder. Add 0.5 mL of phosphoric acid, Analysis
dilute with water to volume, and filter. Samples: Standard solution and Sample solution
Sample solution: Transfer 25.0 mL of the filtered Sample Calculate the percentage of C16H19ClN2 C4H4O4 in the
stock solution to a 100-mL volumetric flask, and dilute with portion of Tablets taken:
water to volume.
Chromatographic system Result = (rU/rS) (CS/CU) 100
Mode: LC rU = chlorpheniramine peak response from the
Detector: UV 280 nm Sample solution
Column: 4.6-mm 15-cm; packing L7 rS = peak response from the Standard solution
Flow rate: 1 mL/min CS = concentration of USP Chlorpheniramine Maleate
Injection volume: 10 L RS in the Standard solution (mg/mL)
System suitability CU = nominal concentration of chlorpheniramine
Sample: Standard solution maleate in the Sample solution (mg/mL)
Suitability requirements Acceptance criteria: 90.0%110.0%
Tailing factor: NMT 2.0 for the acetaminophen peak DEXTROMETHORPHAN HYDROBROMIDE
Relative standard deviation: NMT 2.0% from replicate Mobile phase, Pseudoephedrine hydrochloride stock
injections solution, Pseudoephedrine hydrochloride solution,
Analysis System suitability solution, and Chromatographic
Samples: Standard solution and Sample solution system: Proceed as directed in the Assay for
Calculate the percentage of C8H9NO2 in the portion of Chlorpheniramine maleate.
Tablets taken: Standard stock solution: 0.6 mg/mL of USP
Dextromethorphan Hydrobromide RS
Result = (rU/rS) (CS/CU) 100 Standard solution 0.06 mg/mL of USP Dextromethorphan
Hydrobromide RS in 0.1% phosphoric acid, from Standard
rU = acetaminophen peak response from the Sample stock solution
solution
Sample solution: Transfer an equivalent to 6 mg of Acetaminophen and Codeine Phosphate

dextromethorphan hydrobromide, from powdered Tablets Capsules
(NLT 20), to a 100-mL volumetric flask. Add 10 mL of
methanol, and sonicate to disperse the powder. Add 0.4 mL (Comment on this Monograph)id=m258=Acetaminophen and
of phosphoric acid, and dilute with water to volume. Codeine Phosphate Capsules=A-Monos.pdf)
Analysis DEFINITION
Samples: Standard solution and Sample solution Acetaminophen and Codeine Phosphate Capsules contain NLT
Calculate the percentage of C18H25NO HBr H2O in the 90.0% and NMT 110.0% of the labeled quantities of
portion of Tablets taken: acetaminophen (C8H9NO2) and codeine phosphate (C18H21NO3
H3PO4 1/2H2O).
Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
IDENTIFICATION
rU = dextromethorphan peak response from the A. The retention times of the Sample solution correspond to
Sample solution those of the Standard solution, as obtained in the Assay.
rS = peak response from the Standard solution B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
CS = concentration of USP Dextromethorphan Standard solution: 12 mg/mL each of USP Acetaminophen
Hydrobromide RS in the Standard solution RS and USP Codeine Phosphate RS in methanol
(mg/mL) Sample solution: Transfer a portion of Capsule contents
CU = nominal concentration of dextromethorphan equivalent to 12 mg of codeine phosphate to a separator,
hydrobromide in the Sample solution (mg/mL) and add 5 mL of water, 1 mL of ammonium hydroxide, and
Mr1 = molecular weight of dextromethorphan 5 mL of methylene chloride. Shake for 1 min, and allow the
hydrobromide monohydrate, 370.33 layers to separate. Use the clear, lower layer as the Sample
Mr2 = molecular weight of anhydrous solution.
dextromethorphan hydrobromide, 352.32 Developing solvent system: Methanol and ammonium
Acceptance criteria: 90.0%110.0% hydroxide (49:1)
PERFORMANCE TESTS (See Chromatography 621, Thin-Layer Chromatography.)
DISSOLUTION, Procedure for a Pooled Sample 711 Mode: TLC
Test 1 Adsorbent: 0.25-mm layer of chromatographic silica gel
Medium: Water; 900 mL mixture
Apparatus 2: 50 rpm Application volume: 10 L
Time: 45 min Analysis
Sample solution: Mix 9.0 mL of a filtered portion of the Samples: Standard solution and Sample solution
solution with 1.0 mL of 1% phosphoric acid solution Develop the chromatogram in the Developing solvent system
Analysis: Determine the amounts of acetaminophen, until the solvent front has moved three-fourths of the
chlorpheniramine maleate, and dextromethorphan length of the plate. Locate the spots on the plate by
hydrobromide dissolved, using the Analyses set forth in the examination under short-wavelength UV light.
Assay for Acetaminophen, Assay for Chlorpheniramine Acceptance criteria: The RF values of the two principal
maleate, and Assay for Dextromethorphan hydrobromide, spots of the Sample solution correspond to those of the
respectively, making any necessary volumetric adjustments. Standard solution.
Tolerances: NLT 75% (Q) of the labeled amounts of
C8H9NO2, C16H19ClN2 C4H4O4, and C18H25NO HBr H2O ASSAY
are dissolved. PROCEDURE
Test 2: If the product complies with this test, the labeling Solution A: Dissolve 2.04 g of monobasic potassium
indicates that it meets USP Dissolution Test 2. phosphate in 950 mL of water. Add 2 mL of triethylamine,
Medium: 0.1 M hydrochloric acid; 900 mL adjust with phosphoric acid to a pH of 2.35, and dilute with
Apparatus, Time, Sample solution, Analysis, and water to 1000 mL.
Tolerances: Proceed as directed for Test 1. Mobile phase: Methanol and Solution A (8:92)
Test 3: If the product complies with this test, the labeling Codeine phosphate standard stock solution: 0.3 mg/mL
indicates that it meets USP Dissolution Test 3. of USP Codeine Phosphate RS in Mobile phase
Medium: pH 5.8 phosphate buffer (see Reagents, Standard solution: Transfer 30 mg of USP Acetaminophen
Indicators, and SolutionsBuffer Solutions); 900 mL RS and 100J mL of Codeine phosphate standard stock solution
Apparatus, Time, Sample solution, Analysis, and (J being the ratio of the labeled amount, in mg, of codeine
Tolerances: Proceed as directed for Test 1. phosphate hemihydrate to that of acetaminophen) to a 100-
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements mL volumetric flask, dilute with Mobile phase to volume, and
mix. This solution contains 0.3 mg/mL of acetaminophen
ADDITIONAL REQUIREMENTS and 0.3J mg/mL of codeine phosphate hemihydrate.
PACKAGING AND STORAGE: Preserve in tight containers, and Sample stock solution: Remove as completely as possible
store at controlled room temperature. the contents of NLT 20 Capsules, weigh, and mix. Transfer a
LABELING: The label states the name and quantity of each portion of the combined contents equivalent to 300 mg of
active ingredient and indicates its function (or purpose) in acetaminophen to a 100-mL volumetric flask, add 75 mL of
the article. When more than one Dissolution Test is given, the Mobile phase, and sonicate for 10 min. Dilute with Mobile
labeling states the Dissolution Test used only if Test 1 is not phase to volume.
used. Sample solution: Dilute 5.0 mL of the Sample stock solution
USP REFERENCE STANDARDS 11 with Mobile phase to 50 mL and pass a portion of the
USP Acetaminophen RS solution through a 1-m filter.
USP Chlorpheniramine Maleate RS Chromatographic system
USP Dextromethorphan Hydrobromide RS (See Chromatography 621, System Suitability.)
USP Pseudoephedrine Hydrochloride RS
Mode: LC rU = peak response from the Sample solution

Detector: UV 214 nm rS = peak response from the Standard solution
Column: 4.6-mm 25-cm; 5-m packing L1 CS = concentration of USP Acetaminophen RS in the
Flow rate: 1.5 mL/min Standard solution (mg/mL)
Injection size: 30 L F = dilution factor, 1000
System suitability Calculate the quantity, in mg/mL, of C18H21NO3 H3PO4
Sample: Standard solution 1
/2H2O in the Capsule taken:
Resolution: NLT 2.0 between acetaminophen and Result = (rU/rS) CS (Mr1/Mr2)
codeine phosphate
Relative standard deviation: NMT 2.0% and 3.0%, rU = peak response from the Sample solution
respectively, for acetaminophen and codeine phosphate rS = peak response from the Standard solution
Analysis CS = concentration of USP Codeine Phosphate RS in
Samples: Standard solution and Sample solution the Standard solution (mg/mL)
Calculate the percentage of C8H9NO2 in the Capsules: Mr1 = molecular weight of codeine phosphate
hemihydrate, 406.37
Result = (rU/rS) (CS/CU) 100 Mr2 = molecular weight of anhydrous codeine, 397.37
Acceptance criteria: Meet the requirements
CS = concentration of USP Acetaminophen RS in the PACKAGING AND STORAGE: Preserve in tight, light-resistant
Standard solution (mg/mL) containers, and store at controlled room temperature.
CU = nominal concentration of acetaminophen in the USP REFERENCE STANDARDS 11
Sample solution (mg/mL) USP Acetaminophen RS
Calculate the percentage of C18H21NO3 H3PO4 1/2H2O in USP Codeine Phosphate RS
the Capsules:
Acetaminophen and Codeine Phosphate
rU = peak response from the Sample solution Oral Solution
rS = peak response from the Standard solution (Comment on this Monograph)id=m260=Acetaminophen and
CS = concentration of USP Codeine Phosphate RS in Codeine Phosphate Oral Solution=A-Monos.pdf)
the Standard solution (mg/mL)
CU = nominal concentration of codeine phosphate DEFINITION
hemihydrate in the Sample solution (mg/mL) Acetaminophen and Codeine Phosphate Oral Solution contains
Mr1 = molecular weight of codeine phosphate NLT 90.0% and NMT 110.0% of the labeled quantities of
hemihydrate, 406.37 acetaminophen (C8H9NO2) and codeine phosphate
Mr2 = molecular weight of anhydrous codeine hemihydrate (C18H21NO3 H3PO4 1/2H2O).
phosphate, 397.37
Acceptance criteria: 90.0%110.0% of the labeled IDENTIFICATION
amounts of C8H9NO2 and C18H21NO3 H3PO4 1/2H2O A. The retention times of the Sample solutions correspond to
those of the Standard solutions, as obtained in the Assay for
PERFORMANCE TESTS Acetaminophen and the Assay for Codeine phosphate.
DISSOLUTION 711 B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
Medium: 0.01 N hydrochloric acid; 900 mL Standard solution: 12 mg/mL each of USP Acetaminophen
Apparatus 2: 50 rpm RS and USP Codeine Phosphate RS in methanol
Time: 30 min Sample solution: Transfer a volume of Oral Solution,
Analysis: Determine the amounts of C8H9NO2 and equivalent to 12 mg of codeine phosphate, to a separator,
C18H21NO3 H3PO4 1/2H2O dissolved by using the method add 1 mL of ammonium hydroxide, and 5 mL of methylene
set forth in the Assay, except use 0.01 N hydrochloric acid chloride, shake for 1 min, and allow the layers to separate.
to prepare the Codeine phosphate standard stock solution, Use the clear, lower layer as the Sample solution.
and make any other necessary volumetric adjustments. Developing solvent system: Methanol and ammonium
Tolerances: NLT 75% (Q) of the labeled quantities of hydroxide (49:1)
C8H9NO2 and C18H21NO3 H3PO4 1/2H2O is dissolved. Chromatographic system
UNIFORMITY OF DOSAGE UNITS 905 (See Chromatography 621, Thin-Layer Chromatography.)
Procedure for content uniformity Mode: TLC
Solution A, Mobile phase, Codeine phosphate standard Adsorbent: 0.25-mm layer of chromatographic silica gel
stock solution, Standard solution, and Chromatographic mixture
system: Prepare as directed in the Assay. Application volume: 10 L
Sample stock solution: Transfer the contents of 1 Capsule Analysis
to a 100-mL volumetric flask, add 75 mL of Mobile phase, Samples: Standard solution and Sample solution
and sonicate for 10 min. Dilute with Mobile phase to Develop the chromatogram in the Developing solvent system
volume. until the solvent front has moved three-fourths of the
Sample solution: Dilute 5.0 mL of the Sample stock solution length of the plate. Locate the spots on the plate by
with Mobile phase to 50 mL and pass a portion of the examination under short-wavelength UV light.
solution through a 1-m filter. Acceptance criteria: The RF values of the two principal
Analysis spots of the Sample solution correspond to those of the
Samples: Standard solution and Sample solution Standard solution.
Calculate the quantity, in mg, of C8H9NO2 in the Capsule
taken:
Result = (rU/rS) CS F
ASSAY CS = concentration of USP Codeine Phosphate RS in

ACETAMINOPHEN the Standard solution (mg/mL)
Mobile phase: Methanol and water (3:7) CU = nominal concentration of codeine phosphate in
Standard solution: 0.48 mg/mL of USP Acetaminophen RS the Sample solution (mg/mL)
in Mobile phase Mr1 = molecular weight of codeine phosphate
Sample solution: Equivalent of 0.48 mg/mL of hemihydrate, 406.37
acetaminophen in Mobile phase Mr2 = molecular weight of anhydrous codeine
Chromatographic system phosphate, 397.37
(See Chromatography 621, System Suitability.) Acceptance criteria: 90.0%110.0%
Mode: LC
Detector: UV 280 nm PERFORMANCE TESTS
Column: 3.9-mm 30-cm; packing L1 UNIFORMITY OF DOSAGE UNITS 905 (for Oral Solution
Flow rate: 2.0 mL/min packaged in single-unit containers): Meets the
Injection size: 10 L requirements
System suitability DELIVERABLE VOLUME 698 (for Oral Solution packaged in
Sample: Standard solution multiple-unit containers): Meets the requirements
Column efficiency: NLT 1000 theoretical plates SPECIFIC TESTS
determined from the analyte peak PH 791: 4.06.1
Tailing factor: NMT 1.5 for the analyte peak ALCOHOL DETERMINATION, Method II 611 (test if alcohol is
Relative standard deviation: NMT 2.0% present): 90.0%120.0% of the labeled quantity of
Analysis C2H5OH, acetone being used as the internal standard
Calculate the percentage of C8H9NO2 in the Oral Solution PACKAGING AND STORAGE: Preserve in tight, light-resistant
taken: containers, and store at controlled room temperature.
Result = (rU/rS) (CS/CU) 100 USP Acetaminophen RS
rU = acetaminophen peak response from the Sample USP Codeine Phosphate RS
solution
CS = concentration of USP Acetaminophen RS in the Acetaminophen and Codeine Phosphate
Standard solution (mg/mL) Oral Suspension
CU = nominal concentration of acetaminophen in the
Sample solution (mg/mL) (Comment on this Monograph)id=m265=Acetaminophen and
Acceptance criteria: 90.0%110.0% Codeine Phosphate Oral Suspension=A-Monos.pdf)
CODEINE PHOSPHATE
Mobile phase: Dissolve 4.44 g of docusate sodium in 1000 DEFINITION
mL of a mixture of methanol, tetrahydrofuran, phosphoric Acetaminophen and Codeine Phosphate Oral Suspension is a
acid, and water (600:40:1:360) with stirring, and pass suspension of Acetaminophen and Codeine Phosphate in a
through a membrane filter having a 0.45-m or finer suitable aqueous vehicle. It contains NLT 90.0% and NMT
porosity. 110.0% of the labeled amounts of acetaminophen (C8H9NO2)
Solution A: Methanol and water (3:7) and codeine phosphate hemihydrate (C18H21NO3 H3PO4
Standard solution: 0.12 mg/mL of USP Codeine Phosphate
1
/2H2O).
RS in Solution A IDENTIFICATION
Sample solution: Equivalent of 0.12 mg/mL of A. The retention times of the major peaks of the Sample
acetaminophen in Solution A solution correspond to those of the Standard solution, as
Chromatographic system obtained in the Assay.
(See Chromatography 621, System Suitability.) B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
Mode: LC Standard solution: 12 mg/mL each of USP Acetaminophen
Detector: UV 280 nm RS and USP Codeine Phosphate RS in methanol
Column: 3.9-mm 30-cm; packing L1 Sample solution: Transfer a volume of Oral Suspension,
Flow rate: 1.5 mL/min equivalent to 12 mg of codeine phosphate, to a separator,
Injection size: 10 L add 1 mL of ammonium hydroxide, and 5 mL of methylene
System suitability chloride, shake for 1 min, and allow the layers to separate.
Sample: Standard solution Use the clear, lower layer as the Sample solution.
Suitability requirements Developing solvent system: Methanol and ammonium
Column efficiency: NLT 1500 theoretical plates hydroxide (49:1)
determined from the analyte peak Chromatographic system
Tailing factor: NMT 2.0 for the analyte peak (See Chromatography 621, Thin Layer Chromatography.)
Relative standard deviation: NMT 3.0% Mode: TLC
Analysis Adsorbent: 0.25-mm layer of chromatographic silica gel
Samples: Standard solution and Sample solution mixture
Calculate the percentage of C18H21NO3 H3PO4 1/2H2O in Application volume: 10 L
the Oral Solution taken: Analysis
Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 Develop the chromatogram in the Developing solvent system
rU = codeine phosphate peak response from the until the solvent front has moved three-fourths of the
Sample solution length of the plate. Locate the spots on the plate by
rS = peak response from the Standard solution examination under short-wavelength UV light.
Acceptance criteria: The RF values of the two principal CU = nominal concentration of acetaminophen in the
spots from the Sample solution correspond to those from the Sample solution (mg/mL)
Standard solution. Acceptance criteria: 90.0%110.0%
Calculate the percentage of C18H21NO3 H3PO4 1/2H2O in
ASSAY the Oral Suspension taken:
PROCEDURE
Mobile phase: Dissolve 4.9 g of monobasic potassium Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
phosphate in 900 mL of water, adjust with phosphoric acid
to a pH of 3.9, and add 216 mg of sodium 1- rU = peak response from the Sample solution
octanesulfonate. Add 100 mL of acetonitrile. rS = peak response from the Standard solution
Solution A: Methanol and 0.01 N sodium hydroxide CS = concentration of USP Codeine Phosphate RS in
(30:70) the Standard solution (mg/mL)
Codeine phosphate standard stock solution: 0.5 mg/mL CU = nominal concentration of codeine phosphate
of USP Codeine Phosphate RS in Mobile phase hemihydrate in the Sample solution (mg/mL)
Standard stock solution: Transfer a quantity of 5J mg of Mr1 = molecular weight ratio of codeine phosphate
USP Acetaminophen RS, J being the ratio of the labeled hemihydrate, 406.37
amount, in mg, of acetaminophen to the labeled amount, in Mr2 = molecular weight ratio of anhydrous codeine
mg, of codeine phosphate hemihydrate, and 10.0 mL of phosphate, 397.37
Codeine phosphate standard stock solution to a 100-mL Acceptance criteria: 90.0%110.0%
volumetric flask, and dissolve in and dilute with Solution A to
volume. PERFORMANCE TESTS
Standard solution: Dilute 10.0 mL of the Standard stock UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
solution to 50 mL with Mobile phase (0.01 mg/mL of USP for Oral Suspension packaged in single-unit containers
Codeine Phosphate RS and 0.01J mg/mL of USP DELIVERABLE VOLUME 698: Meets the requirements for Oral
Acetaminophen RS). Suspension packaged in multiple-unit containers
Sample stock solution: Transfer a measured volume of
well-mixed Oral Suspension, equivalent to 50 mg of SPECIFIC TESTS
acetaminophen, to a 100-mL volumetric flask. Add 50 mL of PH 791: 4.06.1
Solution A, and mix by mechanical means for 30 min. Dilute ADDITIONAL REQUIREMENTS
with Solution A to volume. [NOTEFoaming may be PACKAGING AND STORAGE: Preserve in tight, light-resistant
minimized by adding a few drops of acetonitrile before containers, and store at controlled room temperature.
diluting with Solution A to volume.] Centrifuge a portion of USP REFERENCE STANDARDS 11
this mixture. USP Acetaminophen RS
Sample solution: Dilute 10.0 mL of the clear supernatant USP Codeine Phosphate RS
from the Sample stock solution to 50 mL with Mobile phase.
System suitability stock solution: 0.02 mg/mL of sodium
benzoate and 0.03 mg/mL of methylparaben in Solution A
System suitability solution: To 10.0 mL of the System Acetaminophen and Codeine Phosphate
suitability stock solution add 10.0 mL of Standard stock Tablets
solution, and dilute to 50 mL with Mobile phase.
Chromatographic system (Comment on this Monograph)id=m270=Acetaminophen and
(See Chromatography 621,System Suitability.) Codeine Phosphate Tablets=A-Monos.pdf)
Mode: LC DEFINITION
Detector: UV 220 nm Acetaminophen and Codeine Phosphate Tablets contain NLT
Column: 4.6-mm 15-cm; 5-m packing L11 90.0% and NMT 110.0% of the labeled amounts of
Flow rate: 2 mL/min acetaminophen (C8H9 NO2) and codeine phosphate
Injection size: 20 L hemihydrate (C18H21NO3 H3PO4 1/2H2O).
System suitability
Sample: System suiltability solution and Standard solution IDENTIFICATION
[NOTEThe relative retention times for acetaminophen, A. The retention times of the major peaks of the Sample
benzoate, codeine, and methylparaben are about 0.25, solution correspond to those of the Standard solution, as
0.5, 1.0, and 1.3, respectively.] obtained in the Assay.
Suitability requirements B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
Resolution: NLT 2 between each pair of adjacent peaks, Standard solution: 12 mg/mL each of USP Acetaminophen
System suitability solution RS and USP Codeine Phosphate RS in methanol
Tailing factor: NMT 2 for each analyte peak, Standard Sample solution: Transfer a quantity of finely powdered
solution Tablets, equivalent to 12 mg of codeine phosphate, to a
Column efficiency: NLT 500 theoretical plates, Standard separator, add 5 mL of water, 1 mL of ammonium
solution hydroxide, and 5 mL of methylene chloride, shake for 1
Relative standard deviation: NMT 2.0%, Standard min, and allow the layers to separate. Use the clear, lower
solution layer as the Sample solution.
Analysis Developing solvent system: Methanol and ammonium
Samples: Standard solution and Sample Solution hydroxide (49:1)
Calculate the percentage of C8H9NO2 in the Oral Chromatographic system
Suspension taken: (See Chromatography 621, Thin Layer Chromatography.)
Mode: TLC
Result = (rU/rS) (CS/CU) 100 Adsorbent: 0.25-mm layer of chromatographic silica gel
mixture
CS = concentration of USP Acetaminophen RS in the
Application volume: 10 L rS = peak response from the Standard solution

Analysis CS = concentration of USP Codeine Phosphate RS in
Develop the chromatogram in the Developing solvent system CU = nominal concentration of codeine phosphate
until the solvent front has moved three-fourths of the hemihydrate in the Sample solution (mg/mL)
length of the plate. Locate the spots on the plate by Mr1 = molecular weight of codeine phosphate
examination under short-wavelength UV light. hemihydrate, 406.37
Acceptance criteria: The RF values of the two principal Mr2 = molecular weight of anhydrous codeine
spots from the Sample solution correspond to those from the phosphate, 397.37
Standard solution. Acceptance criteria: 90.0%110.0%
ASSAY PERFORMANCE TESTS
PROCEDURE DISSOLUTION 711
Solution A: Dissolve 2.04 g of monobasic potassium Medium: 0.01 N hydrochloric acid; 900 mL
phosphate in about 950 mL of water. Add 2 mL of Apparatus 2: 50 rpm
triethylamine, adjust with phosphoric acid to a pH of 2.35, Time: 30 min
and dilute with water to 1000 mL. Analysis: Determine the amounts of C8H9NO2 and
Mobile phase: Methanol and Solution A (8:92) C18H21NO3 H3PO4 1/2H2O dissolved by employing the
Codeine phosphate standard stock solution: 0.3 mg/mL method set forth in the Assay, except to use 0.01 N
of USP Codeine Phosphate RS in Mobile phase hydrochloric acid to prepare the Codeine phosphate standard
Standard solution: Transfer 30 mg of USP Acetaminophen stock solution and to make any other necessary volumetric
RS and 100 J mL of Codeine phosphate standard stock adjustments.
solution, J being the ratio of the labeled amount, in mg, of Tolerances: NLT 75% (Q) of the labeled amounts of
codeine phosphate hemihydrate to that of acetaminophen, C8H9NO2 and C18H21NO3 H3PO4 1/2H2O is dissolved.
to a 100-mL volumetric flask, and dilute with Mobile phase UNIFORMITY OF DOSAGE UNITS 905
to volume. This solution contains 0.3 mg/mL of Procedure for content uniformity
acetaminophen and 0.3J mg/mL of codeine phosphate Solution A, Mobile phase, Codeine phosphate standard
hemihydrate. stock solution, Standard solution, and Chromatographic
Sample stock solution: Transfer an equivalent to 300 mg of system: Prepare as directed in the Assay.
acetaminophen, from a portion of finely powdered Tablets Sample stock solution: Transfer 1 Tablet to a 100-mL
(NLT 20), to a 100-mL volumetric flask, add 75 mL of Mobile volumetric flask, add 75 mL of Mobile phase, and sonicate
phase, and sonicate for 10 min. Dilute with Mobile phase to for 10 min. Dilute with Mobile phase to volume.
volume. Sample solution: Dilute 5.0 mL of the Sample stock
Sample solution: Dilute 5.0 mL of the Sample stock solution solution with Mobile phase to 50 mL and pass a portion of
with Mobile phase to 50 mL, and pass a portion of the the solution through a suitable 1-m filter.
solution through a suitable 1-m filter. Analysis
Chromatographic system Samples: Standard solution and Sample solution
(See Chromatography 621, System Suitability.) Calculate the quantity, in mg, of C8H9NO2 in the Tablet
Mode: LC taken:
Detector: UV 214 nm
Column: 4.6-mm 25-cm; 5-m packing L1 Result = (rU/rS) CS F
Flow rate: 1.5 mL/min
Injection size: 30 L rU = peak response from the Sample solution
System suitability rS = peak response from the Standard solution
Sample: Standard solution CS = concentration of USP Acetaminophen RS in the
Suitability requirements Standard solution (mg/mL)
Resolution: NLT 2.0 between acetaminophen and F = dilution factor for the Tablet taken, 1000
codeine phosphate Calculate the quantity, in mg/mL, of C18H21NO3 H3PO4
Relative standard deviation: NMT 2.0% and 3.0%,
1
/2H2O in the Tablet taken:
respectively, for acetaminophen and codeine phosphate
Analysis Result = (rU/rS) CU (Mr1/Mr2) F
Samples: Standard solution and Sample Solution
Calculate the percentage of C8H9NO2 in the Tablets taken: rU = peak response from the Sample solution
Result = (rU/rS) (CS/CU) 100 CU = concentration of USP Codeine Phosphate RS in
rU = peak response from the Sample solution F = dilution factor for the Tablet taken, 1000
rS = peak response from the Standard solution Mr1 = molecular weight of codeine phosphate
CS = concentration of USP Acetaminophen RS in the hemihydrate, 406.37
Standard solution (mg/mL) Mr2 = molecular weight of anhydrous codeine
CU = nominal concentration of acetaminophen in the phosphate, 397.37
Sample solution (mg/mL) Acceptance criteria: Meet the requirements
Calculate the percentage of C18H21NO3 H3PO4 1/2H2O in ADDITIONAL REQUIREMENTS
the Tablets taken: PACKAGING AND STORAGE: Preserve in tight, light-resistant
containers, and store at controlled room temperature.
Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 USP REFERENCE STANDARDS 11
USP Acetaminophen RS
rU = peak response from the Sample solution USP Codeine Phosphate RS
Acetaminophen, Dextromethorphan Acceptance criteria: 90.0%110.0%

Hydrobromide, Doxylamine Succinate, DEXTROMETHORPHAN HYDROBROMIDE
and Pseudoephedrine Hydrochloride Solution A: 6.8 mg/mL of monobasic potassium phosphate
in water
Oral Solution Mobile phase: Acetonitrile and Solution A (9:11)
(Comment on this Monograph)id=m275=Acetaminophen, Standard solution: 0.1 mg/mL of USP Dextromethorphan
Dextromethorphan Hydrobromide, Doxylamine Succinate, and Hydrobromide RS, 0.04 mg/mL USP Doxylamine Succinate
Pseudoephedrine Hydrochloride Oral Solution=A-Monos.pdf) RS, and 0.2 mg/mL USP Pseudoephedrine Hydrochloride RS
in Mobile phase
DEFINITION Sample solution: Equivalent to 0.1 mg/mL of
Acetaminophen, Dextromethorphan Hydrobromide, Doxylamine dextromethorphan hydrobromide from a volume of Oral
Succinate, and Pseudoephedrine Hydrochloride Oral Solution Solution in Mobile phase
contains NLT 90.0% and NMT 110.0% of the labeled amounts Chromatographic system
of Acetaminophen (C8H9NO2), Dextromethorphan (See Chromatography 621, System Suitability.)
Hydrobromide (C18H25NO HBr H2O), Doxylamine Succinate Mode: LC
(C17H22N2O C4H6O4), and Pseudoephedrine Hydrochloride Detector: UV 220 nm
(C10H15NO HCl). Column: 4.6-mm 25-cm; packing L9
IDENTIFICATION Injection size: 10 L
A. The retention time of the major peak for acetaminophen System suitability
from the Sample solution corresponds to that of the Standard Sample: Standard solution
solution, as obtained in the Assay for Acetaminophen. [NOTEThe relative retention times for pseudoephedrine,
B. The retention time of the major peak for dextromethorphan, and doxylamine are 0.38, 0.65, and
dextromethorphan from the Sample solution corresponds to 1.0, respectively.]
that of the Standard solution, as obtained in the Assay for Suitability requirements
Dextromethorphan Hydrobromide. Column efficiency: NLT 500 theoretical plates for the
C. The retention time of the major peak for doxylamine dextromethorphan, doxylamine, and pseudoephedrine
from the Sample solution corresponds to that of the Standard peaks
solution, as obtained in the Assay for Doxylamine Succinate. Tailing factor: NMT 2.5 for the dextromethorphan,
D. The retention time of the major peak for doxylamine, and pseudoephedrine peaks
pseudoephedrine from the Sample solution corresponds to Relative standard deviation: NMT 2.0% for
that of the Standard solution, as obtained in the Assay for dextromethorphan, doxylamine, and pseudoephedrine
Pseudoephedrine Hydrochloride. Analysis
ASSAY Samples: Standard solution and Sample solution
ACETAMINOPHEN Calculate the quantity, in percentage, of C18H25NO HBr
Mobile phase: Methanol and water (11:9) H2O in the portion of Oral Solution taken:
Standard solution: 0.2 mg/mL of USP Acetaminophen RS
in Mobile phase Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
Sample solution: Equivalent to 0.2 mg/mL of rU = peak response from the Sample solution
Acetaminophen from a volume of Oral Solution in Mobile rS = peak response from the Standard solution
phase CS = concentration of USP Dextromethorphan
Chromatographic system Hydrobromide RS in the Standard solution
(See Chromatography 621, System Suitability.) (mg/mL)
Mode: LC CU = concentration of dextromethorphan
Detector: UV 254 nm hydrobromide in the Sample solution (mg/mL)
Column: 4.6-mm 25-cm; packing L1 Mr1 = molecular weight of dextromethorphan
Flow rate: 1 mL/min hydrobromide monohydrate, 370.33
Injection size: 10 L Mr2 = molecular weight of anhydrous
System suitability dextromethorphan hydrobromide, 352.32
Sample: Standard solution Acceptance criteria: 90.0%110.0%
Suitability requirements DOXYLAMINE SUCCINATE
Column efficiency: NLT 500 theoretical plates [NOTESolution A, Mobile phase, Standard solution, and
determined from the analyte peak Chromatographic system: proceed as directed in the Assay
Tailing factor: NMT 2.0 for the acetaminophen peak for Dextromethorphan Hydrobromide.]
Relative standard deviation: NMT 2.0% Sample solution: Equivalent to 0.04 mg/mL of doxylamine
Analysis succinate from a volume of Oral Solution in Mobile phase
Samples: Standard solution and Sample Solution Analysis: Proceed as directed for Procedure in the Assay for
Calculate the quantity, in percentage, of C8H9NO2 in each Dextromethorphan Hydrobromide
mL of the Oral Solution taken: Calculate the quantity, in percentage, of C17H22N2O C4H6O4
Result = (rU/rS) (CS/CU) 100 in the portion of Oral Solution taken:
rU = peak response from the Sample solution Result = (rU/rS) (CS/CU) 100
rS = peak response from the Standard solution rU = peak response from the Sample solution
CS = concentration of USP Acetaminophen RS in the rS = peak response from the Standard solution
Standard solution (mg/mL) CS = concentration of USP Doxylamine Succinate RS in
CU = concentration of acetaminophen in the Sample the Standard solution (mg/mL)
solution (mg/mL)
CU = concentration of doxylamine succinate in the ASSAY

Sample solution (mg/mL) ACETAMINOPHEN
Acceptance criteria: 90.0%110.0% Mobile phase: Methanol and water (2:3)
PSEUDOEPHEDRINE HYDROCHLORIDE Internal standard solution: 8.0 mg/mL of guaifenesin in a
[NOTESolution A, Mobile phase, Standard solution, and mixture of water and methanol (4:1)
Chromatographic system: proceed as directed in the Assay Standard solution: Transfer 50 mg of USP Acetaminophen
for Dextromethorphan Hydrobromide.] RS to a 100-mL volumetric flask. Dissolve in 2.5 mL of
Sample solution: Equivalent to 0.2 mg/mL of methanol, and dilute with water to volume. Transfer 2.0 mL
pseudoephedrine hydrochloride from a volume of Oral of this solution to a 50-mL volumetric flask, add 5.0 mL of
Solution in Mobile phase Internal standard solution, and dilute with Mobile phase to
Analysis: Proceed as directed for Procedure in the Assay for volume.
Dextromethorphan Hydrobromide. Sample solution: Weigh and finely powder NLT 20 Tablets.
Calculate the quantity, in percentage, of C10H15NO HCl in Transfer a portion of the powder, equivalent to 500 mg of
the portion of Oral Solution taken: acetaminophen, to a 100-mL volumetric flask, add 25 mL of
methanol, and shake by mechanical means for 10 min.
Result = (rU/rS) (CS/CU) 100 Dilute with water to volume. Transfer 10.0 mL of this
solution to a 100-mL volumetric flask, and dilute with water
rU = peak response from the Sample solution to volume. Transfer 2.0 mL of this solution to a 50-mL
rS = peak response from the Standard solution volumetric flask, add 5.0 mL of Internal standard solution,
CS = concentration of USP Pseudoephedrine and dilute with Mobile phase to volume.
Hydrochloride RS in the Standard solution Chromatographic system
(mg/mL) (See Chromatography 621, System Suitability.)
CU = concentration of pseudoephedrine hydrochloride Mode: LC
in the Sample solution (mg/mL) Detector: UV 254 nm
Acceptance criteria: 90.0%110.0% Column: 4.6-mm 15-cm; 5-m packing L1
Column temperature: 35 0.5
PERFORMANCE TESTS Flow rate: 1 mL/min
UNIFORMITY OF DOSAGE UNITS 905: For oral solution Injection size: 10 L
packaged in single-unit containers, it meets the System suitability
requirements. Sample: Standard solution
DELIVERABLE VOLUME 698: For oral solution packaged in [NOTEThe relative retention times for acetaminophen and
multiple-unit containers, it meets the requirements. guaifenesin are 0.5 and 1.0, respectively.]
SPECIFIC TESTS Suitability requirements
MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED Resolution: NLT 6.0 between the analyte and internal
MICROORGANISMS 62: The total bacterial count does not standard peaks
exceed 100 cfu/g, the total combined molds and yeasts Column efficiency: NLT 1000 theoretical plates
count does not exceed 10 cfu/g, and it meets the determined from the analyte peak
requirements of the tests for absence of Salmonella species Tailing factor: NMT 2.0 for the analyte peak
and Escherichia coli. Relative standard deviation: NMT 2.5% of the peak
PH 791: 4.56.3 response ratios for replicate injections
ALCOHOL DETERMINATION, Method II (if present) 611: Analysis
90.0%110.0% of the labeled amount of C2H5OH Samples: Standard solution and Sample solution
Calculate the percentage of C8H9NO2 in the portion of
ADDITIONAL REQUIREMENTS Tablets taken:
store at controlled room temperature. Result = (RU/RS) (CS/CU) 100
USP Acetaminophen RS RU = peak response ratio of acetaminophen to the
USP Dextromethorphan Hydrobromide RS internal standard peak from the Sample solution
USP Doxylamine Succinate RS RS = peak response ratio of acetaminophen to the
USP Pseudoephedrine Hydrochloride RS internal standard peak from the Standard
solution
CS = concentration of USP Acetaminophen RS in the
Acetaminophen and Diphenhydramine CU = nominal concentration of acetaminophen in the
Citrate Tablets Sample solution (mg/mL)
DIPHENHYDRAMINE CITRATE
(Comment on this Monograph)id=m280=Acetaminophen and Mobile phase: Methanol, water, and glacial acetic acid
Diphenhydramine Citrate Tablets=A-Monos.pdf) (61:38:1) containing 1.0813 g of sodium 1-octanesulfonate
DEFINITION in each 1000 mL of solution
Acetaminophen and Diphenhydramine Citrate Tablets contain Solution A: Methanol and water (1:1)
NLT 90.0% and NMT 110.0% of the labeled amounts of Internal standard solution: 8.0 mg/mL of xylometazoline
acetaminophen (C8H9NO2) and diphenhydramine citrate hydrochloride
(C17H21NO C6H8O7). Standard solution: Transfer 38 mg of USP
Diphenhydramine Citrate RS to a 100-mL volumetric flask
IDENTIFICATION containing 500 mg of acetaminophen. Add 5.0 mL of
The retention times of the major peaks of the Sample Internal standard solution and 50 mL of Solution A, and mix
solutions, obtained in the Assay for Acetaminophen and in the until solution is complete. Dilute with Solution A to volume.
Assay for Diphenhydramine Citrate, relative to the retention Sample solution: Weigh and finely powder NLT 20 Tablets.
times of the respective internal standards, correspond to Transfer a portion of the powder, equivalent to 38 mg of
those of the respective Standard solutions. diphenhydramine citrate, to a 100-mL volumetric flask, add
65 mL of Solution A, and shake by mechanical means for 15 DEFINITION

min. Add 5.0 mL of Internal standard solution, and dilute Acetaminophen, Diphenhydramine Hydrochloride, and
with Solution A to volume. Pseudoephedrine Hydrochloride Tablets contain NLT 90.0%
Chromatographic system and NMT 110.0% of the labeled amounts of acetaminophen
(See Chromatography 621, System Suitability.) (C8H9NO2), diphenhydramine hydrochloride (C17H21NO HCl),
Mode: LC and pseudoephedrine hydrochloride (C10H15NO HCl).
Detector: UV 265 nm
Column: 3.9-mm 30-cm; packing L1 IDENTIFICATION
Column temperature: 35 0.5 A. The retention time of the major peak for acetaminophen
Flow rate: 1.5 mL/min in the Sample solution corresponds to that in the Standard
Injection size: 50 L solution, as obtained in the Assay.
System suitability B. The retention time of the major peak for
Sample: Standard solution diphenhydramine in the Sample solution corresponds to that
[NOTEThe relative retention times for acetaminophen, in the Standard solution, as obtained in the Assay.
diphenhydramine citrate, and xylometazoline C. The retention time of the major peak for
hydrochloride are 0.3, 0.7, and 1.0, respectively.] pseudoephedrine in the Sample solution corresponds to that
Suitability requirements in the Standard solution, as obtained in the Assay.
Resolution: NLT 2.5 between the analyte and internal
standard peaks ASSAY
Column efficiency: NLT 1000 theoretical plates ACETAMINOPHEN
determined from the analyte peak Solution A: Transfer 6.8 g of monobasic potassium
Tailing factor: NMT 1.7 for the analyte peak phosphate to a 1000-mL volumetric flask, and add water to
Relative standard deviation: NMT 2.0% of the peak dissolve. Add 2.0 mL of triethylamine, and dilute with water
response ratios for replicate injections to volume. Adjust with phosphoric acid to a pH of 4.0.
Analysis Diluent: Acetonitrile and Solution A (11:89)
Samples: Standard solution and Sample solution Mobile phase: Acetonitrile and Solution A (3:47)
Calculate the percentage of C17H21NO C6H8O7 in the Standard solution: 25 g/mL of USP Acetaminophen RS,
portion of Tablets taken: 12.5 g/mL of USP Diphenhydramine Hydrochloride RS, and
30 g/mL of USP Pseudoephedrine Hydrochloride RS in
Result = (RU/RS) (CS/CU) 100 Diluent
Sample stock solution: Weigh and finely powder NLT 20
RU = peak response ratio from the Sample solution Tablets, and transfer a portion of the powder, equivalent to
RS = peak response ratio from the Standard solution 500 mg of acetaminophen, to a 100-mL volumetric flask.
CS = concentration of USP Diphenhydramine Citrate Add 75 mL of Diluent, shake, and sonicate for 15 min. Cool
RS in the Standard solution (mg/mL) to room temperature, dilute with Diluent to volume, and
CU = nominal concentration of diphenhydramine mix.
citrate in the Sample solution (mg/mL) Sample solution: Dilute an accurately measured volume of
Acceptance criteria: 90.0%110.0% the Sample stock solution, stepwise if necessary, with Diluent
to obtain a solution having a concentration of 25 g/mL of
PERFORMANCE TESTS acetaminophen.
DISSOLUTION 711, Procedure for a Pooled Sample Chromatographic system
Medium: Water; 900 mL (See Chromatography 621, System Suitability.)
Apparatus 2: 50 rpm Mode: LC
Time: 45 min Detector: UV 220 nm
Analysis: Determine the amount of C8H9NO2 and of Column: 4.6-mm 15-cm; packing L10
C17H21NO C6H8O7 dissolved, using the Analysis set forth in Flow rate: 2 mL/min
the Assay for Acetaminophen and the Assay for Injection size: 20 L
Diphenhydramine Citrate, respectively, making any necessary System suitability
volumetric adjustments. Sample: Standard solution
Tolerances: NLT 75% (Q) of the labeled amounts of Suitability requirements
C8H9NO2 and C17H21NO C6H8O7 is dissolved. Column efficiency: NLT 3000 theoretical plates
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements determined from the acetaminophen, diphenhydramine,
for Content Uniformity with respect to acetaminophen and to and pseudoephedrine peaks
diphenhydramine citrate Tailing factor: NMT 2.0 for the acetaminophen,
diphenhydramine, and pseudoephedrine peaks
ADDITIONAL REQUIREMENTS Relative standard deviation: NMT 2.0% determined
PACKAGING AND STORAGE: Preserve in tight containers, and from the acetaminophen, diphenhydramine hydrochloride,
store at controlled room temperature. and pseudoephedrine hydrochloride responses for
USP REFERENCE STANDARDS 11 replicate injections
USP Acetaminophen RS Analysis
USP Diphenhydramine Citrate RS Samples: Standard solution and Sample Solution
Tablets taken:
Acetaminophen, Diphenhydramine Result = (rU/rS) (CS/CU) 100
Hydrochloride, and Pseudoephedrine
Hydrochloride Tablets rU = peak response from the Sample solution
(Comment on this Monograph)id=m285=Acetaminophen, rS = peak response from the Standard solution
Diphenhydramine Hydrochloride, and Pseudoephedrine CS = concentration of USP Acetaminophen RS in the
Hydrochloride Tablets=A-Monos.pdf) Standard solution (g/mL)
CU = nominal concentration of acetaminophen in the Sample solution A: Combine equal volumes of the filtered
Sample solution (g/mL) solutions, and use the pooled sample.
Acceptance criteria: 90.0%110.0% Sample solution B: 5.0 mL of Sample solution A to a 100-
DIPHENHYDRAMINE HYDROCHLORIDE mL volumetric flask
Solution A, Diluent, Mobile phase, Standard solution, and Dilute with Mobile phase to volume.
Chromatographic system: Proceed as directed in the Analysis: Using Sample solution A and the Standard solution,
Assay for Acetaminophen. and making any necessary volumetric adjustments, proceed
Sample stock solution: Weigh and finely powder NLT 20 as directed in the Assay for Diphenhydramine hydrochloride
Tablets, and transfer a portion of the powder, equivalent to and the Assay for Pseudoephedrine Hydrochloride, and
12.5 mg of diphenhydramine hydrochloride, to a 100-mL determine the amounts of C17H21NO HCl and C10H15NO
volumetric flask. Add 75 mL of Diluent, shake, and sonicate HCl dissolved. Using Sample solution B and the Standard
for 15 min. Cool to room temperature, dilute with Diluent to solution, and making any necessary volumetric adjustments,
volume, and mix. proceed as directed in the Assay for Acetaminophen, and
Sample solution: Dilute a volume of Sample stock solution, determine the amount of C8H9NO2 dissolved.
stepwise if necessary, with Diluent to obtain a solution Tolerances: NLT 75% (Q) of the labeled amounts of
having a concentration of 12.5 g/mL of diphenhydramine. C8H9NO2, C17H21NO HCl, and C10H15NO HCl is dissolved.
Analysis For tablets labeled as chewable:
Samples: Standard solution and Sample Solution Medium: pH 5.8 phosphate buffer (see Reagents,
Calculate the percentage of C17H21NO HCl in the portion Indicators, and SolutionsBuffer Solutions); 900 mL
of Tablets taken: Apparatus 2: 75 rpm
Time: 45 min
Result = (rU/rS) (CS/CU) 100 Tolerances: NLT 75% (Q) of the labeled amounts of
C8H9NO2, C17H21NO HCl and C10H15NO HCl is dissolved.
rU = peak response from the Sample solution UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
CS = concentration of USP Diphenhydramine ADDITIONAL REQUIREMENTS
Hydrochloride RS in the Standard solution PACKAGING AND STORAGE: Preserve in tight containers, and
(g/mL) store at controlled room temperature.
CU = nominal concentration of diphenhydramine USP REFERENCE STANDARDS 11
hydrochloride in the Sample solution USP Acetaminophen RS
(g/mL) USP Diphenhydramine Hydrochloride RS
Acceptance criteria: 90.0%110.0% USP Pseudoephedrine Hydrochloride RS
PSEUDOEPHEDRINE HYDROCHLORIDE
Solution A, Diluent, Mobile phase, Standard solution, and
Chromatographic system: Proceed as directed in the
Assay for Acetaminophen. Acetaminophen and Pseudoephedrine
Sample stock solution: Weigh and finely powder NLT 20 Hydrochloride Tablets
Tablets. Transfer a portion of the powder, equivalent to 30 (Comment on this Monograph)id=m290=Acetaminophen and
mg of pseudoephedrine hydrochloride, to a 100-mL Pseudoephedrine Hydrochloride Tablets=A-Monos.pdf)
volumetric flask, add about 75 mL of Diluent, shake, and
sonicate for 15 min. Cool to room temperature, and dilute DEFINITION
with Diluent to volume. Acetaminophen and Pseudoephedrine Hydrochloride Tablets
Sample solution: Dilute a volume of Sample stock solution, contain NLT 90.0% and NMT 110.0% of the labeled amounts
stepwise if necessary, with Diluent to obtain a solution of acetaminophen (C8H9NO2) and pseudoephedrine
having a concentration of 30 g/mL of pseudoephedrine hydrochloride (C10H15NO HCl).
hydrochloride.
Calculate the percentage of C10H15NO HCl in the portion of IDENTIFICATION
Tablets taken: The retention times of the acetaminophen and
pseudoephedrine peaks of the Sample solution correspond to
Result = (rU/rS) (CS/CU) 100 those of the Standard solution, as obtained in the Assay.
rU = peak response from the Sample solution ASSAY

rS = peak response from the Standard solution PROCEDURE
CS = concentration of USP Pseudoephedrine Diluent: Acetonitrile and water (1:9)
Hydrochloride RS in the Standard solution Solution A: 0.005 M ethanesulfonic acid and 0.05 M
(g/mL) monobasic potassium phosphate
CU = nominal concentration of pseudoephedrine Mobile phase: Acetonitrile and Solution A (1:9)
hydrochloride in the Sample solution (g/mL) Adjust with 5 N sodium hydroxide or 1 N hydrochloric acid
Acceptance criteria: 90.0%110.0% to a pH of 4.6.
Pseudoephedrine hydrochloride standard stock solution:
PERFORMANCE TESTS USP Pseudoephedrine Hydrochloride RS in Diluent (0.6
DISSOLUTION, Procedure for a Pooled Sample 711 mg/mL)
Medium: pH 5.8 phosphate buffer (see Reagents, Indicators, Standard solution: Transfer 6J mg of USP Acetaminophen
and SolutionsBuffer Solutions); 900 mL RS to a 100-mL volumetric flask, J being the ratio of the
Apparatus 2: 50 rpm labeled quantity (mg) of acetaminophen to the labeled
Time: 45 min quantity (mg) of pseudoephedrine hydrochloride in each
Analysis: Determine the amounts of C8H9NO2, C17H21NO Tablet. Add 2.0 mL of 1 N hydrochloric acid and 20 mL of
HCl, and C10H15NO HCl dissolved by employing the Diluent, and mix to dissolve. Add 10.0 mL of
following method. Pseudoephedrine hydrochloride stock standard solution, and
Solution A, Diluent, Standard solution, Mobile phase, and dilute with Diluent to volume. This solution contains 0.06J
Assay for Acetaminophen.
mg/mL of USP Acetaminophen RS and 0.06 mg/mL of USP Calculate the percentage of C8H9NO2 and C10H15NO HCl
Pseudoephedrine Hydrochloride RS. dissolved:
pseudoephedrine hydrochloride, from finely powdered Result = (rU/rS) V (CS/L) 100
tablets (NLT 20), to a 500-mL volumetric flask, add 10.0 mL
of 1 N hydrochloric acid and 100 mL of Diluent, and rU = peak response of the corresponding analyte of
sonicate for 30 min, with occasional shaking. Allow to cool, the Sample solution
and dilute with Diluent to volume. Pass a portion of this rS = peak response of the corresponding analyte of
solution through a glass fiber filter, and use the filtrate. the Standard solution
Chromatographic system V = volume of Medium, 900 mL
(See Chromatography 621, System Suitability.) CS = concentration of the appropriate USP Reference
Mode: LC Standard in the Standard solution (mg/mL)
Detector: UV 214 nm L = label amount of the corresponding analyte in a
Column: 4.6-mm 25-cm; base-deactivated or end- Tablet (mg)
capped packing L1 Tolerances: NLT 75% (Q) of the labeled amounts of
Flow rate: 3 mL/min C8H9NO2 and C10H15NO HCl is dissolved.
Injection size: 10 L For tablets labeled as chewable
System suitability Medium: pH 5.8 phosphate buffer (see Reagents,
Sample: Standard solution Indicators, and SolutionsBuffer Solutions); 900 mL
[NOTEThe relative retention times for acetaminophen and Apparatus 2: 75 rpm
pseudoephedrine are about 0.55 and 1.0, respectively. Time: 45 min
The retention time for acetaminophen is NLT 2 min.] Standard solution, Sample solution, Chromatographic
Suitability requirements system, and Analysis: Proceed as directed above in
Resolution: NLT 3.5 between acetaminophen and Procedure for a Pooled Sample.
pseudoephedrine Tolerances: NLT 75% (Q) of the labeled amounts of
Tailing factor: NMT 2.0 for the pseudoephedrine peak C8H9NO2 and C10H15NO HCl is dissolved.
Relative standard deviation: NMT 2.0% for replicate UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
injections
Analysis ADDITIONAL REQUIREMENTS
Samples: Standard solution and Sample solution PACKAGING AND STORAGE: Preserve in tight containers, and
Calculate the percentage of C8H9NO2 and C10H15NO HCl store at controlled room temperature.
in the portion of Tablets taken: USP REFERENCE STANDARDS 11
USP Acetaminophen RS
Result = (rU/rS) (CS/CU) 100 USP Pseudoephedrine Hydrochloride RS
rU = peak response for the corresponding analyte of

the Sample solution Acetazolamide
rS = peak response for the corresponding analyte of
the Standard solution (Comment on this Monograph)id=m320=Acetazolamide=A-
CS = concentration of the appropriate USP Reference Monos.pdf)
Standard in the Standard solution (mg/mL)
CU = nominal concentration of the appropriate analyte
in the Sample solution (mg/mL)
PERFORMANCE TESTS
DISSOLUTION, Procedure for a Pooled Sample 711
Medium: pH 5.8 phosphate buffer (see Reagents, Indicators,
and SolutionsBuffer Solutions); 900 mL C4H6N4O3S2 222.25
Apparatus 2: 50 rpm Acetamide, N-[5-(aminosulfonyl)-1,3,4-thiadiazol-2-yl]-;
Time: 45 min N-(5-Sulfamoyl-1,3,4-thiadiazol-2-yl)acetamide [59-66-5].
Determine the amount of C8H9NO2 and C10H15NO HCl
dissolved by employing the following method. DEFINITION
Mobile phase: Proceed as directed in the Assay. Acetazolamide contains NLT 98.0% and NMT 102.0% of
Standard solution: L/900 mg/mL of USP Pseudoephedrine C4H6N4O3S2, calculated on the anhydrous basis.
Hydrochloride RS and LJ/900 mg/mL of USP Acetaminophen IDENTIFICATION
RS in Medium A. INFRARED ABSORPTION 197K
[NOTEL is the labeled quantity, in mg, of B. PROCEDURE
pseudoephedrine hydrochloride in each Tablet; and J is Sample solution: 20 mg/mL in 1 N sodium hydroxide
the ratio of the labeled quantity, in mg, of acetaminophen Analysis: To 5 mL of Sample solution, add 5 mL of a
to the labeled quantity, in mg, of pseudoephedrine solution made by dissolving 100 mg of hydroxylamine
hydrochloride in each Tablet.] hydrochloride and 80 mg of cupric sulfate in 10 mL of
Sample solution: Sample per Dissolution 711; filter. water. Heat the resulting pale yellow solution on a steam
Chromatographic system and System suitability: Proceed bath for 5 min.
as directed in the Assay. Acceptance criteria: A clear, bright yellow solution is
Analysis produced. No heavy precipitate or dark brown color results
Samples: Standard solution and Sample solution after the mixing or heating.
[NOTEInject 20 L of the Samples, and measure the
responses for the acetaminophen and pseudoephedrine
peaks.]
USP 32 Official Monographs / Acetazolamide 41
ASSAY Acetazolamide for Injection

PROCEDURE (Comment on this Monograph)id=m350=Acetazolamide for
Standard solution: 20 mg/mL of USP Acetazolamide RS in Injection=A-Monos.pdf)
pyridine
Sample solution: 20 mg/mL of Acetazolamide in pyridine DEFINITION
Blank: Pyridine Acetazolamide for Injection is prepared from Acetazolamide
Spectrometric conditions with the aid of Sodium Hydroxide. It is suitable for parenteral
Mode: IR use. The contents of each container, when constituted as
Analytical wavelength: 7.38 m (1350 cm 1) directed in the labeling, yield a solution containing NLT 95.0%
Cell: 0.1 mm and NMT 110.0% of the labeled amount of acetazolamide
Analysis (C4H6N4O3S2).
Samples: Standard solution and Sample Solution
Calculate the quantity, as a percentage, of C4H6N4O3S2 in IDENTIFICATION
the portion taken: A. PROCEDURE
Sample: Dissolve 500 mg in 5 mL of water, add 2 drops of
Result = (AU/AS) (CS/CU) 100 hydrochloric acid, and allow the mixture to stand for about
15 min. Filter through a fine sintered-glass funnel, wash with
AU = absorbance of the Sample solution several small portions of water, and dry in vacuum over
AS = absorbance of the Standard solution silica gel for 3 h: the crystals meet the requirements of the
CS = concentration of USP Acetazolamide RS in the following tests.
Standard solution (mg/mL) Analysis 1: Infrared Absorption 197K
CU = concentration of Sample solution (mg/mL) Analysis 2
Acceptance criteria: 98.0%102.0% Sample solution: 100 mg/mL of the Sample in 1N sodium
hydroxide
IMPURITIES To 5 mL of the Sample solution, add 5 mL of a solution
Inorganic Impurities made by dissolving 100 mg of hydroxylamine
RESIDUE ON IGNITION 281: NMT 0.1% hydrochloride and 80 mg of cupric sulfate in 10 mL of
CHLORIDE AND SULFATE, Chloride 221: Digest 1.5 g with 75 water. Mix, and heat the resulting pale yellow solution on
mL of water at about 70 for 5 min. Cool to room a steam bath for 5 min.
temperature, and filter. Acceptance criteria: A clear, bright yellow solution is
Acceptance criteria: A 25-mL portion of the filtrate shows produced. No heavy precipitate or dark brown color results
no more chloride than corresponds to 0.10 mL of 0.020 N after the mixing or heating.
hydrochloric acid (0.014%) B. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the
CHLORIDE AND SULFATE, Sulfate 221: A 25-mL portion of the requirements
filtrate prepared in the test for Chloride shows no more
sulfate than corresponds to 0.20 mL of 0.020 N sulfuric acid ASSAY
(0.04%). PROCEDURE
SELENIUM 291: 30 ppm, a 200-mg specimen being used Standard stock solution: 100 g/mL of USP Acetazolamide
HEAVY METALS, Method II 231: NMT 20 ppm RS in sodium hydroxide solution (1 in 100)
Organic Impurities Standard solution: Standard stock solution and 0.1 N
PROCEDURE: ORDINARY IMPURITIES 466 hydrochloric acid (1:9)
Standard solution and Sample solution: Acetone and Sample stock solution: 500 g/mL of acetazolamide from
methanol (1:1) Acetazolamide for Injection, in water
Eluant: n-Propyl alcohol and 1 N ammonium hydroxide [NOTEDissolve the contents of 1 container of
(22:3) Acetazolamide for Injection in a measured volume of
Visualization: 1 water corresponding to the volume of solvent specified in
the labeling. Dilute a portion of this solution to obtain the
SPECIFIC TESTS Sample stock solution.]
WATER DETERMINATION, Method I 921: NMT 0.5% Sample solution: Sample stock solution, 1 N hydrochloric
SILVER-REDUCING SUBSTANCES acid, and water (1:5:44)
Sample: 5 g Blank: 0.1 N hydrochloric acid
Analysis: Thoroughly wet the Sample with alcohol. Add 125 Spectrometric conditions
mL of water, 10 mL of nitric acid, and 5.0 mL of 0.1 N silver Mode: UV
nitrate VS. Stir with a mechanical stirrer for 30 min. Filter, Analytical wavelength: 265 nm
add 5 mL of ferric ammonium sulfate TS to the filtrate, and Analysis
titrate with 0.1 N ammonium thiocyanate VS to a reddish- Samples: Standard solution and Sample Solution
brown endpoint. Calculate the percentage of C4H6N4O3S2 in the portion of
Acceptance criteria: NLT 4.8 mL of 0.1 N ammonium Acetazolamide for Injection taken:
thiocyanate is required, calculated on the anydrous basis.
Result = (AU/AS) (CS/CU) 100
PACKAGING AND STORAGE: Preserve in tight containers, and AU = absorbance of the Sample solution
store at room temperature. AS = absorbance of the Standard solution
USP Acetazolamide RS
42 Acetazolamide / Official Monographs USP 32
CS = concentration of USP Acetazolamide RS in the Sample solution: 250 g/mL of acetazolamide from
Standard solution (g/mL) Acetazolamide Oral Suspension in Mobile phase
CU = nominal concentration of acetazolamide in the [NOTEAgitate the container of Oral Suspension for 30 min
Sample solution (g/mL) on a rotating mixer, remove a 5-mL sample, and store in
Acceptance criteria: 95.0%110.0% a clear glass vial at 70 until analyzed. At the time of
analysis, remove the Sample solution from the freezer,
PERFORMANCE TESTS allow to reach room temperature, and mix with a vortex
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements mixer for 30 s. Pipet 1.0 mL of the Sample solution to a
100-mL volumetric flask, and dilute with Mobile phase to
SPECIFIC TESTS volume.]
PH 791: 9.010.0, in a freshly prepared solution (1 in 10) Chromatographic system
BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.5 USP (See Chromatography 621, System Suitability.)
Endotoxin Unit/mg of acetazolamide. Mode: LC
INJECTIONS 1, Constituted Solutions: At the time of use, it Detector: UV 254 nm
meets the requirements. Column: 4.6-mm 25-cm; 5-m packing L1
STERILITY TESTS 71: It meets the requirements. Flow rate: 2 mL/min
COMPLETENESS OF SOLUTION 641: 100 mg/mL in carbon Injection size: 20 L
dioxidefree water dissolves to yield a clear solution. System suitability
INJECTIONS 1, Labeling: It meets the requirements. Sample: Standard solution
ADDITIONAL REQUIREMENTS [NOTEThe relative retention time for the acetazolamide
PACKAGING AND STORAGE: Preserve as described under peak is about 3 min.]
Injections 1, Containers for Sterile Solids, preferably of Type Suitability requirements
III glass, and store at room temperature. Relative standard deviation: Acetazolamide, NMT 1.1%
USP REFERENCE STANDARDS 11 for replicate injections of the Standard solution
USP Acetazolamide RS Analysis
USP Endotoxin RS Samples: Standard solution and Sample Solution
Calculate the percentage of C4H6N4O3S2 in the portion of
Acetazolamide Oral Suspension taken:
Acetazolamide Oral Suspension Result = (rU/rS) (CS/CU) 100

(Comment on this Monograph)id=m1376=Acetazolamide Oral rU = peak response from the Sample solution
Suspension=A-Monos.pdf) rS = peak response from the Standard solution
DEFINITION CS = concentration of USP Acetazolamide RS in the
Acetazolamide Oral Suspension contains NLT 90.0% and NMT Standard solution (g/mL)
110.0% of the labeled amount of acetazolamide (C4H6N4O3S2). CU = nominal concentration of acetazolamide in the
Prepare Acetazolamide Oral Suspension, 25 mg/mL, as follows. Sample solution (g/mL)
(See Pharmaceutical CompoundingNonsterile Preparations Acceptance criteria: 90.0%110.0%
795.) (See also Acetazolamide Oral Solution.) SPECIFIC TESTS
PH 791: 4.05.0 (Vehicle for Oral Solution and Vehicle for
Acetazolamide 2.5 g Oral Suspension), and 3.13.9 (Cherry Syrup)
Vehicle: a mixture of Vehicle for Oral A sufficient quantity
Solution, NF (regular or sugar-free), and
PACKAGING AND STORAGE: Preserve in tight, light-resistant
Vehicle for Oral Suspension, NF (1:1), or
containers. Store at controlled room temperature, or in a
Cherry Syrup, NF
cold place.
To make 100 mL LABELING: Label it to state that it is to be well shaken
before use, and to state the beyond-use date.
[NOTEIf Tablets are used instead of bulk powder, the BEYOND-USE DATE: 60 days after the day on which it was
preparation becomes a suspension and should be labeled as compounded
such.] USP REFERENCE STANDARDS 11
If using Tablets, place the Tablets in a mortar and comminute USP Acetazolamide RS
the Tablets to a fine powder, or add Acetazolamide powder.
Add about 20 mL of the Vehicle, and mix to a uniform paste.
Add the Vehicle in small portions almost to volume, and mix
thoroughly after each addition. Transfer the contents of the Acetazolamide Tablets
mortar, stepwise and quantitatively, to a calibrated bottle. Add (Comment on this Monograph)id=m360=Acetazolamide
enough liquid Vehicle to bring to final volume, and mix well. Tablets=A-Monos.pdf)
ASSAY DEFINITION
PROCEDURE Acetazolamide Tablets contain NLT 95.0% and NMT 105.0% of
Mobile phase: Dissolve 4.1 g of anhydrous sodium acetate the labeled amount of acetazolamide (C4H6N4O3S2).
in 950 mL of water, and add 20 mL of methanol and 30 mL
of acetonitrile. Adjust with glacial acetic acid to a pH of 4.0. IDENTIFICATION
Standard stock solution: 0.5 mg/mL of USP Acetazolamide Sample stock solution: Equivalent to 10 mg/mL of
RS in 0.5 N sodium hydroxide solution and water (1.0:9.0) acetazolamide in acetone
Standard solution: 250 g/mL from Standard stock solution Sample: Filter the Sample stock solution, and add sufficient
in water solvent hexane to the filtrate to cause formation of a heavy,
USP 32 Official Monographs / Glacial 43
white precipitate. Collect the precipitate on a medium- Acceptance criteria: 95.0%105.0%

porosity, sintered-glass funnel, and dry with suction.
A. INFRARED ABSORPTION 197K: Use the Sample. PERFORMANCE TESTS
B. PROCEDURE: Dissolve 100 mg of Sample in 5 mL of 1 N DISSOLUTION 711
sodium hydroxide, add 5 mL of a solution made by Medium: 0.01 N hydrochloric acid; 900 mL
dissolving 100 mg of hydroxylamine hydrochloride and 80 Apparatus 1: 100 rpm
mg of cupric sulfate in 10 mL of water. Heat the resulting Time: 60 min
pale yellow solution on a steam bath for 5 min: a clear, Detector: UV 265 nm
bright yellow solution is produced. Standard solution: USP Acetazolamide RS in the same
Acceptance criteria: No heavy precipitate or dark brown Medium
color results after the mixing or heating. Sample solutions: Sample per Dissolution 711.
Analysis: Determine the amount of C4H6N4O3S2 dissolved
ASSAY by using UV absorption on filtered portions of the Sample
PROCEDURE solution suitably diluted with Medium, if necessary, in
Mobile phase: 4.1 g of anhydrous sodium acetate in 950 comparison with a Standard solution having a known
mL. Add 20 mL of methanol and 30 mL of acetonitrile. concentration of USP Acetazolamide RS.
Adjust with glacial acetic acid to a pH of 4.0 0.05. Acceptance criteria: NLT 75% (Q) of the labeled amount
Internal standard stock solution: 10 mg/mL of sulfadiazine of C4H6N4O3S2 is dissolved
in 0.5 N sodium hydroxide UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Internal standard solution: 1 mg/mL from Internal
standard stock solution in water ADDITIONAL REQUIREMENTS
Standard stock solution: Dissolve 25 mg of USP PACKAGING AND STORAGE: Preserve in tight containers, and
Acetazolamide RS in 2.5 mL of 0.5 N sodium hydroxide. store at a controlled room temperature.
Dilute with water to 25 mL. USP REFERENCE STANDARDS 11
Standard solution: Transfer 10.0 mL of Standard USP Acetazolamide RS
acetazolamide stock solution, 10.0 mL of Internal standard
solution, and 10 mL of 0.5 N sodium hydroxide to a 100-mL
volumetric flask. Dilute with water to volume. Glacial Acetic Acid
Sample stock solution: Transfer a portion of the powder
equivalent to 100 mg acetazolamide. Add 10 mL of 0.5 N (Comment on this Monograph)id=m440=Glacial Acetic Acid=A-
sodium hydroxide, sonicate for 5 min, cool to room Monos.pdf)
temperature, and dilute with water to 100 mL. Filter a
portion of this solution, discarding the first 20 mL of the
filtrate.
Sample solution: Transfer 10.0 mL of Sample stock solution,
10.0 mL of Internal standard solution, and 10 mL of 0.5 N
sodium hydroxide to a 100-mL volumetric flask, and dilute
with water to volume.
(See Chromatography 621, System Suitability.) C2H4O2 60.05
Mode: LC Acetic acid [64-19-7].
Detector: UV 254 nm DEFINITION
Column: 4.6-mm 25-cm; packing L1 Glacial Acetic Acid contains NLT 99.5% and NMT 100.5%, by
Flow rate: 2 mL/min weight, of C2H4O2.
System suitability IDENTIFICATION
Sample: Standard solution IDENTIFICATION TESTSGENERAL, Acetate 191: Meets the
[NOTEThe relative retention times for acetazolamide and requirements
sulfadiazine are about 0.7 and 1.0, respectively.] Sample solution: Glacial Acetic Acid and water (1:2)
Resolution: NLT 2.0 between the analyte and internal ASSAY
standard peaks PROCEDURE
Relative standard deviation: NMT 1.0% for the ratios of Sample solution: Measure 2 mL of Glacial Acetic Acid into
the analyte peak response to the internal standard peak a glass-stoppered flask, previously tared while containing
response about 20 mL of water, and weigh again to obtain the
Analysis weight of the substance under assay.
Samples: Standard solution and Sample solution Analysis: Add 20 mL of water, then add phenolphthalein
Calculate the quantity, as a percentage, of C4H6N4O3S2 in TS. Titrate with 1 N sodium hydroxide VS. Each mL of 1 N
the portion of Tablets taken: sodium hydroxide is equivalent to 60.05 mg of C2H4O2.
IMPURITIES
RU = peak response ratios of the analyte peak to the Inorganic Impurities
internal standard peak obtained from the LIMIT OF NONVOLATILE RESIDUE: Evaporate 20 mL in a tared
Sample solution dish, and dry at 105 for 1 h: the weight of the residue does
RS = peak response ratios of the analyte peak to the not exceed 1.0 mg.
internal standard peak obtained from the HEAVY METALS 231: NMT 5 ppm
Standard solution Sample solution: To the residue obtained in the test for
CS = concentration of USP Acetazolamide RS in Limit of Nonvolatile Residue add 8 mL of 0.1 N hydrochloric
Standard solution (mg/mL) acid, warm gently until solution is complete, dilute with
CU = nominal concentration of acetazolamide in the water to 100 mL, and use 20 mL.
44 Glacial / Official Monographs USP 32
CHLORIDE AND SULFATE, Chloride 221 Acetic Acid Otic Solution

Sample solution: Dilute 1.0 mL with 20 mL of water. (Comment on this Monograph)id=m460=Acetic Acid Otic
Analysis: Add 5 drops of silver nitrate TS. Solution=A-Monos.pdf)
Acceptance criteria: No opalescence is produced.
CHLORIDE AND SULFATE, Sulfate 221 DEFINITION
Sample: Dilute 1.0 mL with 10 mL of water. Acetic Acid Otic Solution is a solution of Glacial Acetic Acid in a
Analysis: Add 1 mL of barium chloride TS. suitable nonaqueous solvent. It contains NLT 85.0% and NMT
Acceptance criteria: No turbidity is produced. 130.0% of the labeled amount of C2H4O2.
Organic Impurities
PROCEDURE: READILY OXIDIZABLE SUBSTANCES IDENTIFICATION
Sample solution: Dilute 2.0 mL in a glass-stoppered vessel A. PROCEDURE
with 10 mL of water. Sample solution: Acetic Acid Otic Solution and water (1:2)
Analysis: Add 0.10 mL of 0.10 N potassium Analysis: Adjust 10 mL of the Sample solution with 1 N
permanganate. sodium hydroxide to a pH of 7. Add ferric chloride TS.
Acceptance criteria: The pink color is not changed to Acceptance criteria: A deep red color is produced, and it is
brown within 2 h. destroyed by the addition of hydrochloric acid.
B. Warm it with sulfuric acid and alcohol: ethyl acetate,
SPECIFIC TESTS recognizable by its characteristic odor, is evolved.
CONGEALING TEMPERATURE 651: NLT 15.6
ASSAY
ADDITIONAL REQUIREMENTS PROCEDURE
PACKAGING AND STORAGE: Preserve in tight containers, and Sample solution: Transfer a quantity of Otic Solution,
store at room temperature. containing 100 mg of glacial acetic acid, to a 250-mL
conical flask, and add 5 mL of saturated sodium chloride
solution and 40 mL of water. Add 3 drops of
phenolphthalein TS.
Acetic Acid Irrigation Analysis: Titrate with 0.1 N sodium hydroxide VS to a faint
(Comment on this Monograph)id=m450=Acetic Acid pink endpoint. Each mL of 0.1 N sodium hydroxide is
Irrigation=A-Monos.pdf) equivalent to 6.005 mg of C2H4O2.
DEFINITION
Acetic Acid Irrigation is a sterile solution of Glacial Acetic Acid in SPECIFIC TESTS
Water for Injection. It contains, in each 100 mL, NLT 237.5 PH 791: 2.04.0, when diluted with an equal volume of
mg and NMT 262.5 mg of C2H4O2. water
IDENTIFICATION ADDITIONAL REQUIREMENTS
PROCEDURE PACKAGING AND STORAGE: Preserve in tight containers, and
Evaporate 100 mL to about 10 mL: the resulting solution store at controlled room temperature.
meets the requirements for Identification TestsGeneral,
Acetate 191.
ASSAY Acetohexamide
PROCEDURE (Comment on this Monograph)id=m510=Acetohexamide=A-
Analysis: Pipet 50 mL of Irrigation into a 150-mL conical Monos.pdf)
flask, add 2 drops of phenolphthalein TS, and titrate with
0.1 N sodium hydroxide VS. Each mL of 0.1 N sodium
hydroxide is equivalent to 6.005 mg of C2H4O2.
Acceptance criteria: 237.5262.5 mg of C2H4O2
SPECIFIC TESTS
PH 791: 2.83.4
BACTERIAL ENDOTOXINS TEST 85 It contains NMT 0.5
Endotoxin Unit/mL. C15H20N2O4S 324.40
OTHER REQUIREMENTS: It meets the requirements under Benzenesulfonamide, 4-acetyl-N-[[cyclohexylamino]carbonyl]-;
Injections 1, except that the container in which it is 1-[(p-Acetylphenyl)sulfonyl]-3-cyclohexylurea [968-81-0].
packaged may be designed to empty rapidly and may
exceed 1000 mL in capacity. DEFINITION
Acetohexamide contains NLT 97.0% and NMT 101.0% of
ADDITIONAL REQUIREMENTS C15H20N2O4S, calculated on the dried basis.
PACKAGING AND STORAGE: Preserve in single-dose containers,
preferably of Type I or Type II glass, and store at controlled
room temperature. It may be packaged in suitable plastic
containers.
USP Endotoxin RS
USP 32 Official Monographs / Acetohydroxamic 45
IDENTIFICATION Sample solution: Quantitatively dilute Sample stock solution

A. INFRARED ABSORPTION 197K 3 (1 in 10) in water (10 g/mL).
B. ULTRAVIOLET ABSORPTION 197U Blank: 0.01 N sodium hydroxide
Solution: 10 g/mL in 0.01 N sodium hydroxide Spectrometric conditions
Analytical wavelength: 247 nm Mode: UV
Acceptance criteria: Absorptivities, calculated on the dried Analytical wavelength: 247 nm
basis, do not differ by more than 3.0%. Cell: 1 cm
Analysis
ASSAY Samples: Standard solution and Sample Solution
PROCEDURE Calculate the percentage of C15H20N2O4S in the Tablets
Sample solution: Dissolve about 300 mg of Acetohexamide taken:
in 40 mL of dimethylformamide.
Analysis: To the Sample solution, add 5 drops of thymol Result = (AU/AS) (CS/CU) (100/L)
blue TS, and titrate, using a magnetic stirrer, with 0.1 N
sodium methoxide VS to a blue endpoint. Perform a blank AU = absorbance of the Sample solution
determination, and make any necessary correction. Each mL AS = absorbance of the Standard solution
of 0.1 N sodium methoxide is equivalent to 32.44 mg of CS = concentration of the Standard solution (g/mL)
C15H20N2O4S. CU = concentration of the Sample solution (tablet/mL)
Acceptance criteria: 97.0%101.0% L = label claim (mg/tablet)
IMPURITIES
Inorganic Impurities PERFORMANCE TESTS
RESIDUE ON IGNITION 281: NMT 0.1% DISSOLUTION 711
SELENIUM 291: 30 ppm; a 200-mg specimen mixed with Medium: pH 7.6 phosphate buffer (see pH 791); 900 mL
200 mg of magnesium oxide being used Apparatus 1: 100 rpm
HEAVY METALS, Method II 231: NMT 20 ppm Time: 60 min
Detector: UV 245 nm
SPECIFIC TESTS Standard solution: USP Acetohexamide RS in Medium
MELTING RANGE OR TEMPERATURE 741: 182.2187 Sample solution: Sample per Dissolution 711; dilute with
LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT Medium to a concentration that is similar to that of the
1.0% of its weight. Standard solution.
Analysis: Determine the amount of C15H20N2O4S dissolved,
ADDITIONAL REQUIREMENTS by using Medium as the blank, in comparison with the
PACKAGING AND STORAGE: Preserve in well-closed containers. Standard solution.
USP REFERENCE STANDARDS 11 Tolerances: NLT 75% (Q) of the labeled amount of
USP Acetohexamide RS C15H20N2O4S is dissolved.
Acetohexamide Tablets ADDITIONAL REQUIREMENTS

PACKAGING AND STORAGE: Preserve in well-closed containers.
(Comment on this Monograph)id=m518=Acetohexamide USP REFERENCE STANDARDS 11
Tablets=A-Monos.pdf) USP Acetohexamide RS
DEFINITION
Acetohexamide Tablets contain NLT 93.0% and NMT 107.0%
of the labeled amount of C15H20N2O4S. Acetohydroxamic Acid
IDENTIFICATION (Comment on this Monograph)id=m540=Acetohydroxamic
A. INFRARED ABSORPTION 197K Acid=A-Monos.pdf)
Sample preparation: Evaporate 20.0 mL of Sample stock
solution 2 in the Assay to dryness, use the residue in the
procedure.
ASSAY
PROCEDURE
Standard solution: 10 g/mL of USP Acetohexamide RS
Sample stock solution 1: Weigh and finely powder NLT 20
Tablets. Transfer a portion of powder equivalent to 500 mg C2H5NO2 75.07
of acetohexamide to a 100-mL volumetric flask, add 60 mL N-Acetyl hydroxyacetamide;
of 0.1 N sodium hydroxide, shake for 30 min, dilute with Acetohydroxamic acid [546-88-3].
water to volume and filter, discarding the first 20 mL of the DEFINITION
filtrate (5 mg/mL). Acetohydroxamic Acid, dried over phosphorus pentoxide for 16
Sample stock solution 2: Transfer 20.0 mL of Sample stock h, contains NLT 98.0% and NMT 101.0% of C2H5NO2.
solution 1 to a separator, add 2 mL of 3 N hydrochloric acid,
extract with four 40-mL portions of chloroform, filtering IDENTIFICATION
each portion through chloroform-washed paper into a 200- A. INFRARED ABSORPTION 197K
mL volumetric flask. Dilute with chloroform to volume (0.5 B. PROCEDURE
mg/mL). Sample solution: 20 mg/mL in water
Sample stock solution 3: Evaporate 20.0 mL of Sample Analysis: To 10 mL of Sample solution, add 2 drops of
stock solution 2 to dryness, transfer the residue, with the aid potassium permanganate TS.
of 0.1 N sodium hydroxide, to a 100-mL volumetric flask, Acceptance criteria: The pink color of the permanganate
and dilute with 0.1 N sodium hydroxide to volume (0.1 disappears.
mg/mL).
46 Acetohydroxamic / Official Monographs USP 32
ASSAY the best-fit straight line from the fluorescence intensities

PROCEDURE of the three Standard solutions versus the hydroxylamine
Solution A: 20 mg/mL of ferric chloride in 0.1 N hydrochloride concentrations, in g/mL. From the best-fit
hydrochloric acid straight line, determine the concentration, in g/mL, of
Standard solution: 500 g/mL of USP Acetohydroxamic hydroxylamine hydrochloride in the Sample solution.
Acid RS in 0.1 N hydrochloric acid Calculate the percentage of hydroxylamine in the portion
Sample solution: 500 g/mL of Acetohydroxamic Acid in of Acetohydroxamic Acid taken:
0.1 N hydrochloric acid
Blank: 0.1 N hydrochloric acid Result = (CU/C) (Mr1/Mr2) 100
Analysis: Transfer 10.0 mL each of the Standard solution,
the Sample solution, and Blank, to separate 100-mL CU = measured concentration of hydroxylamine
volumetric flasks. To each flask add 50 mL of 0.1 N hydrochloride in the Sample solution (g/mL)
hydrochloric acid and 10.0 mL of Solution A, and dilute with C = concentration of Acetohydroxamic Acid in the
0.1 N hydrochloric acid to volume. [NOTEWithout delay, Sample solution (g/mL)
concomitantly determine the absorbances.] Mr1 = molecular weight of hydroxylamine, 33.03
Spectrometric conditions Mr2 = molecular weight of hydroxylamine
Mode: UV hydrochloride, 69.50
Analytical wavelength: 502 nm Acceptance criteria: NMT 0.5%
Analysis
Samples: Standard solution and Sample solution SPECIFIC TESTS
Calculate the percentage of C2H5NO2 in the portion of LOSS ON DRYING 731: Dry it over phosphorus pentoxide for
Acetohydroxamic Acid taken: 16 h: it loses NMT 1.0% of its weight.
COMPLETENESS OF SOLUTION 641: A 1.0-g portion dissolves
Result = (AU/AS) (CS/CU) 100 in 10 mL of water to yield a clear solution
COLOR OF SOLUTION
AU = absorbance of the Sample solution Sample solution: 200 mg/mL
AS = absorbance of the Standard solution Blank: Water
CS = concentration of USP Acetohydroxamic Acid RS Spectrometric conditions
in the Standard solution (g/mL) Mode: Spectroscopy
CU = concentration of Acetohydroxamic Acid in the Cell: 1 cm
Sample solution (g/mL) Analytical wavelength: Between 400 and 750 nm
Acceptance criteria: 98.0%101.0% Analysis
Sample: Sample solution
IMPURITIES Measure the absorbance.
Inorganic Impurities Acceptance criteria: NMT 0.050
HEAVY METALS, Method I 231: NMT 20 ppm ADDITIONAL REQUIREMENTS
Sample solution: 40 mg/mL in a mixture of 1 N acetic PACKAGING AND STORAGE: Preserve in tight containers, and
acid and water (2:23) store in a cool, dry place.
Organic Impurities USP REFERENCE STANDARDS 11
PROCEDURE: LIMIT OF HYDROXYLAMINE USP Acetohydroxamic Acid RS
Solution A: Dissolve 1.36 g of monobasic potassium
phosphate in 950 mL of water, adjust with 1 M potassium
hydroxide to a pH of 7.4, and dilute with water to 1000 Acetohydroxamic Acid Tablets
mL.
Solution B: 1 mg/mL of pyridoxal 5-phosphate (Comment on this Monograph)id=m548=Acetohydroxamic Acid
monohydrate in Solution A in a low-actinic flask [NOTE Tablets=A-Monos.pdf)
Prepare fresh before use.]
Standard stock solution: 2.0 mg/mL of hydroxylamine DEFINITION
hydrochloride Acetohydroxamic Acid Tablets contain NLT 90.0% and NMT
Standard solutions: Transfer 5.0, 10.0, and 15.0 mL of 110.0% of the labeled amount of C2H5NO2.
the Standard stock solution, to separate 100-mL volumetric IDENTIFICATION
flasks, and dilute with water to volume. Tablets produce a purple color when mixed with an acidic
Sample solution: Transfer 1500 mg of Acetohydroxamic solution of ferric chloride.
Acid, previously dried, to a 100-mL beaker, and dissolve in
a sufficient amount of water to cover the electrode of a ASSAY
calibrated pH meter (60 mL). While stirring, adjust with PROCEDURE
0.05 M potassium hydroxide to a pH of 7.4. Transfer the Solution A: 20 mg/mL of ferric chloride in 0.1 N
contents of the beaker, with the aid of small portions of hydrochloric acid
water, to a 100-mL volumetric flask, and dilute with water Standard solution: 500 g/mL of USP Acetohydroxamic
to volume. Acid RS in 0.1 N hydrochloric acid
Blank: Water Sample solution: Transfer an equivalent to 500 mg of
Analysis: Transfer 2.0 mL of each Standard solution, the acetohydroxamic acid, from finely powdered Tablets (NLT
Sample solution, and Blank into separate 100-mL volumetric 20), to a 1000-mL volumetric flask, add 500 mL of 0.1 N
flasks. To each flask, add 4.0 mL of Solution B. After 8 min, hydrochloric acid, and shake for 1 min. Dilute with 0.1 N
accurately timed, dilute the contents of each flask with hydrochloric acid to volume, and mix. Filter, discarding the
Solution A to volume. first 40 mL of the filtrate. Use the clear filtrate.
Immediately determine the fluorescence intensities of the Blank: 0.1 N hydrochloric acid
solutions from the Standard solutions and the Sample Analysis: Transfer 10.0 mL each of the Standard solution,
solution in a fluorometer at an excitation wavelength of the Sample solution, and Blank, to separate 100-mL
350 nm and an emission wavelength of 450 nm, setting volumetric flasks. To each flask add 50 mL of 0.1 N
the instrument to zero with the reagent blank. Determine
USP 32 Official Monographs / Acetylcholine 47
hydrochloric acid and 10.0 mL of Solution A. Dilute with 0.1 350 nm and an emission wavelength of 450 nm, setting
N hydrochloric acid to volume. [NOTEWithout delay, the instrument to zero with the reagent blank. Determine
concomitantly determine the absorbances.] the best-fit straight line from the fluorescence intensities
Spectrometric conditions of the three Standard solutions versus the hydroxylamine
Mode: UV hydrochloride concentrations, in g/mL. From the best-fit
Analytical wavelength: 502 nm straight line, determine the concentration, in g/mL, of
Analysis hydroxylamine hydrochloride in the Sample solution.
Samples: Standard solution and Sample solution Calculate the percentage of H3NO in the portion of Tablets
Calculate the percentage of C2H5NO2 in the portion of taken:
Tablets taken:
Result = (CS/CU) (Mr1/Mr2) 100
CS = concentration of USP Hydroxylamine
AU = absorbance of the Sample solution Hydrochloride RS in the Standard solution
AS = absorbance of the Standard solution (g/mL)
CS = concentration of USP Acetohydroxamic Acid RS CU = nominal concentration of hydroxylamine
in the Standard solution (g/mL) hydrochloride in the Sample solution (g/mL)
CU = nominal concentration of acetohydroxamic acid Mr1 = molecular weight of hydroxylamine, 33.03
in the Sample solution (g/mL) Mr2 = molecular weight of hydroxylamine
Acceptance criteria: 90.0%110.0% hydrochloride, 69.50
Acceptance criteria: NMT 0.5%
PERFORMANCE TESTS
DISSOLUTION 711, Procedure for a Pooled Sample ADDITIONAL REQUIREMENTS
Medium: 0.01 N hydrochloric acid; 900 mL PACKAGING AND STORAGE: Preserve in tight containers.
Apparatus 1: 100 rpm USP REFERENCE STANDARDS 11
Time: 30 min USP Acetohydroxamic Acid RS
Sample solution: Sample per Dissolution 711.
Standard solution: USP Acetohydroxamic Acid RS in the
same Medium
Analysis: Determine the amount of C2H5NO2 dissolved, as Acetylcholine Chloride
directed in the Analysis under the Assay, using a filtered (Comment on this Monograph)id=m680=Acetylcholine
portion of the Sample solution in comparison with a Standard Chloride=A-Monos.pdf)
solution having a known concentration of USP
Acetohydroxamic Acid RS.
Tolerances: NLT 85% (Q) of the labeled amount of
C2H5NO2 is dissolved.
IMPURITIES
Organic Impurities
PROCEDURE: LIMIT OF HYDROXYLAMINE C7H16ClNO2 181.66
Solution A: Dissolve 1.36 g of monobasic potassium Ethanaminium, 2-(acetyloxy)-N, N, N-trimethyl-, chloride;
phosphate in 950 mL of water, adjust with 1 M potassium Choline acetate (ester) chloride. [60-31-1].
hydroxide to a pH of 7.4, and dilute with water to 1000
mL. DEFINITION
Solution B: 1 mg/mL of pyridoxal 5-phosphate Acetylcholine Chloride contains NLT 98.0% and NMT 102.0%
monohydrate in Solution A in a low-actinic flask [NOTE of C7H16ClNO2, calculated on the dried basis.
Prepare fresh before use.]
Standard stock solution: 2.0 mg/mL of hydroxylamine IDENTIFICATION
hydrochloride in water A. INFRARED ABSORPTION 197K
Standard solutions: Transfer 5.0, 10.0, and 15.0 mL of B. PROCEDURE
the Standard stock solution, to separate 100-mL volumetric Sample solution: 100 mg/mL in water
flasks, and dilute with water to volume. Analysis: To 5 mL of Sample solution, add 5 mL of silver
Sample solution: Transfer an equivalent to 1500 mg of nitrate TS: a white, curdy precipitate, which is soluble in
acetohydroxamic acid, from finely powdered Tablets (NLT ammonium hydroxide but insoluble in nitric acid, is formed.
20), to a 50-mL stoppered centrifuge tube. Add 30.0 mL of ASSAY
water, shake for 2 min, and centrifuge. Pipet 15.0 mL of PROCEDURE
the clear solution into a 50-mL beaker, add just enough Sample: 400 mg of Acetylcholine Chloride
water to cover the electrode of a calibrated pH meter, and Analysis: Dissolve in 15 mL of water in a glass-stoppered
while stirring, adjust with 0.5 M potassium hydroxide to a conical flask, add 40.0 mL of 0.1 N sodium hydroxide VS,
pH of 7.4. Transfer the contents of the beaker, with the aid and heat on a steam bath for 30 min. Insert the stopper,
of small portions of water, to a 50-mL volumetric flask, and allow to cool, add phenolphthalein TS, and titrate the excess
dilute with water to volume. alkali with 0.1 N sulfuric acid VS. Perform a blank
Blank: Water determination (see Residual Titrations under Titrimetry 541).
Analysis: Transfer 2.0 mL of each Standard solution, the Each mL of 0.1 N sodium hydroxide is equivalent to 18.17
Sample solution, and Blank into separate 100-mL volumetric mg of C7H16ClNO2.
flasks. To each flask, add 4.0 mL of Solution B. After 8 min, Acceptance criteria: 98.0%102.0%
accurately timed, dilute the contents of each flask with
Solution A to volume. OTHER COMPONENTS
Immediately determine the fluorescence intensities of the CONTENT OF CHLORIDE: Transfer 280 mg to a porcelain
solutions from the Standard solutions and the Sample casserole, and add 140 mL of water and 1 mL of
solution in a fluorometer at an excitation wavelength of
48 Acetylcholine / Official Monographs USP 32
dichlorofluorescein TS. Titrate with 0.1 N silver nitrate VS ASSAY

until the silver chloride flocculates and the mixture acquires PROCEDURE
a faint pink color. Each mL of 0.1 N silver nitrate is Mobile phase: Add 1.03 g of sodium 1-heptanesulfonate to
equivalent to 3.545 mg of Cl. NLT 19.3% and NMT 19.8% a mixture of 900 mL of water and 10 mL of methanol. Mix,
of Cl, calculated on the dried basis. then add sufficient glacial acetic acid and ammonium
hydroxide, if necessary, to adjust the solution to a pH of 4.0.
IMPURITIES Add 50 mL of acetonitrile, then add water to make 1000
Inorganic Impurities mL. [NOTESlight variation of the amount of acetonitrile
RESIDUE ON IGNITION 281: NMT 0.2% may be required to improve resolution or adjust retention
time.]
SPECIFIC TESTS System suitability solution: 0.2% solution of each of
MELTING RANGE OR TEMPERATURE, Class I 741: 149152 acetylcholine chloride and choline chloride
ACIDITY: Dissolve 100 mg in 10 mL of recently boiled water, Standard solution: A quantity of USP Acetylcholine
and add at once 1 drop of bromothymol blue TS: NMT 0.50 Chloride RS in Mobile phase, to obtain a solution having a
mL of 0.010 N sodium hydroxide is required in order to known concentration equal to that of the acetylcholine
produce a color change. chloride in the Sample solution
LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT Sample solution: Transfer the contents of 1 container of
1.0% of its weight. Acetylcholine Chloride for Ophthalmic Solution to a 10-mL
ADDITIONAL REQUIREMENTS volumetric flask with the aid of Mobile phase, and add
PACKAGING AND STORAGE: Preserve in a tight container, and Mobile phase to volume.
store at controlled room temperature. Chromatographic system
USP REFERENCE STANDARDS 11 (See Chromatography 621, System Suitability.)
USP Acetylcholine Chloride RS Mode: LC
Detector: Refractive index
Column: 3.9 mm 30 cm; packing L1
Flow rate: 2 mL/min
Acetylcholine Chloride for Ophthalmic Injection size: 50 L
Solution System suitability
Samples: System suitability solution and Standard solution
(Comment on this Monograph)id=m700=Acetylcholine Chloride Suitability requirements
for Ophthalmic Solution=A-Monos.pdf) Resolution: NLT 2.0 between acetylcholine chloride and
DEFINITION choline chloride, System suitability solution
Acetylcholine Chloride for Ophthalmic Solution is a sterile Relative standard deviation: NMT 3.5%, Standard
mixture of Acetylcholine Chloride with Mannitol or other solution
suitable diluent, prepared by freeze-drying. Each container Analysis
contains NLT 90.0% and NMT 115.0% of the labeled amount Samples: Standard solution and Sample Solution
of acetylcholine chloride (C7H16ClNO2). Calculate the percentage of C7H16ClNO2 in the container
taken:
IDENTIFICATION
A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Result = (rU/rS) (CS/CU) 100
Adsorbent: 0.25-mm layer of aluminum oxide
Standard solution: 10 mg/mL of USP Acetylcholine rU = peak response of the Sample solution
Chloride RS rS = peak response of the Standard solution
Sample solution: 10 mg/mL of acetylcholine chloride CS = concentration of USP Acetylcholine Chloride RS
Spray reagent A: 5 mg/mL of cobaltous chloride in a in the Standard solution (mg/mL)
mixture of alcohol and water (1:1) [NOTEThis solution is CU = nominal concentration of acetylcholine chloride
freshly prepared.] in the Sample solution (mg/mL)
Spray reagent B: 10 mg/mL of potassium ferrocyanide in a Acceptance criteria: 90.0%115.0%
mixture of alcohol and water (1:1) PERFORMANCE TESTS
Application volume: 2 L UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Developing solvent system: Upper layer obtained by
mixing water, butyl alcohol, and glacial acetic acid (5:4:1) SPECIFIC TESTS
[NOTEAllow to separate completely.] STERILITY TESTS 71: It meets the requirements.
Analysis ACIDITY: Equivalent to 100 mg of acetylcholine chloride in
Samples: Standard solution and Sample solution 10 mL of recently boiled water
Proceed as directed for Chromatography 621, Thin-Layer Add at once 1 drop of bromothymol blue TS: NMT 0.50
Chromatography. Develop the chromatogram, without mL of 0.010 N sodium hydroxide is required in order to
delay, in a vapor-saturated chamber. Allow the solvent produce a color change.
front to move about 10 cm beyond the initial spotting WATER DETERMINATION, Method I 921: Perform the titration
line. Immediately spray the plate with Spray reagent A. Dry in the original container, observing precautions against
the plate as before, and immediately spray the plate with contact with water or moist atmosphere. Adjust the
Spray reagent B. concentration of the reagent so that the titration volume
Acceptance criteria: The RF value and color of the principal approaches but does not exceed the capacity of the
spot obtained from the Sample solution correspond to those container. Titrate to an amber color that persists for 15 s
obtained from the Standard solution. after mixing.
B. PROCEDURE Acceptance criteria: NMT 1.0%
Sample solution: 10 mg/mL of acetylcholine chloride CONSTITUTED SOLUTION: At the time of use, it meets the
Analysis: To 2 mL of Sample solution, add 1 drop of nitric
acid and 1 mL of silver nitrate TS. requirements for Injections 1, Constituted Solutions.
Acceptance criteria: A curdy, white precipitate, soluble in
an excess of 6 N ammonium hydroxide, is formed.
USP 32 Official Monographs / Acetylcysteine 49
ADDITIONAL REQUIREMENTS RU = internal standard ratio of the Sample solution

PACKAGING AND STORAGE: Preserve in tight containers as RS = internal standard ratio of the Standard solution
described under Injections 1, Containers for Sterile Solids, CS = concentration of USP Acetylcysteine RS in the
and store at controlled room temperature. Standard solution (mg/mL)
USP REFERENCE STANDARDS 11 CU = concentration of Acetylcysteine in the Sample
USP Acetylcholine Chloride RS solution (mg/mL)
IMPURITIES
Acetylcysteine Inorganic Impurities
(Comment on this Monograph)id=m750=Acetylcysteine=A- RESIDUE ON IGNITION 281
Monos.pdf) Sample: 2 g
Analysis: Transfer the Sample to a tared fused silica dish,
heat on a hot plate until thoroughly charred, cool, add 1
mL of sulfuric acid, and heat gently until fuming ceases.
Ignite at 600 until the carbon is consumed.
HEAVY METALS, Method II 231: NMT 10 ppm
[CAUTIONExercise care because explosion may occur.]
[NOTEIn a dropwise manner, wet the Sample with 2 mL of
C5H9NO3S 163.20 nitric acid, and proceed as directed for the Sample solution.]
L-Cysteine, N-acetyl-;
N-Acetyl-L-cysteine [616-91-1]. SPECIFIC TESTS
OPTICAL ROTATION, Specific Rotation 781S: +21 to +27
DEFINITION pH 7.0 Buffer solution: Mix 29.5 mL of 1 N sodium
Acetylcysteine contains NLT 98.0% and NMT 102.0% of hydroxide, 50 mL of 1 M monobasic potassium phosphate,
C5H9NO3S, calculated on the dried basis. and sufficient water to make 100 mL, and using a pH meter,
adjust to a pH of 7.0 0.1 by adding more of either solution
IDENTIFICATION as necessary.
INFRARED ABSORPTION 197K Sample solution: In a 25-mL volumetric flask, mix 1.25 g
with 1 mL of edetate disodium solution (1 in 100), add 7.5
ASSAY mL of sodium hydroxide solution (1 in 25), and mix to
PROCEDURE dissolve. Dilute with pH 7.0 Buffer solution to volume.
Mobile phase: 6.8 mg/mL of monobasic potassium PH 791: 2.02.8, in a solution (1 in 100)
phosphate LOSS ON DRYING 731: Dry it at a pressure of about 50 mm
Pass through a membrane filter having a 0.45-m porosity, of mercury at 70 for 4 h: it loses NMT 1.0% of its weight.
and degas. Adjust with phosphoric acid to a pH of 3.0.
Solution A: 0.5 mg/mL of sodium metabisulfite ADDITIONAL REQUIREMENTS
Internal standard solution: 5 mg/mL of USP L- PACKAGING AND STORAGE: Preserve in tight containers, and
Phenylalanine RS in freshly-prepared Solution A store at controlled room temperature.
Standard stock solution: 10 mg/mL of USP Acetylcysteine USP REFERENCE STANDARDS 11
RS in Solution A USP Acetylcysteine RS
Standard solution: 0.5 mg/mL of USP Acetylcysteine RS USP L-Phenylalanine RS
Pipet 10.0 mL of Standard stock solution and 10.0 mL of
Internal standard solution to a 200-mL volumetric flask, and
dilute with Solution A to volume.
Sample stock solution: 10 mg/mL of Acetylcysteine in Acetylcysteine Solution
Solution A (Comment on this Monograph)id=m780=Acetylcysteine
Sample solution: 0.5 mg/mL of Acetylcysteine Solution=A-Monos.pdf)
Pipet 10.0 mL of Sample stock solution and 10.0 mL of
Internal standard solution to a 200-mL volumetric flask, and DEFINITION
dilute with Solution A to volume. Acetylcysteine Solution is a sterile solution of Acetylcysteine in
Chromatographic system water, prepared with the aid of Sodium Hydroxide. It contains
Mode: LC NLT 90.0% and NMT 110.0% of the labeled amount of
Detector: UV 214 nm acetylcysteine (C5H9NO3S).
Column: 3.9-mm 30-cm; packing L1 IDENTIFICATION
Flow rate: 1.5 mL/min INFRARED ABSORPTION 197K
Injection size: 5 L Sample solution: Place 10 mL in a suitable beaker, and
System suitability adjust to a pH of 2 (pH indicator paper) using 3 N
Sample: Standard solution hydrochloric acid. Add up to 2 g of finely powdered sodium
[NOTEThe relative retention times for acetylcysteine and L- chloride, in two portions of 200 mg each initially, and then
phenylalanine are about 0.5 and 1.0, respectively.] in smaller portions of 25 mg, stirring after each addition
Suitability requirements until the sodium chloride dissolves and a precipitate is
Resolution: NLT 6 between acetylcysteine and DL- formed. [NOTEThe precipitate appears as a very fine
phenylalanine powder, and the solution turns cloudy. If no precipitate
Relative standard deviation: NMT 2.0% forms, add an additional drop of 3 N hydrochloric acid, and
Analysis stir until the precipitate forms.] Allow to stand at room
Samples: Standard solution and Sample solution temperature for 15 min, and collect the residue by suction
Calculate the percentage of C5H9NO3S in the portion of filtration. Use the acetylcysteine so obtained, after being
Acetylcysteine taken: dried at a pressure of 50 mm of mercury at 70 for 4 h.
50 Acetylcysteine / Official Monographs USP 32
ASSAY Acetylcysteine and Isoproterenol

PROCEDURE Hydrochloride Inhalation Solution
Mobile phase: 6.8 mg/mL monobasic potassium phosphate
in water (Comment on this Monograph)id=m810=Acetylcysteine and
Adjust with phosphoric acid to a pH of 3.0 Isoproterenol Hydrochloride Inhalation Solution=A-Monos.pdf)
Solution A: 0.5 mg/mL of sodium metabisulfite solution in DEFINITION
water Acetylcysteine and Isoproterenol Hydrochloride Inhalation
Internal standard solution: 5 mg/mL of USP L- Solution is a sterile solution of Acetylcysteine and Isoproterenol
Phenylalanine RS in Solution A Hydrochloride in water. It contains NLT 90.0% and NMT
Standard stock solution: 10 mg/mL of USP Acetylcysteine 110.0% of the labeled amount of acetylcysteine (C5H9NO3S),
RS in Solution A and NLT 90.0% and NMT 115.0% of the labeled amount of
Standard solution: 0.5 mg/mL of USP Acetylcysteine RS in isoproterenol hydrochloride (C11H17NO3 HCl).
Solution A, from Standard stock solution, Internal standard
solution and Solution A (1:1:18) IDENTIFICATION
Sample stock solution: Equivalent to 10 mg/mL of A. INFRARED ABSORPTION 197K
Acetylcysteine, from volume of solution in Solution A Sample solution: Place 2 mL in a 10-mL beaker, and adjust
Sample solution: 0.5 mg/mL of acetylcysteine in Solution A, with 3 N hydrochloric acid to a pH of 3 (pH indicator
from Sample stock solution, Internal standard solution, and paper). Add 500 mg to 1 g of finely powdered sodium
Solution A (1:1:18) chloride, in two portions of 200 mg each initially, and then
Chromatographic system in smaller portions of 25 mg, stirring after each addition
(See Chromatography 621, System Suitability.) until the sodium chloride dissolves and a precipitate is
Mode: LC formed. Allow to stand at room temperature for 15 min,
Detector: UV 214 nm and collect the residue by suction filtration. Use the
Column: 3.9-mm 30-cm; packing L1 acetylcysteine so obtained, after being dried at a pressure of
Flow rate: 1.5 mL/min 50 mm of mercury at 70 for 4 h.
Injection size: 5 L B. PROCEDURE
System suitability Ferro-citrate solution and Buffer solution: Prepare as
Sample: Standard solution directed under Epinephrine Assay 391.
Suitability requirements Sample solution: Equivalent to 0.26 mg of isoproterenol
Resolution: NLT 6 between acetylcysteine and DL- hydrochloride from a volume of Inhalation Solution, in a test
phenylalanine tube with 3 mL of 0.1 M mercuric chloride
Relative standard deviation: NMT 2.0% Analysis: Add 100 L of Ferro-citrate solution and 1.0 mL of
Analysis Buffer solution
Samples: Standard solution and Sample solution Acceptance criteria: The presence of isoproterenol
[NOTEThe relative retention times for acetylcysteine and hydrochloride is confirmed by the development of a purple
L-phenylalanine are about 0.5 and 1.0, respectively.] color.
Calculate the percentage of C5H9NO3S in each mL of the
solution taken: ASSAY
ACETYLCYSTEINE
Result = (RU/RS) (CS/CU) 100 Mobile phase: 6.8 mg/mL of monobasic potassium
phosphate in water
RU = ratio of the peak response of acetylcysteine to Adjust with phosphoric acid to a pH of 3.0
that of L-phenylalanine of the Sample solution Solution A: 0.5 mg/mL of sodium metabisulfite solution in
RS = ratio of the peak response of acetylcysteine to water
that of L-phenylalanine of the Standard solution Internal standard solution: 5 mg/mL of USP L-
CS = concentration of USP Acetylcysteine RS in the Phenylalanine RS in Solution A
Standard solution (mg/mL) Standard stock solution: 10 mg/mL of USP Acetylcysteine
CU = nominal concentration of acetylcysteine in the RS in Solution A
Sample solution (mg/mL) Standard solution: 0.5 mg/mL of USP Acetylcysteine RS in
Acceptance criteria: 90.0%110.0% Solution A, from Standard stock solution, Internal standard
solution, and Solution A (1:1:18)
SPECIFIC TESTS Sample stock solution: Equivalent to 10 mg/mL of
PH 791: 6.07.5 acetylcysteine, from volume of Inhalation Solution in Solution
STERILITY 71: Meets the requirements A
ADDITIONAL REQUIREMENTS Sample solution: 0.5 mg/mL of acetylcysteine in Solution A,
PACKAGING AND STORAGE: Preserve in single-unit or multiple- from Sample stock solution, Internal standard solution, and
unit tight containers that effectively exclude oxygen, and Solution A (1:1:18)
store at controlled room temperature. Chromatographic system
USP Acetylcysteine RS
USP L-Phenylalanine RS
USP 32 Official Monographs / Acitretin 51
Mode: LC RU = ratio of the peak responses of isoproterenol

Detector: UV 214 nm hydrochloride to acetaminophen from the
Column: 3.9-mm 30-cm; packing L1 Sample solution
Flow rate: 1.5 mL/min RS = ratio of the peak responses of USP Isoproterenol
Injection size: 5 L Hydrochloride RS to acetaminophen from the
System suitability Standard solution
Sample: Standard solution CS = concentration of USP Isoproterenol
[NOTEThe relative retention times for acetylcysteine and Hydrochloride RS in the Standard solution
L-phenylalanine are about 0.5 and 1.0, respectively.] (mg/mL)
Suitability requirements CU = nominal concentration of isoproterenol
Resolution: NLT 6 between acetylcysteine and DL- hydrochloride in the Sample solution (mg/mL)
phenylalanine Acceptance criteria: 90.0%115.0%
Analysis SPECIFIC TESTS
Samples: Standard solution and Sample solution PH 791: 6.07.0
Calculate the percentage of C5H9NO3S in each mL of the STERILITY TESTS 71: It meets the requirements.
Inhalation Solution taken: COLOR AND CLARITY
Standard solution: 0.100 N iodine VS and water (1:249)
Result = (RU/RS) (CS/CU) 100 Sample solution: A portion of the Inhalation Solution
Analysis: Visually analyze the Sample solution in a suitable
RU = ratio of the peak response of acetylcysteine to clear glass test tube against a white background: it is not
that of L-phenylalanine from the Sample pinkish and it contains no precipitate. If any yellow color is
solution observed in the Sample solution, concomitantly determine
RS = ratio of the peak response of acetylcysteine to the absorbances of the Sample solution and the Standard
that of L-phenylalanine from the Standard solution in 1-cm cells with a suitable spectrophotometer set
solution at 460 nm.
CS = concentration of USP Acetylcysteine RS in the Acceptance criteria: The absorbance of the Sample solution
Standard solution (mg/mL) does not exceed that of the Standard solution.
CU = nominal concentration of acetylcysteine in the
Sample solution (mg/mL) ADDITIONAL REQUIREMENTS
Acceptance criteria: 90.0%110.0% PACKAGING AND STORAGE: Preserve in single-dose or
ISOPROTERENOL HYDROCHLORIDE multiple-dose containers, preferably of Type I glass, tightly
Solution A: 13.6 mg/mL of monobasic potassium closed with a glass or polyethylene closure, and store at
phosphate in water controlled room temperature.
Mobile phase: Methanol and Solution A (1:49) LABELING: The label indicates that the Inhalation Solution is
Internal standard solution: 150 mg of acetaminophen in a not to be used if its color is pinkish or darker than slightly
500-mL volumetric flask. Add 5 mL of glacial acetic acid, yellow or if it contains a precipitate.
and dilute with water to volume. USP REFERENCE STANDARDS 11
Standard stock solution: 0.15 mg/mL of USP Isoproterenol USP Acetylcysteine RS
Hydrochloride RS, in 0.05 M sodium metabisulfite USP Isoproterenol Hydrochloride RS
Standard solution: 0.6 mg/mL of USP Isoproterenol USP L-Phenylalanine RS
Hydrochloride RS, from Standard stock solution, Internal
standard solution, and 0.2 M acetic acid (2:2:1)
Sample solution: Equivalent to 1.5 mg of isoproterenol Acitretin
hydrochloride from a volume of Inhalation Solution, and 10
mL of Internal standard solution to a 25-mL volumetric flask. (Comment on this Monograph)id=m850=Acitretin=A-
Add dilute glacial acetic acid (1 in 100) to volume. Monos.pdf)
Mode: LC
Detector: UV 280 nm
Flow rate: 2 mL/min
Injection size: 25 L C21H26O3 326.43
System suitability 2,4,6,8-Nonatetraenoic acid, 9-(4-methoxy-2,3,6-
Sample: Standard solution trimethylphenyl)-3,7-dimethyl-, (all-E)-;
[NOTEThe relative retention times for isoproterenol (all-E)-9-(4-Methoxy-2,3,6-trimethylphenyl)-3,7-
hydrochloride and acetaminophen are about 0.5 and 1.0, dimethyl-2,4,6,8-nonatetraenoic acid [55079-83-9].
respectively.]
Suitability requirements DEFINITION
Resolution: NLT 6 between isoproterenol hydrochloride Acitretin contains NLT 98.0% and NMT 102.0% of C21H26O3,
and acetaminophen calculated on the dried basis.
Relative standard deviation: NMT 2.0% [CAUTIONAcitretin is a teratogen. Great care should be taken
Analysis when handling to avoid inhalation of dust or contact with
Samples: Standard solution and Sample solution skin.]
Calculate the percentage of C11H17NO3 HCl in each mL of [NOTEUse low actinic glassware and perform all tests under
the Inhalation Solution taken: yellow and subdued light.]
52 Acitretin / Official Monographs USP 32
IDENTIFICATION Dilute quantitatively with alcohol.

A. INFRARED ABSORPTION 197K [NOTEStore the solution at 4 prior to injection.]
B. The retention time of the major peak of the Sample Sample solution: 0.25 mg/mL of Acitretin in
solution corresponds to that of the Standard solution, as tetrahydrofuran, and alcohol (1:19)
obtained in the Assay. [NOTEStore the solution at 4 prior to injection.]
System suitability
ASSAY (See Chromatography 621.)
PROCEDURE Sample: Standard solution
Mobile phase: Alcohol, glacial acetic acid, and water [NOTEThe relative retention times for acitretin related
(92:0.3:8) compound A, acitretin, and acitretin related rompound B
System suitability solution: In a 200-mL volumetric flask, are 0.78, 1.0, and 1.61, respectively.]
dissolve 2.0 mg each of USP Acitretin RS and USP Tretinoin Suitability requirements
RS in tetrahydrofuran. Dilute with alcohol to volume. Pipet Resolution: NLT 1.5 between acitretin related
5.0 mL of this solution into a 200-mL volumetric flask, and compound A and acitretin; NLT 1.5 between acitretin
dilute quantitatively with alcohol. related rompound B and acitretin
[NOTEStore the solution at 4 prior to injection.] Relative standard deviation: NMT 10.0% for acitretin
Standard solution: 0.1 mg/mL of USP Acitretin RS in related compound A and NMT 10.0% for acitretin
alcohol related compound B
[NOTEUse tetrahydrofuran to dissolve USP Acitretin RS Analysis
before diluting with alcohol. The final concentration of Samples: Standard solution and Sample solution
tetrahydrofuran in the preparation will be 2%. Store the Calculate the percentage of acitretin related compound A
solution at 4 prior to injection.] and acitretin related compound B in the portion of
Sample stock solution: 0.25 mg/mL of Acitretin in Acitretin taken:
tetrahydrofuran and alcohol (1:19)
Sample solution: 0.1 mg/mL of Acitretin from Sample stock Result = (rU/rS) (CS/CU) 100
solution in alcohol
[NOTEStore the solution at 4 prior to injection.] rU = peak response of the relevant impurity in the
Chromatographic system chromatogram of the Sample solution
Mode: LC rS = peak response of the relevant impurity in the
Detector: UV 360 nm chromatogram of the Standard solution
Column: 4-mm 25-cm; packing L1 CS = concentration of USP Acitretin Related
Flow rate: 0.6 mL/min Compound A RS or USP Acitretin Related
Injection size: 10 L Compound B RS in the Standard solution
System suitability (g/mL)
(See Chromatography 621, System Suitability.) CU = concentration of Acitretin in the Sample solution
Sample: System suitability solution (g/mL)
[NOTEThe relative retention times for tretinoin and for Calculate the percentage of impurities other than acitretin
acitretin are 0.84 and 1.0, respectively.] related compounds A and B in the portion of Acitretin
Suitability requirements taken
Resolution: NLT 2.0 tretinoin and acitretin in System
suitability solution Result = (rU/rS) (CS/CU) 100
Relative standard deviation: NMT 1.0% Acitretin
Analysis rU = peak response of each individual unspecified
Samples: Standard solution and Sample solution impurity in the Sample solution
Calculate the percentage of C21H26O3 in the portion of rS = peak response of USP Acitretin RS in the
Acitretin taken: Standard solution
CS = concentration of USP Acitretin RS in the
Result = (rU/rS) (CS/CU) 100 Standard solution (g/mL)
CU = concentration of Acitretin in the Sample solution
rU = peak response from the Sample solution (g/mL)
rS = peak response from the Standard solution Acceptance criteria: NMT 0.3% of acitretin related
CS = concentration of USP Acitretin RS in the Standard compound A, NMT 0.3% of acitretin related compound B;
solution (mg/mL) NMT 0.1% of any individual unspecified impurity, and
CU = concentration of Acitretin in the Sample solution NMT 0.4% of total unspecified impurities
(mg/mL) Total impurities: NMT 1.0%
SPECIFIC TESTS
IMPURITIES LOSS ON DRYING 731: Dry it in vacuum at a pressure not
Inorganic Impurities exceeding 19 mm of mercury at 100 for 4 h: it loses NMT
RESIDUE ON IGNITION 281: NMT 0.1% 0.2% of its weight.
HEAVY METALS, Method I 231: NMT 20 ppm
Organic Impurities ADDITIONAL REQUIREMENTS
PROCEDURE PACKAGING AND STORAGE: Preserve in tight containers,
Mobile phase and Chromatographic system: Proceed as protected from light. Store at controlled room temperature.
directed in the Assay. USP REFERENCE STANDARDS 11
Standard solution: 0.8 g/mL each of USP Acitretin RS, USP Acitretin RS
USP Acitretin Related Compound A RS, and USP Acitretin USP Acitretin Related Compound A RS
Related Compound B RS in tetrahydrofuran USP Acitretin Related Compound B RS
USP 32 Official Monographs / Acitretin 53
USP Tretinoin RS Mode: LC

Detector: UV 365 nm
Column: 4.6-mm 15-cm; 5-m packing L1
Flow rate: 1 mL/min
Acitretin Capsules Injection size: 25 L
(Comment on this Monograph)id=m854=Acitretin Capsules=A- System suitability
Monos.pdf) Samples: Standard solution and System suitability solution
[NOTEThe relative retention times for acitretin related
DEFINITION compound A, acitretin, and 9-cis isomer are 0.84, 1.0, and
Acitretin Capsules contain NLT 90.0% and NMT 110.0% of the 1.09, respectively.]
labeled amount of acitretin (C21H26O3). Suitability requirements
[CAUTIONAcitretin is a teratogen. Great care should be taken Resolution: NLT 3.0 between acitretin related compound
when handling to avoid inhalation of dust or contact with A and acitretin; NLT 1.8 between the 9-cis isomer and
skin.] acitretin
[NOTEUse low-actinic glassware and perform all tests under Relative standard deviation: NMT 2.0%
yellow and subdued light. Make all injections within 1 h of Analysis
Sample solution.] Samples: Standard solution and Sample solution
Calculate the percentage of C21H26O3 in the portion of
IDENTIFICATION Capsules taken:
A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
Standard solution: 10 mg/mL of USP Acitretin RS in Result = (rU/rS) (CS/CU) 100
tetrahydrofuran
Sample solution: Equivalent to 20 mg of acitretin from rU = peak response from the Sample solution
Capsules rS = peak response from the Standard solution
Grind to a fine powder, then triturate for 30 s with 2 mL of CS = concentration of USP Acitretin RS in the Standard
tetrahydrofuran. Transfer the suspension to a 12-mL conical solution (mg/mL)
centrifuge tube, and centrifuge to obtain a clear CU = nominal concentration of the Sample solution
supernatant. (Capsules/mL)
Application volume: 10 L Acceptance criteria: 90.0%110.0%
Developing solvent system: Chloroform and methanol
(4:1) IMPURITIES
Analysis Organic Impurities
Samples: Standard solution and Sample solution LIMIT OF DEGRADATION PRODUCTS
Procedure: Proceed as directed in the chapter, and then Diluent, Mobile phase, System suitability solution, and
air-dry. Spray the plate with a saturated solution of Chromatographic system: Proceed as directed in the
antimony trichloride in chloroform (25 g in 100 mL) Assay.
followed by concentrated sulfuric acid, and then locate the Sample solution: Use the Sample solution, prepared as
spots. directed in the Assay.
Analysis
ASSAY Samples: Standard solution and Sample solution
PROCEDURE Calculate the percentage of each degradation product in
Diluent: Methanol and tetrahydrofuran (13:10) the portion of Capsules taken:
Mobile phase: Methanol, alcohol, glacial acetic acid, and
water (74:5:0.5:21) Result = (rU/rT) 100
System suitability solution: Transfer 2 mL of the Standard
solution to a clear 4-mL glass vial. After sealing the vial with rU = peak response for each individual impurity
a teflon-lined silicone septum and cap, place the vial on its rT = sum of the responses of all of the peaks
side in a light chamber, expose it to 400 foot-candles of Acceptance criteria: NMT 0.5% of acitretin related
fluorescent light for 5 min, and then completely wrap the compound A, NMT 0.4% of any individual unspecified
vial with aluminum foil. impurity, and NMT 0.8% of total unspecified impurities is
[NOTEExposure to the fluorescent light allows for the found.
formation of two degradation products: acitretin related
compound A and the 9-cis isomer [(E,E,Z,E)-9-(4- PERFORMANCE TESTS
methoxy-2,3,6-trimethylphenyl)-3,7-dimethyl-2,4,6,8- DISSOLUTION 711
nonatetraenoic acid.] Medium: 3% sodium lauryl sulfate in deaerated water, pH
Standard solution: 0.1 mg/mL of USP Acitretin RS in water 9.6 to 10.0; 900 mL
and Diluent (2:23) Apparatus 1: 100 rpm
[NOTEBefore make-up to final volume sonicate for 5 min.] Time: 30 min
Sample solution: Carefully separate and place both halves Determine the amount of acitretin (C21H26O3) dissolved
of 10 Capsules into a 100-mL volumetric flask. Stopper and employing the following method.
shake the flask to remove the fill. Add 8 mL of water while Sample solutions: Sample per 711 Dissolution. Dilute with
rinsing any fill from the neck of the flask. Place the flask in a Medium to a concentration that is similar to the Standard
water bath set at 45 for 10 min, shaking initially and at 5- solution. Use portions of the solution under test passed
min intervals up to 10 min. Place the resulting suspension in through a suitable 0.45-m filter.
an ultrasonic bath for 15 min. Dilute with Diluent to volume, Standard solution: 14 mg of USP Acitretin RS in alcohol,
and sonicate for 5 additional min. Cool to room and Medium (1:9). For Capsules labeled to contain 10 mg,
temperature and, if necessary, dilute with Diluent to volume. transfer 20.0 mL of this solution to a 50-mL volumetric flask,
Filter the suspension, and use the clear filtrate. and dilute with Medium to volume.
Chromatographic system Capsule shell solution: 6 clean empty-shell Capsules in 900
(See Chromatography 621, System Suitability.) mL of Medium
54 Acitretin / Official Monographs USP 32
Analysis Dissolve both in a volume of 0.1 N sodium hydroxide of

Samples: Standard solution, Blank, Capsule shell solution, NMT 10% of the final solution volume, and then dilute the
and Sample solution solution to volume with water.
Determine the amount of C21H26O3 dissolved by employing System suitability solution B: 0.7 g/mL of guanine
UV absorption at the wavelength of maximum absorbance Dissolve in a volume of 0.1 N sodium hydroxide of NMT
at about 347 nm, using 2-mm cells, in comparison with 10% of the final solution volume, and then dilute to
the appropriate Standard solution. Use the Medium as a volume with water.
blank. Determine the absorbance of the Capsule shell Guanine standard stock solution: Transfer 8.75 mg of
solution under the same conditions. guanine to a 500-mL volumetric flask. Dissolve in 50 mL of
Calculate the amount of C21H26O3 dissolved: 0.1 N sodium hydroxide, and dilute with water to volume.
Guanine standard solution: 0.7 g/mL guanine in 0.01 N
Result = [(AU ACS)/AS] (CS/CU) 100 sodium hydroxide from Guanine standard stock solution
Standard stock solution: Dissolve 25 mg of USP Acyclovir
AU = absorbance of the Sample solution RS in 5 mL of 0.1 N sodium hydroxide in a 50-mL
ACS = capsule shell correction, calculated as shown volumetric flask, and dilute with water to volume.
below Standard solution: 0.1 mg/mL of USP Acyclovir RS in 0.01
AS = absorbance of the appropriate Standard solution N sodium hydroxide from Standard stock solution
CS = concentration of the appropriate Standard Sample stock solution: Dissolve 100 mg of Acyclovir in 20
solution (mg/mL) mL of 0.1 N sodium hydroxide in a 200-mL volumetric flask,
CU = concentration of the Sample solution and dilute with water to volume.
(Capsules/mL) Sample solution: 0.1 mg/mL Acyclovir in 0.01 N sodium
The Capsule shell correction, ACS, is calculated as follows: hydroxide from Sample stock solution
ACS = ACSS/N (See Chromatography 621, System Suitability.)
Mode: LC
ACSS = absorbance of the Capsule shell solution Detector: UV 254 nm
N = number of Capsule shells used to prepare the Column: 4.6-mm 25-cm; packing L1
Capsule shell solution Flow rate: 3 mL/min
Tolerances: NLT 85% (Q) of the labeled amount of acitretin Injection size: 20 L
(C21H26O3) is dissolved. System suitability
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Samples: System suitability solution A and System suitability
ADDITIONAL REQUIREMENTS solution B
PACKAGING AND STORAGE: Preserve in well-closed, light- Suitability requirements
resistant containers. Resolution: NLT 2.0 between acyclovir and guanine from
USP REFERENCE STANDARDS 11 System suitability solution A
USP Acitretin RS Tailing factor: NMT 2, System suitability solution A
Relative standard deviation: NMT 2.0% for replicate
injections of acyclovir for System suitability solution A; NMT
2.0% for replicate injections of System suitability solution B
Acyclovir Analysis
(Comment on this Monograph)id=m890=Acyclovir=A- Samples: Standard solution, Guanine standard solution, and
Monos.pdf) Sample solution
Calculate the percentage of guanine in the portion of
Acyclovir taken:

rS = peak response from the Guanine standard solution
CS = concentration of guanine in the Guanine standard
C8H11N5O3 225.20 solution (g/mL)
6H-Purin-6-one, 2-amino-1,9-dihydro-9-[(2- CU = concentration of the Sample solution (g/mL)
hydroxyethoxy)methyl]-; Acceptance criteria: NMT 0.7% of guanine
9-[(2-Hydroxyethoxy)methyl]guanine [59277-89-3]. Calculate the percentage of C8H11N5O3 in the portion of
Acyclovir taken:
DEFINITION
Acyclovir contains NLT 98.0% and NMT 101.0% of C8H11N5O3, Result = (rU/rS) (CS/CU) 100
calculated on the anhydrous basis.
IDENTIFICATION rS = peak response from the Standard solution
A. INFRARED ABSORPTION 197K CS = concentration of USP Acyclovir RS in the
B. The retention time of the major peak of the Sample Standard solution (mg/mL)
solution corresponds to that of the Standard solution, as CU = concentration of the Sample solution (mg/mL)
obtained in the Assay.
ASSAY
LIMIT FOR GUANINE
Mobile phase: Glacial acetic acid in water (1 in 1000)
System suitability solution A: 0.1 mg/mL each of USP
Acyclovir RS and guanine
USP 32 Official Monographs / Acyclovir 55
Acceptance criteria: 98.0%101.0% Analysis: Separately inject the Standard solution and the
Sample solution into the chromatograph, record the
IMPURITIES chromatograms, and measure the peak responses.
Organic Impurities Calculate the percentage of C8H11N5O3 in the Capsules
PROCEDURE: ORDINARY IMPURITIES 466 taken:
Sample solution: 10 mg/mL in dimethyl sulfoxide
Standard solutions: 0.01, 0.05, 0.1, and 0.2 mg/mL of Result = (rU/rS) (CS/CU) 100
USP Acyclovir RS in dimethyl sulfoxide
Eluant: Chloroform, methanol, and ammonium hydroxide rU = peak response of the Sample solution
(80:20:2) rS = peak response of the Standard solution
Visualization: 1 CS = concentration of USP Acyclovir RS in the
Application volume: 5 L Standard solution (mg/mL)
Acceptance criteria: NMT 1% CU = nominal concentration of the Sample solution
(mg/mL)
SPECIFIC TESTS Acceptance criteria: 93.0%107.0%
WATER DETERMINATION, Method I 921: NMT 6.0%
IMPURITIES
ADDITIONAL REQUIREMENTS Organic Impurities
PACKAGING AND STORAGE: Preserve in tight containers, and PROCEDURE
store at room temperature. Protect from light and moisture. [NOTEMobile phase, Sample solution, and Chromatographic
USP REFERENCE STANDARDS 11 system: proceed as directed in the Assay.]
USP Acyclovir RS Analysis: Inject the Sample solution into the chromatograph,
record the chromatograms, and measure the peak
responses.
Acyclovir Capsules Calculate the percentage of each impurity in the portion of
Capsules taken:
(Comment on this Monograph)id=m893=Acyclovir Capsules=A-
Monos.pdf) Result = (rU/rT) 100
DEFINITION rU = peak response for each impurity
Acyclovir Capsules contain NLT 93.0% and NMT 107.0% of the rT = sum of the responses for all the peaks
labeled amount of acyclovir (C8H11N5O3). Acceptance criteria
Guanine: NMT 2.0%
IDENTIFICATION Any individual impurity: NMT 0.5%
The retention time of the major peak of the Sample solution
corresponds to that of the Standard solution, as obtained in PERFORMANCE TESTS
the Assay. DISSOLUTION 711
Medium: 0.1 N hydrochloric acid; 900 mL
ASSAY Apparatus 1: 100 rpm
PROCEDURE Time: 45 min
Mobile phase: 0.02 M acetic acid Detector: UV 254 nm
System suitability solution A: 0.1 mg/mL each of USP Sample solutions: Sample per Dissolution 711. Dilute with
Acyclovir RS and guanine in 0.1 N sodium hydroxide Medium to a concentration that is similar to the Standard
System suitability solution B: 2.0 g/mL of guanine in 0.1 solution.
N sodium hydroxide Standard solution: USP Acyclovir RS in Medium
Standard solution: 0.1 mg/mL of USP Acyclovir RS in 0.1 N Analysis: Determine the amount of C8H11N5O3 dissolved
sodium hydroxide from UV absorption at the wavelength of maximum
Sample solution: Transfer the contents of Capsules absorption on filtered portions of the solution under test.
equivalent to 10 mg of acyclovir (NLT 10 Capsules) to a Tolerances: NLT 75% (Q) of the labeled amount of
100-mL volumetric flask, dissolve in 10 mL of 0.1 N sodium C8H11N5O3 is dissolved.
hydroxide, dilute to volume with water, and filter. UNIFORMITY OF DOSAGE UNITS 905
Chromatographic system Acceptance criteria: Meet the requirements for Content
(See Chromatography 621, System Suitability.) Uniformity
Mode: LC
Detector: UV 254 nm ADDITIONAL REQUIREMENTS
Column: 4.2-mm 25-cm; packing L1 PACKAGING AND STORAGE: Preserve in tight containers. Store
Flow rate: 1.5 mL/min between 15 and 25. Protect from light and moisture.
Injection size: 20 L USP REFERENCE STANDARDS 11
System suitability USP Acyclovir RS
Sample: System suitability solution A and System suitability
solution B
[NOTEThe relative retention times for System suitability
solution A for guanine and acyclovir are about 0.6 and Acyclovir for Injection
1.0, respectively.] (Comment on this Monograph)id=m894=Acyclovir for
Suitability requirements Injection=A-Monos.pdf)
Resolution: NLT 2.0 between guanine and acyclovir for
System suitability solution A DEFINITION
Relative standard deviation: NMT 2.0% for replicate Acyclovir for Injection contains NLT 90.0% and NMT 110.0% of
injections of acyclovir using System suitability solution A the labeled amount of acyclovir (C8H11N5O3).
injections of System suitability solution B
56 Acyclovir / Official Monographs USP 32
IDENTIFICATION System suitability solution: 0.5 g/mL each of purine and

The retention time of the major peak of the Sample solution USP Acyclovir RS in Solution A
corresponds to that of the Standard solution, as obtained in Acyclovir standard solution: 5 g/mL of USP Acyclovir RS
the Assay. in Solution A
Guanine solution: Dissolve 50 mg of guanine in 50 mL of
ASSAY 0.1 N sodium hydroxide in a 500-mL volumetric flask, and
PROCEDURE bring the solution to volume with water.
Mobile phase: 0.02 M acetic acid Standard solution A: 0.5 g/mL of Acyclovir standard
System suitability solution A: 0.1 mg/mL each of USP solution in Solution A
Acyclovir RS and guanine in 0.1 N sodium hydroxide Standard solution B: 5 g/mL of Guanine solution in
System suitability solution B: 2.0 g/mL of guanine in 0.1 Solution A
N sodium hydroxide Sample solution: 0.5 mg/mL of acyclovir in Solution A,
Standard solution: 0.1 mg/mL of USP Acyclovir RS in 0.1 N prepared by constituting NLT 10 vials of Acyclovir for
sodium hydroxide Injection and diluting in Solution A
Sample solution: Constitute 1 vial of Acyclovir for Injection Chromatographic system
with water. Transfer an amount, equivalent to 10 mg of (See Chromatography 621, System Suitability.)
acyclovir, to a 100-mL volumetric flask, and dilute with Mode: LC
water to volume. Detector: UV 254 nm
Chromatographic system Column: 4.6-mm 25-cm; packing L1
(See Chromatography 621, System Suitability.) Flow rate: 1 mL/min
Detector: UV 254 nm System suitability
Column: 4.2-mm 25-cm; packing L1 Samples: System suitability solution, Standard solution A,
Flow rate: 1.5 mL/min and Standard solution B
Injection size: 20 L [NOTETypical retention times for guanine and acyclovir of
System suitability Standard solution A and Standard solution B are 5.8 min
Samples: System suitability solution A and System suitability and 14 min, respectively.]
solution B Suitability requirements
[NOTEThe relative retention times for guanine and Resolution: NLT 2.0 between purine and acyclovir,
acyclovir are 0.6 and 1.0, respectively, in the System System suitability solution
suitability solution A.] Relative standard deviation: NMT 1% for the acyclovir
Suitability requirements peak area and the guanine peak area using replicate
Resolution: NLT 2.0 between guanine and acyclovir, injections of Standard solution A and Standard solution B
System suitability solution A Analysis:
Relative standard deviation: NMT 2.0% for replicate Calculate the percentage of guanine in the Acyclovir for
injections for the acyclovir peak, System suitability solution Injection taken:
A
Relative standard deviation: NMT 2.0% for replicate Result = (rU/rS) (CS/CU) 100
injections, System suitability solution B
Analysis rU = peak response for guanine, if present, in the
Calculate the percentage of C8H11N5O3 in the portion of Sample solution
Acyclovir for Injection taken: rS = peak response of guanine in the Standard
solution
Result = (rU/rS) (CS/CU) 100 CS = concentration of guanine in the Standard
solution (mg/mL)
rU = peak response of the Sample solution CU = concentration of acyclovir in the Sample solution
rS = peak response of the Standard solution (mg/mL, based on the label claim)
CS = concentration of USP Acyclovir RS in the Acceptance criteria: NMT 1.0% guanine
Standard solution (mg/mL) Calculate the percentage of each other impurity in the
CU = concentration of the Sample solution (mg/mL) Acyclovir for Injection taken:
IMPURITIES
Organic Impurities rU = peak response for each impurity
PROCEDURE rS = peak response of acyclovir in the Standard
Solution A: 0.17 M acetic acid and methanol (125:8) solution
Make adjustments if necessary. CS = concentration of USP Acyclovir RS in the
Solution B: Methanol Standard solution (mg/mL)
Mobile phase: Use variable mixtures of Solution A and CU = concentration of acyclovir in the Sample solution
Solution B as directed for Chromatographic system. Make (mg/mL, based on the label claim)
adjustments to either solution as necessary. Acceptance criteria: NMT 0.15% for any peak having a
relative retention time of about 0.7 compared to the
Time (min) Solution A (%) Solution B (%) acyclovir peak; NMT 0.5% for any other individual
0 100 0
impurity; and NMT 1.0% for the total of all other
impurities
15 100 0
45 65 35
46 100 0
56 100 0
USP 32 Official Monographs / Acyclovir 57

PH 791: 11.012.5, 50 mg/mL of acyclovir rS = peak response from the Standard solution
WATER DETERMINATION Method I 921: NMT 5.5% CS = concentration of USP Acyclovir RS in the
STERILITY TESTS 71: Meets the requirements Standard solution (mg/mL)
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements CU = concentration of acyclovir in the Sample solution
BACTERIAL ENDOTOXINS TEST 85 NMT 0.174 USP Endotoxin (mg/mL)
Unit/mg of acyclovir Acceptance criteria: 90.0%110.0%
OTHER REQUIREMENTS: Meets the requirements under
Injections 1, Labeling. PERFORMANCE TESTS
MINIMUM FILL 755: Meets the requirements
PACKAGING AND STORAGE: Preserve in tight containers. Store IMPURITIES
between 15 and 25. Protect from light. Organic Impurities
USP REFERENCE STANDARDS 11 PROCEDURE: LIMIT OF GUANINE
USP Acyclovir RS [NOTEFor Mobile phase, Sample solution, and
USP Endotoxin RS Chromatographic system, proceed as directed in the Assay.]
Standard solution: 2.0 g/mL of guanine in 0.1 M sodium
hydroxide
Analysis
Acyclovir Ointment Samples: Standard solution and Sample solution
(Comment on this Monograph)id=m895=Acyclovir Calculate the percentage of guanine in the portion of
Ointment=A-Monos.pdf) Ointment taken:
DEFINITION Result = (rU/rS) (CS/CU) 100

Acyclovir Ointment contains NLT 90.0% and NMT 110.0% of
the labeled amount of acyclovir (C8H11N5O3), in a suitable rU = peak response of guanine from the Sample
ointment base. solution
rS = peak response of guanine from the Standard
The retention time of the major peak of the Sample solution CS = concentration of guanine in the Standard solution
corresponds to that of the Standard solution, as obtained in (mg/mL)
the Assay. CU = concentration of acyclovir in the Sample solution
(mg/mL)
ASSAY Acceptance criteria: NMT 2.0%
PROCEDURE
Mobile phase: 0.02 M acetic acid SPECIFIC TESTS
System suitability solution A: 0.1 mg/mL each of USP MICROBIAL ENUMERATION TESTS 61, and TESTS FOR SPECIFIED
Acyclovir RS and guanine in 0.1 N sodium hydroxide MICROORGANISMS 62
System suitability solution B: 2.0 g/mL of guanine in 0.1 Acceptance criteria: Meets the requirements of the tests for
N sodium hydroxide absence of Staphylococcus aureus and Pseudomonas
Standard solution: 0.1 mg/mL of USP Acyclovir RS in 0.1 N aeruginosa
sodium hydroxide
Sample solution: Transfer an amount of Ointment, ADDITIONAL REQUIREMENTS
equivalent to 10 mg of acyclovir, to a 100-mL volumetric PACKAGING AND STORAGE: Preserve in tight containers. Store
flask, dissolve in and dilute with 0.1 N sodium hydroxide to between 15 and 25 in a dry place.
volume. USP REFERENCE STANDARDS 11
Chromatographic system USP Acyclovir RS
Mode: LC
Detector: UV 254 nm Acyclovir Oral Suspension
Flow rate: 3 mL/min (Comment on this Monograph)id=m898=Acyclovir Oral
Injection size: 20 L Suspension=A-Monos.pdf)
System suitability
Samples: System suitability solution A and System suitability DEFINITION
solution B Acyclovir Oral Suspension contains NLT 90.0% and NMT
[NOTEThe relative retention times for guanine and 110.0% of the labeled amount of acyclovir (C8H11N5O3).
acyclovir, are about 0.6 and 1.0, respectively, in System IDENTIFICATION
suitability solution A.] The retention time of the major peak of the Sample solution
Suitability requirements corresponds to that of the Standard solution, as obtained in
Resolution: NLT 2.0 between guanine and acyclovir, the Assay.
System suitability solution A
Relative standard deviation: NMT 2.0% for replicate ASSAY
injections for the acyclovir peak, System suitability solution PROCEDURE
A; NMT 2.0% for replicate injections, System suitability Mobile phase: 0.02 M acetic acid
solution B System suitability solution A: 0.1 mg/mL each of USP
Analysis Acyclovir RS and guanine in 0.1 N sodium hydroxide
Samples: Standard solution and Sample solution System suitability solution B: 2.0 g/mL of guanine in 0.1
Calculate the quantity, in percentage, of C8H11N5O3 in the N sodium hydroxide
portion of Ointment taken: Standard solution: 0.1 mg/mL of USP Acyclovir RS in 0.1 N
sodium hydroxide
58 Acyclovir / Official Monographs USP 32
Sample stock solution: Transfer an amount of well-shaken Acceptance criteria: NMT 2.0%
Oral Suspension equivalent to 200 mg of acyclovir to a 200-
mL volumetric flask, add 100 mL of 0.1 N sodium SPECIFIC TESTS
hydroxide, shake by mechanical means for 15 min, and MICROBIAL ENUMERATION TESTS 61, and TESTS FOR SPECIFIED
sonicate, if necessary, to dissolve the Oral Suspension MICROORGANISMS 62
completely. Dilute to volume with 0.1 N sodium hydroxide. Acceptance criteria: Its total count does not exceed 10
Sample solution: Transfer 10.0 mL of the Sample stock cfu/mL, and it meets the requirements of the tests for
solution to a 100-mL volumetric flask, and dilute to volume absence of Salmonella species and Escherichia coli.
with water. PH 791
Chromatographic system Acceptance criteria: Between 4.5 and 7.0
(See Chromatography 621, System Suitability).
Mode: LC PERFORMANCE TESTS
Detector: UV 254 nm UNIFORMITY OF DOSAGE UNITS 905
Column: 4.6-mm 25-cm; packing L1 Acceptance criteria: Meets the requirements for Oral
Flow rate: 3 mL/min Suspension packaged in single-unit containers
Injection size: 20 L DELIVERABLE VOLUME 698
System suitability Acceptance criteria: Meets the requirements for Oral
Sample: System suitability solution A and System suitability Suspension packaged in single-unit containers
solution B ADDITIONAL REQUIREMENTS
[NOTEThe relative retention times for guanine for PACKAGING AND STORAGE: Preserve in tight containers. Store
acyclovir are about 0.6 and 1.0, respectively, in System between 15 and 25. Protect from light.
suitability solution A.] USP REFERENCE STANDARDS 11
Suitability requirements USP Acyclovir RS
Resolution: NLT 2.0 between guanine and acyclovir for
System suitability solution A
injections for the acyclovir peak using System suitability Acyclovir Tablets
solution A (Comment on this Monograph)id=m900=Acyclovir Tablets=A-
Relative standard deviation: NMT 2.0% for replicate Monos.pdf)
injections of System suitability solution B
Analysis: Separately inject the Standard solution and the DEFINITION
Sample solution into the chromatograph, record the Acyclovir Tablets contain NLT 90.0% and NMT 110.0% of the
chromatograms, and measure the peak responses. labeled amount of acyclovir (C8H11N5O3).
Calculate the percentage of C8H11N5O3 in the portion of
Oral Suspension taken: IDENTIFICATION
Result = (rU/rS) (CS/CU) 100 corresponds to that of the Standard solution, as obtained in
the Assay.
rS = peak response from the Standard solution ASSAY
CS = concentration of USP Acyclovir RS in the PROCEDURE
Standard solution (mg/mL) Mobile phase: 0.02 M acetic acid
CU = nominal concentration of acyclovir in the Sample System suitability solution A: 0.1 mg/mL each of USP
solution (mg/mL) Acyclovir RS and guanine in 0.1 N sodium hydroxide
Acceptance criteria: 90.0%110.0% System suitability solution B: 2.0 g/mL of guanine in 0.1
N sodium hydroxide
IMPURITIES Standard solution: 0.1 mg/mL of USP Acyclovir RS in 0.1 N
Organic Impurities sodium hydroxide
PROCEDURE: LIMIT OF GUANINE Sample solution: Transfer an amount of finely powdered
[NOTEMobile phase, Sample solution, and Chromatographic Tablets equivalent to 10 mg of acyclovir (NLT 10 Tablets) to
system: proceed as directed in the Assay.] a 100-mL volumetric flask, dissolve in 10 mL of 0.1 N
Standard solution: 2.0 g/mL of guanine in 0.1 M sodium sodium hydroxide, dilute with water to volume, and filter.
hydroxide Chromatographic system
Analysis: Separately inject the Standard solution and the (See Chromatography 621, System Suitability.)
Sample solution into the chromatograph, record the Mode: LC
chromatograms, and measure the peak responses. Detector: UV 254 nm
Calculate the percentage of guanine in the portion of Oral Column: 4.6-mm 25-cm; packing L1
Suspension taken: Column temperature: 40
Result = (rU/rS) (CS/CU) 100 Injection size: 20 L
System suitability
rU = peak response for guanine from the Sample Sample: System suitability solution A, and System suitability
solution solution B
rS = peak response for guanine from the Standard [NOTEFor System suitability solution A, the relative
solution retention times for guanine and acyclovir are about 0.6
CS = concentration of guanine in the Standard solution and 1.0, respectively.]
(mg/mL)
CU = nominal concentration of acyclovir in the Sample
solution (mg/mL)
USP 32 Official Monographs / Adenine 59
Suitability requirements Adenine

Resolution: NLT 2.0 between guanine and acyclovir for (Comment on this Monograph)id=m930=Adenine=A-
System suitability solution A Monos.pdf)
Relative standard deviation: NMT 2.0% for acyclovir
peak for multiple injections of System suitability solution A;
NMT 2.0% for multiple injections of System suitability
solution B
Analysis
Calculate the quantity, as a percentage, of C8H11N5O3 in
the portion of Tablets taken:
C5H5N5 135.13
Result = (rU/rS) (CS/CU) 100 1H-Purin-6-amine;
1,6-Dihydro-6-iminopurine [73-24-5].
rS = peak response from the Standard solution DEFINITION
CS = concentration of USP Acyclovir RS in the Adenine contains NLT 98.0% and NMT 102.0% of C5H5N5,
Standard solution (mg/mL) calculated on the dried basis.
CU = nominal concentration of acyclovir in the
Sample solution (mg/mL) IDENTIFICATION
Acceptance criteria: 90.0%110.0% INFRARED ABSORPTION 197K
PERFORMANCE TESTS ASSAY
DISSOLUTION 711 PROCEDURE
Medium: 0.1 N hydrochloric acid; 900 mL 0.1 N Perchloric acid: Standardize 0.1 N perchloric acid VS
Apparatus 2: 50 rpm as follows: 300 mg of potassium biphthalate to a 150-mL
Time: 45 min beaker, and dissolve in 80 mL of a mixture of 100 mL of
Detector: UV 254 nm glacial acetic acid and 300 mL of acetic anhydride, by
Standard solution: USP Acyclovir RS in Medium stirring. Titrate with the perchloric acid solution. Each 20.42
Sample solutions: Sample per Dissolution 711. Dilute with mg of potassium biphthalate is equivalent to 1 mL of 0.1 N
Medium to a concentration that is similar to the Standard perchloric acid.
solution. Sample: 200 mg of Adenine
Analysis: Determine the amount of C8H11N5O3 dissolved Analysis: Dissolve in 80 mL of a mixture of 100 mL of
from UV absorption at the wavelength on filtered portions glacial acetic acid and 300 mL of acetic anhydride by
of the solution under test. stirring. Titrate with 0.1 N Perchloric acid. Perform a blank
Tolerances: NLT 80% (Q) of the labeled amount of determination, and make any necessary correction. Each mL
C8H11N5O3 is dissolved. of 0.1 N Perchloric acid is equivalent to 13.514 mg of
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements C5H5N5.
for Weight Variation Acceptance criteria: 98.0%102.0%
IMPURITIES OTHER COMPONENTS
Organic Impurities NITROGEN CONTENT, Method II 461: 50.2%53.4% is found,
PROCEDURE calculated on the dried basis
[NOTEFor Mobile phase, Standard solution, Sample solution,
and Chromatographic system, proceed as directed in the IMPURITIES
Assay.] Inorganic Impurities
Analysis RESIDUE ON IGNITION 281: NMT 0.1%
Sample: Sample solution HEAVY METALS, Method II 231: NMT 10 ppm
Calculate the percentage of each impurity in the portion of Organic Impurities
Tablets taken: PROCEDURE
Solution A: Dissolve 4.54 g of monobasic potassium
Result = 100(rI/rS) phosphate in water to make 500 mL of solution. Dissolve
4.73 g of anhydrous dibasic sodium phosphate in water to
rI = peak response for each impurity make 500 mL of solution. Mix 38.9 mL of the monobasic
rS = sum of the responses for all of the peaks potassium phosphate solution with 61.1 mL of the dibasic
Acceptance criteria sodium phosphate solution. Adjust, if necessary, by the
Individual impurities: NMT 2.0% for guanine, and NMT dropwise addition of the dibasic sodium phosphate solution
0.5% for any other impurity to a pH of 7.0.
Standard stock solution: Dissolve a suitable quantity of
ADDITIONAL REQUIREMENTS USP Adenine RS in hot water, cool, and dilute
PACKAGING AND STORAGE: Preserve in tight containers. Store quantitatively with water to obtain a solution having a
between 15 and 25. Protect from light and moisture. known concentration of 0.19 mg/mL.
USP Acyclovir RS
60 Adenine / Official Monographs USP 32
Standard solutions: Pipet 5-mL portions into three 100- Acceptance criteria: 99.0%101.0%
mL volumetric flasks, dilute with 0.10 N hydrochloric acid,
0.010 N sodium hydroxide, and Solution A, respectively. IMPURITIES
Sample stock solution: Dissolve a suitable quantity of Inorganic Impurities
Adenine in hot water, cool, and dilute quantitatively with RESIDUE ON IGNITION 281: NMT 0.1%
water to obtain a solution having a known concentration of HEAVY METALS, Method II 231: NMT 10 ppm
0.19 mg/mL. Organic Impurities
Sample solutions: Pipet 5-mL portions into three 100-mL Solution A: 6.8 g/L of potassium hydrogen sulfate and 3.4
volumetric flasks, dilute with 0.10 N hydrochloric acid, g/L of tetrabutylammonium hydrogen sulfate in water.
0.010 N sodium hydroxide, and Solution A, respectively. Adjust with 2 N potassium hydroxide to a pH of 6.5.
Blank: Water Mobile phase: Solution A and (1 in 10,000) sodium azide
Mode: Spectrometry solution (3:2)
Analytical wavelength: 220320 nm System suitability solution: 0.2 mg/mL each of Adenosine
Cell: 1 cm and inosine in Mobile phase
Analysis Sample solution: 1.0 mg/mL of Adenosine in Mobile phase
Samples: Standard solutions and Sample solutions Chromatographic system
Acceptance criteria: The respective absorptivities, (See Chromatography 621, System Suitability.)
calculated on the dried basis, at the wavelengths of Mode: LC
maximum absorbance, for each pair of corresponding Detector: UV 254 nm
solutions do not differ by more than 2.0%. Column: 4.6-mm 25-cm; 5-m packing L1
LOSS ON DRYING 731: Dry it at 110 for 4 h: it loses NMT System suitability
1.0% of its weight. Samples: System suitability solution
ADDITIONAL REQUIREMENTS Resolution: NLT 9.0 between adenosine and inosine,
PACKAGING AND STORAGE: Preserve in well-closed containers. System suitability solution
USP REFERENCE STANDARDS 11 Tailing factor: NMT 2.5, System suitability solution
USP Adenine RS Relative standard deviation: NMT 2.0%, System
suitability solution
[NOTEChromatograph the Sample solution, and adjust the
Adenosine run time to at least twice the retention time of the major
peak.]
(Comment on this Monograph)id=m938=Adenosine=A- Analysis
Monos.pdf) Samples: Sample solution
Determine the percentage of each impurity in the portion of
Adenosine taken.
Individual impurities: NMT 0.1% each of guanosine,
inosine, and uridine, and NMT 0.2% of adenine
Total impurities: NMT 0.5%
SPECIFIC TESTS
C10H13N5O4 267.25 OPTICAL ROTATION, Specific Rotation 781S: 68 to 72
9--D-Ribofuranosyladenine; Sample solution: 20 mg/mL in sodium hydroxide solution
6-Amino-9--D-ribofuranosyl-9-H-purine [58-61-7]. (1 in 20), determined on a Sample previously dried at 105
for 2 h
DEFINITION ACIDITY OR ALKALINITY: Suspend 1000 mg in 20 mL of
Adenosine contains NLT 99.0% and NMT 101.0% of carbon dioxide-free water. Stir for 30 s, and pass through a
C10H13N5O4, calculated on the dried basis. coarse filter. To each of two 10-mL portions of the filtrate,
IDENTIFICATION add 0.1 mL of bromocresol purple TS.
Acceptance criteria: NMT 0.3 mL of 0.01 N sodium
INFRARED ABSORPTION 197M hydroxide is required to produce a blue-violet color in one
ASSAY portion. NMT 0.1 mL of 0.01 N hydrochloric acid is required
PROCEDURE to produce a yellow color in the other portion.
Sample: 200 mg of Adenosine previously dried at 105 for LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT
2h 0.5% of its weight.
Analysis: Dissolve in 50 mL of glacial acetic acid and titrate LIMIT OF AMMONIA
with 0.1 N perchloric acid VS. Perform a blank Sample solution: Suspend 0.5 g in 10 mL of water. Stir for
determination, and make any necessary correction. Each mL 30 s, and pass through a coarse filter. Dilute the filtrate with
of 0.1 N perchloric acid is equivalent to 26.72 mg of water to 15 mL, and use the filtrate.
C10H13N5O4.
USP 32 Official Monographs / Adenosine 61
Standard stock solution: Dilute 1 mL of ammonium anticipated final volume of the Standard solution before the
chloride solution (314 mg in 1000 mL) with 100 mL of addition of the warm water.]
water. System sensitivity solution: Standard solution and water
Standard solution: Standard stock solution and water (2:13) (3:197)
Analysis: To the Sample solution and the Standard solution Sample stock solution: Equivalent to 0.3 mg/mL of
add 0.3 mL of alkaline mercuric-potassium iodide TS, cap adenosine from volume of Injection, in water [NOTEReserve
the test tubes, and allow to stand for 5 min. a portion of this stock solution for use in the test for Organic
Acceptance criteria: The Sample solution does not exhibit a Impurities.]
more intense yellow color than that of the Standard solution Sample solution: 0.03 mg/mL of adenosine, from Sample
(NMT 0.0004% ammonia). stock solution and water (1:9)
LIMIT OF CHLORIDE Chromatographic system
Sample solution: Suspend 0.2 g in 10 mL of water. Stir for (See Chromatography 621, System Suitability.)
30 s, pass through a coarse filter, and use the filtrate. Mode: LC
Standard solution: Dilute 1 mL of sodium chloride solution Detector: UV 254 nm
(231 mg in 1000 mL) with 100 mL of water. Column: 3.9-mm 30-cm; packing L1
Analysis: To the Sample solution and 10 mL of the Standard Flow rate: 2.5 mL/min
solution, add 1 mL of nitric acid and 1 mL of silver nitrate Injection size: 10 L
TS, dilute each solution with water to 40 mL. Allow the System suitability
solutions to stand for 5 min, protected from light. Sample: System suitability solution and Standard solution
Acceptance criteria: When viewed against a dark [NOTEChromatograph the System sensitivity solution and
background, the Sample solution is not more turbid than the adjust the run time to 21/2 times the retention time of
Standard solution (NMT 0.007% chloride). adenosine.]
LIMIT OF SULFATE Suitability requirements
Sample solution: Suspend 0.75 g in 15 mL of water. Stir Resolution: NLT 6.0 between adenosine and inosine,
for 30 s, pass through a coarse filter, and use the filtrate. System suitability solution
Standard solution: Add 0.15 mL of 0.020 N sulfuric acid to Tailing factor: NMT 2.0 for the adenosine peak, System
15 mL of water. suitability solution
Analysis: To the Sample solution and the Standard solution Relative standard deviation: NMT 1.5%, Standard
add 2 mL of barium chloride TS and 1 mL of 3 N solution
hydrochloric acid, dilute each solution with water to 30 mL, Analysis
and mix. Allow the solutions to stand for 5 min. Samples: Standard solution and Sample solution
Acceptance criteria: The Sample solution is not more turbid Calculate the percentage of C10H13N5O4 in each mL of the
than the Standard solution (NMT 0.02% sulfate). Injection taken:
containers, and store at controlled room temperature. rU = peak responses from the Sample solution
USP REFERENCE STANDARDS 11 rS = peak responses from the Standard solution
USP Adenosine RS CS = concentration of USP Adenosine RS in the
CU = nominal concentration of adenosine in the
Adenosine Injection Acceptance criteria: 90.0%110.0%
(Comment on this Monograph)id=m940=Adenosine
Injection=A-Monos.pdf) IMPURITIES
Organic Impurities
DEFINITION PROCEDURE
Adenosine Injection is a sterile solution of Adenosine in Water Mobile phase, System suitability solution, Standard
for Injection. It may contain Sodium Chloride. It contains NLT solution, System sensitivity solution, and
90.0% and NMT 110.0% of the labeled amount of adenosine Chromatographic system: Proceed as directed in the
(C10H13N5O4). Assay.
Sample solution: Use the Sample stock solution reserved
IDENTIFICATION from the Assay.
The retention time of the adenosine peak of the Sample Analysis
solution corresponds to that of the Standard solution, as Sample: Sample solution
obtained in the Assay. Calculate the percentage of each impurity in the volume
of Injection taken:
ASSAY
PROCEDURE Result = (ri/rs) 100
Mobile phase: Dissolve 2.0 g of monobasic potassium
phosphate in 800 mL of water. Add 5 mL of 1.0 M ri = peak response for each impurity
tetrabutylammonium dihydrogen phosphate solution, dilute rs = sum of the responses of all of the peaks
with water to 980 mL, and mix. Add 20 mL of acetonitrile, Acceptance criteria
mix, filter, and degas. Make adjustments if necessary. Individual impurity: NMT 1.0%
System suitability solution: 0.03 mg/mL of each adenosine Total impurities: NMT 1.5%
and inosine from USP Adenosine RS and inosine dissolved in
warm water (50 to 55), and diluted with water SPECIFIC TESTS
Standard solution: 0.03 mg/mL of USP Adenosine RS PH 791: 4.57.5
dissolved in warm water (50 to 55) and diluted with water PARTICULATE MATTER IN INJECTIONS 788: It meets the
[NOTEIf sodium chloride is present in the Injection, add requirements for small-volume injections.
0.01 mL of sodium chloride solution (0.9 in 100)/mL of the
62 Adenosine / Official Monographs USP 32
BACTERIAL ENDOTOXINS TEST 85: When the product is used not to be treated with any toxic, sleep-inducing, or narcosis-
for rapid intravenous injection, it contains NMT 11.62 USP producing compounds, and are not to be treated with any
Endotoxin Units/mg of adenosine. When the product is used compound that would be irritating to the respiratory tract
for continuous peripheral intravenous infusion, it contains when the Medical Air is used. [NOTEReduce the container
NMT 5.95 USP Endotoxin Units/mg of adenosine. pressure by means of a regulator. Measure the gases with a
OTHER REQUIREMENTS: It meets the requirements under gas volume meter downstream from the detector tube to
Injections 1. minimize contamination or change of the specimens.]
The various detector tubes called for in the respective tests
ADDITIONAL REQUIREMENTS are listed under Reagents, Indicators, and SolutionsReagent
PACKAGING AND STORAGE: Preserve in tight, single-dose Specifications.
containers, preferably of Type I glass, and store at controlled LABELING: Where it is piped directly from the collecting tank
room temperature. to the point of use, label each outlet Medical Air.
USP Adenosine RS
USP Endotoxin RS
Alanine
(Comment on this Monograph)id=m1130=Alanine=A-
Medical Air Monos.pdf)
(Comment on this Monograph)id=m1000=Medical Air=A-
Monos.pdf)
DEFINITION
Medical Air is a natural or synthetic mixture of gases consisting
largely of nitrogen and oxygen. It contains NLT 19.5% and
NMT 23.5%, by volume, of O2.
C3H7NO2 89.09
ASSAY L-Alanine [56-41-7].
PROCEDURE
Analysis: Determine the oxygen concentration of Medical DEFINITION
Air using an electrochemical cell analyzer readable to 0.1% Alanine contains NLT 98.5% and NMT 101.5% of C3H7NO2, as
of oxygen and calibrated with ambient air to an accuracy of L-alanine, calculated on the dried basis.
0.2% of oxygen.
[NOTEThe instrument uses the variations of electric IDENTIFICATION
current produced by the interaction of oxygen with an INFRARED ABSORPTION 197K
electrochemical cell to display the oxygen strength of a
confined sample or an in-line flow of the gas. This current ASSAY
generates a signal proportional to the oxygen PROCEDURE
concentration, which is displayed on a meter.] Sample: 80 mg of Alanine
Acceptance criteria: 19.5%23.5% Analysis: Dissolve the Sample in a mixture of glacial acetic
acid and formic acid (50:3). Titrate with 0.1 N perchloric
IMPURITIES acid VS. Perform a blank determination (see Titrimetry
Inorganic Impurities 541). Each mL of 0.1 N perchloric acid is equivalent to
CARBON DIOXIDE: Pass 1000 50 mL through a carbon 8.909 mg of C3H7NO2.
dioxide detector tube at the rate specified for the tube: the Acceptance criteria: 98.5%101.5%
indicator change corresponds to NMT 500 ppm.
CARBON MONOXIDE: Pass 1000 50 mL through a carbon IMPURITIES
monoxide detector tube at the rate specified for the tube: Inorganic Impurities
the indicator change corresponds to NMT 10 ppm. RESIDUE ON IGNITION 281: NMT 0.15%
SULFUR DIOXIDE: Pass 1050 50 mL through a sulfur dioxide CHLORIDE AND SULFATE, Chloride 221: A 1.0-g portion
detector tube at the rate specified for the tube: the indicator shows chloride NMT corresponds to 0.70 mL of 0.020 N
change corresponds to NMT 5 ppm. hydrochloric acid (0.05%).
LIMIT OF NITRIC OXIDE AND NITROGEN DIOXIDE: Pass 550 50 CHLORIDE AND SULFATE, Sulfate 221: A 1.0-g portion shows
mL through a nitric oxidenitrogen dioxide detector tube at sulfate NMT corresponds to 0.30 mL of 0.020 N sulfuric acid
the rate specified for the tube: the indicator change (0.03%).
corresponds to NMT 2.5 ppm. IRON 241: NMT 30 ppm
WATER AND OIL: Support 1 container in an inverted position HEAVY METALS, Method I 231: NMT 15 ppm
(with the valve at the bottom) for 5 min. Cautiously open Organic Impurities
the valve slightly, maintaining the container in an inverted PROCEDURE
position. Vent the gas with a barely audible flow against a Adsorbent: 0.25-mm layer of chromatographic silica gel
stainless steel mirror for a few s: no liquid is discernible on mixture
the mirror. Sample solution: 10 mg/mL of Alanine
Standard solution: 0.05 mg/mL of USP L-Alanine RS
SPECIFIC TESTS [NOTEThis solution has a concentration equivalent to
ODOR: Carefully open the container valve to produce a 0.5% of that of the Sample solution.]
moderate flow of gas. Do not direct the gas stream toward System suitability solution: 0.4 mg/mL each of USP L-
the face, but deflect a portion of the stream toward the Alanine RS and USP Glycine RS
nose: no appreciable odor is discernible. Spray reagent: 2 mg/mL of ninhydrin in a mixture of
ADDITIONAL REQUIREMENTS butyl alcohol and 2 N acetic acid (19:1)
PACKAGING AND STORAGE: Preserve in cylinders or in a low- Developing solvent system: Butyl alcohol, glacial acetic
pressure collecting tank. Containers used for Medical Air are acid, and water (3:1:1)
USP 32 Official Monographs / Albendazole 63
Application volume: 5 L necessary. Cool and titrate with 0.1 N perchloric acid VS to
Analysis a potentiometricUSP32 endpoint (see Titrimetry 541)USP32.
Samples: Sample solution, Standard solution, and System Perform a blank determination. Each mL of 0.1 N perchloric
suitability solution acid is equivalent to 26.53 mg of C12H15N3O2S.
Proceed as directed for Chromatography 621, Thin-Layer Acceptance criteria: 98.0%102.0%
Chromatography. After air-drying the plate, repeat the
development process. After air-drying a second time, IMPURITIES
spray with Spray reagent, and heat to 100105 for 15 Inorganic Impurities
min. Examine the plate under white light. The RESIDUE ON IGNITION 281: NMT 0.2%
chromatogram obtained from the System suitability Organic Impurities
solution exhibits two clearly separated spots. Any PROCEDURE
secondary spot of the Sample solution is not larger or Standard stock solution: 5 mg/mL of USP Albendazole
more intense than the principal spot of the Standard RS in glacial acetic acid
solution. Standard solution: 0.05 mg/mL of USP Albendazole RS in
Acceptance criteria glacial acetic acid, from Standard solution A
Individual impurities: NMT 0.5% Sample solution: 10 mg/mL in glacial acetic acid
Total impurities: NMT 2.0% Chromatographic system
SPECIFIC TESTS Mode: TLC
OPTICAL ROTATION, Specific Rotation 781S: +13.7 to Adsorbant: 0.25-mm layer of silica gel
+15.1 Application volume: 10 L
Sample solution: 100 mg/mL in 6 N hydrochloric acid Developing solvent system: Chloroform, ether, and
PH 791: 5.57.0, in a solution (1 in 20) glacial acetic acid (60:10:10)
LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT Visualization: Short-wavelength UV light
0.2% of its weight. Analysis: Proceed as directed for Chromatography 621,
Thin-Layer Chromatography.
ADDITIONAL REQUIREMENTS Samples: Standard stock solution, Standard solution, and
PACKAGING AND STORAGE: Preserve in tight containers, and Sample solution
store at a controlled room temperature. Develop the chromatogram in the Developing solvent
USP REFERENCE STANDARDS 11 system until the solvent front has moved about three-
USP L-Alanine RS fourths of the length of the plate. Remove the plate from
USP Glycine RS the developing chamber, mark the solvent front, allow
the solvent to evaporate from the plate, and examine the
plate under short-wavelength UV light.
Albendazole Acceptance criteria: No spot, other than the principal
spot of the Sample solution, is larger or more intense than
(Comment on this Monograph)id=m1150=Albendazole=A- the principal spot of the Standard solution (0.5%).
Monos.pdf)
SPECIFIC TESTS
LOSS ON DRYING 731: Dry a sample at 105 for 4 h: it loses
NMT 0.5% of its weight.
C12H15N3O2S 265.33 USP Albendazole RS
Carbamic acid, [5-(propylthio)-1H-benzimidazol-2-yl]-, methyl
ester;
Methyl 5-(propylthio)-2-benzimidazolecarbamate
[54965-21-8]. Albendazole Oral Suspension
(Comment on this Monograph)id=m1158=Albendazole Oral
DEFINITION Suspension=A-Monos.pdf)
Albendazole contains NLT 98.0% and NMT 102.0% of
C12H15N3O2S, calculated on the dried basis. DEFINITION
Albendazole Oral Suspension is Albendazole in an aqueous
IDENTIFICATION vehicle. It contains one or more preservatives and dispersing or
A. INFRARED ABSORPTION 197M suspending agents. It contains NLT 90.0% and NMT 110.0%
B. The RF value of the principal spot of the Sample solution of the labeled amount of albendazole (C12H15N3O2S).
corresponds to that of the principal spot of the Standard
solution, as obtained in the test for Organic Impurities. IDENTIFICATION
ULTRAVIOLET ABSORPTION 197U
ASSAY Sample stock solution: 1.0 mg/mL of albendazole from a
quantity of Suspension, in a mixture of methanol and
Change to read: hydrochloric acid (99:1) [NOTEFilter the mixture, if
necessary, to obtain a clear solution.]
Sample solution: Sample stock solution and 0.1 N sodium
PROCEDURE hydroxide (1:99)
Sample: 250 mg of Albendazole
Analysis: Transfer the Sample to a suitable flask and dissolve
in 100 mL of glacial acetic acid, warming gently if
64 Albendazole / Official Monographs USP 32
ASSAY the Assay with Acidified methanol, prepared as directed for

PROCEDURE Dissolution, to obtain solutions containing 10 g/mL
Solution A: Methanol and hydrochloric acid (99:1) albendazole.
Solution B: 13.75 mg/mL of monobasic sodium phosphate B. The retention time of the major peak for albendazole of
Mobile phase: Methanol and Solution B (3:2) the Sample solution corresponds to that of the Standard
Standard stock solution: 1.0 mg/mL of USP Albendazole solution, as obtained in the Assay.
RS in Solution A
Standard solution: 100 g/mL of USP Albendazole RS, ASSAY
from Standard stock solution in Mobile phase PROCEDURE
Sample stock solution: Equivalent to 1.0 mg/mL of Mobile phase: Dissolve 0.50 g of monobasic ammonium
albendazole from a volume of Oral Suspension, in Solution A phosphate in 400 mL of water. Add 600 mL of methanol
Sample solution: 100 g/mL of albendazole from Sample and filter, discarding the first 15 mL of filtrate. Degas the
stock solution in Mobile phase [NOTEFilter, if necessary, to clear filtrate before use.
obtain a clear solution.] Solution A: Methanol and sulfuric acid (99:1)
Chromatographic system Internal standard solution: Transfer 150 mg of USP
(See Chromatography 621, System Suitability.) Parbendazole RS to a 50-mL volumetric flask, dissolve with
Mode: LC shaking in 5 mL of Solution A and 25 mL of methanol, then
Detector: UV 308 nm dilute with methanol to volume.
Column: 4-mm 25-cm; packing L1 Standard stock solution: Transfer 100 mg of USP
Flow rate: 2 mL/min Albendazole RS to a 50-mL volumetric flask, dissolve with
Injection size: 20 L shaking in 5 mL of Solution A and 25 mL of methanol, then
System suitability dilute with methanol to volume.
Sample: Standard solution Standard solution: Transfer 5.0 mL of Standard stock
Suitability requirements solution and 5.0 mL of Internal standard solution to a 50-mL
Column efficiency: NLT 2000 theoretical plates volumetric flask, and dilute with methanol to volume.
Tailing factor: NMT 2.0 Sample stock solution: Transfer finely powdered Tablets
Relative standard deviation: NMT 2.0% (NLT 20 Tablets) equivalent to 100 mg of albendazole to a
Analysis 50-mL volumetric flask, add 5 mL of Solution A and 20 mL
Samples: Standard solution and Sample solution of methanol, and shake by mechanical means for about 15
Calculate the percentage of C12H15N3O2S in each mL of the min. Dilute to volume with methanol and filter, discarding
Oral Suspension taken: the first 15 mL of the filtrate (2 mg/mL).
Sample solution: Transfer 5.0 mL of the Sample stock
Result = (rU/rS) (CS/CU) 100 solution and 5.0 mL of the Internal standard solution to a 50-
mL volumetric flask, and dilute with methanol to volume.
rU = peak response from the Sample solution Chromatographic system
rS = peak response from the Standard solution (See Chromatography 621, System Suitability.)
CS = concentration of USP Albendazole RS in the Mode: LC
Standard solution (mg/mL) Detector: UV 254 nm
CU = nominal concentration of albendazole in the Column: 4.6-mm 25-cm; 5-m packing L1
Sample solution (mg/mL) Flow rate: 2 mL/min
Acceptance criteria: 90.0%110.0% Injection size: 20 L
System suitability
SPECIFIC TESTS Sample: Standard solution
PH 791: 4.55.5 Suitability requirements
Resolution: NLT 2.0 between the albendazole peak and
ADDITIONAL REQUIREMENTS the parbendazole peak
PACKAGING AND STORAGE: Preserve in tight containers, and Column efficiency: NLT 1000 theoretical plates
store at controlled room temperature. Tailing factor: NMT 2.0
LABELING: Label it to indicate that it is for veterinary use Relative standard deviation: NMT 2.0% for replicate
only. injections
USP REFERENCE STANDARDS 11 Analysis: [NOTEUse peak heights where peak responses
USP Albendazole RS are indicated.]
Calculate the quantity, in percentage, of C12H15N3O2S in the
Albendazole Tablets portion of Tablets taken:
(Comment on this Monograph)id=m1162=Albendazole Result = (RU/RS) (CS/CU) 100
Tablets=A-Monos.pdf)
RU = peak response ratio of the albendazole peak to
DEFINITION the parbendazole peak from the Sample solution
Albendazole Tablets contain NLT 90.0% and NMT 110.0% of RS = peak response ratio of the albendazole peak to
the labeled amount of albendazole (C12H15N3O2S). the parbendazole peak from the Standard
A. ULTRAVIOLET ABSORPTION 197U CS = concentration of USP Albendazole RS in the
Solution: Dilute a portion of the clear filtrate used to Standard solution (mg/mL)
prepare the Sample solution and a portion of the stock CU = nominal concentration of albendazole in the
solution used to prepare the Standard solution prepared in Sample solution (mg/L)
USP 32 Official Monographs / Albumin 65
Acceptance criteria: 90.0%110.0% ADDITIONAL REQUIREMENTS

PERFORMANCE TESTS store at controlled room temperature.
DISSOLUTION 711 LABELING: Tablets intended for veterinary use only are so
Medium: 0.1 N hydrochloric acid; 900 mL labeled.
Time: 30 min USP Albendazole RS
[NOTEDetermine the amount of C12H15N3O2S dissolved USP Parbendazole RS
using the following Analysis.]
Acidified methanol: Methanol and hydrochloric acid (49:1)
Standard stock solution: Dissolve 90 mg of USP
Albendazole RS in 10 mL of Acidified methanol in a 250-mL Albumin Human
volumetric flask with shaking. Dilute with 0.1 N hydrochloric (Comment on this Monograph)id=m1180=Albumin Human=A-
acid to volume. Monos.pdf)
Standard solution: 5.0 mL of the Standard stock solution
diluted to 200-mL with 0.1 N sodium hydroxide DEFINITION
Sample solution: Sample per Dissolution 711. Transfer Albumin Human conforms to the regulations of the federal
10.0 mL of a filtered portion of the Sample to a 250-mL Food and Drug Administration concerning biologics (640.80 to
volumetric flask, and dilute with 0.1 N sodium hydroxide to 640.86) (see Biologics 1041). It is a sterile, nonpyrogenic
volume. preparation of serum albumin obtained by fractionating
Blank: 0.1 N sodium hydroxide material (source blood, plasma, serum, or placentas) from
Analysis: Concomitantly determine the absorbances of the healthy human donors, the source material being tested for
Sample solution and the Standard solution at the wavelengths the absence of hepatitis B surface antigen. It is made by a
of maximum and minimum absorbance at about 308 nm process that yields a product that is safe for intravenous use.
and 350 nm against the Blank. NLT 96% of its total protein is albumin. It is a solution
Calculate the quantity, in mg, of C12H15N3O2S dissolved: containing, in each 100 mL, either 25 g of serum albumin
osmotically equivalent to 500 mL of normal human plasma, or
Result = 22.5C(AU/AS) 20 g equivalent to 400 mL, or 5 g equivalent to 100 mL, or 4
g equivalent to 80 mL thereof, and contains NLT 93.75% and
C = concentration of USP Albendazole RS in the NMT 106.25% of the labeled amount in the case of the
Standard solution (g/mL) solution containing 4 g in each 100 mL, and NLT 94.0% and
AU = difference in absorbance between 308 nm and NMT 106.0% of the labeled amount in the other cases. It
350 nm from the Sample solution contains no added antimicrobial agent, but may contain
AS = difference in absorbance between 308 nm and sodium acetyltryptophanate with or without sodium caprylate
350 nm from Standard solution as a stabilizing agent. It has a sodium content of NLT 130 mEq
Tolerances: NLT 80% (Q) of the labeled amount is /L and NMT 160 mEq/L. It has a heme content such that the
dissolved. absorbance of a solution, diluted to contain 1% of protein, in
UNIFORMITY OF DOSAGE UNITS, Content Uniformity 905 a 1-cm holding cell, measured at a wavelength of 403 nm, is
Acidified methanol: Prepare as directed in Dissolution. NMT 0.25. It meets the requirements of the test for heat
Standard stock solution: Prepare as directed in Dissolution. stability and for pH.
Standard solution: Prepare as directed in Dissolution.
Sample stock solution: Place 1 Tablet in a 500-mL ADDITIONAL REQUIREMENTS
volumetric flask, add about 300 mL of Acidified methanol, PACKAGING AND STORAGE: Preserve in tight containers, and
shake by mechanical means for about 30 min, and dilute store at the temperature recommended by the manufacturer
with Acidified methanol to volume. Filter a portion of this or indicated on the label.
solution, discarding the first 20 mL of the filtrate. EXPIRATION DATE: The expiration date is not later than 5
Sample solution: Transfer 4.0 mL of the Sample stock years after issue from manufacturers cold storage (5, 3
solution to a 200-mL volumetric flask, and dilute with 0.1 N years) if labeling recommends storage between 2 and 10;
sodium hydroxide to volume. not later than 3 years after issue from manufacturers cold
Blank: 0.1 N sodium hydroxide storage (5, 3 years) if labeling recommends storage at
Analysis: Concomitantly determine the absorbances of the temperatures not higher than 37; and not later than 10
Standard solution and the Sample solution at the wavelengths years after date of manufacture if in a hermetically sealed
of maximum and minimum absorbance at about 308 nm metal container and labeling recommends storage between
and 350 nm against the Blank. 2 and 10.
Calculate the quantity, in mg, of C12H15N3O2S in the Tablet LABELING: Label it to state that it is not to be used if it is
taken: turbid and that it is to be used within 4 h after the container
is entered. Label it also to state the osmotic equivalent in
Result = 25C(AU/AS) terms of plasma, the sodium content, and the type of source
material (venous plasma, placental plasma, or both) from
C = concentration of USP Albendazole RS in the which it was prepared. Label it also to indicate that
Standard solution (g/mL) additional fluids are needed when the 20-g/100-mL or 25-
AU = difference in absorbance between 308 nm and g/100-mL product is administered to a markedly dehydrated
350 nm from the Sample solution patient.
AS = difference in absorbance between 308 nm and
350 nm from Standard solution
66 Albuterol / Official Monographs USP 32
Albuterol SPECIFIC TESTS

(Comment on this Monograph)id=m1195=Albuterol=A- WATER DETERMINATION, Method I 921: NMT 0.5%
Monos.pdf) ADDITIONAL REQUIREMENTS
PACKAGING AND STORAGE: Preserve in well-closed, light-
resistant containers.
USP Albuterol RS
C13H21NO3 239.31
Albuterol Sulfate
1,3-Benzenedimethanol, 1-[[(1,1- (Comment on this Monograph)id=m1210=Albuterol Sulfate=A-
dimethylethyl)amino]methyl]-4-hydroxy-; Monos.pdf)
1-[(tert-Butylamino)methyl]-4-hydroxy-m-xylene-,-diol (C13H21NO3)2 H2SO4 576.70
[18559-94-9]. 1,3-Benzenedimethanol, 1-[[(1,1-
dimethylethyl)amino]methyl]-4-hydroxy-, sulfate (2:1) (salt);
DEFINITION 1-[(tert-Butylamino)methyl]-4-hydroxy-m-xylene-,-diol
Albuterol contains NLT 98.5% and NMT 101.0% of C13H21NO3, sulfate (2:1) (salt) [51022-70-9].
DEFINITION
IDENTIFICATION Albuterol Sulfate contains NLT 98.5% and NMT 101.0% of
A. INFRARED ABSORPTION 197K (C13H21NO3)2 H2SO4, calculated on the anhydrous basis.
B. ULTRAVIOLET ABSORPTION 197U
Sample solution: 80 g/mL in 0.1 N hydrochloric acid IDENTIFICATION
ASSAY B. ULTRAVIOLET ABSORPTION 197U
PROCEDURE Sample solution: 80 g/mL in 0.1 N hydrochloric acid
Sample solution: 8 mg/mL of Albuterol in glacial acetic C. IDENTIFICATION TESTSGENERAL, Sulfate 191
acid Sample solution: Shake an amount of sample equivalent to
Analysis: To 50 mL of Sample solution, add 2 drops of 4 mg of albuterol with 10 mL of water, and filter.
crystal violet TS, and titrate with 0.1 N perchloric acid VS. Acceptance criteria: Meets the requirements of the tests
Perform a blank determination, and make any necessary D. The retention time of the major peak of the Sample
correction. Each mL of 0.1 N perchloric acid is equivalent to solution corresponds to that of the Standard solution, as
23.93 mg of C13H21NO3. obtained in the Assay.
ASSAY
IMPURITIES PROCEDURE
Inorganic Impurities Solution A: 3.85 mg/mL of ammonium acetate
RESIDUE ON IGNITION 281: NMT 0.1% Mobile phase: Isopropanol, Solution A, and water [(5
Organic Impurities 1):30:65], filtered and degassed. Adjust dropwise with acetic
PROCEDURE acid to a pH of 4.5 0.3.
Standard solution: 0.10 mg/mL of USP Albuterol RS in Standard solution: 0.6 mg/mL of USP Albuterol Sulfate RS
methanol Sample solution: 0.6 mg/mL of Albuterol Sulfate
Sample solution: 20 mg/mL of Albuterol in methanol Chromatographic system
See Chromatography 621, Thin-Layer Chromatography Mode: LC
Mode: TLC Detector: UV 276 nm
Adsorbent: 0.25-mm layer of chromatographic silica gel Column: 4.6-mm 20-cm; packing L10
Application volume: 10 L Flow rate: 2.0 mL/min
Developing solvent system: Methyl isobutyl ketone, Injection size: 10 L
isopropyl alcohol, ethyl acetate, ammonium hydroxide, System suitability
and water (50:45:35:3:18) Sample: Standard solution
Visualization: Iodine vapor Suitability requirements
Analysis Resolution: NLT 1.5 between albuterol and 4-[2-[(1,1-
Samples: Standard solution and Sample solution dimethylethyl)amino]-1-hydroxyethyl]-2-methylphenol
Proceed as directed for Chromatography 621, Thin-Layer sulfate
Chromatography, applying aliquots of the Standard Relative standard deviation: NMT 1.5%
solution and the Sample solution. Develop in the Analysis:
Developing solvent system until the solvent front has Sample: Standard solution and Sample solution
moved three-fourths the length of the plate. Remove the Calculate the quantity, in percentage, of (C13H21NO3)2
plate from the developing chamber, air-dry, and expose H2SO4 in the portion of Albuterol Sulfate taken:
it to iodine vapor.
Acceptance criteria: Any spot, other than the principal Result = (rU/rS) (CS/CU) 100
spot, obtained from the Sample solution is not greater in
size and intensity than the spot produced by the Standard rU = peak response from the Sample solution
solution (0.5%), and the sum of the impurities is not rS = peak response from the Standard solution
greater than 2.0%.
USP 32 Official Monographs / Albuterol 67
CS = concentration of USP Albuterol Sulfate RS in the Diluent: Methanol and water (2:3)
Standard solution (mg/mL) Mobile phase: Methanol and Solution B (2:3)
CU = concentration of the Sample solution (mg/mL) Standard stock solution: 0.12 mg/mL of USP Albuterol
Acceptance criteria: NLT 98.5% and NMT 101.0% Sulfate RS in a mixture of Solution A and methanol
[NOTETo USP Albuterol Sulfate RS, add a portion of
IMPURITIES Solution A corresponding to 60% of the final solution
Inorganic Impurities volume. Sonicate for 5 min, and dilute to final volume
RESIDUE ON IGNITION 281 with methanol.]
Acceptance criteria: NMT 0.1% Standard solution: 0.03 mg/mL of USP Albuterol Sulfate RS
Organic Impurities from Standard stock solution, in Diluent
PROCEDURE Sample solution: To the whole Tablets, add a portion of
Adsorbent: 0.25-mm layer of chromatographic silica gel Solution A corresponding to 60% of the final solution
Standard solution: 0.10 mg/mL of USP Albuterol Sulfate volume, shake by mechanical means for 45 min, sonicate for
RS 10 min, allow to cool to room temperature, and dilute to
Sample solution: 20 mg/mL of Albuterol Sulfate final volume with methanol equivalent to 25 g/mL of
Application volume: 10 L albuterol from a number of whole Tablets in a mixture of
Developing solvent system: Methyl isobutyl ketone, Solution A and methanol. Pass through a suitable 0.45-m or
isopropyl alcohol, ethyl acetate, ammonium hydroxide and finer porosity fliter.
water (50:45:35:3:18) Chromatographic system
Visualization: Iodine vapor (See Chromatography 621, System Suitability.)
Analysis: Proceed as directed in Chromatography 621, Mode: LC
Thin-layer Chromatography, applying aliquots of the Detector: UV 276 nm
Standard solution and the Sample solution. Develop in the Column: 4.6-mm 15-cm; packing L1
Developing solvent system until the solvent front has moved Flow rate: 1.5 mL/min
three-fourths the length of the plate. Remove the plate Injection size: 25 L
from the developing chamber, air-dry, and expose it to System suitability
iodine vapor. Sample: Standard solution
Acceptance criteria: Any spot, other than the principal Suitability requirements
spot, obtained from the Sample solution is not greater in Column efficiency: NLT 800 theoretical plates
size and intensity than the spot produced by the Standard determined from the analyte peak
solution (0.5%), and the sum of the impurities is not Tailing factor: NMT 2.5 for the analyte peak
greater than 2.0%. Relative standard deviation: NMT 2.0%
Analysis
SPECIFIC TESTS Samples: Standard solution and Sample Solution
WATER DETERMINATION, Method I 921: NMT 0.5% Calculate the quantity, as a percentage, of C13H21NO3 in
ADDITIONAL REQUIREMENTS the portion of Tablets taken:
resistant containers. Result = (rU/rS) (CS/CU) N (Mr1/Mr2) 100
USP Albuterol Sulfate RS rS = peak response from the Standard solution
CS = concentration of USP Albuterol Sulfate RS in the
Albuterol Tablets CU = nominal concentration of albuterol in the Sample
solution (mg/mL)
(Comment on this Monograph)id=m1218=Albuterol Tablets=A- N = number of molecules of albuterol released from
Monos.pdf) each molecule of albuterol sulfate, 2
DEFINITION Mr1 = molecular weight of albuterol, 239.31
Albuterol Tablets contain an amount of albuterol sulfate Mr2 = molecular weight of albuterol sulfate, 576.70
[(C13H21NO3)2 H2SO4] equivalent to NLT 90.0% and NMT Acceptance criteria: 90.0%110.0%
110.0% of the labeled amount of albuterol (C13H21NO3). PERFORMANCE TESTS
IDENTIFICATION DISSOLUTION 711, Procedure for a Pooled Sample
A. The RF value of the principal spot obtained from the Medium: Water; 500 mL
Sample solution corresponds to that obtained from Standard Apparatus 2: 50 rpm
solution A obtained as directed in the Procedure for Organic Time: 30 min
Impurities. Determine the amount of C13H21NO3 dissolved using the
B. IDENTIFICATION TESTSGENERAL, Sulfate 191: Shake a following method. [NOTEFor the Mobile phase, Standard
quantity of the powdered tablets equivalent to 4 mg of solution, and Chromatographic system, prepare as directed
albuterol with 10 mL, and filter. The filtrate so obtained in the Assay.]
meets the requirements of the test. Analysis: Inject a suitable volume (about 100 L) of a
portion of the Sample solution, previously passed through a
ASSAY 0.45-m nylon filter, into the chromatograph. Calculate the
PROCEDURE quantity of C13H21NO3 dissolved by comparing this peak
Solution A: Dilute 20 mL of glacial acetic acid to 2 L. response with the major peak response similarly obtained on
Solution B: 1.13 g of sodium 1-hexanesulfonate in 1200 chromatographing the Standard solution previously diluted, if
mL of water. Add 12 mL of glacial acetic acid. necessary, with a mixture of water and methanol (6:4) to
68 Albuterol / Official Monographs USP 32
obtain a Standard solution having a known concentration of Alclometasone Dipropionate

USP Albuterol Sulfate RS approximately corresponding to the (Comment on this Monograph)id=m1225=Alclometasone
concentration of the Sample solution. Dipropionate=A-Monos.pdf)
C13H21NO3 is dissolved.
IMPURITIES
Organic Impurities
PROCEDURE
Adsorbent: 0.25-mm layer of chromatographic silica gel
Standard solution A: Prepare a solution of USP Albuterol C28H37ClO7 521.04
Sulfate RS having a known concentration of 0.580 mg/mL Pregna-1,4-diene-3,20-dione, 7-chloro-11-hydroxy-16-
equivalent to 0.483 mg of albuterol. methyl-17,21-bis(1-oxopropoxy)-, (7,11,16)-;
Standard solution B: Prepare a solution of USP Albuterol 7-Chloro-11,17,21-trihydroxy-16-methylpregna-1,4-
Sulfate RS having a known concentration of 0.218 mg/mL diene-3,20-dione 17,21-dipropionate [66734-13-2].
equivalent to 0.183 mg of albuterol.
Standard solution C: Prepare a solution of USP Albuterol DEFINITION
Sulfate RS having a known concentration of 0.073 mg/mL Alclometasone Dipropionate contains NLT 97.0% and NMT
equivalent to 0.061 mg of albuterol. 102.0% of C28H37ClO7, calculated on the dried basis.
Sample solution: Place a quantity of finely powdered
Tablets, equivalent to 48 mg of albuterol, into a suitable IDENTIFICATION
container. Add 60 mL of diluted alcohol (1 in 2), and shake A. INFRARED ABSORPTION 197M
by mechanical means for 30 min. Filter the mixture, and B. The retention time of the major peak of the Sample
wash the filter with small portions of alcohol, combining solution corresponds to that of the Standard solution, both
this with the filtrate. Evaporate the filtrate to dryness under relative to the Internal standard solution, as obtained in the
reduced pressure at a temperature below 40. Dissolve the Assay.
residue as completely as possible in 2 mL.
Application volume: 10-L aliquots (in two successive ASSAY
portions of 5 L, allowing the solvent to evaporate PROCEDURE
between applications) Solution A: 6.80 mg/mL of monobasic potassium
Developing solvent system: Methyl isobutyl ketone, phosphate (0.05 M)
isopropyl alcohol, ethyl acetate, ammonium hydroxide, and Mobile phase: Methanol and Solution A (2:1)
water (50:45:35:3:18) Internal standard solution: 2 mg/mL of betamethasone
Spray reagent A: 3-Methyl-2-benzothiazolinone hydrazone dipropionate in methanol
hydrochloride TS Standard stock solution: 1.2 mg/mL of USP Alclometasone
Spray reagent B: Ammoniacal potassium ferricyanide TS Dipropionate RS in methanol
Analysis Standard solution: 4.0 mL Standard stock solution and 4.0
Samples: Sample solution, Standard solution A, Standard mL Internal standard solution. Dilute with methanol to 25
solution B, and Standard solultion C mL.
Proceed as directed for Chromatography 621, Thin-layer [NOTEThis solution contains approximately 0.2 mg/mL of
Chromatography. Develop the chromatograms until the USP Alclometasone Dipropionate RS.]
solvent front has moved about 17 cm. Spray the plate Sample stock solution: 1.2 mg/mL of Alclometasone
first with Spray reagent A, and then Spray reagent B, and Dipropionate in methanol
finally again with Spray reagent A. Examine the plate and Sample solution: 4 mL Sample stock solution and 4 mL
estimate the responses of any secondary spots observed Internal standard solution. Dilute with methanol to 25 mL.
in the lane of the Sample solution by comparison with Chromatographic system
those of Standard solutions A, B, and C. (See Chromatography 621, System Suitability.)
Acceptance criteria: No major secondary spot is greater in Mode: LC
size or intensity than the principal spot produced by Detector: UV 254 nm
Standard solution A (2.0%). No other secondary spot is Column: 4-mm 30-cm; packing L1
greater in size or intensity than the principal spot produced Flow rate: 1.2 mL/min
by Standard solution B (0.75%). No more than two other Injection size: 10 L
secondary spots are equal in size or intensity than the System suitability
principal spot produced by Standard solution C (0.25%). Sample: Standard solution
The sum of the intensities of all secondary spots obtained [NOTEThe relative retention times for alclometasone
from the Sample solution corresponds to NMT 3.5%. dipropionate and betamethasone dipropionate are about
0.7 and 1.0, respectively.]
ADDITIONAL REQUIREMENTS Suitability requirements
PACKAGING AND STORAGE: Preserve in tight, light-resistant Resolution: NLT 3.0 between the analyte and the Internal
containers, and store at a controlled room temperature. standard solution peaks
USP Albuterol Sulfate RS
USP 32 Official Monographs / Alclometasone 69
Relative standard deviation: NMT 2% ADDITIONAL REQUIREMENTS

Analysis PACKAGING AND STORAGE: Preserve in tight containers, and
Samples: Standard solution and Sample solution store at a controlled room temperature.
Calculate the percentage of C28H37ClO7 in the portion USP REFERENCE STANDARDS 11
taken: USP Alclometasone Dipropionate RS
rU = peak height ratio from the Sample solution Alclometasone Dipropionate Cream
rS = peak height ratio from the Standard solution (Comment on this Monograph)id=m1227=Alclometasone
CS = concentration of USP Alclometasone Dipropionate Cream=A-Monos.pdf)
Dipropionate RS in the Standard solution
(mg/mL) DEFINITION
CU = concentration of the Sample solution (mg/mL) Alclometasone Dipropionate Cream contains NLT 90.0% and
Acceptance criteria: 97.0%102.0% NMT 110.0% of the labeled amount of alclometasone
dipropionate (C28H37ClO7) in a suitable cream base.
IMPURITIES
Inorganic Impurities IDENTIFICATION
RESIDUE ON IGNITION 281: NMT 0.1% A. The retention time of the major peak of the Sample
HEAVY METALS, Method II 231: NMT 30 ppm solution corresponds to that of the Standard solution, both
Organic Impurities relative to the Internal standard solution, as obtained in the
PROCEDURE Assay.
Adsorbent: 0.25-mm layer of chromatographic silica gel B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
mixture Adsorbent: 0.25-mm layer of chromatographic silica gel
Diluent: Dilute 250 mL methanol with dichloromethane to mixture
500 mL. Standard solution: 0.08 mg/mL in methanol
Standard stock solution: 250 10 g/mL of USP Sample solution: Place a quantity of Cream, equivalent to
Alclometasone Dipropionate RS in Diluent 1.25 mg of alclometasone dipropionate, in a 50-mL
Standard solutions: Dilute volumes of Standard stock centrifuge tube, and add 15 mL of methanol. Insert a
solution quantitatively with Diluent to obtain Standard stopper securely into the tube, and place the tube in a
solutions, designated below by letter, having the following water bath maintained at 60 until the semisolid
concentrations. components melt. Remove the tube from the bath, shake
vigorously until the specimen components resolidify, and
Standard Concentration Percentage place the tube in an icemethanol bath for 15 min. Remove
Solution Dilution (g/mL) (%) the tube from the bath, and centrifuge at 2500 rpm for 5
min. Transfer the clear supernatant to a vial, and allow this
A (4 in 5) 200 2.0 Sample solution to equilibrate to room temperature.
B (3 in 5) 150 1.5 Application volume: 20 L
C (2 in 5) 100 1.0 Visualization: Short-wavelength UV light
Developing solvent system: Chloroform and acetone (7:1)
D (3 in 10) 75 0.75 Analysis: Standard solution and Sample solution
E (2 in 10) 50 0.50 [NOTEDry the applications with the aid of a stream of
F (1 in 10) 25 0.25 nitrogen.]
Proceed as directed for Chromatography 621, Thin-Layer
Sample solution: 10 0.4 mg/mL of Alclometasone Chromatography until the solvent has moved about three-
Dipropionate in Diluent fourths the length of the plate. Observe the plate under
Application volume: 25 L short-wavelength UV light.
Visualization: Short-wavelength UV light Acceptance criteria: The RF value of the principal spot
Developing solvent system: Chloroform and acetone obtained from the Sample solution corresponds to that
(7:1) obtained from the Standard solution.
Analysis ASSAY
Samples: Standard solutions AF and Sample solution PROCEDURE
Proceed as directed for Chromatography 621, Thin-Layer Solution A: 6.80 mg/mL of monobasic potassium
Chromatography. Develop the chromatograms until the phosphate (0.05 M)
solvent front has moved about three-fourths the length Mobile phase: Methanol and Solution A (2:1)
of the plate. Observe the plate under short-wavelength Internal standard solution: 0.4 mg/mL of betamethasone
UV light, and compare the intensities of any secondary dipropionate in methanol
spots observed in the chromatogram of the Sample Standard stock solution: 0.25 mg/mL of USP
solution with those of the principal spots in the Alclometasone Dipropionate RS in methanol
chromatograms of the Standard solutions. Standard solution: Transfer about 25 mg of USP
Acceptance criteria: The sum of the intensities of Alclometasone Diproprionate RS to a 100-mL volumetric
secondary spots obtained from the Sample solutions flask, add methanol to volume, and mix. Transfer 5.0 mL of
corresponds to NMT 3.0% of related compounds. this solution to a small stoppered flask, add 5.0 mL of
SPECIFIC TESTS methanol and 5.0 mL of Internal standard solution, and mix
OPTICAL ROTATION, Specific Rotation 781S: +21 to +25 to obtain a Standard solution having a known concentration
Sample solution: 30 mg/mL in dioxane of about 0.08 mg of USP Alclometasone Dipropionate RS
LOSS ON DRYING 731: Dry it in vacuum at a pressure not per mL.
exceeding 5 mm of mercury at 105 for 3 h: it loses NMT Sample solution: Transfer a quantity of Cream, equivalent
0.5% of its weight. to 1.25 mg of alclometasone dipropionate, to a 50-mL
70 Alclometasone / Official Monographs USP 32
centrifuge tube. Add 5.0 mL of Internal standard solution, relative to the Internal standard solution, as obtained in the
and add 10.0 mL of methanol. Insert a stopper securely into Assay.
the tube, and place it in a water bath maintained at 60 B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
until the semisolid components melt. Remove the tube from Adsorbent: 0.25-mm layer of chromatographic silica gel
the bath, shake vigorously until the specimen components mixture
resolidify, and return the tube to the 60 water bath until Standard solution: 0.25 mg/mL USP Alclometasone
the semisolid components melt. Remove the tube from the Dipropionate RS in methanol
bath, shake vigorously until the specimen components Sample solution: Place a quantity of Ointment, equivalent
resolidify, and place the tube in an icemethanol bath for 15 to 1.25 mg of alclometasone dipropionate, in a 50-mL
min. Remove the tube from the bath, and centrifuge at centrifuge tube, add 10 mL of 2,2,4-trimethylpentane, insert
2500 rpm for 5 min. Transfer the clear supernatant to a a stopper securely into the tube, and disperse the specimen
small stoppered flask, and allow this Sample solution to using a vortex mixer. Add 5.0 mL of a solution of methanol
equilibrate to room temperature. in water (45 in 50), insert the stopper securely, shake
Chromatographic system vigorously for 2 min, and centrifuge at 2500 rpm for 3 min.
(See Chromatography 621, System Suitability.) Remove the lower, aqueous alcohol phase, and transfer this
Mode: LC Sample solution to a stoppered vial.
Detector: UV 254 nm Application volume: 20 L
Column: 4-mm 30-cm; packing L1 Developing solvent system: Chloroform and acetone (7:1)
Flow rate: 1.2 mL/min Analysis
Injection size: 20 L Samples: Standard solution and Sample solution
System suitability [NOTEDry the applications with the aid of a stream of
Sample: Standard solution nitrogen.]
[NOTEThe relative retention times for alclometasone Proceed as directed for Chromatography 621, Thin-Layer
dipropionate and betamethasone dipropionate are about Chromatography until the solvent front has moved about
0.7 and 1.0, respectively.] three-fourths of the length of the plate. Observe the plate
Suitability requirements under short-wavelength UV light.
Resolution: NLT 3.0 between the analyte and internal Acceptance criteria: The RF value of the principal spot
standard peaks obtained from the Sample solution corresponds to that
Relative standard deviation: NMT 2% obtained from the Standard solution.
Analysis: Standard solution and Sample solution
Calculate the quantity, as a percentage, of C28H37ClO7 in the ASSAY
portion of Cream taken: PROCEDURE
Solution A: 6.80 mg/mL of monobasic potassium
Result = (RU/RS) (CS/CU) 100 phosphate (0.05 M)
Solution B: Dilute 450 mL of methanol to 500 mL with
RU = peak height ratio from the Sample solution water.
RS = peak height ratio from the Standard solution Mobile phase: Methanol and Solution A (2:1)
CS = concentration of USP Alclometasone Internal standard solution: 0.15 mg/mL of betamethasone
Dipropionate RS in the Standard solution dipropionate in Solution B
(mg/mL) Standard stock solution: 0.1 mg/mL of USP Alclometasone
CU = nominal concentration of alclometasone Dipropionate RS in Solution B
dipropionate in the Sample solution (mg/mL) Standard solution: 0.05 mg/mL of USP Alclometasone
Acceptance criteria: 90.0%110.0% Dipropionate RS obtained from a Standard stock solution and
Internal standard solution (1:1)
SPECIFIC TESTS Sample solution: Transfer a quantity of Ointment,
MICROBIAL ENUMERATION TESTS 61, and TESTS FOR SPECIFIED equivalent to 0.5 mg of alclometasone dipropionate, to a
MICROORGANISMS 62: Meets the requirements of the tests 50-mL centrifuge tube, add 10 mL of 2,2,4-
for absence of Staphylococcus aureus and Pseudomonas trimethylpentane, insert a stopper securely into the tube,
aeruginosa. and disperse the specimen using a vortex mixer. Add 5.0
MINIMUM FILL 755: Meets the requirements mL of Internal standard solution and 5.0 mL of Solution B,
insert the stopper securely, shake vigorously for 2 min, and
ADDITIONAL REQUIREMENTS centrifuge at 2500 rpm for 3 min. Remove the lower,
PACKAGING AND STORAGE: Preserve in collapsible tubes or aqueous alcohol phase, and transfer this Sample solution to a
tight containers, and store at a controlled room temperature. stoppered vial.
USP REFERENCE STANDARDS 11 Chromatographic system
USP Alclometasone Dipropionate RS (See Chromatography 621, System Suitability.)
Mode: LC
Detector: UV 254 nm
Alclometasone Dipropionate Ointment Column: 4-mm 30-cm; packing L1
(Comment on this Monograph)id=m1230=Alclometasone Injection size: 20 L
Dipropionate Ointment=A-Monos.pdf) System suitability
DEFINITION Sample: Standard solution
Alclometasone Dipropionate Ointment contains NLT 90.0% and [NOTEThe relative retention times for alclometasone
NMT 110.0% of the labeled amount of alclometasone dipropionate and betamethasone dipropionate are about
dipropionate (C28H37ClO7), in a suitable ointment base. 0.7 and 1.0, respectively.]
IDENTIFICATION Resolution: NLT 3.0 between the analyte and internal
A. The retention time of the major peak of the Sample standard peaks
solution corresponds to that of the Standard solution, both
USP 32 Official Monographs / Alcohol 71
Relative standard deviation: NMT 2% examined. Dilute 100 L of the solution to 10.0 mL with
Analysis Sample solution A.
Samples: Standard solution and Sample solution Standard solution C: Dilute 150 L of acetal to 50.0 mL
Calculate the quantity, as a percentage, of C28H37ClO7 in the with the substance to be examined. Dilute 100 L of the
portion of Ointment taken: solution to 10.0 mL with Sample solution A.
Standard solution D: Dilute 100 L of benzene to 100.0
Result = (RU/RS) (CS/CU) 100 mL with Sample solution A. Dilute 100 L of this solution to
50.0 mL with Sample solution A.
RU = peak height ratio from the Sample solution Chromatographic system
RS = peak height ratio from the Standard solution (See Chromatography 621, System Suitability.)
CS = concentration of USP Alclometasone Mode: GC
Dipropionate RS in the Standard solution Detector: Flame ionization
(mg/mL) Column: 0.32-mm 30-m fused silica capillary column
CU = nominal concentration of alclometasone bonded with a 1.8-m layer of phase G43
dipropionate in the Sample solution (mg/mL) Split ratio: 1:20
Acceptance criteria: 90.0%110.0% Temperature
Column:
SPECIFIC TESTS
MICROBIAL ENUMERATION TESTS 61, and TESTS FOR SPECIFIED
MICROORGANISMS 62: Meets the requirements of the tests Time Temperature
for absence of Staphylococcus aureus and Pseudomonas (min) ()
aeruginosa 0 40
MINIMUM FILL 755: Meets the requirements 12 40
ADDITIONAL REQUIREMENTS 32 240
PACKAGING AND STORAGE: Preserve in collapsible tubes or 42 240
tight containers, and store at a controlled room temperature.
USP REFERENCE STANDARDS 11 Detector: 280
USP Alclometasone Dipropionate RS Injection port: 200
Linear velocity: 35 cm/min
Carrier gas: Helium
Alcohol Injection size: 1.0 L
System suitability
(Comment on this Monograph)id=m1238=Alcohol=A- Sample: Standard solution B
Monos.pdf) Suitability requirements
Resolution: Between the first major peak (acetaldehyde)
and the second major peak (methanol) is NLT 1.5 in the
Standard solution B
Analysis
Samples: Standard solution A, Standard solution C,
Standard solution D, Sample solution A and Sample solution
B
C2H6O 46.07 Calculate the concentration of methanol in Sample solution
Ethanol; A: NMT half the area of the corresponding peak in the
Ethyl alcohol [64-17-5]. chromatogram obtained with Standard solution A (200
ppm).
DEFINITION Calculate the sum of the contents of acetaldehyde and
Alcohol contains NLT 92.3% and NMT 93.8%, by weight, acetal (ppm), expressed as acetaldehyde:
corresponding to NLT 94.9% and NMT 96.0%, by volume, at
15.56, of C2H5OH. Result (ppm) = (10 AE)/(AT AE) + (30 CE)/(CT CE)
IDENTIFICATION AE = area of the acetaldehyde peak obtained with

A. PROCEDURE: It meets the requirements of the test for Sample solution A
Specific Gravity 841. AT = area of the acetaldehyde peak obtained with
B. INFRARED ABSORPTION 197F or 197S: Neat Standard solution B
CE = area of the acetal peak obtained with Sample
IMPURITIES solution A
Inorganic Impurities CT = area of the acetal peak obtained with Standard
LIMIT OF NONVOLATILE RESIDUE: Evaporate 100 mL in a tared solution C: NMT 10 ppm, expressed as
dish on a water bath, and dry at 100 to 105 for 1 h: the acetaldehyde
weight of the residue is NMT 2.5 mg. Calculate the content of benzene (ppm):
Organic Impurities
PROCEDURE: VOLATILE IMPURITIES Result (ppm) = (2BE)/(BT BE)
Sample solution A: Substance to be examined
Sample solution B: Add 150 L of 4-methylpentan-2-ol to BE = area of the benzene peak obtained with Sample
500.0 mL of the substance to be examined. solution A
Standard solution A: Dilute 100 L of methanol to 50.0 BT = area of the benzene peak obtained with
mL with the substance to be examined. Dilute 5.0 mL of Standard solution D: NMT 2 ppm
the solution to 50.0 mL with Sample solution A. If necessary, the identity of benzene can be confirmed
Standard solution B: Dilute 50 L of methanol and 50 L using another suitable chromatographic system
of acetaldehyde to 50.0 mL with the substance to be (stationary phase with a different polarity).
72 Alcohol / Official Monographs USP 32
The total of all other impurities in the chromatogram ACIDITY OR ALKALINITY

obtained with Sample solution B: NMT area of the peak Phenolphthalein solution: 1 mg/mL of phenolphthalein in
due to 4-methylpentan-2-ol in the chromatogram a mixture of alcohol and water (4:1)
obtained with Sample solution B (300 ppm). Analysis: To 20 mL of alcohol, add 20 mL of freshly boiled
Disregard any peaks that are 0.03 times the area of the and cooled water and 0.1 mL of Phenolphthalein solution.
peak corresponding to 4-methylpentan-2-ol in the The solution is colorless. Add 1.0 mL of 0.01 N sodium
chromatogram obtained with Sample solution B (9 ppm). hydroxide. The solution is pink (30 ppm, expressed as acetic
acid).
SPECIFIC TESTS COLOR OF SOLUTION
SPECIFIC GRAVITY 841: 0.8120.816 at 15.56, indicating Standard stock solution: Ferric chloride CS, cobaltous
92.3%93.8%, by weight, or 94.9%96.0%, by volume, of chloride CS, cupric sulfate CS, and dilute hydrochloric acid
C2H5OH (3:3:2.4:1.6) (10 g/L)
ULTRAVIOLET ABSORPTION Standard solution: Transfer 1.0 mL of Standard stock
Analytical wavelength: 200 to 400 nm solution to a 100-mL volumetric flask, and dilute with dilute
Cell: 5 cm hydrochloric acid (10 g/L).
Acceptance criteria: Maximum absorbance 0.40 at 240 [NOTEPrepare the Standard solution immediately before
nm, 0.30 between 250 and 260 nm, and 0.10 between 270 use.]
and 340 nm. Examine between 235 and 340 nm, using Sample solution: Substance to be examined
water as the compensation liquid. The absorption curve is Analysis: Transfer a sufficient portion of the Sample solution
smooth. to a test tube of colorless, transparent, neutral glass with a
CLARITY OF SOLUTION flat base and an internal diameter of 15 to 25 mm to obtain
[NOTEThe Sample solution is to be compared to Reference a depth of 40 mm. Similarly transfer portions of Standard
suspension A and to water in diffused daylight 5 min after solution and water to separate, matching test tubes.
preparation of Reference suspension A.] Compare the Sample solution, Standard solution, and water
Hydrazine solution: 10 mg/mL of hydrazine sulfate in in diffused daylight, viewing vertically against a white
water [NOTEAllow to stand for 4 to 6 h.] background.
Methenamine solution: Transfer 2.5 g of methenamine to (See Spectrophotometry and Light-Scattering 851, Visual
a 100-mL glass-stoppered flask, add 25.0 mL of water, insert Comparison.)
the glass stopper, and mix to dissolve. Acceptance criteria: The Sample solution has the
Primary opalescent suspension: Transfer 25.0 mL of appearance of water or is not more intensely colored than
Hydrazine solution to the Methenamine solution in the 100- the Standard solution.
mL glass-stoppered flask. Mix, and allow to stand for 24 h.
[NOTEThis suspension is stable for 2 months, provided it is ADDITIONAL REQUIREMENTS
stored in a glass container free from surface defects. The PACKAGING AND STORAGE: Preserve in tight containers,
suspension must not adhere to the glass and must be well protected from light.
mixed before use.] USP REFERENCE STANDARDS 11
Opalescence standard: Transfer 15.0 mL of the Primary USP Alcohol RS
opalescent suspension to a 1000-mL volumetric flask, and
dilute with water to volume. [NOTEThis suspension should
not be used beyond 24 h after preparation.]
Reference suspension A: Transfer 5.0 mL of the Dehydrated Alcohol
Opalescence standard to a 100-mL volumetric flask, and (Comment on this Monograph)id=m1240=Dehydrated
dilute with water to volume. Alcohol=A-Monos.pdf)
Reference suspension B: Transfer 10.0 mL of the
Opalescence standard to a second 100-mL volumetric flask,
and dilute with water to volume.
Sample solution A: Substance to be examined
Sample solution B: Dilute 1.0 mL of Sample solution A to
20 mL with water, and allow to stand for 5 min before
testing.
Analysis: Transfer a sufficient portion of Sample solution A
and Sample solution B to separate test tubes of colorless, C2H6O 46.07
transparent, neutral glass with a flat base and an internal Ethanol;
diameter of 15 to 25 mm to obtain a depth of 40 mm. Ethyl alcohol [64-17-5].
Similarly transfer portions of Reference suspension A, Reference
suspension B, and water to separate matching test tubes. DEFINITION
Compare Sample solution A, Sample solution B, Reference Dehydrated Alcohol contains NLT 99.2%, by weight,
suspension A, Reference suspension B, and water in diffused corresponding to NLT 99.5%, by volume, at 15.56, of
daylight, viewing vertically against a black background. C2H5OH.
(See Spectrophotometry and Light-Scattering 851, Visual IDENTIFICATION
Comparison.) A. SPECIFIC GRAVITY 841: NMT 0.7962 at 15.56,
[NOTEThe diffusion of light must be such that Reference indicating NLT 99.2% of C2H5OH, by weight
suspension A can readily be distinguished from water, and B. INFRARED ABSORPTION 197S or 197F: Neat
that Reference suspension B can readily be distinguished
from Reference suspension A.] SPECIFIC TESTS
Acceptance criteria: Sample solution A and Sample solution ULTRAVIOLET ABSORPTION
B show the same clarity as that of water or their opalescence (See Spectroscopy and Light-Scattering 851
is not more pronounced than that of Reference suspension A.
Analytical wavelength: 235340 nm COLOR OF SOLUTION

Cell: 5 cm Standard stock solution: Ferric chloride CS, cobaltous
Blank: Water chloride CS, cupric sulfate CS, and dilute hydrochloric acid
Acceptance criteria (10 mg/mL) (3:3:2.4:1.6)
Absorbance: NMT 0.40 at 240 nm; NMT 0.30 between Standard solution: 1.0 mL of Standard stock solution, and
250 and 260 nm; and NMT 0.10 between 270 and 340 dilute to 100 mL with dilute hydrochloric acid (10 mg/mL)
nm [NOTEPrepare this immediately before use.]
[NOTEThe absorption curve is smooth.] Sample solution: Substance to be examined
LIMIT OF NONVOLATILE RESIDUE Analysis
Sample: 100 mL of Dehydrated Alcohol Samples: Sample solution,Standard solution, and water.
Analysis: Evaporate the sample in a tared dish on a water Transfer a sufficient portion of each Sample solution to
bath, and dry at 100105 for 1 h. individual test tubes of colorless, transparent, neutral glass
Acceptance criteria: The weight of the residue is NMT 2.5 with a flat base and an internal diameter of 1525 mm to
mg. obtain a depth of 40 mm. Compare the Samples in
CLARITY OF SOLUTION diffused daylight, viewing vertically against a white
[NOTEThe Sample solution is to be compared to Reference background (see Spectrophotometry and Light-Scattering
suspension A and to water in diffused daylight 5 min after 851, Visual Comparison).
preparation of Reference suspension A.] Acceptance criteria: The Sample solution has the
Hydrazine solution: 10 mg/mL of hydrazine sulfate appearance of water or is not more intensely colored than
[NOTEAllow to stand for 46 h.] the Standard solution.
Reference suspension: 2.5 g of Methenamine and add VOLATILE IMPURITIES
25.0 mL to a 100-mL glass-stoppered flask. Insert the glass- Sample solution A: Substance to be examined
stopper, and mix to dissolve. Add 25.0 mL of Hydrazine Sample solution B: 150 L of 4-methylpentan-2-ol to
solution, mix, and allow to stand for 24 h. [NOTEThis 500.0 mL of the substance to be examined
suspension is stable for 2 months, provided it is stored in a Standard solution A: 0.2 L/mL of methanol in Sample
glass container free from surface defects. The suspension solution A.
must not adhere to the glass and must be well mixed before Standard solution B: 0.01 L/mL of methanol and 0.01
use.] L/mL of acetaldehyde in Sample solution A.
Standard stock suspension: Dilute 15.0 mL of Reference Standard solution C: 0.03 L/mL of acetal in Sample
suspension to 1000 mL. [NOTEThis suspension should not solution A.
be used beyond 24 h after preparation.] Standard solution D: 2 nL/mL of benzene in Sample
Standard suspension A: Dilute 5.0 mL of Standard stock solution A.
suspension to 100 mL. Chromatographic system
Standard suspension B: Dilute 10.0 mL of Standard stock Mode: GC
suspension to 100 mL. Detector: Flame ionization
Sample solution A: Substance to be examined Column: 0.32-mm 30-m fused silica capillary column
Sample solution B: 1.0 mL of Sample solution A diluted to bonded with a 1.8-m layer of phase G43
20 mL Split ratio: 1:20
Blank: Water. Temperature: See the temperature program table below.
[NOTEAllow to stand for 5 min before testing.]
Analysis Time
Samples: Sample solution A, Sample solution B, Standard (min) Temperature
suspension A, Standard suspension B, and Blank. Transfer a
sufficient portion of Sample solution A and Sample solution B Injection port 200
to separate test tubes of colorless, transparent, neutral glass Detector port 280
with a flat base and an internal diameter of 1525 mm to Column 0 40
obtain a depth of 40 mm. Similarly transfer portions of
Standard suspension A, Standard suspension B, and Blank to 12 40
separate matching test tubes. Compare samples in diffused 32 240
daylight, viewing vertically against a black background (see 42 240
Spectrophotometry and Light-Scattering 851, Visual
Comparison). Flow rate: 35 cm/s
Acceptance criteria: Sample solution A and Sample solution Carrier gas: Helium
B show the same clarity as that of water, or their Injection size: 1 L
opalescence is not more pronounced than that of Standard System suitability
suspension A. Sample: Standard solution B
[NOTEThe diffusion of light must be such that Standard Suitability requirements
suspension A can readily be distinguished from water, and Resolution: NLT 1.5 between the first major peak
that Standard suspension B can readily be distinguished (acetaldehyde) and the second major peak (methanol)
from Standard suspension A.] Analysis 1
ACIDITY OR ALKALINITY Samples: Sample solution A, Sample solution B, Standard
Phenolphthalein solution: Dissolve 0.1 g of solution A, Standard solution B, Standard solution C, and
phenolphthalein in 80 mL alcohol, and dilute to 100 mL Standard Solution D
with water. Acceptance criteria 1: The area of the methanol peak
Analysis: To 20 mL of Dehydrated Alcohol add 20 mL of from Sample solution A is NMT half the area of the
freshly boiled and cooled water and 0.1 mL of corresponding peak from Standard solution A (200 ppm).
phenolphthalein solution. The solution is colorless. Add 1.0
mL of 0.01 N sodium hydroxide.
Acceptance criteria: The solution is pink (30 ppm,
expressed as acetic acid).
Analysis 2 IMPURITIES
Calculate the sum of the contents of acetaldehyde and Organic Impurities
acetal, expressed as acetaldehyde, using the following METHANOL
formula: Analysis: To 1 drop of sample add 1 drop of dilute
phosphoric acid (1 in 20) and 1 drop of 50 g/mL
Result = (10 AE)/(AT AE) + (30 CE)/(CT CE) potassium permanganate solution. Mix, allow to stand for
1 min, and add 50 g/mL sodium metabisulfite solution,
AE = area of the acetaldehyde peak from Sample dropwise, until the permanganate color is discharged. If a
solution A brown color remains, add 1 drop of dilute phosphoric acid
AT = area of the acetaldehyde peak from Standard (1 in 20). To the colorless solution, add 5 mL of freshly
solution B prepared chromotropic acid TS, and heat in a water bath
CE = area of the acetal peak from Sample solution A at 60 for 10 min.
CT = area of the acetal peak from Standard solution C Acceptance criteria: No violet color appears.
Acceptance criteria 2: NMT 10 ppm, expressed as ALDEHYDES AND OTHER FOREIGN ORGANIC SUBSTANCES
acetaldehyde [NOTEAll glassware used in this test should be thoroughly
Analysis 3 cleaned with hydrochloric acid, then rinsed with water and
Calculate the content of benzene using the following finally with a volume of dehydrated alcohol injection.]
formula: Sample: 20 mL
Analysis: Place the Sample in a glass-stoppered cylinder,
Result = (2BE)/(BT BE) cool the contents to approximately 15, and add, by pipet,
0.10 mL of 0.10 N potassium permanganate, noting
BE = area of the benzene peak from Sample solution A accurately the time of addition. Mix at once by inverting
BT = area of the benzene peak from Standard solution the stoppered cylinder, and allow it to stand at 15 for 5
D min.
If necessary, the identity of benzene can be confirmed using Acceptance criteria: The pink color does not entirely
another suitable chromatographic system (stationary phase disappear.
with a different polarity). LIMIT OF ACETONE AND ISOPROPYL ALCOHOL
Acceptance criteria 3: NMT 2 ppm Sample: 1.0 mL
General acceptance criteria: The sum of all other Solution A: 1 g/mL of furfural
impurities from Sample solution B is NMT the area of the Analysis: To the Sample add 1 mL of water, 1 mL of a
peak due to 4-methylpentan-2-ol from Sample solution B saturated solution of dibasic sodium phosphate, and 3 mL
(300 ppm). [NOTEDisregard any peaks that are 0.03 times of a saturated solution of potassium permanganate. Warm
the area of the peak corresponding to 4-methylpentan-2-ol the mixture to 4550, and allow to stand until the
fromSample solution B (9 ppm).] permanganate color is discharged. Add 3 mL of 2.5 N
ADDITIONAL REQUIREMENTS sodium hydroxide, and pass, without washing, through a
PACKAGING AND STORAGE: Preserve in tight containers, sintered-glass filter. Prepare a control containing 1 mL of
protected from light. the saturated solution of dibasic sodium phosphate, 3 mL
USP REFERENCE STANDARDS 11 of 2.5 N sodium hydroxide, and 80 g of acetone in 9 mL.
USP Dehydrated Alcohol RS To each solution, add 1 mL of Solution A, and allow to
stand for 10 min, then to 1.0 mL of each solution, add 3
mL of hydrochloric acid.
Acceptance criteria: Any pink color produced by the
Dehydrated Alcohol Injection Sample is not more intense than that produced by the
(Comment on this Monograph)id=m1250=Dehydrated Alcohol control.
Injection=A-Monos.pdf) SPECIFIC TESTS
DEFINITION SPECIFIC GRAVITY 841
Dehydrated Alcohol Injection is Dehydrated Alcohol suitable for Acceptance criteria: NMT 0.8035 at 15.56, indicating NLT
parenteral use. 96.8%, by weight, of C2H5OH
ACIDITY
IDENTIFICATION Sample: 50 mL
A. PROCEDURE Analysis: To the Sample, in a glass-stoppered flask, add 50
Sample: Injection mL of recently boiled water, phenolphthalein TS, and titrate
Analysis: Mix 5 drops of Sample in a small beaker with 1 with 0.020 N sodium hydroxide to a pink color that persists
mL of 1 /mL potassium permanganate and 5 drops of 2 N for 30 s.
sulfuric acid, and cover the beaker immediately with a filter Acceptance criteria: NMT 10.0 mL of 0.020 N sodium
paper moistened with a solution recently prepared by hydroxide is required.
dissolving 0.1 g of sodium nitroferricyanide and 0.25 g of AMYL ALCOHOL AND NONVOLATILE, CARBONIZABLE SUBSTANCES
piperazine in 5 mL of water. Sample: 25 mL
Acceptance criteria: An intense blue color is produced on Analysis: Allow the Sample to evaporate spontaneously from
the filter paper, the color becoming paler after a few a porcelain dish, carefully protected from dust, until the
minutes. surface of the dish is barely moist.
B. PROCEDURE Acceptance criteria: No red or brown color is produced
Sample solution: 0.1 mL/mL of Injection in water. immediately upon the addition of a few drops of sulfuric
Analysis: To 5 mL of the Sample solution, add 1 mL of 1.0 N acid.
sodium hydroxide, then slowly (over a period of 3 min) add LIMIT OF NONVOLATILE RESIDUE
2 mL of 0.1 N iodine. Sample: 40 mL
Acceptance criteria: The odor of iodoform develops, and a Analysis: Evaporate the Sample in a tared dish on a water
yellow precipitate is formed within 30 min. bath, and dry at 105 for 1 h.
Acceptance criteria: The weight of the residue does not Mode: UV

exceed 1 mg. Analytical Wavelength: 410 nm
WATER-INSOLUBLE SUBSTANCES Cell: 1 cm
Sample: Injection Analysis:
Analysis: Dilute a portion of Sample with an equal volume Samples: Standard solution, Sample solution, and Solution A.
of water. Transfer 10.0 mL each of the samples to individual 250-mL
Acceptance criteria: The mixture is clear and remains clear separators, add 40 mL of Solution A, 10 mL of Solution B,
for 30 min after cooling to 10. and 60 mL of chloroform. Shake the separators vigorously
ULTRAVIOLET ABSORBANCE for 2 min, allow to stand for 15 min, then withdraw the
(See Spectroscopy and Light-Scattering 851.) chloroform layers through chloroform-washed cotton into
Analytical wavelength: 235340 nm 100-mL volumetric flasks. Repeat the extraction with 20
Cell: 1 cm mL of chloroform, adding the filtered chloroform extracts
Blank: Water to the respective volumetric flasks, and dilute with
Acceptance criteria: Absorbance is NMT 0.08 at 240 nm; chloroform to volume. Without delay, concomitantly
and NMT 0.02 between 270 and 340 nm. [NOTEThe curve determine the absorbances of the solutions. [NOTEUse
through these points is smooth.] the solution prepared from Solution A as the Blank.]
OTHER REQUIREMENTS: Meets the requirements for Injections Calculate the quantity, in mg, of denatonium benzoate
1 (C28H34N2O3 H2O) in 100 mL of Rubbing Alcohol taken:
ADDITIONAL REQUIREMENTS Result = (AU/AS) CS D
PACKAGING AND STORAGE: Preserve in tight, single-dose
containers, preferably of Type I glass, and store at controlled AU = absorbance of the Sample solution
room temperature. The container may contain an inert gas AS = absorbance of the Standard solution
in the headspace. CS = concentration of USP Denatonium Benzoate RS
in the Standard solution (mg/mL)
D = sample dilution factor, 25
Acceptance criteria: NLT 1.40 mg of denatonium
Rubbing Alcohol benzoate/100 mL of Rubbing Alcohol
(Comment on this Monograph)id=m1270=Rubbing Alcohol=A- SUCROSE OCTAACETATE
Monos.pdf) Sample solution: Using 50 mL of 70% alcohol, transfer the
residue obtained in the test for Limit of Nonvolatile Residue to
DEFINITION a 500-mL conical flask.
Rubbing Alcohol and all preparations under the classification of Analysis: Neutralize the Sample solution with 0.1 N sodium
Rubbing Alcohols are manufactured in accordance with the hydroxide VS, using phenolphthalein TS as the indicator.
requirements of the U.S. Treasury Department, Bureau of Add 25.0 mL of 0.1 N sodium hydroxide, attach an air
Alcohol, Tobacco, and Firearms, Formula 23-H (8 parts by condenser to the flask, and reflux on a steam bath for 1 h.
volume of acetone, 1.5 parts by volume of methyl isobutyl Remove from the steam bath, cool quickly, and titrate the
ketone, and 100 parts by volume of ethyl alcohol) being used. excess alkali with 0.1 N sulfuric acid VS, using
It contains NLT 68.5% and NMT 71.5% by volume of phenolphthalein TS as the indicator. Perform a blank
dehydrated alcohol, the remainder consisting of water and the determination (see Titrimetry 541, Residual Titrations). Each
denaturants, with or without color additives, and perfume oils. mL of 0.1 N sodium hydroxide is equivalent to 8.483 mg of
Rubbing Alcohol contains, in each 100 mL, NLT 355 mg of sucrose octaacetate (C28H38O19).
sucrose octaacetate or NLT 1.40 mg of denatonium benzoate. Acceptance criteria: NLT 335 mg of sucrose
The preparation may be colored with one or more color octaacetate/100 mL of Rubbing Alcohol
additives, listed by the FDA for use in drugs. A suitable
stabilizer may be added. Rubbing Alcohol complies with the IMPURITIES
requirements of the U.S. Treasury Department, Bureau of Organic Impurities
Alcohol, Tobacco, and Firearms. PROCEDURE: METHANOL
[NOTERubbing Alcohol is packaged, labeled, and sold in Sample solution: 0.50 mL of sample diluted to 1.0 mL
accordance with the regulations issued by the U.S. Treasury Analysis: To 0.50 mL of the Sample solution add 1 drop of
Department, Bureau of Alcohol, Tobacco, and Firearms.] dilute phosphoric acid (1 in 20) and 1 drop of 50 mg/mL
potassium permanganate solution. Mix, allow to stand for
ASSAY 1 min, and add 50 mg/mL of sodium metabisulfite
PROCEDURE solution, dropwise, until the permanganate color is
[NOTEA sample must meet either the requirement for the discharged. If a brown color remains, add 1 drop of dilute
Assay for Denatonium Benzoate or the requirement for the phosphoric acid (1 in 20). To the colorless solution add 5
Assay for Sucrose Octaacetate.] mL of freshly prepared chromotropic acid TS, and heat in a
DENATONIUM BENZOATE water bath at 60 for 10 min.
Solution A: 9.23 mg/mL of anhydrous dibasic sodium Acceptance criteria: No violet color appears.
phosphate in water. Adjust with saturated citric acid solution
to a pH of 4 0.1 before final dilution. SPECIFIC TESTS
Solution B: Bromophenol blue in chloroform (1 in 1000) SPECIFIC GRAVITY 841
Standard solution: 50 g/mL of USP Denatonium Benzoate Acceptance criteria: 0.86910.8771 at 15.56 (the U.S.
RS Government standard temperature for alcohol
Sample solution: Dissolve the residue obtained in the test determination) for Rubbing Alcohol manufactured with
for Limit of Nonvolatile Residue in 50.0 mL of water. specially denatured alcohol Formula 23-H
(See Spectoscopy and Light-Scattering 851.)
LIMIT OF NONVOLATILE RESIDUE A = length of the polarimeter tube (mm)

Where the denaturant is denatonium benzoate R = observed rotation (degrees)
Sample: 200.0 mL Acceptance criteria: 95.0%105.0%
Analysis: Evaporate the Sample, transferred in convenient
portions, in a suitable tared dish on a steam bath, and dry IMPURITIES
the residue at 105 for 1 h. Inorganic Impurities
Acceptance criteria: The weight of the residue is not less HEAVY METALS 231
than 2.8 mg. (Retain the residue for the Assay for Sample: Equivalent to 4.0 g of dextrose adjusted to 25 mL
Denatonium Benzoate.) by evaporation or by addition of water, as necessary.
Where the denaturant is sucrose octaacetate Acceptance criteria: NMT 5 C ppm, where C is the
Sample: 25.0 mL labeled amount, in g, of C6H12O6 H2O per mL of Injection
Analysis: Evaporate the Sample in a suitable tared dish on Organic Impurities
a steam bath, and dry the residue at 105 for 1 h. PROCEDURE: LIMIT OF 5-HYDROXYMETHYLFURFURAL AND RELATED
Acceptance criteria: The weight of the residue is NLT 89 SUBSTANCES
mg. (Retain the residue for the Assay for Sucrose Sample solution: 2 mg/mL of dextrose
Octaacetate.) Spectrometric conditions
(See Spectroscopy and Light-scattering 851.)
ADDITIONAL REQUIREMENTS Mode: UV
PACKAGING AND STORAGE: Preserve in tight containers, Analytical wavelength: 284 nm
remote from fire, and store at controlled room temperature. Cell: 1 cm
LABELING: Label it to indicate that it is flammable. Blank: Water
USP REFERENCE STANDARDS 11 Analysis:
USP Denatonium Benzoate RS Sample: Sample solution
Acceptance criteria: The absorbance is NMT 0.25.
SPECIFIC TESTS
Alcohol in Dextrose Injection PH 791: 3.56.5
(Comment on this Monograph)id=m1320=Alcohol in Dextrose Sample solution: A portion of Dextrose injection to which
Injection=A-Monos.pdf) 0.30 mL of saturated potassium chloride solution has been
added for each 100 mL and which previously has been
DEFINITION diluted with water, if necessary, to a concentration of NMT
Alcohol in Dextrose Injection is a sterile solution of Alcohol and 5% of dextrose
Dextrose in Water for Injection. It contains NLT 90.0% and BACTERIAL ENDOTOXINS TEST 85: NMT 0.5 USP Endotoxin
NMT 110.0% of the labeled amount of alcohol (C2H5OH), and unit/mL
NLT 95.0% and NMT 105.0% of the labeled amount of OTHER REQUIREMENTS: Meets the requirements under
dextrose (C6H12O6 H2O). Injections 1
A. PROCEDURE PACKAGING AND STORAGE: Preserve in tight, single-dose
Sample solution: 50 L/mL of Dextrose Injection in water containers, preferably of Type I or Type II glass, and store at
Analysis: Add a few drops of the Sample solution to 5 mL of a controlled room temperature.
hot alkaline cupric tartrate TS. LABELING: The label states the total osmolarity of the
Acceptance criteria: A copious red precipitate of cuprous solution expressed in mOsmol/L.
oxide is formed. USP REFERENCE STANDARDS 11
B. SPECIFIC GRAVITY 841: NMT 0.7962 at 15.56, USP Dehydrated Alcohol RS
indicating NLT 99.2% of C2H5OH by weight USP Endotoxin RS
C. INFRARED ABSORPTION 197S or 197F: Meets the
requirements
ASSAY Alendronate Sodium
ALCOHOL DETERMINATION, Method 1Distillation Method 611 (Comment on this Monograph)id=m1324=Alendronate
Sample: 50.0 mL Sodium=A-Monos.pdf)
DEXTROSE
Sample: Amount equivalent to 25 g of dextrose
Analysis: Transfer the Sample to a 100-mL volumetric flask.
Add 0.2 mL of 6 N ammonium hydroxide, dilute with water
to volume, and mix. Determine the angular rotation in a
suitable polarimeter tube (see Optical Rotation 781).
Calculate the percentage (g/100 mL) of C6H12O6 H2O
taken: C4H12NNaO7P2 3H2O 325.12
Phosphonic acid, (4-amino-1-hydroxybutylidene) bis-,
Result = [(Mr1/Mr2)/Rmid] 100/A R 100 monosodium salt, trihydrate;
Sodium trihydrogen (4-amino-1-
Mr1 = molecular weight of dextrose monohydrate, hydroxybutylidene)diphosphonate, trihydrate [121268-17-5].
198.17
Mr2 = molecular weight of anhydrous dextrose, 180.16 DEFINITION
Rmid = midpoint of the specific rotation range for Alendronate Sodium contains NLT 98.0% and NMT 102.0% of
anhydrous dextrose (degrees), 52.9 C4H12NNaO7P2, calculated on the dried basis.
USP 32 Official Monographs / Alendronate 77

A. INFRARED ABSORPTION 197M
B. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the IMPURITIES
requirements of the flame test Inorganic Impurities
HEAVY METALS, Method III 231: NMT 10 ppm
ASSAY Organic Impurities
PROCEDURE PROCEDURE
Solution A: 14.7 mg/mL of sodium citrate dihydrate and Diluent: Prepare as directed in the Assay.
7.05 mg/mL of anhydrous dibasic sodium phosphate Solution A: 2.94 mg/mL of sodium citrate dihydrate and
[NOTEAdjust solution to a pH of 8 with phosphoric acid.] 1.42 mg/mL of anhydrous dibasic sodium phosphate
Solution B: 19.1 mg/mL of sodium borate [NOTEAdjust solution to a pH of 8 with phosphoric acid,
Solution C: 0.5 mg/mL of 9-fluorenylmethyl chloroformate and then pass through a filter having a 0.5-m or finer
in acetonitrile porosity.]
[NOTEPrepare this solution fresh just before use.] Solution B: Prepare as directed in the Assay.
Mobile phase: Acetonitrile, methanol, and Solution A Solution C: 4 mg/mL of 9-fluorenylmethyl chloroformate
(25:5:70) in acetonitrile
Diluent: 29.4 mg/mL of sodium citrate dihydrate [NOTEPrepare this solution fresh just before use.]
Standard stock solution: 0.1 mg/mL of USP Alendronate Solution D: Acetonitrile and Solution A (3:17)
Sodium RS in Diluent Solution E: Acetonitrile and Solution A (7:3)
Standard solution: Transfer 5.0 mL of the Standard stock Mobile phase: See the gradient table below.
solution to a 50-mL polypropylene, screw-cap centrifuge
tube containing 5 mL of Solution B. Add 5 mL of Solution C, Time Solution D Solution E
and shake for 30 s. Allow to stand at room temperature for (min) (%) (%)
25 min. Add 25 mL of methylene chloride, and shake
vigorously for 1 min. Centrifuge for 510 min. Use a portion 0 100 0
of the clear upper aqueous layer. 15 50 50
Reagent blank: Transfer 5.0 mL of Diluent to a 50-mL 25 0 100
polypropylene, screw-cap centrifuge tube containing 5 mL
of Solution B. Add 5 mL of Solution C, and shake for 30 s. 27 100 0
Allow to stand at room temperature for 25 min. Add 25 mL 32 100 0
of methylene chloride, and shake vigorously for 1 min.
Centrifuge for 510 min. Use a portion of the clear upper Standard stock solution: 0.6 mg/mL of USP Alendronate
aqueous layer. Sodium RS in Diluent
Sample stock solution: 0.1 mg/mL of Alendronate Sodium Standard solution A: Transfer 5.0 mL of the Standard
in Diluent stock solution to a 50-mL polypropylene, screw-cap
Sample solution: Transfer 5.0 mL of the Sample stock centrifuge tube containing 5 mL of Solution B. Add 5 mL of
solution to a 50-mL polypropylene, screw-cap centrifuge acetonitrile and 5 mL of Solution C, and shake for 45 s.
tube containing 5 mL of Solution B. Add 5 mL of Solution C, Allow to stand at room temperature for 30 min. Add 20
and shake for 30 s. Allow to stand at room temperature for mL of methylene chloride, and shake vigorously for 1 min.
25 min. Add 25 mL of methylene chloride, and shake Centrifuge for 5 to 10 min, and use a portion of the clear
vigorously for 1 min. Centrifuge for 510 min. Use a portion upper aqueous layer.
of the clear upper aqueous layer. Standard solution B: Dilute the Standard stock solution to
Chromatographic system 0.6 g/mL with Diluent. Transfer 5 mL to a 50-mL
(See Chromatography 621, System Suitability.) polypropylene, screw-cap centrifuge tube containing 5 mL
Mode: LC of Solution B. Add 5 mL of acetonitrile and 5 mL of Solution
Detector: UV 266 nm C, and shake for 45 s. Allow to stand at room temperature
Column: 4.1-mm 25-cm; packing L21 for 30 min. Add 20 mL of methylene chloride, and shake
Column temperature: 35 vigorously for 1 min. Centrifuge for 5 to 10 min, and use a
Flow rate: 1.2 mL/min portion of the clear upper aqueous layer.
Injection size: 10 L Blank: Transfer 5.0-mL of Diluent to a 50-mL
System suitability polypropylene, screw-cap centrifuge tube containing 5 mL
Sample: Standard solution of Solution B. Add 5 mL of acetonitrile and 5 mL of Solution
Suitability requirements C, and shake for 45 s. Allow to stand at room temperature
Column efficiency: NLT 1500 theoretical plates for 30 min. Add 20 mL of methylene chloride, and shake
Tailing factor: NMT 1.5 vigorously for 1 min. Centrifuge for 5 to 10 min, and use a
Relative standard deviation: NMT 2.0% for replicate portion of the clear upper aqueous layer.
injections Sample stock solution: 0.6 mg/mL of Alendronate
Analysis Sodium in Diluent
Samples: Standard solution, Sample solution, and Reagent Sample solution: Transfer 5.0-mL of Sample stock solution
blank to a 50-mL polypropylene, screw-cap centrifuge tube
Calculate the percentage of C4H12NNaO7P2 in the portion containing 5 mL of Solution B. Add 5 mL of acetonitrile and
of Alendronate Sodium taken: 5 mL of Solution C, and shake for 45 s. Allow to stand at
room temperature for 30 min. Add 20 mL of methylene
Result = (rU/rS) (CS/CU) 100 chloride, and shake vigorously for 1 min. Centrifuge for 5
to 10 min, and use a portion of the clear upper aqueous
rU = peak area from the Sample solution layer.
rS = peak area from the Standard solution Chromatographic system
CS = concentration of anhydrous sodium alendronate (See Chromatography 621, System Suitability.)
in the Standard stock solution
(mg/mL)
CU = concentration of the Sample stock solution
(mg/mL)
78 Alendronate / Official Monographs USP 32
Mode: LC Diluent: 29.4 mg/mL of sodium citrate dihydrate

Detector: UV 266 nm Standard stock solution: 0.03 mg/mL of anhydrous USP
Column: 4.1-mm 25-cm; packing L21 Alendronate Sodium RS in Diluent
Column temperature: 45 Standard solution: Transfer 5.0 mL of the Standard stock
Flow rate: 1.8 mL/min solution to a 50-mL polypropylene screw-cap centrifuge tube
Injection size: 20 L containing 5 mL of Solution B, and mix for 3 min. Add 4 mL
System suitability of Solution C, and agitate for 30 s. Allow the solution to
Samples: Standard solution A and Standard solution B stand at room temperature for 25 min. Add 25 mL of
Suitability requirements methylene chloride, and agitate for 40 s. Centrifuge the
Tailing factor: NMT 2.0 for the main peak, Standard mixture for 10 min. Use the clear upper aqueous layer.
solution A Sample stock solution: Transfer NLT 10 Tablets to a 1000-
Signal-to-noise ratio: NLT 3 for Standard solution B for mL volumetric flask. Add 500 mL of Diluent, shake by
the peak at the locus corresponding to the main peak of mechanical means for 30 min, and sonicate for 5 min.
Standard solution A Dilute with Diluent to volume, and centrifuge a portion of
Analysis this solution. Quantitatively dilute a portion of the clear
Samples: Sample solution and Blank supernatant to a concentration equivalent to 0.020.03
[NOTEDisregard any peak corresponding to those mg/mL of alendronate sodium.
obtained from the Blank.] Sample solution: Transfer 5.0 mL of the Sample stock
Calculate the percentage of each impurity in the portion solution to a 50-mL polypropylene screw-cap centrifuge tube
of Alendronate Sodium taken: containing 5 mL of Solution B, and mix for 3 min. Add 4 mL
of Solution C, and agitate for 30 s. Allow the solution to
Result = (rU/rT) 100 stand at room temperature for 25 min. Add 25 mL of
methylene chloride, and agitate for 40 s. Centrifuge the
rU = area of each impurity peak mixture for 10 min. Use the clear upper aqueous layer.
rT = sum of all impurity peaks and the main peak Blank: Transfer 5 mL of Diluent to a 50-mL polypropylene
Acceptance criteria screw-cap centrifuge tube containing 5 mL of Solution B,
Individual impurities: NMT 0.1% and mix for 3 min. Add 4 mL of Solution C, and agitate for
Total impurities: NMT 0.5% 30 s. Allow the solution to stand at room temperature for
25 min. Add 25 mL of methylene chloride, and agitate for
SPECIFIC TESTS 40 s. Centrifuge the mixture for 10 min. Use the clear upper
LOSS ON DRYING 731: Dry it at a pressure of NMT 5 mm of aqueous layer.
mercury at 140 to constant weight: it loses NLT 16.1% and Chromatographic system
NMT 17.1% of its weight. (See Chromatography 621, System Suitability.)
ADDITIONAL REQUIREMENTS Mode: LC
PACKAGING AND STORAGE: Preserve in well-closed containers, Detector: UV 266 nm
and store at room temperature. Column: 4.1-mm 25-cm; packing L21
USP REFERENCE STANDARDS 11 Column temperature: 35
USP Alendronate Sodium RS Flow rate: 1 mL/min
System suitability
Alendronate Sodium Tablets Suitability requirements
(Comment on this Monograph)id=m1327=Alendronate Sodium Capacity factor: NLT 2.0
Tablets=A-Monos.pdf) Relative standard deviation: NMT 2.0% for replicate
injections
DEFINITION Analysis
Alendronate Sodium Tablets contain an amount of Alendronate Samples: Standard solution, Sample solution, and Blank
Sodium equivalent to NLT 90.0% and NMT 110.0% of the Calculate the percentage of the label claim in the portion
labeled amount of alendronic acid (C4H13NO7P2). of C4H13NO7P2 taken:
IDENTIFICATION Result = (rU/rS) (CS/CU) F 100/L

corresponds to that of the Standard solution, as obtained in rU = peak area from the Sample solution
the Assay. rS = peak area from the Standard solution
CS = concentration of anhydrous USP Alendronate
ASSAY Sodium RS in the Standard stock solution
PROCEDURE (mg/mL)
Solution A: 14.7 mg/mL of sodium citrate dihydrate and CU = concentration of the Tablet in the Sample stock
7.05 mg/mL of anhydrous dibasic sodium phosphate solution (Tablets/mL)
[NOTEAdjust pH to 8.0 with phosphoric acid before F = molecular weight conversion factor
bringing the solution to volume.] (C4H13NO7P2/C4H12NNaO7P2), 0.919
Solution B: 38.1 mg/mL of sodium borate L = label claim (mg/Tablet)
Solution C: 1 mg/mL of 9-fluorenylmethyl chloroformate Acceptance criteria: C4H12NNaO7P2 equivalent to
in acetonitrile 90.0%110.0% of the labeled amount of C4H13NO7P2
[NOTEPrepare this solution fresh just before use.]
Mobile phase: Acetonitrile, methanol, and Solution A
(20:5:75)
USP 32 Official Monographs / Alfentanil 79
PERFORMANCE TESTS Test 2

If the product complies with this test, the labeling indicates
that the product meets USP Dissolution Test 2.
Change to read: Medium: Water; 900 mL
Apparatus 2: 50 rpm
DISSOLUTION 711 Time: 30 min
Test 13 Determine the amount of C4H12NNaO7P2 3H2O dissolved as
Medium: Water; 900 mL Tolerances: NLT 80% (Q) of the labeled amount of
Apparatus 2: 50 rpm alendronate sodium (C4H12NNaO7P2 3H2O) is dissolved.3
Time: 15 min
Determine the amount of C4H13NO7P2 dissolved by using the UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
following method. ADDITIONAL REQUIREMENTS
Solution A and Mobile Phase: Proceed as directed in the PACKAGING AND STORAGE: Preserve in tight containers. Store
Assay. between 15 and 30.
Diluent: 176.4 mg/mL of sodium citrate in Medium
Solution B: Dissolve 6.2 g of boric acid in approximately
950 mL of water. Adjust with 1 N sodium hydroxide to a Change to read:
pH of 9.0, and dilute with water to 1 L.
Solution C: 0.5 mg/mL of 9-fluorenylmethyl chloroformate
in acetonitrile LABELING: The labeling indicates weekly dosing where
[NOTEPrepare this solution fresh.] appropriate. When more than one Dissolution test is given, the
Standard stock solution: USP Alendronate Sodium RS in labeling states the test used only if Test 1 is not used.3
Medium to make a concentration equivalent to dissolving 1 USP REFERENCE STANDARDS 11
Tablet of sample in 900 mL of the same Medium USP Alendronate Sodium RS
Standard solution: Transfer 5.0 mL of the Standard stock
solution to a 50-mL polypropylene screw-cap centrifuge
tube containing 1.0 mL of Diluent and 5.0 mL of Solution B,
and mix for 3 min. Add 4.0 mL of Solution C, and agitate Alfentanil Hydrochloride
for 30 s. Allow the solution to stand at room temperature (Comment on this Monograph)id=m1350=Alfentanil
for 25 min. Add 25 mL of methylene chloride, and agitate Hydrochloride=A-Monos.pdf)
for 40 s. Centrifuge the mixture for 5 min. Use a portion of
the clear, upper aqueous layer.
Blank: Transfer 5 mL of water to a 50-mL polypropylene
screw-cap centrifuge tube containing 1.0 mL of Diluent and
5.0 mL of Solution B, and mix for 3 min. Add 4.0 mL of
Solution C, and agitate for 30 s. Allow the solution to stand
at room temperature for 25 min. Add 25 mL of methylene
chloride, and agitate for 40 s. Centrifuge the mixture for 5
min. Use a portion of the clear, upper aqueous layer. C21H32N6O3 HCl H2O 470.99
Sample stock solution: Withdraw a portion of the solution C21H32N6O3 HCI 452.98
under test, centrifuge immediately, and use a portion of Propanamide, N-[1-[2-(4-ethyl-4,5-dihydro-5-oxo-1H-tetrazol-1-
the clear upper supernatant. yl)ethyl]-4-(methoxymethyl)-4-piperidinyl]-N-phenyl,
Sample solution: Transfer 5.0 mL of the Sample stock monohydrochloride, monohydrate;
solution to a 50-mL polypropylene screw-cap centrifuge N-[1-[2-(4-Ethyl-5-oxo-2-tetrazolin-1-yl)-ethyl]-4-
tube containing 1.0 mL of Diluent and 5.0 mL of Solution B, (methoxymethyl)-4-piperidyl]propionanilide
and mix for 3 min. Add 4.0 mL of Solution C, and agitate monohydrochloride monohydrate [70879-28-6].
for 30 s. Allow the solution to stand at room temperature Anhydrous [69049-06-5].
for 25 min. Add 25 mL of methylene chloride, and agitate
for 40 s. Centrifuge the mixture for 5 min. Use a portion of DEFINITION
the clear, upper aqueous layer. Alfentanil Hydrochloride contains NLT 98.0% and NMT 102.0%
Chromatographic system and System suitabilitity: of C21H32N6O3 HCl, calculated on the anhydrous basis.
Proceed as directed in the Assay. [CAUTIONHandle Alfentanil Hydrochloride with great care
Analysis because it is a potent opioid analgesic. Great care should be
Samples: Standard solution and Sample solution taken to prevent inhaling particles of Alfentanil Hydrochloride
Calculate the percentage of C4H13NO7P2 dissolved: and exposing the skin to it.]
Result = (rU/rS) CS F V 100/L IDENTIFICATION
INFRARED ABSORPTION 197K
rU = peak area from the Sample solution
rS = peak area from the Standard solution ASSAY
CS = concentration of USP Alendronate Sodium RS in PROCEDURE
the Standard solution (mg/mL) Sample solution: 11.7 mg/mL of Alfentanil Hydrochloride
F = molecular weight conversion factor in glacial acetic acid.
(C4H13NO7P2/C4H12NNaO7P2), 0.919 Analysis: To 30 mL of Sample solution add 3 mL of mercuric
V = volume of the Medium, 900 mL acetate TS and 3 drops of p-naphtholbenzein TS, and titrate
L = Tablet label claim (mg) with 0.1 N perchloric acid VS to a green endpoint. Perform
Tolerances: NLT 80% (Q) of the labeled amount of a blank determination, and make any necessary correction.
C4H13NO7P2 is dissolved; for tablets labeled for weekly Each mL of 0.1 N perchloric acid is equivalent to 45.298 mg
dosing, NLT 75% (Q) of the labeled amount of C4H13NO7P2 of C21H32N6O3 HCl.
is dissolved.
80 Alfentanil / Official Monographs USP 32
Acceptance criteria: 98.0%102.0% Visualizing agent: Dragendorffs reagent

Analysis
IMPURITIES Samples: Standard solution and Sample solution
Inorganic Impurities Proceed as directed in the chapter.
RESIDUE ON IGNITION 281: NMT 0.1% Acceptance criteria: Meets the requirements
Organic Impurities B. The retention time of the major peak from the Sample
PROCEDURE solution corresponds to that from the Standard solution, as
Mobile phase: Acetonitrile and 0.01 M obtained in the Assay.
tetrabutylammonium hydrogen sulfate (86:14)
Standard solution: 0.54 mg/mL of USP Alfentanil ASSAY
Hydrochloride RS in Mobile phase PROCEDURE
Sample solution: 0.54 mg/mL of Alfentanil Hydrochloride Mobile phase: Acetonitrile and 0.01 M
in Mobile phase tetrabutylammonium hydrogen sulfate (86:14)
Chromatographic system Standard solution: 0.54 mg/mL of USP Alfentanil
(See Chromatography 621, System Suitability.) Hydrochloride RS in saline TS
Mode: LC Sample solution: 0.50 mg/mL of Alfentanil from volume of
Detector: UV 235 nm Injection in saline TS
Column: 4.6-mm 25-cm; contains spherical 5-m Chromatographic system
packing L1 (See Chromatography 621, System Suitability.)
Flow rate: 2 mL/min Mode: LC
Injection size: 25 L Detector: UV 235 nm
System suitability Column: 4.6-mm 25-cm; contains spherical 5-m
Sample: Standard solution packing L1
Suitability requirements Flow rate: 2 mL/min
Column efficiency: NLT 5400 theoretical plates Injection size: 25 L
Tailing factor: NMT 1.3 System suitability
Relative standard deviation: NMT 1.0% for replicate Sample: Standard solution
injections Suitability requirements
Analysis Column efficiency: NLT 5400 theoretical plates
Sample: Sample solution Tailing factor: NMT 1.3
Calculate the percentage of each impurity in the portion Relative standard deviation: NMT 1.0% for replicate
of Alfentanil Hydrochloride taken: injections
Analysis
Result = (rU/rT) 100 Samples: Standard solution and Sample solution
Calculate the percentage of C21H32N6O3 in each mL of the
rU = response of each impurity peak Injection taken:
rT = sum of all of the peaks
Acceptance criteria Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
Any single impurity: NMT 0.5%
Total impurities: NMT 1.0% rU = peak response from the Sample solution
SPECIFIC TESTS CS = concentration of USP Alfentanil Hydrochloride RS
WATER DETERMINATION, Method I 921: NMT 4.0% in the Standard solution (mg/mL)
CU = concentration of alfentanil in the Sample solution
ADDITIONAL REQUIREMENTS (mg/mL)
PACKAGING AND STORAGE: Preserve in tight containers, and Mr1 = molecular weight for alfentanil, 416.52
store at a controlled room temperature. Mr2 = molecular weight for alfentanil hydrochloride,
USP REFERENCE STANDARDS 11 452.98
USP Alfentanil Hydrochloride RS Acceptance criteria: 90.0%110.0%
IMPURITIES
Alfentanil Injection Organic Impurities
PROCEDURE
(Comment on this Monograph)id=m1355=Alfentanil Mobile phase: Acetonitrile and 0.01 M
Injection=A-Monos.pdf) tetrabutylammonium hydrogen sulfate (86:14)
Standard solution: 0.54 mg/mL of USP Alfentanil
DEFINITION Hydrochloride RS in saline TS
Alfentanil Injection is a sterile solution of Alfentanil Sample solution: 0.50 mg/mL of Alfentanil from volume
Hydrochloride in Water for Injection. It contains an amount of of Injection in saline TS
Alfentanil Hydrochloride equivalent to NLT 90.0% and NMT Chromatographic system
110.0% of the labeled amount of C21H32N6O3. (See Chromatography 621, System Suitability.)
[CAUTIONHandle Alfentanil Injection with great care because it Mode: LC
is a potent opioid analgesic.] Detector: UV 235 nm
IDENTIFICATION Column: 4.6-mm 25-cm; contains spherical 5-m
A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 packing L1
Standard solution: 0.54 mg/mL of USP Alfentanil
Hydrochloride RS
Sample solution: 0.5 mg/mL of alfentanil in water
Application volume: 200 L
Developing solvent system: Chloroform, methanol, and
formic acid (85:10:5)
USP 32 Official Monographs / Alfuzosin 81
Flow rate: 2 mL/min acid. Perform a blank determination, and make any
Injection size: 25 L necessary correction (see Titrimetry 541). Each mL of 0.1 M
System suitability perchloric acid titrant is equivalent to 42.59 mg of
Sample: Standard solution C19H27N5O4 HCl.
Column efficiency: NLT 5400 theoretical plates IMPURITIES
Tailing factor: NMT 1.3 Inorganic Impurities
Relative standard deviation: NMT 1.0% for replicate RESIDUE ON IGNITION 281: NMT 0.1%
injections Organic Impurities
Analysis PROCEDURE
Sample: Sample solution Solution A: 58.5 mM perchloric acid. Adjust with 2 M
Calculate the percentage of each impurity in the portion sodium hydroxide to a pH of 3.5 before final dilution.
of Injection taken: Mobile phase: Acetonitrile, tetrahydrofuran, and Solution A
(20:1:80)
Result = (rU/rT) 100 System suitability solution: 0.4 mg /mL of USP Alfuzosin
Hydrochloride System Suitability Mixture RS in Mobile phase
rU = response of each impurity peak Sample solution A: 0.40 mg/mL of Alfuzosin
rT = sum of all of the peaks Hydrochloride in Mobile phase
Acceptance criteria: The sum of all impurities is NMT Sample solution B: 0.40 g/mL of Alfuzosin
2.0%. Hydrochloride, from Sample solution A, in Mobile phase
SPECIFIC TESTS (See Chromatography 621, System Suitability.)
PH 791: 4.06.0 Mode: LC
PARTICULATE MATTER IN INJECTIONS 788: Meets the Detector: UV 254 nm
requirements for small-volume injections Column: 4.6-mm 15-cm; 5 m packing L1
BACTERIAL ENDOTOXINS TEST 85: NMT 10 USP Endotoxin Flow rate: 1.5 mL/min
Units/mL Injection size: 10 L
OTHER REQUIREMENTS: Meets the requirements under System suitability
Injections 1. Sample: System suitability solution
ADDITIONAL REQUIREMENTS Peak-to-valley ratio: NLT 5 for impurity A and alfuzosin
PACKAGING AND STORAGE: Preserve in tight, single-dose or Calculate the peak-to-valley ratio:
multiple-dose containers, preferably of Type I glass, and store
at controlled room temperature. Result = (HP/HV)
USP Alfentanil Hydrochloride RS HP = height of impurity A peak above baseline
USP Endotoxin RS HV = lowest point between the impurity A peak and
the alfuzosin peak.
Analysis
Alfuzosin Hydrochloride Samples: Sample solution A and Sample solution B
Calculate the percentage of each impurity in the portion
(Comment on this Monograph)id=m891=Alfuzosin of Alfuzosin Hydrochloride taken:
Hydrochloride=A-Monos.pdf)
rU = peak response of each impurity from Sample
solution A
rS = peak response from Sample solution B
CS = concentration of Sample solution B (mg/mL)
CU = concentration of the Sample solution A (mg/mL)
Acceptance criteria
C19H27N5O4 HCl 425.91 Individual impurities: See Impurity Table 1.
2-Furancarboxamide, ()-N-[3-[(4-amino-6,7-dimethoxy-2- [NOTEDisregard any peaks less than 0.05%.]
quinazolinyl)methylamino]propyl]tetrahydro-, Total impurities: NMT 0.30 %
monohydrochloride;
()-N-[3-[(4-Amino-6,7-dimethoxy-2-
Impurity Table 1
quinazolinyl)methylamino]propyl]tetrahydro-2-furamide
monohydrochloride [81403-68-1]. Relative Acceptance
Name Retention Time Criteria NMT %
DEFINITION
Impurity Aa 1.2 b
Alfuzosin Hydrochloride contains NLT 99.0% and NMT 101.0%
of C19H27N5O4 HCl, calculated on the anhydrous basis. Impurity D c 0.5 0.20
IDENTIFICATION a N-[3-[(4-Amino-6,7-dimethoxyquinazolin-2-
A. INFRARED ABSORPTION 197K yl)(methyl)amino]propyl]furan-2-carboxamide.
b Impurity A, a component of USP Alfuzosin System Suitability Mixture RS,
B. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the
requirements is not a specified impurity.
c N-(4-Amino-6,7-dimethoxyquinazolin-2-yl)-N-methylpropane-1,3-diamine.
ASSAY
PROCEDURE
Sample: 300 mg
Analysis: Dissolve the Sample in 80 mL of glacial acetic acid
and acetic anhydride (1:1). Titrate with 0.1 M perchloric
82 Alfuzosin / Official Monographs USP 32
Impurity Table 1 (continued) a suitable electrode system (see Titrimetry 541). Each mL of
Relative Acceptance
0.1 M sodium hydroxide is equivalent to 15.81 mg of
Name Retention Time Criteria NMT %
C4H6N4O3.
Acceptance criteria: NLT 98.5% and NMT 101.0%
Alfuzosin 1.0
Any other individual, 0.10 IMPURITIES
unidentified impurity Inorganic Impurities
a N-[3-[(4-Amino-6,7-dimethoxyquinazolin-2- Organic Impurities
yl)(methyl)amino]propyl]furan-2-carboxamide. PROCEDURE
b Impurity A, a component of USP Alfuzosin System Suitability Mixture RS,
Adsorbent: Cellulose
is not a specified impurity. Standard solution A: 1 mg/mL of USP Allantoin RS in
c N-(4-Amino-6,7-dimethoxyquinazolin-2-yl)-N-methylpropane-1,3-diamine.
methanol and water (1:1)
SPECIFIC TESTS Urea stock solution: 1 mg/mL of USP Urea RS
OPTICAL ROTATION 781: 10 to +10 Standard solution B: 0.1 mg/mL in methanol, from Urea
Sample solution: 20 mg/mL in carbon dioxide-free water stock solution
WATER DETERMINATION, Method I 921: NMT 0.5% Standard solution C: Standard solution A and Standard
solution B (1:1)
ADDITIONAL REQUIREMENTS Sample solution A: Transfer 0.10 g of Allantoin to a 10-mL
PACKAGING AND STORAGE: Preserve in tight containers. volumetric flask, add 5 mL of water, dissolve by heating,
Protect from light and moisture, and store at room and allow to cool. Dilute with methanol to volume.
temperature. [NOTEUse immediately after preparation.]
USP REFERENCE STANDARDS 11 Sample solution B: Transfer 1 mL of Sample solution A to a
USP Alfuzosin Hydrochloride RS 10-mL volumetric flask, and dilute with a mixture of
USP Alfuzosin System Suitability Mixture RS methanol and water (1:1) to volume.
Spray reagent: 5 mg/mL of p-
dimethylaminobenzaldehyde in a mixture of methanol and
Allantoin hydrochloric acid (3:1)
Application volume: 5 or 10 L for Sample solution A
(Comment on this Monograph)id=m1400=Allantoin=A- Developing solvent system: Butyl alcohol, glacial acetic
Monos.pdf) acid, and water (60:15:25)
Analysis
Samples: Standard solution A, Standard solution B,
Standard solution C, Sample solution A, and Sample solution
B
Proceed as directed for Chromatography 621, Thin-Layer
Chromatography. Develop the chromatogram until the
solvent front has moved about 10 cm. Spray the plate
with Spray reagent, dry in a current of hot air, and after
C4H6N4O3 158.12 30 min examine under visible light.
Urea, (2,5-dioxo-4-imidazolidinyl)- ; Acceptance criteria: Any spot from Sample solution A,
Allantoin [97-59-6]. except for the principal spot, is not more intense than the
DEFINITION spot from Standard solution B: NMT 0.5% of any individual
Allantoin contains NLT 98.5% and NMT 101.0% of C4H6N4O3. impurity. [NOTEThe test is not valid unless the principal
spots from Standard solution C are clearly separated.]
IDENTIFICATION
A. INFRARED ABSORPTION 197K SPECIFIC TESTS
B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201: OPTICAL ROTATION, Angular Rotation 781A
The RF value of the principal spot from Sample solution A Sample solution: 10 mg/mL, in carbon dioxide-free water
corresponds to that from Standard solution A, as described in [NOTEReserve a portion of this solution for use in the test
Organic Impurities. for Acidity.]
C. PROCEDURE Acceptance criteria: 10 to +10
Sample solution: Dissolve 20 mg of Allantoin in 2 mL of 1 ACIDITY OR ALKALINITY
M sodium hydroxide. Heat to boiling, allow to cool, and Sample: Use the Sample solution retained from the test for
add 1 mL of 2 M hydrochloric acid. Angular Rotation.
Analysis: To 0.1 mL of Sample solution add 0.1 mL of 100 Analysis: To 5 mL of the Sample add 5 mL of water, 0.1 mL
mg/mL of potassium bromide solution, 0.1 mL of 20 mg/mL of methyl red TS, and 0.2 mL of 0.01 M sodium hydroxide.
of resorcinol solution, and 3 mL of sulfuric acid. Heat on a Acceptance criteria: A yellow color is observed. Solution
water bath for 510 min. turns red upon addition of 0.4 mL of 0.01 M hydrochloric
Acceptance criteria: A dark blue color, which turns red acid.
after cooling and pouring into about 10 mL of water, is LOSS ON DRYING 731: Dry a sample at 105 to constant
observed. weight: it loses NMT 0.1% of its weight.
REDUCING SUBSTANCES
ASSAY Sample solution: 1.0 g of Allantoin in 10 mL of water,
Procedure shaken for 2 min, and filtered
Sample: 120 mg Analysis: To the Sample solution add 1.5 mL of 0.02 M
Analysis: Transfer the Sample to a 100-mL beaker, dissolve potassium permanganate.
by stirring in 40 mL of water, and titrate with 0.1 M sodium Acceptance criteria: The solution remains violet for at least
hydroxide. Determine the endpoint potentiometrically, using 10 min.
USP 32 Official Monographs / Allopurinol 83

USP REFERENCE STANDARDS 11 Detector: UV 230 nm
USP Allantoin RS Column: 4.6-mm 25-cm; packing L1
USP Urea RS Flow rate: 1.8 mL/min
System suitability
Allopurinol [NOTEThe relative retention times for allopurinol related
(Comment on this Monograph)id=m1430=Allopurinol=A- compound B, for allopurinol related compound C, and for
Monos.pdf) allopurinol are about 0.7, 0.8, and 1.0, respectively.]
Resolution: NLT 1.1 between allopurinol related
compound B and allopurinol related compound C and
NLT 6.0 between allopurinol related compound C and
allopurinol, System suitability solution
injections, Standard solution
Analysis
C5H4N4O 136.11 Samples: Standard solution and Sample solution
4H-Pyrazolo[3,4-d]pyrimidin-4-one, 1,5-dihydro-; Calculate the percentage of C5H4N4O in the portion of
1,5-Dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one; Allopurinol taken:
1H-Pyrazolo[3,4-d]pyrimidin-4-ol. [315-30-0].
DEFINITION
Allopurinol contains NLT 98.0% and NMT 102.0% of C5H4N4O, rU = peak response from the Sample solution
calculated on the dried basis. rS = peak response from the Standard solution
CS = concentration of USP Allopurinol RS in Standard
IDENTIFICATION solution (mg/mL)
INFRARED ABSORPTION 197K CU = concentration of Sample solution (mg/mL)
ASSAY
PROCEDURE IMPURITIES
[NOTEStore and inject the System suitability solution, the
Standard solution, and the Sample solution at 8, using a
cooled autosampler.] Change to read:
Mobile phase: 1.25 mg/mL of monobasic potassium
phosphate Organic Impurities
System suitability stock solution 1: 50 g/mL of USP PROCEDURE 1
Allopurinol RS in Mobile phase. [NOTEStore and inject the Standard solution and the
[NOTEDissolve the reference standard in a small amount Sample solution at 8, using a cooled autosampler.]
of 0.1 N sodium hydroxide before diluting with Mobile Solution A: 1.25 mg/mL of monobasic potassium
phase.] phosphate in water
System suitability stock solution 2: 50 g/mL of USP Solution B: Methanol
Allopurinol Related Compound B RS in Mobile phase. Diluent: Solution A and Solution B (9:1)
[NOTEDissolve the reference standard in a small amount Allopurinol related compound F solution: Weigh 5 mg
of 0.1 N sodium hydroxide before diluting with Mobile of USP Allopurinol Related Compound F RS into a suitable
phase.] flask. Add 2.0 mL of 0.1 N sodium hydroxide, promptly
System suitability stock solution 3: 50 g/mL of USP sonicate with swirling for 13 min to dissolve, add 80 mL
Allopurinol Related Compound C RS in Mobile phase. of Diluent, and sonicate for an additional 5 min.USP32
[NOTEDissolve the reference standard in a small amount
of 0.1 N sodium hydroxide before diluting with Mobile Standard stock solution: Transfer 5 mg of each of USP
phase.] Allopurinol RS, USP Allopurinol Related Compound A RS,
System suitability solution: Transfer 1.0 mL of each of the USP Allopurinol Related Compound B RS, USP Allopurinol
three System suitability stock solutions to a 100-mL Related Compound C RS, USP Allopurinol Related
volumetric flask, and dilute with Mobile phase to volume. Compound D RS, andUSP32 USP Allopurinol Related
Standard stock solution: 0.5 mg/mL of USP Allopurinol RS, Compound E RS USP32 to a 100-mL volumetric flask. Add
prepared by dissolving the USP Allopurinol RS in a small 2.0 mL of 0.1 N sodium hydroxide to dissolve, add the
volume of 0.1 N sodium hydroxide and immediately diluting entire volume of Allopurinol related compound F solution to
to the appropriate volume with Mobile phase. the flask, rinse the flask in which Allopurinol related
Standard solution: 80 g/mL of USP Allopurinol RS from compound F solution that was prepared with a small
the Standard stock solution in Mobile phase. amount of Diluent, and add the rinsings to the
Sample stock solution: Dissolve 50 mg of Allopurinol in solutionUSP32. Sonicate for an additional 5 min. Dilute with
5.0 mL of 0.1 N sodium hydroxide in a 100-mL volumetric Diluent to volume. [NOTEThis solution is stable for 48 h
flask. Immediate dilute with Mobile phase to volume. when stored at 8.]
Sample solution: 80 g/mL of Allopurinol from the Sample Standard solution: 0.5 g/mL of allopurinol, and
stock solution in Mobile phase. allopurinol related compounds A, B C, D, E, and F from the
Chromatographic system Standard stock solution in Diluent.
84 Allopurinol / Official Monographs USP 32
Sample solution: Transfer 25 mg of Allopurinol to a 100- Impurity Table 1 (continued)

mL volumetric flask. Add 5.0 mL of 0.1 N sodium Relative Relative Acceptance
hydroxide to dissolve, promptly sonicate with swirling for Retention Response Criteria
NMT 1 min, add 80 mL of Diluent, and sonicate for an Name Time Factor NMT %
additional 5 min. Dilute with Diluent to volume.
Mobile phase: Use gradient table below. below. Allopurinol related 0.81 0.2
compound Bb
Time Solution A Solution B Allopurinol 1.0
(min) (%) (%) Allopurinol related 4.4 0.2
0 90 10 compound Dd
30 70 30 Allopurinol related 4.8 0.2
compound Ee
35 70 30
Allopurinol related 6.5 0.2
36 90 10
compound Ff
46 90 10 a 3-Amino-1H-pyrazole-4-carboxamide.
b 5-(Formylamino)-1H-pyrazole-4-carboxamide.
Chromatographic system c 5-(4H-1,2,4-Triazol-4-yl)-1H-pyrazole-4-carboxamide.
(See Chromatography 621, System Suitability.) d Ethyl-5-amino-1H-pyrazole-4-carboxylate.
Mode: LC e Ethyl-5-(formylamino)-1H-pyrazole-4-carboxylate.
Detector: UV 220 nm f Ethyl-(E/Z)-3-(2-carbethoxy-2-cyanoethenyl)amino-1H-pyrazole-4-
Column: 4.6-mm 25-cm; 5-m packing L1 carboxylate.
Column temperature: 30
Flow rate: 1 mL/min Procedure 2: Limit of Hydrazine [NOTEUnder the

Injection size: 40 L following conditions, any hydrazine present in the sample

System suitability will react with benzaldehyde to form benzalazine.]
Sample: Standard solution Mobile phase: Hexane and isopropyl alcohol (95:5)
Suitability requirements Solution A: 85 mg/mL sodium hydroxide [NOTE
Resolution: NLT 0.8 between allopurinol related Alternatively, a commercially available 2 N sodium
compounds C and B hydroxide solution can be used.]
Tailing factor: NMT 1.5 for the allopurinol peak Diluent: Methanol and Solution A (1:1)
Analysis Solution B: 40 mg/mL benzaldehyde in Diluent
Samples: Standard solution and Sample solution Standard stock solution: 2.0 g/mL hydrazine sulfate in
Calculate the percentage of each impurity in the portion Diluent [NOTESonicate, as necessary, to facilitate
of Allopurinol taken: dissolution.]
Standard solution: Mix 5.0 mL of the Standard stock
Result = (rU/rS) (CS/CU) 100 solution and 4 mL of Solution B. Allow to stand for 2.5 h at
room temperature. Add 5.0 mL of hexane, and shake for 1
rU = peak response for each individual impurity from min. Allow the layers to separate, and use the upper
the Sample solution (hexane) layer.
rS = peak response for each individual impurity from Sample stock solution: Dissolve 250.0 mg of Allopurinol
the Standard solution [NOTEFor unspecified in 5.0 mL of Diluent.
impurities, rS is the peak response for the Sample solution: To the entire volume of Sample stock
allopurinol peak from the Standard solution.] solution add 4 mL of Solution B, mix, and allow to stand for
CS = concentration of each individual impurity in the 2.5 h at room temperature. Add 5.0 mL of hexane, and
Standard solution (mg/mL) shake for 1 min. Allow the layers to separate, and use the
CU = concentration of Allopurinol in the Sample upper (hexane) layer.
solution (mg/mL) Blank solution: Mix 5.0 mL of Diluent and 4 mL of
Acceptance criteria Solution B, and allow to stand for 2.5 h at room
Individual specified impurities: see Impurity Table I. temperature. Add 5.0 mL of hexane, and shake for 1 min.
Individual unspecified impurities: NMT 0.1% Allow the layers to separate, and use the upper (hexane)
Total impurities: NMT 1.0% layer.
Impurity Table 1 (See Chromatography 621, System Suitability.)
Mode: LC
Relative Relative Acceptance Detector: UV 310 nm
Retention Response Criteria Column: 4.6-mm 25-cm; 5-m packing L10
Name Time Factor NMT % Column temperature: 30
Allopurinol related 0.62 0.2 Flow rate: 1.5 mL/min
compound Aa Injection size: 20 L
Allopurinol related 0.79 0.2
System suitability
compound Cc
[NOTEThe relative retention times for benzalazine and
a 3-Amino-1H-pyrazole-4-carboxamide. benzaldehyde are about 0.8 and 1.0, respectively.]
b 5-(Formylamino)-1H-pyrazole-4-carboxamide. Suitability requirements
c 5-(4H-1,2,4-Triazol-4-yl)-1H-pyrazole-4-carboxamide. Resolution: NLT 2.0 between benzalazine and
d Ethyl-5-amino-1H-pyrazole-4-carboxylate. benzaldehyde
e Ethyl-5-(formylamino)-1H-pyrazole-4-carboxylate. Relative standard deviation: NMT 15.0% for the
f Ethyl-(E/Z)-3-(2-carbethoxy-2-cyanoethenyl)amino-1H-pyrazole-4- benzalazine peak
carboxylate.
USP 32 Official Monographs / Allopurinol 85
Analysis LABELING: Label it to state that it is to be shaken well before

Samples: Standard solution and Sample solution use and that it is to be discarded after 60 days. Label it to
Calculate the amount, in ppm, of hydrazine in the portion state that it is to be kept out of the reach of children. Label
of Allopurinol taken: it to indicate the nominal content of allopurinol in the Oral
Suspension.
Result = (rU/rS) (CS/CT) (Mr1/Mr2) F BEYOND-USE DATE: NMT 60 days after preparation
rU = peak response for the benzalazine peak from the
Sample solution
rS = peak response for the benzalazine peak from the Allopurinol Tablets
Standard solution (Comment on this Monograph)id=m1460=Allopurinol
CS = concentration of hydrazine sulfate in the Tablets=A-Monos.pdf)
Standard stock solution (g/mL)
CT = concentration of allopurinol in the Sample stock DEFINITION
solution (mg/mL) Allopurinol Tablets contain NLT 93.0% and NMT 107.0% of the
Mr1 = molecular weight of hydrazine, 32.05 labeled amount of C5H4N4O.
Mr2 = molecular weight of hydrazine sulfate, 130.12
F = unit conversion factor (from g/mg to ppm), IDENTIFICATION
1000 INFRARED ABSORPTION 197K
Acceptance criteria: NMT 10 ppm of hydrazineUSP32 Sample: Extract a quantity of finely powdered Tablets
equivalent to 50 mg of allopurinol by trituration with 10 mL
SPECIFIC TESTS of 0.1 N sodium hydroxide. Filter, acidify the filtrate with 1
LOSS ON DRYING 731: Dry a sample in vacuum at 105 for N acetic acid, collect the precipitated allopurinol (allow
5 h: it loses NLT 0.5% of its weight. 1015 min for sufficient precipitation to occur), wash the
precipitate with 3 mL of dehydrated alcohol, in portions,
ADDITIONAL REQUIREMENTS and finally wash with 4 mL of anhydrous ethyl ether. Allow
PACKAGING AND STORAGE: Preserve in well-closed containers. to dry in air for 15 min, then dry at 105 for 3 h.
Store at room temperature.
USP REFERENCE STANDARDS 11 ASSAY
USP Allopurinol RS PROCEDURE
USP Allopurinol Related Compound A RS [NOTEDo not allow the Mobile phase to remain in the
USP Allopurinol Related Compound B RS column overnight. After performing the procedure, flush
USP Allopurinol Related Compound C RS the system with water for NLT 20 min, and then flush with
USP Allopurinol Related Compound D RS methanol for 20 min.]
USP Allopurinol Related Compound E RS Mobile phase: 0.05 M monobasic ammonium phosphate
USP Allopurinol Related Compound F RS Internal standard solution: Dissolve 50 mg of
hypoxanthine in 10 mL of 0.1 N sodium hydroxide, shake
by mechanical means until dissolved (about 10 min), and
dilute with water to 50 mL. [NOTEPrepare on the day of
Allopurinol Oral Suspension use.]
(Comment on this Monograph)id=m385=Allopurinol Oral Standard stock solution: Transfer 50 mg of USP Allopurinol
Suspension=A-Monos.pdf) RS to a 50-mL volumetric flask. Add 10 mL of 0.1 N sodium
hydroxide, shake by mechanical means until dissolved
DEFINITION (about 10 min), and dilute with water to volume. [NOTE
Allopurinol Oral Suspension contains NLT 90.0% and NMT Prepare on the day of use.]
110.0% of the labeled amount of allopurinol (C5H4N4O). Standard solution: Transfer 4.0 mL of the Standard stock
Prepare Allopurinol Oral Suspension at a 2% concentration, for solution to a 200-mL volumetric flask. Add 2.0 mL of the
example, as follows (see Pharmaceutical Compounding Internal standard solution, and dilute with Mobile phase to
Nonsterile Preparations 795). volume.
Sample stock solution: Transfer an amount of finely
Allopurinol 2g powdered Tablets (NLT 20 Tablets) equivalent to 50 mg of
Glycerin 5 mL allopurinol to a 500-mL volumetric flask. Add 10 mL of 0.1
N sodium hydroxide, shake by mechanical means for 10
Vehicle for Oral Suspension, NF 45 mL min, and add water to volume. [NOTEFrom this point,
Vehicle for Oral Solution, NF A sufficient quantity conduct the remainder of the Assay without delay.] Filter,
To make 100 mL rejecting the first 10 mL of the filtrate.
Sample solution: Transfer 4.0 mL of the Sample stock
Select the number of Tablets that contain the specified amount solution to a 200-mL volumetric flask. Add 2.0 mL of Internal
of allopurinol and calculate the quantity of each ingredient standard solution, and dilute with Mobile phase to volume.
required for the total amount to be prepared. Count/weigh/ Chromtographic system
measure each ingredient. Thoroughly pulverize the tablets. (See Chromatography 621, System Suitability.)
Mix the powdered Allopurinol Tablets and Glycerin to form a Mode: LC
smooth paste, incorporate the Vehicle for Oral Suspension, add Detector: UV 254 nm
sufficient Vehicle for Oral Solution to volume, and mix well. Column: 4-mm 30-cm; packing L1
Adjust the pH, if necessary. Package, and label. Flow rate: 1.5 mL/min
SPECIFIC TESTS System suitability
PH 791: An apparent pH between 6.5 and 7.5 Sample: Standard solution
[NOTEThe relative retention times for hypoxanthine and
ADDITIONAL REQUIREMENTS allopurinol are about 0.6 and 1.0, respectively.]
PACKAGING AND STORAGE: Package in a tight container, and
86 Allopurinol / Official Monographs USP 32

Resolution: NLT 5 between the analyte and internal Allyl Isothiocyanate contains NLT 93.0% and NMT 105.0% of
standard C4H5NS.
Relative standard deviation: NMT 3.0% [CAUTIONAllyl Isothiocyanate is a potent lachrymator, with a
Analysis pungent irritating odor. Care should be taken to protect the
Samples: Standard solution and Sample solution eyes, to prevent inhalation of fumes, and to avoid tasting.]
Calculate the quantity of C5H4N4O in the portion of Tablets
taken: IDENTIFICATION
INFRARED ABSORPTION 197F: The spectrum exhibits
Result = (RU/RS) (CS/CU) 100 pronounced peaks at about 700cm, 950cm, 980cm, 1300cm,
1340cm, 1350cm, 1410cm, 1420cm, 1650cm, 2100cm, and
RU = peak response ratio of allopurinol to 2200cm.
hypoxanthine from the Sample solution
RS = peak response ratio of allopurinol to ASSAY
hypoxanthine from the Standard solution PROCEDURE
CS = concentration of USP Allopurinol RS in the Sample solution: Dilute a 4-mL sample to 100 mL in a
Standard solution (mg/mL) volumetric flask with alcohol.
CU = nominal concentration of allopurinol in the Analysis: Transfer 5.0 mL of Sample solution to a 100-mL
Sample solution (mg/mL) conical flask. Add 50.0 mL of 0.1 N silver nitrate VS and 5
Acceptance criteria: 93.0%107.0% mL of ammonia TS. Connect the flask to a reflux condenser,
heat on a water bath for 1 h, and allow to cool to room
PERFORMANCE TESTS temperature. Disconnect the flask from the condenser,
DISSOLUTION 711 transfer the contents of the conical flask to a 100-mL
Medium: 0.01 N hydrochloric acid; 900 mL volumetric flask with the aid of water, and dilute with water
Apparatus 2: 75 rpm to volume. Pass through a dry filter, discarding the first 10
Time: 45 min mL of the filtrate. To 50.0 mL of the subsequent filtrate add
Detector: UV 250 nm 5 mL of nitric acid and 2 mL of ferric ammonium sulfate TS,
Sample solution: Sample per Dissolution 711. Dilute with and titrate the excess silver nitrate with 0.1 N ammonium
Medium to a concentration that is similar to that of the thiocyanate VS. Perform a blank determination, using 5 mL
Standard solution. of alcohol in place of the Sample solution, and make any
Standard stock solution: Transfer 40 mg of USP Allopurinol necessary correction. Each mL of 0.1 N silver nitrate is
RS to a 200-mL volumetric flask. Add 10 mL of 0.1 N equivalent to 4.958 mg of C4H5NS.
sodium hydroxide, sonicate for about 2 min, and shake by Acceptance criteria: 93.0%105.0%
mechanical means for about 10 min. Dilute with Medium to
volume. SPECIFIC TESTS
Standard solution: Dilute the Standard stock solution with SPECIFIC GRAVITY 841: 1.0131.020
Medium to obtain a solution having a concentration similar REFRACTIVE INDEX 831: 1.5271.531 at 20
to that expected in the solution under test. DISTILLING RANGE, Method I 721: 148154
Analysis: Determine the amount of C5H4N4O dissolved by LIMIT OF PHENOLS
using UV absorption at the wavelength of maximum Sample solution: Dilute a 1-mL sample with 4 mL of
absorbance at about 250 nm on filtered portions of the alcohol.
solution under test, suitably diluted with Medium, in Analysis: Add 1 drop of ferric chloride TS to the Sample
comparison to the Standard solution. solution.
Tolerances: NLT 75% (Q) of the labeled amount of Acceptance criteria: A blue color is not produced
C5H4N4O. immediately.
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
USP Allopurinol RS Aloe
(Comment on this Monograph)id=m1540=Aloe=A-Monos.pdf)
Allyl Isothiocyanate DEFINITION
(Comment on this Monograph)id=m1480=Allyl Aloe is the dried latex of the leaves of Aloe barbadensis Miller
Isothiocyanate=A-Monos.pdf) (Aloe vera Linne), known in commerce as Curacao Aloe, or of
Aloe ferox Miller and hybrids of this species with Aloe africana
Miller and Aloe spicata Baker, known in commerce as Cape
Aloe (Fam. Liliaceae). Aloe yields NLT 50.0% of water-soluble
extractive.
IDENTIFICATION
A. Powdered Aloe dissolves in nitric acid with effervescence,
forming a reddish-brown to brown-or-green solution.
C4H5NS 99.16
3-Isothiocyanato-1-propene;
Isothiocyanic acid allyl ester [57-06-7].
USP 32 Official Monographs / Alprazolam 87
B. PROCEDURE fragments, the hues of which depend to some extent upon

Sample: 1 g, finely powdered the thickness of the fragments.
Analysis: Mix the Sample with 25 mL of cold water. Shake
the mixture occasionally during 2 h, filter, and wash the
filter and residue with sufficient cold water to make the
filtrate measure 100 mL. Alprazolam
Acceptance criteria: The color of the filtrate, viewed in the (Comment on this Monograph)id=m1640=Alprazolam=A-
bulb of a 100-mL volumetric flask, is dark orange with Monos.pdf)
Curacao Aloe, and greenish yellow with Cape Aloe. The
filtrate darkens on standing. [NOTEReserve the filtrate for
Procedures C and D.]
C. PROCEDURE
Sample: 5 mL of the filtrate obtained in Identification,
Procedure B
Analysis: Add 2 mL of nitric acid to the Sample.
Acceptance criteria: The mixture exhibits a reddish-orange
color with Curacao Aloe, and a reddish-brown color which C17H13ClN4 308.76
changes rapidly to green with Cape Aloe. 4H-[1,2,4]Triazolo[4,3-][1,4]benzodiazepine, 8-chloro-1-
D. PROCEDURE methyl-6-phenyl-;
Sample: 10 mL of the filtrate obtained in Identification, 8-Chloro-1-methyl-6-phenyl-4H-s-triazolo[4,3-]
Procedure B [1,4]benzodiazepine [28981-97-7].
Analysis: Mix the Sample with 2 mL of ammonium
hydroxide. DEFINITION
Acceptance criteria: The mixture exhibits an amber color Alprazolam contains NLT 98.0% and NMT 102.0% of
with Cape Aloe, and a dark amber color with Curacao Aloe. C17H13ClN4.
[CAUTIONCare should be taken to prevent inhaling particles of
ASSAY Alprazolam and exposing the skin to it.]
PROCEDURE
Sample: 2 g IDENTIFICATION
Analysis: Macerate the Sample in 70 mL of water in a A. INFRARED ABSORPTION 197M
suitable flask. Shake the mixture during 8 h at 30-min B. ULTRAVIOLET ABSORPTION 197U
intervals, and allow it to stand for 16 h without shaking. Sample solution: 4 g/mL in alcohol
Filter, and wash the flask and residue with small portions of Analytical wavelength: 220 nm
water, passing the washings through the filter, until the Acceptance criteria: Absorptivities calculated on the dried
filtrate measures 100.0 mL. Evaporate a 50-mL aliquot of the basis, do not differ by more than 3.0%.
filtrate in a tared dish on a steam bath to dryness, and dry ASSAY
at 110 to constant weight. PROCEDURE
Acceptance criteria: NLT 50.0% of the weight of Aloe Diluent: Acetonitrile and water (1:1)
taken Buffer: 1.36 mg/mL of monobasic potassium phosphate
SPECIFIC TESTS (KH2 PO4) in water
WATER DETERMINATION, Method III 921 Mobile phase: Acetonitrile and Buffer (1:1)
Sample: Use a powdered sample. (If the Aloe is not Standard solution: 25 g/mL of USP Alprazolam RS in
powdered, crush it in a mortar until it passes through a no. Diluent
40 sieve, and mix the ground material before weighing the [NOTEThe solution is stable for 48 h at room temperature
sample.) when stored in closed containers.]
Analysis: Dry at 105 for 5 h Sample solution: 25 g/mL of Alprazolam in Diluent.
Acceptance criteria: NMT 12.0% [NOTESonication for 1 min may be used to aid
ARTICLES OF BOTANICAL ORIGIN, Total Ash 561 dissolution. The solution is stable for 48 h at room
Acceptance criteria: NMT 4.0% temperature when stored in closed containers.]
ALCOHOL-INSOLUBLE SUBSTANCES Chromatographic system
Sample: 1 g of powdered Aloe (See Chromatography 621, System Suitability.)
Analysis: Add the Sample to 50 mL of alcohol in a flask. Mode: LC
Heat the mixture to boiling, and maintain at incipient Detector: UV 231 nm
boiling for 15 min, replacing any loss by evaporation. Column: 4.6-mm 25-cm; packing L1
Remove from the heat, and shake the mixture at intervals Flow rate: 1 mL/min
during 1 h. Pass through a small dried and tared filter paper Injection size: 20 L
or a dried and tared filtering crucible, and wash the residue System suitability
on the filter with alcohol until the last washing is colorless. Sample: Standard solution
Dry the residue at 105 to constant weight. Suitability requirements
Acceptance criteria: NMT 10.0% of the weight of Aloe Tailing factor: NMT 2.0
taken Relative standard deviation: NMT 2.0% for replicate
BOTANIC CHARACTERISTICS injections
Curacao aloe: Brownish black, opaque masses. Its fractured Analysis
surface is uneven, waxy, and somewhat resinous. Samples: Standard solution and Sample solution
Cape aloe: Dusky to dark brown irregular masses, the Calculate the quantity, in percentage, of C17H13ClN4 in the
surfaces of which are often covered with a yellowish portion of Alprazolam taken:
powder. Its fracture is smooth and glassy. Result = (rU/rS) (CS/CU) 100
Powdered aloe: Yellow, yellowish brown to olive-brown in
color. When mounted in a bland expressed oil, it appears as rU = peak area from the Sample solution
greenish yellow to reddish brown angular or irregular
88 Alprazolam / Official Monographs USP 32
rS = peak area from the Standard solution Impurity Table 1 (continued)

CS = concentration of USP Alprazolam RS in the Relative Relative Acceptance
Standard solution (mg/mL) Retention Response Criteria,
CU = concentration of alprazolam in the Sample Name Time Factor NMT (%)
solution (mg/mL)
Acceptance criteria: 98.0%102.0% 2-Amino-5- 4.0 1.0 0.15
chlorobenzophenone
IMPURITIES Individual 1.0 0.10
Inorganic Impurities unspecified
RESIDUE ON IGNITION 281 impurity
HEAVY METALS, Method II 231
Acceptance criteria: NMT 20 ppm SPECIFIC TESTS
Organic Impurities LOSS ON DRYING 731: Dry at a pressure of NMT 5 mm of
PROCEDURE mercury at 60 for 16 h: it loses NMT 0.5% of its weight.
Diluent: Prepare as directed in the Assay.
Buffer: Prepare as directed in the Assay. ADDITIONAL REQUIREMENTS
Mobile phase: Prepare as directed in the Assay. PACKAGING AND STORAGE: Preserve in tight containers, and
System suitability solution: 20 g/mL each of USP store at controlled room temperature.
Alprazolam RS, USP Alprazolam Related Compound A RS, USP REFERENCE STANDARDS 11
and USP 2-Amino-5-chlorobenzophenone RS in Diluent USP Alprazolam RS
Sample solution: 0.25 mg/mL of alprazolam in Diluent
[NOTESonication for 1 min may be used to aid
dissolution. When stored in closed containers, the Sample
solution is stable for 24 h at room temperature.] Alprazolam Oral Suspension
Standard solution: 0.25 g/mL of USP Alprazolam RS in (Comment on this Monograph)id=m1377=Alprazolam Oral
Diluent [NOTEWhen stored in closed containers, the Suspension=A-Monos.pdf)
Standard solution is stable for 48 h at room temperature.]
Chromatographic system DEFINITION
(See Chromatography 621.) Alprazolam Oral Suspension contains NLT 90.0% and NMT
Proceed as directed in the Assay. 110.0% of the labeled amount of alprazolam (C17H13ClN4).
System suitability Prepare Alprazolam Oral Suspension 1 mg/mL as follows (see
Samples: Standard solution and System suitability solution Pharmaceutical CompoundingNonsterile Solutions 795).
[NOTEFor relative retention times, see Impurity Table 1.]
Suitability requirements Alprazolam 100 mg
Resolution: NMT 2.0 between alprazolam Related Vehicle: a mixture of Vehicle for Oral A sufficient quantity
compound A and alprazolam, System sutiability solution Solution (regular or sugar-free), NF, and
Relative standard deviation: NMT 5.0, Standard Vehicle for Oral Suspension, NF (1:1)
solution
Analysis To make 100 mL
Calculate the percentage of each impurity in each of the Comminute Tablets in a suitable mortar to a fine powder, or
related compounds and any of the unspecified add Alprazolam powder. Add about 20 mL of the Vehicle, and
impurities: mix until a uniform paste is formed. Add the Vehicle in small
portions almost to volume, and mix thoroughly after each
Result = (rU/rS) (CS/CU) (1/F) 100 addition. Transfer the contents of the mortar, stepwise and
quantitatively, to a calibrated bottle. Add sufficient Vehicle to
rU = peak response for each impurity in the Sample bring to final volume, and mix well.
solution
rS = peak response for alprazolam from the Standard ASSAY
solution PROCEDURE
CS = concentration of USP Alprazolam RS in the Solution A: 0.04 M sodium acetate solution. Adjust with
Standard solution (mg/mL) glacial acetic acid to a pH of 2.4.
CU = concentration of alprazolam in the Sample Mobile phase: Methanol, acetonitrile, and Solution A
solution (45:8:47)
F = relative response factor for the corresponding Standard solution: 20 g/mL of USP Alprazolam RS in
impurity from Impurity Table 1 Mobile phase
Acceptance criteria Sample solution: Agitate the container of Alprazolam Oral
Individual impurities: See Impurity Table 1 Suspension for 30 min on a rotating mixer, remove a 5-mL
Total impurities: NMT 1.0% sample, and store in a clear glass vial at 70 until analyzed.
At the time of analysis, remove the sample from the freezer,
allow it to reach room temperature, and mix with a vortex
Impurity Table 1 mixer for 30 s. Pipet 1.0 mL of the sample into a 50-mL
Relative Relative Acceptance volumetric flask, and dilute with Mobile phase to volume.
Retention Response Criteria, Chromatographic system
Name Time Factor NMT (%) (See Chromatography 621, System Suitability.)
Alprazolam related 0.8 0.76 0.15
compound A
Alprazolam 1.0 1.0
USP 32 Official Monographs / Alprazolam 89
Mode: LC 891, 826, 779, 746, 696, and 658 wavenumbers in the
Detector: UV 230 nm region of 975600 cm1.
Flow rate: 0.6 mL/min ASSAY
System suitability Mobile phase: Acetonitrile, chloroform, butyl alcohol,
Sample: Standard solution glacial acetic acid, and water (850:80:50:0.5:20)
Suitability requirements Internal standard solution: 0.25 mg/mL of triazolam in
Relative standard deviation: NMT 1.4% acetonitrile
Retention time: 10 min Standard stock solution: 0.25 mg/mL of USP Alprazolam
Analysis RS in Internal standard solution
Samples: Standard solution and Sample solution Standard solution: 25 g/mL of USP Alprazolam RS from
Calculate the percentage of C17H13ClN4 in each mL of Oral Standard stock solution in acetonitrile
Suspension taken: Sample solution: Weigh and finely powder NLT 20 Tablets.
Transfer a quantity of the powder, equivalent to about 5 mg
Result = (rU/rS) (CS/CU) 100 of alprazolam, to a 200-mL volumetric flask. Transfer 2 mL
of water and 20 mL of Internal standard solution, shake
rU = peak response from the Sample solution vigorously for 10 min, and dilute with acetonitrile to
rS = peak response from the Standard solution volume.
CS = concentration of USP Alprazolam RS in the Chromatographic system
Standard solution (g/mL) (See Chromatography 621, System Suitability.)
CU = nominal concentration of the Sample solution Mode: LC
(g/mL) Detector: UV 254 nm
Acceptance criteria: 90.0%110.0% Column: 4.6-mm 30-cm; packing L3
Flow rate: 2 mL/min
PH 791: 4.05.0 System suitability
PACKAGING AND STORAGE: Preserve in tight, light-resistant Resolution: NLT 2.0 between the internal standard and
containers. Store at controlled room temperature, or under alprazolam
refrigeration. Relative standard deviation: NMT 2.0% for replicate
LABELING: Label it to state that it is to be well-shaken before injections
use, and to state the beyond-use date. Analysis
Beyond-Use Date: 60 days after the day on which it was Samples: Standard solution and Sample solution
compounded Calculate the quantity, as a percentage, of C17H13ClN4 in
USP REFERENCE STANDARDS 11 the portion of Tablets taken:
USP Alprazolam RS
Alprazolam Tablets RU = peak area response ratio of the alprazolam peak

relative to the internal standard peak from the
(Comment on this Monograph)id=m1647=Alprazolam Sample solution
Tablets=A-Monos.pdf) RS = peak area response ratio of the alprazolam peak
relative to the internal standard peak from the
DEFINITION Standard solution
Alprazolam Tablets contain NLT 90.0% and NMT 110.0% of the CS = concentration of USP Alprazolam RS in the
labeled amount of alprazolam (C17H13ClN4). Standard solution (mg/mL)
IDENTIFICATION CU = nominal concentration of the Sample solution
A. INFRARED ABSORPTION (Tablets/mL)
Sample: Finely powdered Tablets, equivalent to 15 mg of Acceptance criteria: 90.0%110.0%
alprazolam PERFORMANCE TESTS
Analysis: Dissolve the Sample in 10 mL of 10 mg/mL of DISSOLUTION, Procedure for a Pooled Sample 711
sodium carbonate solution. Add 15 mL of chloroform, and Solution A: 80 g/L of monobasic potassium phosphate and
shake vigorously for 30 min. Centrifuge, withdraw the 20 g/L dibasic potassium phosphate in water. Add, with
aqueous layer, and transfer the chloroform to a clean mixing, phosphoric acid or potassium hydroxide solution
container. Add 200 mg of potassium bromide. Evaporate (45 in 100), as necessary to adjust the solution such that,
the chloroform from this mixture to dryness, and dry the when this is diluted 1 in 10 with water, the resulting
dispersion in vacuum at 60 for 24 h. Grind this dispersion solution has a pH of 6.0 0.1.
into a fine powder. Prepare a suitable pellet for testing by Buffer: 1-in-10 dilution of Solution A in water to obtain a
placing 100 mg of dried potassium bromide into a die. solution having a pH of 6.0 0.1
Sprinkle 20 mg of the finely ground alprazolampotassium Medium: Buffer; 500 mL
bromide dispersion onto the dried potassium bromide layer, Apparatus 1: 100 rpm
and cover with another specimen of 100 mg of dried Time: 30 min
potassium bromide. [NOTEDetermine the amount of C17H13ClN4 dissolved
Acceptance criteria: The IR absorption spectrum of the using the following method.]
potassium bromide dispersion so obtained exhibits maxima Mobile phase: Acetonitrile, tetrahydrofuran, and Buffer
characteristic of alprazolam, as compared to that of a similar (35:5:60)
preparation of USP Alprazolam RS, at the following Standard stock solution: 0.05 mg/mL of USP Alprazolam
wavenumbers: at 1609, 1578, 1566, 1539, 1487, and 1379 RS in methanol
wavenumbers in the region of 16501300 cm1; at 932,
90 Alprazolam / Official Monographs USP 32
Standard solution: Add 50 mL of Solution A and 250 mL of V = volume of Internal standard solution used to
water to a 500-mL flask. Add to the flask 5.0 mL of Standard prepare the Sample solution
stock solution for every 0.25 mg of alprazolam contained in L = label claim (mg/Tablet)
the Tablet being assayed. Dilute with water to volume. Acceptance criteria: Meet the requirements of the test for
Sample solution: Sample per 711 Dissolution. Dilute with Content Uniformity
Medium to a concentration that is similar to the Standard
solution. ADDITIONAL REQUIREMENTS
Chromatographic system PACKAGING AND STORAGE: Preserve in tight, light-resistant
(See Chromatography 621, System Suitability.) containers, and store at controlled room temperature.
Mode: LC USP REFERENCE STANDARDS 11
Detector: UV 254 nm USP Alprazolam RS
Flow rate: 1 mL/min
System suitability Alprostadil
Suitability requirements (Comment on this Monograph)id=m1658=Alprostadil=A-
Column efficiency: NLT 500 theoretical plates Monos.pdf)
injections
Analysis
Samples: Filtered portions of the solution from the
Dissolution vessel and Standard solution
Calculate the quantity of C17H13ClN4 dissolved based on the
peak responses from the solution under test and the
Standard solution.
Tolerances: NLT 80% (Q) of the labeled amount of C17 H13 C20H34O5 354.48
ClN4 is dissolved. Prost-13-en-1-oic acid, 11,15-dihydroxy-9-oxo-,
UNIFORMITY OF DOSAGE UNITS 905 (11,13E, 15S)-;
Mobile phase: Acetonitrile, chloroform, butyl alcohol, (1R, 2R, 3R)-3-Hydroxy-2-[(E)-(3S)-3-hydroxy-1-octenyl]-5-
glacial acetic acid, and water (850:80:50:0.5:20) oxocyclopentane heptanoic acid [745-65-3].
Internal standard solution: 32 g/mL of triazolam in DEFINITION
acetonitrile Alprostadil contains NLT 95.0% and NMT 105.0% of C20H34O5,
Standard solution: 25 g/mL of USP Alprazolam RS in calculated on the anhydrous basis.
Internal standard solution [CAUTIONGreat care should be taken to prevent inhaling
Sample solution: Transfer 1 Tablet to a container. Add 0.4 particles of Alprostadil and exposing the skin to it.]
mL of water directly onto the Tablet, allow the Tablet to
stand for 2 min, and then swirl the container to disperse the IDENTIFICATION
Tablet. For every 0.25 mg of alprazolam contained in the INFRARED ABSORPTION 197M
Tablet, add 10.0 mL of Internal standard solution to the
container. Shake, and centrifuge if necessary. ASSAY
Chromatographic system PROCEDURE
(See Chromatography 621, System Suitability.) [NOTEUse freshly prepared solutions.]
Mode: LC Mobile phase: Methanol, acetonitrile, and 0.1 M
Detector: UV 254 nm monobasic potassium phosphate (2:1:2). Adjust with
Column: 4.6-mm 30-cm; packing L3 phosphoric acid to a pH of 3.0.
Flow rate: 2 mL/min Internal standard solution: 0.05 mg/mL of ethylparaben in
Injection size: 20 L methanol and water (9:1)
System suitability Standard stock solution: 0.3 mg/mL of USP Alprostadil RS
Sample: Standard solution in methanol and water (9:1)
Suitability requirements Standard solution: Internal standard solution and Standard
Resolution: NLT 2.0 between the internal standard and stock solution (1:2)
alprazolam System suitability stock solution: 4.5 g/mL of
Relative standard deviation: NMT 2.0% for replicate prostaglandin A1 from USP Prostaglandin A1 RS in Standard
injections solution
Analysis System suitability solution: Internal standard solution and
Samples: Standard solution and Sample solution System suitability stock solution (1:2)
Calculate the percentage of C17H13ClN4 in the Tablet taken: Sample stock solution: 0.3 mg/mL of Alprostadil in a
mixture of methanol and water (9:1)
Result = (RU/RS) C V 100/L Sample solution: Combine 1.0 mL of Internal standard
solution and 2.0 mL of Sample stock solution.
RU = peak area response ratio of the alprazolam peak Chromatographic system
relative to the internal standard peak from the (See Chromatography 621, System Suitability.)
Sample solution Mode: LC
RS = peak area response ratio of the alprazolam peak Detector: Photodiode array or equivalent capable of
relative to the internal standard peak from the detecting UV wavelengths of 200300 nm
Standard solution
C = concentration of USP Alprazolam RS in the
Standard solution
USP 32 Official Monographs / Alprostadil 91
Analytical wavelength: 200 nm Sample solution: 2 mg/mL of Alprostadil in alcohol

Column: 4.6-mm 25-cm; packing L1 Blank: Alcohol
Column temperature: 40 Spectrometric conditions
Flow rate: 1 mL/min (See Spectrophotometetry and Light-Scattering 851.)
Injection size: 20 L Mode: Graphite furnace atomic absorption
System suitability spectrophotometer (equipped with a rhodium hollow-
Sample: System suitability solution cathode lamp)
Suitability requirements Analytical wavelength: 343.5 nm
Resolution: NLT 7.5 between prostaglandin A1 and Drying temperature: 100
alprostadil, and NLT 2.0 between prostaglandin A1 and Ashing temperature: 1000
ethylparaben Atomization temperature: 2800
Relative standard deviation: NMT 2.0%, determined Injection size: 20 L
from the peak area ratio of alprostadil to ethylparaben for Analysis
replicate injections Samples: Standard solution and Sample solution
Analysis Calculate the percentage of Rh in the portion of
Samples: Standard solution and Sample solution Alprostadil taken:
Calculate the percentage of C20H34O5 in the portion taken:
AU = absorbance of the Sample solution
RU = peak area ratio of Alprostadil to ethylparaben in AS = absorbance of the Standard solution
the Sample solution CS = concentration of rhodium in the Standard
RS = peak area ratio of alprostadil to ethylparaben in solution (mg/mL)
the Standard solution CU = concentration of Alprostadil in the Sample
CS = concentration of USP Alprostadil RS in the solution (mg/mL)
Standard solution (mg/mL) Acceptance criteria: NMT 20 ppm
CU = concentration of Alprostadil in the Sample Organic Impurities
solution (mg/mL) PROCEDURE 1: LIMIT OF FOREIGN PROSTAGLANDINS
Acceptance criteria: 95.0%105.0% [NOTEUse freshly prepared solutions. ]
Mobile phase: Methanol, acetonitrile, and 0.1 M
IMPURITIES monobasic potassium phosphate (2:1:2). Adjust with
Inorganic Impurities phosphoric acid to a pH of 3.0.
RESIDUE ON IGNITION 281 Standard solution: 6 g/mL of USP Alprostadil RS,15
Sample: 0.3 g g/mL of USP Prostaglandin A1 RS, and 6 g/mL of USP
Acceptance criteria: NMT 0.5% Prostaglandin B1 RS in methanol and water (9:1)
LIMIT OF CHROMIUM Sample solution: 3.0 mg/mL of Alprostadil in methanol
Standard stock solution: 3.04 g/mL of chromium and water (9:1)
trichloride in 0.05 M nitric acid Chromatographic system
Standard solution: 20 ng/mL of chromium (Cr) in alcohol (See Chromatography 621, System Suitability.)
by diluting 2 mL of Standard stock solution to 100 mL with Mode: LC
alcohol Detector: Photodiode array detector or equivalent
Sample solution: 1 mg/mL of Alprostadil in alcohol capable of detecting UV wavelengths of 200300 nm
Blank: Alcohol Analytical wavelengths: Prostaglandin A1 at 224 nm,
Spectrometric conditions prostaglandin B1 at 280 nm, and all other foreign
(See Spectrophotometetry and Light-Scattering 851.) prostaglandin impurities at 200 nm.
Mode: Graphite furnace atomic absorption Column: 4.6-mm 25-cm; packing L1
spectrophotometer (equipped with a chromium hollow- Column temperature: 40
cathode lamp) Flow rate: 1 mL/min
Analytical wavelength: 357.9 nm Injection size: 20 L
Drying temperature: 100 System suitability
Ashing temperature: 1000 Sample: Standard solution
Atomization temperature: 2700 Suitability requirements
Injection size: 20 L Resolution: NLT 7.5 between prostaglandin A1 and
Analysis alprostadil
Samples: Standard solution and Sample solution Relative standard deviation: NMT 4.0%, determined
Calculate the percentage of Cr in the portion of C20H34O5 from the peaks at their respective wavelength for
taken: replicate injections
Analysis 1
Result = (AU/AS) (CS/CU) 100 Samples: Standard solution and Sample solution
Calculate the percentage of prostaglandin A1 and
AU = absorbance of the Sample solution prostaglandin B1 in the portion of C20H34O5 taken:
AS = absorbance of the Standard solution
CS = concentration of chromium in the Standard Result = (rU/rS) (CS/CU) 100
solution (mg/mL)
CU = concentration of Alprostadil in the Sample rU = peak response of prostaglandin A1 or
solution (mg/mL) prostaglandin A2 from the Sample solution
Acceptance criteria: NMT 20 ppm rS = peak response of prostaglandin A1 or
LIMIT OF RHODIUM prostaglandin A2 from the Standard solution
Standard stock solution: 100 g/mL of rhodium in 1.2 M CS = concentration of USP Prostaglandin A1 RS or
hydrochloric acid by diluting rhodium chloride hydrate USP Prostaglandin B1 RS in the Standard
Standard solution: 50 ng/mL of rhodium (Rh) in alcohol solution (mg/mL)
from Standard stock solution
92 Alprostadil / Official Monographs USP 32
CU = concentration of Alprostadil in the Sample Relative standard deviation: NMT 2.0%, determined
solution (mg/mL) from the main peak in the Standard solution (multiple
Acceptance criteria 1 injections)
Prostaglandin A1: NMT 1.5% Analysis
Prostaglandin B1: NMT 0.1% Samples: Standard solution and Sample solution
Analysis 2: Calculate the percentage of each impurity Calculate the percentage of each impurity occurring at
occurring at 200 nm and eluting before prostaglandin A1 200 nm and eluting after prostaglandin A1, excluding
in the portion of C20H34O5 taken: prostaglandin B1, in the portion of C20H34O5 taken:
Result = (rU/rS) (CS/CU) 100 Result = (rU/rS) (CS/CU) 100
rU = peak response for each impurity of the Sample rU = peak response for each impurity of the Sample
solution solution
rS = peak response for alprostadil of the Standard rS = peak response for alprostadil of the Standard
solution solution
CS = concentration of USP Alprostadil RS in the CS = concentration of USP Alprostadil RS in the
Standard solution (mg/mL) Standard solution (mg/mL)
CU = concentration of Alprostadil in the Sample CU = concentration of Alprostadil in the Sample
solution (mg/mL) solution (mg/mL)
Acceptance criteria 2: NMT 0.9% of any foreign Acceptance criteria: The sum of the peaks having relative
prostaglandin impurity eluting before prostaglandin A1 retention times of 2.0 and 2.3 is NMT 0.6%; any other
Analysis 3: Calculate the percentage of any impurity foreign prostaglandin impurity eluting after prostaglanin A1
having a relative retention time of 0.6, relative to the is NMT 0.9%.
prostaglandin A1 peak detected at 224 nm, in the portion Total impurities for Procedure 1 and Procedure 2: NMT
of C20H34O5 taken: 2.0 %
Result = (rU/rS) (CS/CU) 100 SPECIFIC TESTS
WATER DETERMINATION, Method I 921
rU = peak response for any impurity having a relative Sample: 0.5 g
retention time of 0.6, relative to the Acceptance criteria: NMT 0.5%
prostaglandin A1 peak, from the Sample
solution ADDITIONAL REQUIREMENTS
rS = peak response for prostaglandin A1 from the PACKAGING AND STORAGE: Preserve in tight containers, and
Standard solution store in a refrigerator.
CS = concentration of USP Prostaglandin A1 RS in the USP REFERENCE STANDARDS 11
Standard solution (mg/mL) USP Alprostadil RS
CU = concentration of Alprostadil in the Sample USP Prostaglandin A1 RS
solution (mg/mL) USP Prostaglandin B1 RS
Acceptance criteria 3: NMT 0.9% of any impurity having
a relative retention time of 0.6, relative to the
prostaglandin A1 peak Alprostadil Injection
PROCEDURE 2: LIMIT OF FOREIGN PROSTAGLANDINS
Mobile phase: Methanol, acetonitrile, and 0.02 M (Comment on this Monograph)id=m1660=Alprostadil
monobasic potassium phosphate (2:1:1). Adjust with Injection=A-Monos.pdf)
phosphoric acid to a pH of 3.
Standard solution: 10 g/mL of USP Alprostadil RS in DEFINITION
acetonitrile and water (1:1) Alprostadil Injection is a sterile solution of Alprostadil in
Sample solution: 5.0 mg/mL of Alprostadil in acetonitrile Dehydrated Alcohol. It contains NLT 90.0% and NMT 115.0%
and water (1:1) of the labeled amount of alprostadil (C20H34O5).
[NOTESonicate if necessary.] IDENTIFICATION
System suitability solution: 6 g/mL of USP Alprostadil INFRARED ABSORPTION 197K
RS, 15 g/mL of USP Prostaglandin A1 RS, and 6 g/mL of Standard: A preparation similar to that of the Sample, using
USP Prostaglandin B1 RS in methanol and water (9:1) USP Alprostadil RS in dehydrated alcohol
Chromatographic system Sample: Dry an amount of Injection, equivalent to 2 mg of
(See Chromatography 621, System Suitability.) alprostadil, on 500 mg of spectroscopic grade potassium
Mode: LC bromide at about 4050 under vacuum. Prepare a pellet
Detector: Photodiode array detector or equivalent, from this mixture.
capable of detecting UV wavelengths of 200300 nm
Analytical wavelengths: 224 nm for prostaglandin A1, ASSAY
280 nm for prostaglandin B1 , and 200 nm for all other PROCEDURE
prostaglandins Mobile phase: Methylene chloride, 1,3-butanediol, and
Column: 4.6-mm 25-cm; packing L1 water (1000:6:0.5)
Flow rate: 1.2 mL/min Internal standard solution: 50 g/mL of ethylparaben in
Injection size: 20 L methylene chloride
System suitability Standard stock solution: 0.5 mg/mL of USP Alprostadil RS
Samples: Standard solution and System suitability solution in dehydrated alcohol
[NOTEThe relative retention times for prostaglandin A1 Standard solution: Gently evaporate a 0.5-mL portion of
and alprostadil in the chromatogram of the System the Standard stock solution to dryness with a stream of
suitability solution are 1.2 and 1.0, respectively.] nitrogen. Add 150 mL of a freshly prepared 5 mg/mL of
Suitability requirements diisopropylethylamine in acetonitrile to the container, rinse
Resolution: NLT 4.0 between prostaglandin A1 and the inside of the container with this solution, and swirl. Cap
alprostadil for the Identification solution
USP 32 Official Monographs / Alteplase 93
and sonicate to dissolve. Heat the container at 45 for 45 Alteplase

min, swirling occasionally. Sonicate again after heating is (Comment on this Monograph)id=m1670=Alteplase=A-
complete. [NOTEIf the entire sample does not dissolve, the Monos.pdf)
specimen should be discarded.] Evaporate the solution using
a stream of nitrogen, add 2.0 mL of Internal standard
solution, and sonicate to dissolve. [NOTEIf incomplete
dissolution is still observed, discard the specimen.]
Sample solution: Pool the contents of several containers of
the Injection, and gently evaporate measured volume,
equivalent to 0.25 mg of alprostadil, to dryness using a
stream of nitrogen. Add 150 L of a freshly prepared
solution of 40 mg/mL -bromo-2-acetonaphthone in C2569H3894N746O781S40 59,007.61
acetonitrile, rinse the inside of the container with this [105857-23-6].
solution, and swirl. Gently evaporate a 0.5-mL portion of the
Standard stock solution to dryness with a stream of nitrogen. DEFINITION
Add 150 mL of a freshly prepared 5 g/mL Alteplase is a highly purified glycosylated serine protease with
diisopropylethylamine in acetonitrile to the container, rinse fibrin-binding properties and plasminogen-specific proteolytic
the inside of the container with this solution, and swirl. Cap activities. It is produced by recombinant DNA synthesis in
and sonicate to dissolve. Heat the container at 45 for 45 mammalian cell culture. It has a biological potency of NLT
min, swirling occasionally. Sonicate again after heating is 90.0% and NMT 115.0% of the potency stated on the label,
complete. [NOTEIf the entire sample does not dissolve, the the potency being 580,000 USP Alteplase Units/mg of protein.
specimen should be discarded.] Evaporate the solution using The presence of host cell DNA and host cell protein impurities
a stream of nitrogen, add 2.0 mL of Internal standard in Alteplase is process specific; the limits of these impurities are
solution, and sonicate to dissolve. [NOTEIf incomplete determined by validated methods.
dissolution is still observed, discard the specimen.]
Chromatographic system IDENTIFICATION
(See Chromatography 621, System Suitability.) A. PROCEDURE
Mode: LC Sample solution: 1.02.5 mg/mL Alteplase
Detector: UV 254 nm Standard solution: Prepare similarly to the Sample solution.
Flow rate: 1.5 mL/min Samples: Standard solution and Sample solution
System suitability To each of three test tubes, transfer 1 mL of 0.5 mg/mL H-
Sample: Standard solution D-isoleucyl-prolyl-arginyl-p-nitroaniline dihydrochloride.
[NOTEThe relative retention times for ethylparaben and Separately transfer 200 L of the Sample solution and 200
alprostadil are about 0.4 and 1.0, respectively.] L of the Standard solution to two of the test tubes, and
Suitability requirements to the third test tube, add 200 L of 0.2 M arginine
Resolution: NLT 9.0 between alprostadil and the internal solution that has been adjusted with phosphoric acid
standard (negative control) to a pH of 7.3. Mix the solutions in the
Relative standard deviation: NMT 2.5% test tubes, and allow to stand for 1 min.
Analysis Acceptance criteria: A yellow color is produced in the
Samples: Standard solution and Sample Solution solutions from the Sample solution and the Standard solution,
Calculate the quantity, as a percentage, of C20H34O5 in the while no yellow color is produced in the negative control.
volume of Injection taken: ASSAY
Result = (RU/RS) (CS/CU) 100/L PROCEDURE FOR BIOLOGICAL POTENCY
Buffer: 1.38 mg/mL of monobasic sodium phosphate, 7.10
RU = peak response ratio of alprostadil to the internal mg/mL of anhydrous dibasic sodium phosphate, 0.20
standard from the Sample solution mg/mL of sodium azide, and 0.10 mg/mL of polysorbate 80
RS = peak response ratio of alprostadil to the internal in water
standard from the Standard solution Human thrombin solution: 33 U.S. Units/mL in Buffer
CS = concentration of USP Alprostadil RS in the [NOTEUnits are in terms of the U.S. Standard Thrombin.]
Standard solution (mg/mL) Human fibrinogen solution: 2 mg/mL of human
CU = concentration of the Sample solution (unit/mL) fibrinogen in Buffer
L = label claim (mg/unit) Human plasminogen solution: 1 mg/mL of human
plasminogen in Buffer
Acceptance criteria: 90.0%115.0% Standard stock solution: 1.0 mg/mL (580,000 USP
SPECIFIC TESTS Alteplase Units) of USP Alteplase RS
BACTERIAL ENDOTOXINS TEST 85: It contains NMT 5 USP Standard solution: Dilute volumes of Standard stock
Endotoxin Units/100 g of alprostadil. solution to obtain a series of five Standard solutions having
STERILITY TESTS 71: It meets the requirements when tested known concentrations ranging from 145 to 9.3 USP
as directed under Test for Sterility of the Product to Be Alteplase Units/mL.
Examined for Membrane Filtration. Sample stock solution: 1.0 mg/mL of Alteplase
WATER DETERMINATION, Method I 921: NMT 0.4% Sample solution: Dilute a volume of Sample stock solution
OTHER REQUIREMENTS: It meets the requirements under with Buffer to obtain a series of dilutions of 1:20,000,
Injections 1. 1:10,000, and 1:5,000.
Analysis: To a set of labeled glass test tubes add 0.5 mL of
ADDITIONAL REQUIREMENTS Human thrombin solution. To separate test tubes add 0.5 mL
PACKAGING AND STORAGE: Preserve in tight, single-dose of each of the Standard solutions and Sample solutions, and
containers, preferably of Type I glass. Store in a refrigerator. store on ice. To a second set of labeled glass tubes, add 20
USP REFERENCE STANDARDS 11 L of Human plasminogen solution and 1 mL of Human
USP Alprostadil RS fibrinogen solution, and store on ice. Beginning with the
USP Endotoxin RS thrombin mixture tubes containing the Standard solution
94 Alteplase / Official Monographs USP 32
with the lowest number of USP Units/mL, record the time, Running buffer: 3.03 mg/mL of
and separately add 200 L of each of the thrombin mixtures tris(hydroxymethyl)aminomethane and 14.26 mg/mL of
to the test tubes containing the plasminogen-fibrinogen glycine in sodium dodecyl sulfate (1 in 1000)
mixture. Using a vortex mixer, intermittently mix the Carboxymethylation buffer: 480 mg/mL of urea, 44
contents of each tube for a total of 15 s, and carefully place mg/mL of tris(hydroxymethyl)aminomethane, and 1.2
into a rack in a 37 circulating water bath. A visually turbid mg/mL of edetic acid in water. Adjust with hydrochloric
clot forms within 30 s, followed by the formation of bubbles acid, if necessary, to a pH of 8.6.
within the clot. Record the clot lysis time (tcl) from the first Gel: Prepare a 10% T (total acrylamide)0.25% C (cross-
addition of the Alteplase solution to the last bubble to rise linked bisacrylamide) resolving gel containing 0.1% sodium
to the surface. dodecyl sulfate, 0.375 M tris(hydroxymethyl)aminomethane
Using a least squares fit, determine the equation of the line hydrochloride, and 0.05 M
using the log values of the standard concentration, in USP tris(hydroxymethyl)aminomethane.
Alteplase Units/mL, versus the log values of their clot lysis Arginine solution: 34.8 mg/mL of arginine in water.
times in seconds taken: Adjust with phosphoric acid to a pH of 7.3.
Standard stock solution: 1 mg/mL of USP Alteplase RS in
log t = m(log US) + b water
Standard solution: 0.25 mg/mL USP Alteplase RS from
t = time to bubble release (s) Standard stock solution in Arginine solution. Heat 0.5 mL of
m = slope of the line this solution with 116 L of SDS buffer and 10 L of 1 M
US = activity of the Standard solution (USP Alteplase dithiothreitol at 80 for 2 min.
Units/mL) Carboxymethylated standard solution: Dilute 1.0 mL of
b = y-intercept of the line Standard stock solution with 1 mL of Carboxymethylation
[NOTEThe correlation coefficient is NLT 0.9900.] buffer, and adjust with 1 M sodium hydroxide to a pH of
From the line equation and using the log of the clot lysis 8.5. Add 20 L of 1 M dithiothreitol, and incubate at 37
time for the Sample solution, calculate the log of the for 60 min. Add 100 L of 1 M iodoacetic acid, and
activity (UA): incubate in the dark for 20 min. Desalt by passing the
solution through a chromatographic column containing
log UA = [(log t)-b]/m fine gel chromatographic packing equilibrated with a buffer
solution containing, in each mL 20 mg of sodium dodecyl
Calculate the alteplase activity in USP Alteplase Units/mL sulfate, 100 mg of glycerol, 1.42 mg of
taken: tris(hydroxymethyl)aminomethane hydrochloride, and 0.85
Result = D(10logU) mg of tris(hydroxymethyl)aminomethane. Collect the
protein fraction of the preparation by elution with the
D = dilution factor for the Sample solution same buffer, and add 20 L of 1 M dithiothreitol. Adjust
Calculate the specific activity in the portion of Alteplase the protein concentration to about 0.2 mg/mL with a
taken: buffer solution containing, in each mL 20 mg of sodium
dodecyl sulfate, 100 mg of glycerol, 1.42 mg of
Result = UA/P tris(hydroxymethyl)aminomethane hydrochloride, 0.85 mg
of tris(hydroxymethyl)aminomethane, 1.06 mg of
P = concentration of protein obtained in the test for dithiothreitol, 0.05 mg of bromophenol blue, and 0.05 mg
Protein Content of xylene cyanole FF.
Acceptance criteria: NLT 90% and NMT 115% of the Sample stock solution, Sample solution, and
potency stated on the label, the potency being 580,000 USP Carboxymethylated sample solution: Using Alteplase,
Alteplase Units/mg of protein proceed as directed for Standard stock solution, Standard
solution, and Carboxymethylated standard solution.
IMPURITIES Molecular weight standard solution: Use a commercially
Organic Impurities available preparation of low molecular weight protein
PROCEDURE standards (10,000 to 100,000 Da) at 2 mg/mL. Mix 990 L
(See Electrophoresis 726.) of Diluted SDS buffer and 10 L of the molecular weight
SDS buffer: 400 mg/mL of glycerol, 5.52 mg/mL of standard mixture.
tris(hydroxymethyl)aminomethane hydrochloride, 3.28 Control solution: Prepare a control solution of bovine
mg/mL of tris(hydroxymethyl)aminomethane, 0.20 mg/mL serum albumin containing 10 g/ mL. For a 10 ng/25 L
of bromophenol blue, and 0.20 mg/mL of xylene cyanole load, mix 600 L of Diluted SDS buffer and 25 L of the
FF in sodium dodecyl sulfate solution (8 in 100) control solution, and heat at 90 for 2 min. For a 2.5 ng
Diluted SDS buffer: Dilute 1 volume of SDS buffer with per 25 L load, mix 594 L of Diluted SDS buffer and 6 L
four volumes of water. of the control solution, and heat at 90 for 2 min.
Ammoniacal silver nitrate solution: Transfer 105 mL of Blank: Mix 500 L of water, 126 L of SDS buffer, and 10
sodium hydroxide solution (0.36 in 100) and 7.0 mL of L of 1 M dithiothreitol.
ammonium hydroxide to a 500-mL volumetric flask, and Analysis
add slowly, with stirring, 20.0 mL of silver nitrate solution Samples: Sample solution, Standard solution,
(20 in 100). Dilute with water to volume. Carboxymethylated sample solution, and Carboxymethylated
[NOTEPrepare this solution immediately before use, and standard solution, Control solution, Molecular weight
protect it from light. This amount of solution is sufficient standard solution, and Blank.
for two slab gels.] Separately apply equal volumes (about 25 L) of the
Citric acidformaldehyde solution: To 500 mL of water Sample solution, Standard solution, Carboxymethylated
add 25 mg of citric acid, 0.25 mL of formaldehyde, and sample solution, and Carboxymethylated standard solution
0.025 mL of methanol, omitting the methanol if the at the 5 g load; apply equal volumes (about 38 L) of
formaldehyde is preserved with methanol. the Standard solution and the Carboxymethylated standard
[NOTEPrepare this solution fresh at the time of use. This solution at the 7.5 g load; and apply the Control
amount of solution is sufficient for two slab gels.]
solutions at the 10 and 2.5 ng load onto separate lanes Mode: LC

of the gel. Apply about 25 L of the Molecular weight Detector: UV 214 nm
standard solution to each side of the gel, and apply about Column: 7.5-mm 60-cm; packing L25
25 L of the Blank onto a separate lane. Apply the Flow rate: 0.5 mL/min
Sample solution and the Standard solution on one half Injection size: 50 L
and the Carboxymethylated sample solution and System suitability
Carboxymethylated standard solution on the other half. Sample: Standard solution
Perform the electrophoresis using a constant current of Suitability requirements
1.31.5 mAmp/cm of gel length and the Running buffer. Resolution: NLT 1.1 between the single-chain and two-
Remove the gel from the apparatus 1020 min after the chain alteplase peaks
tracking dye starts to move. Place the gel in 250 mL of a Analysis
solution of 20% alcohol and 6% glacial acetic acid for Samples: Standard solution and Sample solution
NLT 1 h, and change the solution every 20 min, leaving [NOTEThe major peaks are from single-chain and two-
the gel to soak overnight following the last change. chain alteplase and from higher and lower molecular
Perform silver staining of the gel by placing the gel in weight species.]
250 mL of a solution (1 in 10) in a shallow dish, and Calculate the percentage of single-chain alteplase in the
shake for about 30 min. Replace the glutaraldehyde portion of Alteplase taken:
solution with distilled water, allow the gel to soak for
about 20 min, and then change the water. Repeat for a Result = (rA/rS) 100
total of three washings. Transfer the gel to a dish, and
cover with 250 mL of Ammoniacal silver nitrate solution. rA = peak response for single-chain alteplase
Place the dish on a shaker for about 15 min. Rinse four rS = sum of the responses of all the alteplase peaks
times with 250 mL of water, rocking the dish for 1 min Acceptance criteria: NLT 60%; no peaks or shoulders in
between rinses. Continue rocking to prevent the gel from the Sample solution that are not present in the Standard
sticking and to facilitate washing. solution are found.
Transfer the gel to a clear dish containing 250 mL of Citric PEPTIDE MAPPING
acidformaldehyde solution, and rock the dish. Protein Solution A: 6.9 mg/mL of monobasic sodium phosphate in
bands become visible. When the gel is visibly stained, water, adjusted with phosphoric acid to a pH of 2.85
wash immediately with water, and rinse it repeatedly [NOTEMake adjustments if necessary.]
with water to remove the Citric acidformaldehyde Solution B: Acetonitrile
solution. Rinse the gel for NLT 1 h, and dry. Soak Mobile phase: See the gradient table below.
cellophane membranes in glycerol solution (2 in 100). [NOTEEquilibrate the system before use with 100%
Roll a membrane onto a rigid sheet of plastic. Roll the Solution A.]
gel onto the membrane, and cover with another
membrane. Lay a frame on the edges of the membranes, Time Solution A Solution B
and clamp it to the rigid plastic sheet. Dismantle the (min) (%) (%)
dryer, and cut off excess cellophane when dry (about 24 0 100 0
h). Visually examine the gel under light.
Acceptance criteria: The Sample solution exhibits three 90 70 30
major bands in the region between 66,000 Da and 31,000 120 40 60
Da, corresponding to the major bands from the Standard 130 40 60
solution. The Carboxymethylated sample solution exhibits six
major bands in the region between 92,500 Da and 45,000 Dialysis solution: 480 mg/mL of urea, 44 mg/mL of
Da, corresponding to the major bands from the tris(hydroxymethyl)aminomethane, and 0.88 mg/mL of
Carboxymethylated standard solution. edetic acid in water
System suitability: The 2.5 and 10ng Controls must be [NOTEAdjust with hydrochloric acid to a pH of 8.6.]
visible. The nonreduced Control solutions migrate with an Standard solution: Prepare a solution containing 1.0
apparent molecular weight of slightly less than 66,000 Da, mg/mL of USP Alteplase RS in water. Dialyze 2.0 mL of this
as compared with the molecular weight standards. solution into the Dialysis solution at room temperature for
SPECIFIC TESTS NLT 12 h. Measure the volume of the solution, and transfer
BACTERIAL ENDOTOXINS TEST 85: NMT 1 USP Endotoxin it to a clean test tube. For each mL of solution in the tube,
Unit/mg of alteplase add 10 L of 1 M dithiothreitol. Incubate at room
SINGLE-CHAIN CONTENT temperature for 4 h, then add 25 L of 1 M iodoacetic
Mobile phase: 27.6 g of monobasic sodium phosphate in acid/mL of the solution, and incubate in the dark for 30
1000 mL of sodium dodecyl sulfate solution. Adjust with min. Quench the reaction by adding 50 L of 1 M
sodium hydroxide to a pH of 6.8. dithiothreitol/mL of the solution. Dialyze the solution against
Dithiothreitol solution: 3.12 mg/mL of dithiothreitol in 0.1 M ammonium bicarbonate for 24 h, replacing the 0.1 M
Mobile phase ammonium bicarbonate twice during the dialysis period. To
Standard stock solution: 1 mg/mL of USP Alteplase RS in 2.0 mL of the dialyzed solution, add 20 g of trypsin, and
water incubate for 68 h at room temperature. Again add 20 g
Standard solution: Pipet 1 mL of the Standard stock of trypsin, and incubate for 1618 h for a total of 24 h of
solution into a glass tube, add 3 mL of Dithiothreitol solution, incubation of the trypsin-treated solution.
cap the tube, and invert to mix. Heat for 3 to 5 min at [NOTEStore this Standard solution in a freezer.]
about 80. Sample solution: Using a quantity of Sample, proceed as
Sample stock solution: 1 mg/mL of Alteplase in water directed for the Standard solution.
Sample solution: Pipet 1 mL of the Sample stock solution Chromatographic system
into a glass tube, add 3 mL of Dithiothreitol solution, cap the (See Chromatography 621, System Suitability.)
tube, and invert to mix. Heat for 3 to 5 min at about 80.
Chromatographic System
Mode: LC 95% and NMT 111% of the total protein content stated on
Detector: UV 214 nm the label.
Flow rate: 1 mL/min IDENTIFICATION
Injection size: 100 L A. PROCEDURE
System suitability Arginine solution: 0.2 M Arginine that has been adjusted
Sample: Standard solution with phosphoric acid (negative control) to a pH of 7.3
Suitability requirements Standard solution: Prepare a concentration similar to that
Requirement 1: NLT 1.5 between peaks 6 and 7 as of the Sample solution using USP Alteplase RS.
defined by the USP Alteplase RS Data Sheet Sample solution: 1.02.5 mg/mL of Alteplase in water
Requirement 2: NMT 0.5 min for their baseline widths Analysis
Analysis Samples: Sample solution and Standard solution
Samples: Standard solution, Sample Solution, and a mixture To each of three test tubes transfer 1 mL of 0.5 mg/mL H-
of the Standard solution and the Sample solution (1:1) D-isoleucyl-prolyl-arginyl-p-nitroaniline dihydrochloride
[NOTEMeasure the responses for NLT 20 major peaks as containing 0.5 mg/mL. Separately transfer 200 L of the
defined in the USP Alteplase RS Data Sheet.] Sample solution and 200 L of the Standard solution to
Acceptance criteria: The retention times of corresponding two of the test tubes, and to the third test tube, add 200
peaks from the Standard solution and the Sample solution do L of Arginine solution. Mix the solutions in the three test
not differ by more than 0.4 min, and the peak area ratios tubes, and allow to stand for 1 min.
relative to peak 19 (as shown on the USP Alteplase RS Data Acceptance criteria: A yellow color is produced in the
Sheet) do not differ by more than 20%. No additional solutions containing the Sample solution and the Standard
significant peaks or shoulders are found, a significant peak solution, while no yellow color is produced by the Arginine
or shoulder being defined as one having a peak area solution negative control.
response of NLT 5% of peak 19. B. PEPTIDE MAPPING
PROTEIN CONTENT Solution A: 6.9 mg/mL of monobasic sodium phosphate in
Arginine solution: 34.8 mg/mL of arginine in water. Adjust water, adjusted with phosphoric acid to a pH of 2.85
with phosphoric acid to a pH of 7.3. [NOTEFilter and degas. Make adjustments if necessary.]
Sample stock solution: 1 mg/mL of Alteplase in water Solution B: Acetonitrile
Sample solution: Dilute a volume of Sample stock solution Mobile phase: See the gradient table below.
with a volume of Arginine solution to obtain a solution [NOTEEquilibrate the system before use with 100%
having an absorbance value of 0.51.0 at the wavelength of Solution A.]
maximum absorbance at about 280 nm. Determine the
dilution volume (V). Time Solution A Solution B
Spectrometric conditions (min) (%) (%)
(See Spectroscopy and Light-Scattering 851.) 0 100 0
Mode: UV
Wavelength range: 240500 nm 90 70 30
Analytical wavelengths: 320 nm and peak maxima about 120 40 60
280 nm 130 40 60
Cell: 1 cm
Blank: Arginine solution Dialysis solution: 480 mg/mL of urea, 44 mg/mL of
Analysis tris(hydroxymethyl)aminomethane, and 0.88 mg/mL of
Samples: Sample solution and Blank edetic acid in water.
Calculate the protein content in the portion of Alteplase [NOTEAdjust with hydrochloric acid to a pH of 8.6.]
taken: Standard solution: Prepare a solution containing 1.0
Result = (Amax A320)/1.9 V mg/mL of USP Alteplase RS in water. Dialyze 2.0 mL of this
solution into the Dialysis solution at room temperature for
Amax = absorbance value at the wavelength of NLT 12 h. Measure the volume of the solution, and transfer
maximum absorbance (at about 280 nm) it to a clean test tube. For each mL of solution in the tube,
A320 = absorbance of the Sample solution at 320 nm add 10 L of 1 M dithiothreitol. Incubate at room
V = volume of 0.2 M Arginine solution required to temperature for 4 h, then add 25 L of 1 M iodoacetic
prepare the Sample solution acid/mL of the solution, and incubate in the dark for 30
min. Quench the reaction by the addition of 50 L of 1 M
ADDITIONAL REQUIREMENTS dithiothreitol/mL of the solution. Dialyze the solution against
PACKAGING AND STORAGE: Preserve in tight containers, and 0.1 M ammonium bicarbonate for 24 h, replacing the 0.1 M
store in the frozen state at a temperature of 20 or below. ammonium bicarbonate twice during the dialysis period. To
USP REFERENCE STANDARDS 11 2.0 mL of the dialyzed solution, add 20 g of trypsin, and
USP Alteplase RS incubate for 68 h at room temperature. Again add 20 g
USP Endotoxin RS of trypsin, and incubate for 1618 h for a total of 24 h of
incubation of the trypsin-treated solution. [NOTEStore in a
freezer.]
Sample solution: Prepare a solution containing 1.0 mg/mL
Alteplase for Injection of alteplase in water. Dialyze 2.0 mL of this solution into the
(Comment on this Monograph)id=m1673=Alteplase for Dialysis solution at room temperature for NLT 12 h. Measure
Injection=A-Monos.pdf) the volume of the solution, and transfer it to a clean test
tube. For each mL of solution in the tube, add 10 L of 1 M
DEFINITION dithiothreitol. Incubate at room temperature for 4 h, then
Alteplase for Injection is a sterile lyophilized preparation of add 25 L of 1 M iodoacetic acid/mL of the solution, and
Alteplase. Its biological activity is NLT 90% and NMT 115% of incubate in the dark for 30 min. Quench the reaction by the
that stated on the label in USP Alteplase Units. It contains NLT
addition of 50 L of 1 M dithiothreitol/mL of the solution. plasminogenfibrinogen mixture. Using a vortex mixer,

Dialyze the solution against 0.1 M ammonium bicarbonate intermittently mix the contents of each tube for a total of
for 24 h, replacing the 0.1 M ammonium bicarbonate twice 15 s, and carefully place into a rack in a 37 circulating
during the dialysis period. To 2.0 mL of the dialyzed water bath. A visually turbid clot forms within 30 s,
solution, add 20 g of trypsin, and incubate for 68 h at followed by the formation of bubbles within the clot.
room temperature. Again add 20 g of trypsin, and Record the clot lysis time, tcl, from the first addition of the
incubate for 1618 h for a total of 24 h of incubation of the Alteplase solution to the last bubble to rise to the surface.
trypsin-treated solution. [NOTEStore in a freezer.] Using a least squares fit, determine the equation of the
Chromatographic system line using the log values of the standard concentration, in
(See Chromatography 621, System Suitability.) USP Alteplase Units/mL, versus the log values of their clot
Mode: LC lysis times in s taken:
Detector: UV 214 nm
Column: 4.6-mm 10-cm; packing L1 log t = m(log US) + b
Flow rate: 1 mL/min
Injection size: 100 L t = time to bubble release(s)
System suitability US = activity of the Standard solution (USP Alteplase
Sample: Standard solution Units/mL)
Suitability requirements m = slope of the line
Requirement 1: The resolution between peaks 6 and 7 as b = y-intercept of the line
defined by the USP Alteplase RS Data Sheet is NLT 1.5 [NOTEThe correlation coefficient is NLT 0.9900.]
Requirement 2: The width of the peaks at the baseline From the line equation and using the log of the clot lysis
are NMT 0.5 min. time for the Sample solution, calculate the log of the
Analysis activity, UA:
Samples: Standard solution, Sample Solution,and a mixture
of the Standard solution and the Sample solution (1:1) log UA = [(log t) b]/m
[NOTEMeasure the responses for NLT 20 major peaks as
defined in the USP Alteplase RS Data Sheet.] Calculate the alteplase activity, in USP Alteplase Units/mL:
Acceptance criteria: The retention times of corresponding Result = D(10log U)
peaks from the Standard solution and the Sample solution do
not differ by more than 0.4 min, and the peak area ratios D = dilution factor for the Sample solution
relative to peak 19 (as shown on the USP Alteplase RS Data Calculate the specific activity in the portion of Alteplase:
Sheet) do not differ by more than 20%. No additional
significant peaks or shoulders are found, a significant peak Result = UA/P
or shoulder being defined as one having a peak area
response of NLT 5% of peak 19. P = concentration of protein obtained in the test
Procedure 1: Content of Protein
ASSAY Acceptance criteria: 90.0%115.0%
BIOLOGICAL POTENCY
[NOTEWhen constituted with water, Alteplase for Injection OTHER COMPONENTS
meets this requirement.] PROCEDURE 1: CONTENT OF PROTEIN
Buffer: 1.38 mg/mL of monobasic sodium phosphate, 7.10 (See Spectrophotometry and Light-Scattering 851.)
mg/mL of anhydrous dibasic sodium phosphate, 0.20 Arginine solution: 34.8 mg/mL of arginine
mg/mL of sodium azide, and 0.10 mg/mL of polysorbate 80 [NOTEAdjust with phosphoric acid to a pH of 7.3.]
in water Sample stock solution: 1 mg/mL
Human thrombin solution: 33 U.S. Units/mL in Buffer Sample solution: Dilute a volume of Sample stock solution
[NOTEUnits are in terms of the U.S. Standard Thrombin.] with a volume of Arginine solution to obtain a solution
Human fibrinogen solution: 2 mg/mL of human having an absorbance value of 0.51.0 at the wavelength of
fibrinogen in Buffer maximum absorbance at about 280 nm. Determine the
Human plasminogen solution: 1 mg/mL of human dilution volume, V.
plasminogen in Buffer Spectrometric conditions
Standard stock solution: 1.0 mg/mL (580,000 USP (See Spectrophotometry and Light-Scattering 851.)
Alteplase Units) of USP Alteplase RS Mode: UV
Standard solutions: Dilute volumes of Standard stock Wavelength range: 240500 nm
solution to obtain a series of five Standard solutions having Analytical wavelengths: 320 nm and peak maxima B at
known concentrations ranging from 145 to 9.3 USP about 280 nm
Alteplase Units/mL. Cell: 1 cm
Sample stock solution: 1.0 mg/mL of Alteplase Blank: Arginine solution
Sample solutions: Dilute a volume of Buffer to obtain a Analysis
series of dilutions of 1:20,000, 1:10,000, and 1:5,000. Samples: Sample solution and Blank
Analysis Calculate the protein content in the portion of Alteplase
Samples: Standard solutions and Sample solutions taken:
To a set of labeled glass test tubes, add 0.5 mL of Human
thrombin solution. To separate test tubes, add 0.5 mL of Result = (Amax A320)/F V
each of the Standard solutions and Sample solutions, and
store on ice. To a second set of labeled glass tubes, add Amax = absorbance value at the wavelength of
20 L of Human plasminogen solution and 1 mL of Human maximum absorbance
fibrinogen solution, and store on ice. Beginning with the A320 = absorbance of the Sample solution at 320 nm
thrombin mixture tubes containing the Standard solution F = conversion factor, 1.9
with the lowest number of USP Units/mL, record the time, V = volume of Arginine solution required to prepare
and separately add 200 L of each of the thrombin the Sample solution
mixtures to the test tubes containing the
Acceptance criteria: 95.0%111.0% No peaks or shoulders in the chromatogram of the Sample

PROCEDURE 2: PERCENT MONOMER solution that are not present in the chromatogram of the
Mobile phase: 34.84 mg/mL of arginine, 158.56 mg/mL of Standard solution are found.
ammonium sulfate, and 100 mL/L of isopropyl alcohol in Calculate the percentage of single-chain alteplase in the
water. Adjust with phosphoric acid to a pH of 7.3, degas, portion of Alteplase taken:
and pass through a 0.45-m porosity filter. Make
adjustments if necessary. Result = (rA/rS) 100
System suitability solution: 1.0 mg/mL each of chicken
ovalbumin and bovine gamma globulin rA = peak response for single-chain alteplase
Standard solution: 1 mg/mL of USP Alteplase RS rS = sum of the responses of all of the alteplase peaks
Sample solution: 1 mg/mL Acceptance criteria: NLT 60%
(See Chromatography 621, System Suitability.) PERFORMANCE TESTS
Mode: LC UNIFORMITY OF DOSAGE UNITS 905
Detector: UV 280 nm Acceptance criteria: Meets the requirements for Content
Column: 7.5-mm 30-cm; packing L25 Uniformity
Flow rate: 0.51.0 mL/min SPECIFIC TESTS
Injection size: 50 L CONSTITUTED SOLUTIONS
System suitability Acceptance criteria: At the time of use, it meets the
Sample: System suitability solution and Standard solution requirements for Injections 1, Constituted Solutions.
Suitability requirements PH 791
Resolution: NLT 1.6 between gamma globulin and Sample solution: Constitute as directed in the labeling.
ovalbumin, System suitability solution. Acceptance criteria: 7.17.5
Column efficiency: NLT 1200 theoretical plates, Sample WATER DETERMINATION, Method I 921: NMT 4.0%
solution BACTERIAL ENDOTOXINS 85: NMT 1 USP Endotoxin Unit/mg
Analysis STERILITY TESTS 71
Samples: Sample solution and Standard solution Acceptance criteria: Meets the requirements when tested
Calculate, as a percentage, the monomer in the portion of as directed under Test for Sterility of the Product to Be
Alteplase for Injection taken: Examined, Membrane Filtration.
SAFETY
Result = (rM/rS) 100 Acceptance criteria: Meets the requirements for biologics
rM = peak response for the alteplase monomer as set forth for Biological Reactivity Tests, In Vivo 88, Safety
rS = sum of the responses of all of the alteplase TestsBiologicals.
related peaks ADDITIONAL REQUIREMENTS
Acceptance criteria: NLT 95% PACKAGING AND STORAGE: Preserve in hermetic, light-
PROCEDURE 3: SINGLE-CHAIN CONTENT resistant containers, and store in a refrigerator.
[NOTEWhen constituted with water, Alteplase for Injection LABELING: Label it to state the biological activity in USP
meets this requirement.] Alteplase Units/vial and the amount of protein/vial.
Mobile phase: 27.6 mg/mL of monobasic sodium USP REFERENCE STANDARDS 11
phosphate in sodium dodecyl sulfate solution (1 in 1000). USP Alteplase RS
Adjust with sodium hydroxide to a pH of 6.8, filter, and USP Endotoxin RS
degas. Make adjustments if necessary.
0.02 M dithiothreitol solution: 3.12 mg/mL of
dithiothreitol in Mobile phase
Standard stock solution: 1 mg/mL of USP Alteplase RS Altretamine
Standard solution: Pipet 1 mL of the Standard stock (Comment on this Monograph)id=m1678=Altretamine=A-
solution into a glass tube. Add 3 mL of 0.02 M dithiothreitol Monos.pdf)
solution, cap the tube, and invert to mix. Heat for 35 min
at about 80.
Sample stock solution: 1 mg/mL of Alteplase
Sample solution: Pipet 1 mL of the Sample stock solution
into a glass tube. Add 3 mL of 0.02 M dithiothreitol solution,
cap the tube, and invert to mix. Heat for 35 min at about
80.
(See Chromatography 621, System Suitability.) C9H18N6 210.28
Mode: LC 1,3,5-Triazine-2,4,6-triamine, N,N,N,N,N,N-hexamethyl-;
Detector: UV 214 nm Hexamethylmelamine [645-05-6].
Flow rate: 0.5 mL/min DEFINITION
Injection size: 50 L Altretamine contains NLT 98.0% and NMT 102.0% of C9H18N6,
System suitability calculated on the anhydrous basis.
Suitability requirements IDENTIFICATION
Resolution: NLT 1.1 between the single-chain and two- A. INFRARED ABSORPTION 197K
chain alteplase peaks B. The retention time of the major peak of the Sample
Analysis solution corresponds to that of the Standard solution, as
Samples: Sample solution and Standard solution [NOTEThe obtained in the Assay.
major peaks are from single-chain and two-chain alteplase
and from higher and lower molecular weight species.]
USP 32 Official Monographs / Altretamine 99
ASSAY B. The retention time of the major peak of the Sample

PROCEDURE solution corresponds to that of the Standard solution, as
Buffer: 790 g/mL of ammonium carbonate; if necessary, obtained in the Assay.
adjust to pH of 8.0 0.05 with a solution of formic acid (1
in 10) or ammonium hydroxide (1 in 10). ASSAY
Diluent: Methanol and water (65:35) PROCEDURE
Mobile phase: Methanol and Buffer (65:35) Buffer: 790 mg/L of ammonium carbonate; if necessary,
Standard stock solution: Dissolve 25 mg of USP adjust to pH of 8.0 0.05 with a solution of formic acid (1
Altretamine RS in 35 mL of methanol, and dilute to 50 mL in 10) or ammonium hydroxide (1 in 10).
with water. Diluent: Methanol and water (65:35)
Standard solution: 50 g/mL of Altretamine in Diluent, Mobile phase: Methanol and Buffer (65:35), filtered and
from Standard stock solution degassed
Sample stock solution: Dissolve 25 mg of Sample in 35 mL Standard stock solution: Dissolve 25 mg of USP
of methanol, and dilute to 50 mL with water. Altretamine RS in 35 mL of methanol, and dilute to 50 mL
Sample solution: 50 g/mL of Altretamine in Diluent, from with water.
Sample stock solution Standard solution: 50 g/mL in Diluent from Standard
Chromatographic system stock solution
(See Chromatography 621, System Suitability.) Sample stock solution: Remove as completely as possible
Mode: LC the contents of NLT 20 Capsules, and weigh. Mix the
Detector: UV 227 nm combined contents, and transfer as completely as possible
Column: 4.6-mm 30-cm; packing L1 to a 500-mL volumetric flask. Add 325 mL of methanol, and
Flow rate: 2 mL/min sonicate. Dilute with water to volume.
Injection size: 10 L Sample solution: Transfer a volume of the Sample stock
System suitability solution, equivalent to 10 mg of altretamine, to a 200-mL
Sample: Standard solution volumetric flask and dilute with Diluent to volume.
Suitability requirements Chromatographic system
Tailing factor: NMT 1.5 (See Chromatography 621, System Suitability.)
Relative standard deviation: NMT 2.0% Mode: LC
Analysis Detector: UV 227 nm
Samples: Standard solution and Sample solution Column: 4.6-mm 30-cm; packing L1
Calculate the quantity, as a percentage, of C9H18N6 in the Flow rate: 2 mL/min
portion of Altretamine taken: Injection size: 10 L
System suitability
Result = (rU/rS) (CS/CU) 100 Sample: Standard solution
rU = peak response from the Sample solution Tailing factor: NMT 1.5
rS = peak response from the Standard solution Relative standard deviation: NMT 2.0%
CS = concentration of USP Altretamine RS in Standard Analysis
solution (mg/mL) Sample: Standard solution and Sample solution
CU = concentration of the Sample solution (mg/mL) Calculate the quantity, as a percentage, of C9H18N6 in the
Acceptance criteria: 98.0%102.0% portion of Capsules taken:
IMPURITIES Result = (rU/rS) (CS/CU) 100
RESIDUE ON IGNITION 281: NMT 0.1% rU = peak response from the Sample solution
HEAVY METALS, Method II 231: NMT 40 ppm rS = peak response from the Standard solution
CS = concentration of USP Altretamine RS in the
SPECIFIC TESTS Standard solution (mg/mL)
WATER DETERMINATION, Method I 921: NMT 1% CU = nominal concentration of altretamine in the
ADDITIONAL REQUIREMENTS Acceptance criteria: 90.0%110.0%
store at a controlled room temperature. PERFORMANCE TESTS
USP REFERENCE STANDARDS 11 DISSOLUTION 711
USP Altretamine RS Medium: 0.1 N hydrochloric acid; 900 mL
Apparatus 1: 100 rpm
Time: 30 min
Altretamine Capsules Detector: UV 242 nm
Analysis: Determine the amount of C9H18N6 dissolved from
(Comment on this Monograph)id=m1679=Altretamine UV absorbances on filtered portions of the solution under
Capsules=A-Monos.pdf) test, suitably diluted if necessary with Medium, in
comparison with a standard solution having a known
DEFINITION concentration of USP Altretamine RS in the same Medium.
Altretamine Capsules contain NLT 90.0% and NMT 110.0% of Tolerances: NLT 80% (Q) of the labeled amount of C9H18N6
the labeled amount of altretamine (C9H18N6). is dissolved.
IDENTIFICATION UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
A. INFRARED ABSORPTION 197K ADDITIONAL REQUIREMENTS
Sample: Remove as completely as possible the contents of PACKAGING AND STORAGE: Preserve in tight, light-resistant
5 Capsules, and dissolve, with shaking, in 10 mL of containers, and store at a controlled room temperature.
chloroform. Filter, and evaporate the chloroform solution to
dryness.
100 Altretamine / Official Monographs USP 32
USP REFERENCE STANDARDS 11 Acceptance criteria: 99.0%100.5%

USP Altretamine RS
IMPURITIES
HEAVY METALS, Method I 231
Ammonium Alum Sample solution: Dissolve 1 g in water to make 20 mL
(Comment on this Monograph)id=m1680=Ammonium Alum=A- and add 5 mL of 0.1 N hydrochloric acid. Evaporate the
Monos.pdf) solution in a porcelain evaporating dish to dryness. Treat
the residue with 20 mL of water, and add 50 mg of
AINH4(SO4)2 12H2O 453.33 hydroxylamine hydrochloride. Heat the solution on a steam
AINH4(SO4)2 237.15 bath for 10 min, cool, and dilute with water to 25 mL.
Sulfuric acid, aluminum ammonium salt (2:1:1), dodecahydrate; Analysis: Proceed as directed in General Chapter, except
Aluminum ammonium sulfate (1:1:2), dodecahydrate to add 50 mg of hydroxylamine hydrochloride to the
[7784-26-1]. Standard Preparation.
Anhydrous [7784-25-0]. Acceptance criteria: 20 ppm
IRON
DEFINITION Sample solution: 1 in 150
Ammonium Alum contains NLT 99.0% and NMT 100.5% of Analysis: Add 5 drops of potassium ferrocyanide TS to 20
AlNH4(SO4)2, calculated on the dried basis. mL of the Sample solution.
Acceptance criteria: No blue color is produced
IDENTIFICATION immediately.
A. PROCEDURE
Sample solution: 1 in 20 SPECIFIC TESTS
Analysis: Add 1 N sodium hydroxide dropwise to the LOSS ON DRYING 731
Sample solution. Sample: 2.0 g
Acceptance criteria: A precipitate is formed that dissolves Analysis: Transfer the Sample, in a tared porcelain crucible,
in an excess of the reagent with the evolution of ammonia, to a muffle furnace at 200. Increase the temperature to
recognizable by its odor and its alkaline effect upon 300, and dry at 300 to a constant weight. Cool in a
moistened red litmus paper exposed to the vapor. desiccator, and weigh.
B. IDENTIFICATION TESTSGENERAL, Aluminum 191 and Sulfate Acceptance criteria: The sample loses 45.0%48.0% of its
191: Meets the requirements weight.
Sample solution: 1 in 20
ASSAY
PROCEDURE Potassium Alum
Edetate disodium titrant: 18.6 mg/mL of edetate disodium (Comment on this Monograph)id=m1685=Potassium Alum=A-
in water. Standardize as follows: weigh 2 g of aluminum Monos.pdf)
wire, transfer to a 1000-mL volumetric flask, and add 50 mL
of a mixture of hydrochloric acid and water (1:1). Swirl the AIK(SO4)2 12H2O 474.39
flask to ensure contact of the aluminum and the acid, and AIK(SO4)2 258.21
allow the reaction to proceed until all of the aluminum has Sulfuric acid, aluminum potassium salt (2:1:1), dodecahydrate;
dissolved. Dilute with water to volume. Pipet 10 mL of this Aluminum potassium sulfate (1:1:2) dodecahydrate
solution into a 250-mL beaker. Add, in the order named and [7784-24-9].
with continuous stirring, 25.0 mL of Edetate disodium titrant Anhydrous [10043-67-1].
and 20 mL of acetic acidammonium acetate buffer TS, and
boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 DEFINITION
mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a Potassium Alum contains NLT 99.0% and NMT 100.5% of
bright rose-pink color. Perform a blank determination, AlK(SO4)2, calculated on the dried basis.
substituting 10 mL of water for the aluminum solution, and
make any necessary correction. IDENTIFICATION
Calculate the molarity of the solution taken: A. PROCEDURE
Sample solution: 50 mg/mL in water
Result = W/Ar V Analysis: Add 1 N sodium hydroxide dropwise to the
Sample solution.
W = weight of aluminum (mg) Acceptance criteria: A precipitate is formed that dissolves
Ar = atomic weight of aluminum, 26.98 in an excess of the reagent. Ammonia is not evolved
V = volume of Edetate disodium titrate (mL) (distinction from ammonium alum).
Sample: 800 mg B. PROCEDURE
Analysis: Transfer the Sample to a 400-mL beaker, moisten Analysis: Hold a sample in a nonluminous flame.
with 1 mL of glacial acetic acid, and add 50 mL of water, Acceptance criteria: A violet color is imparted to the flame.
50.0 mL of Edetate disodium titrant and 20 mL of acetic C. PROCEDURE
acidammonium acetate buffer TS. Warm on a steam bath Sample solution: Saturated solution in water
until solution is complete, and boil gently for 5 min. Cool, Analysis: Add 10 mL of sodium bitartrate TS to 5 mL of the
add 50 mL of alcohol and 2 mL of dithizone TS, and titrate Sample solution.
0.05 M zinc sulfate VS to a bright rose-pink color. Perform a Acceptance criteria: A white, crystalline precipitate is
blank determination, and make any necessary correction. formed within 30 min.
Each mL of 0.05 M Edetate disodium titrant is equivalent to
11.86 mg of AlNH4 (SO4)2.
USP 32 Official Monographs / Alumina 101
D. IDENTIFICATION TESTSGENERAL, Aluminum and Sulfate 191 ADDITIONAL REQUIREMENTS

Sample solution: 50 mg/mL in water PACKAGING AND STORAGE: Preserve in tight containers, and
Acceptance criteria: Meets the requirements store at room temperature.
ASSAY
PROCEDURE
Edetate disodium titrant: 18.6 mg/mL of edetate disodium Alumina and Magnesia Oral Suspension
in water. Standardize as follows: Weigh 2 g of aluminum (Comment on this Monograph)id=m1700=Alumina and
wire, transfer to a 1000-mL volumetric flask, and add 50 mL Magnesia Oral Suspension=A-Monos.pdf)
of a mixture of hydrochloric acid and water (1:1). Swirl the
flask to ensure contact of the aluminum and the acid, and DEFINITION
allow the reaction to proceed until all of the aluminum has Alumina and Magnesia Oral Suspension is a mixture containing
dissolved. Dilute with water to volume. Pipet 10 mL of this aluminum hydroxide [Al(OH)3] and Magnesium Hydroxide
solution into a 250-mL beaker and add, in the order named [Mg(OH)2]. It contains the equivalent of NLT 90.0% and NMT
and with continuous stirring, 25.0 mL of Edetate disodium 110.0% of the labeled amounts of aluminum hydroxide
titrant and 20 mL of acetic acidammonium acetate buffer [Al(OH)3] and magnesium hydroxide [Mg(OH)2]. It may
TS, and boil gently for 5 min. Cool, and add 50 mL of contain a flavoring agent, and may contain suitable
alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc antimicrobial agents.
sulfate VS to a bright rose-pink color. Perform a blank
determination, substituting 10 mL of water for the IDENTIFICATION
aluminum solution, and make any necessary correction. A. IDENTIFICATION TESTSGENERAL, Magnesium 191
Calculate the molarity of the solution taken: Sample solution: To a solution of 5 g in 10 mL of 3 N
hydrochloric acid add 5 drops of methyl red TS, heat to
Result = W/Ar V boiling, add 6 N ammonium hydroxide until the color of the
solution changes to deep yellow, then continue boiling for 2
W = weight of aluminum in the portion of solution min, and filter.
taken (mg) Acceptance criteria: The filtrate meets the requirements.
Ar = atomic weight of aluminum (Al), 26.98 B. IDENTIFICATION TESTSGENERAL, Aluminum 191
V = volume of Edetate disodium titrant consumed Sample solution: Wash the precipitate obtained in
(mL) Identification A with 20 mg/mL of hot ammonium chloride
Sample: 800 mg solution, and dissolve the precipitate in hydrochloric acid.
Analysis: Transfer the Sample to a 400-mL beaker, moisten Acceptance criteria: The solution meets the requirements.
with 1 mL of glacial acetic acid, and add 50 mL of water,
50.0 mL of Edetate disodium titrant, and 20 mL of acetic ASSAY
acidammonium acetate buffer TS. Warm on a steam bath ALUMINUM HYDROXIDE
until solution is complete, and boil gently for 5 min. Cool, Edetate disodium titrant: 18.6 mg/mL of edetate disodium
add 50 mL of alcohol and 2 mL of dithizone TS, and titrate in water. Standardize as follows: Weigh 2 g of aluminum
0.05 M zinc sulfate VS to a bright rose-pink color. Perform a wire, transfer to a 1000-mL volumetric flask, and add 50 mL
blank determination, and make any necessary correction. of a mixture of hydrochloric acid and water (1:1). Swirl the
Each mL of 0.05 M Edetate disodium titrant is equivalent to flask to ensure contact of the aluminum and the acid, and
12.91 mg of AlK(SO4)2. allow the reaction to proceed until all of the aluminum has
Acceptance criteria: 99.0% 100.5% dissolved. Dilute with water to volume. Pipet 10 mL of this
solution into a 250-mL beaker and add, in the order named
IMPURITIES and with continuous stirring, 25.0 mL of Edetate disodium
Inorganic Impurities titrant and 20 mL of acetic acidammonium acetate buffer
HEAVY METALS, Method I 231 TS, and boil gently for 5 min. Cool, and add 50 mL of
Sample solution: Dissolve 1 g in water to make 20 mL, alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc
and add 5 mL of 0.1 N hydrochloric acid. Evaporate the sulfate VS to a bright rose-pink color. Perform a blank
solution in a porcelain evaporating dish to dryness. Treat determination, substituting 10 mL of water for the
the residue with 20 mL of water, and add 50 mg of aluminum solution.
hydroxylamine hydrochloride. Heat the solution on a steam Calculate the molarity of the solution taken:
bath for 10 min, cool, and dilute with water to 25 mL.
Analysis: Proceed as directed, except to add 50 mg of Result = W/Ar V
hydroxylamine hydrochloride to the Standard Preparation.
Acceptance criteria: 20 ppm W = weight of aluminum in the portion of solution
IRON 241 taken (mg)
Sample solution: 1 in 150 Ar = atomic weight of aluminum; 26.98
Analysis: Add 5 drops of potassium ferrocyanide TS to 20 V = volume of Edetate disodium titrant consumed
mL of the Sample solution. (mL)
Acceptance criteria: No blue color is produced Sample solution: Transfer a volume of Oral Suspension,
immediately. previously well shaken in its original container, equivalent to
1200 mg of aluminum hydroxide, to a suitable beaker. Add
SPECIFIC TESTS 20 mL of water, stir, and slowly add 10 mL of hydrochloric
LOSS ON DRYING 731 acid. Heat gently, if necessary, to aid solution, cool, and
Sample: 2.0 g filter into a 200-mL volumetric flask. Wash the filter with
Analysis: Transfer the Sample solution in a tared porcelain water into the flask, and add water to volume.
crucible to a muffle furnace at 200. Increase the Analysis: Pipet 10 mL of the Sample solution into a beaker,
temperature to 400, and dry at 400 to constant weight. add 20 mL of water, then add, in the order named and with
Cool in a desiccator, and weigh. continuous stirring, 25.0 mL of Edetate disodium titrant, 20
Acceptance criteria: The sample loses 43.0%46.0% of its mL of acetic acidammonium acetate buffer TS, and heat
weight.
102 Alumina / Official Monographs USP 32
near the boiling point for 5 min. Cool, and add 50 mL of ANC1 = theoretical acid-neutralizing capacity of Al(OH)3,
alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc 0.0385 mEq
sulfate VS until the color changes from green-violet to rose- A = amount of Al(OH)3 in the specimen tested, based
pink. Perform a blank determination, substituting 10 mL of on the labeled quantity (mg)
water for the Sample solution. Each mL of 0.05 M Edetate ANC2 = theoretical acid-neutralizing capacity of Mg(OH)2,
disodium titrant consumed is equivalent to 3.900 mg of 0.0343 mEq
Al(OH)3. M = amount of Mg(OH)2 in the specimen tested,
Acceptance criteria: 90.0%110.0% based on the labeled quantity (mg)
MAGNESIUM HYDROXIDE
Sample solution: Proceed as directed under Assay, ADDITIONAL REQUIREMENTS
Aluminum hydroxide. PACKAGING AND STORAGE: Preserve in tight containers, and
Analysis: Pipet a volume of Sample solution, equivalent to avoid freezing.
40 mg of magnesium hydroxide, into a 400-mL beaker. Add LABELING: Oral Suspension may be labeled to state the
200 mL of water and 20 mL of triethanolamine, and stir. aluminum hydroxide content in terms of the equivalent
Add 10 mL of ammoniaammonium chloride buffer TS and amount of dried aluminum hydroxide gel, on the basis that
3 drops of an eriochrome black indicator solution (prepared each mg of dried gel is equivalent to 0.765 mg of Al(OH)3.
by dissolving 200 mg of eriochrome black T in a mixture of
15 mL of triethanolamine and 5 mL of dehydrated alcohol,
and mixing). Cool the solution to between 3 and 4 by Alumina and Magnesia Tablets
immersion of the beaker in an ice bath, then remove, and
titrate with 0.05 M edetate disodium VS to a blue endpoint. (Comment on this Monograph)id=m1703=Alumina and
Perform a blank determination, substituting 10 mL of water Magnesia Tablets=A-Monos.pdf)
for the Sample solution. Each mL of 0.05 M edetate disodium
consumed is equivalent to 2.916 mg of Mg(OH)2. DEFINITION
Acceptance criteria: 90.0%110.0% Alumina and Magnesia Tablets contain NLT 90.0% and NMT
110.0% of the labeled amounts of aluminum hydroxide
IMPURITIES [Al(OH)3] and magnesium hydroxide [Mg(OH)2].
CHLORIDE AND SULFATE, Chloride 221 IDENTIFICATION
Sample solution: Dissolve 5.0 g in the minimum volume A. IDENTIFICATION TESTSGENERAL, Magnesium 191
of nitric acid required to achieve complete solution, add 1 Sample solution: To a 0.7-g portion of finely powdered
mL of acid in excess, then add water to make 100 mL, and Tablets, add 10 mL of 3 N hydrochloric acid and 5 drops of
filter. methyl red TS, heat to boiling, and add 6 N ammonium
Acceptance criteria: A 10-mL portion of the Sample hydroxide until the color of the solution changes to deep
solution shows no more chloride than corresponds to 1.0 yellow. Continue boiling for 2 min, and filter: the filtrate
mL of 0.020 N hydrochloric acid (0.14%). meets the requirements of the test.
CHLORIDE AND SULFATE, Sulfate 221 B. IDENTIFICATION TESTSGENERAL, Aluminum 191
Sample solution: Dissolve 5.0 g in 5 mL of 3 N Sample solution: Wash the precipitate obtained in
hydrochloric acid, with gentle heating. Cool, add water to Identification test A with hot ammonium chloride solution (1
make 250 mL, and filter. in 50), and dissolve the precipitate in hydrochloric acid: the
Acceptance criteria: A 20-mL portion of the Sample solution meets the requirements of the test.
solution shows no more sulfate than corresponds to 0.40 ASSAY
mL of 0.020 N sulfuric acid (0.1%). ALUMINUM HYDROXIDE
ARSENIC, Method I 211 Edetate disodium titrant: 18.6 mg/mL of edetate disodium
Standard solution: Prepare as directed in Arsenic 211, in water
except prepare it to contain 5 g of arsenic instead of 3 Standardization of titrant: Transfer 2 g of aluminum wire
g. to a 1000-mL volumetric flask, and add 50 mL of a mixture
Sample solution: Oral Suspension, equivalent to 0.5 g of of hydrochloric acid and water (1:1). Swirl the flask to
Al(OH)3, in 20 mL of 7 N sulfuric acid ensure contact of the aluminum and the acid, and allow the
Acceptance criteria: NMT 10 ppm, based on the Al(OH)3 reaction to proceed until all of the aluminum has dissolved.
content Dilute with water to volume. Pipet 10 mL of this solution
HEAVY METALS 231 into a 250-mL beaker, add, in the order named and with
Sample solution: Oral Suspension, equivalent to 0.24 g of continuous stirring, 25 mL of Edetate disodium titrant and 20
Al(OH)3, in 10 mL of 3 N hydrochloric acid with the aid of mL of acetic acidammonium acetate buffer TS, and boil
heat, filter, if necessary, and dilute with water to 25 mL gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL
Acceptance criteria: NMT 83 ppm, based on the Al(OH)3 of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a
content bright rose-pink color. Perform a blank determination,
SPECIFIC TESTS substituting 10 mL of water for the aluminum solution, and
MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED make any necessary correction.
MICROORGANISMS 62 Calculate the molarity of the solution taken:
Acceptance criteria: Its total aerobic microbial count does Result = W/ArV
not exceed 100 cfu/mL, and it meets the requirements for
absence of Escherichia coli. W = weight of aluminum in the portion of solution
PH 791: 7.38.5 taken (mg)
ACID-NEUTRALIZING CAPACITY 301 Ar = atomic weight of aluminum, 26.98
Acceptance criteria: The acid consumed by the minimum V = volume of Edetate disodium titrant consumed
single dose recommended in the labeling is NLT 5 mEq, and (mL)
NLT the number of mEq calculated: Sample solution: Finely powder NLT 20 Tablets. Transfer a
portion of the powder, equivalent to 1200 mg of aluminum
Result = 0.55 (ANC1A) + 0.8 (ANC2M)
hydroxide, to a 150-mL beaker. Add 20 mL of water, stir, Alumina, Magnesia, and Calcium
and slowly add 30 mL of 3 N hydrochloric acid. Heat gently, Carbonate Oral Suspension
if necessary, to aid solution, cool, and filter into a 200-mL
volumetric flask. Wash the filter with water into the flask, (Comment on this Monograph)id=m1710=Alumina, Magnesia,
and add water to volume. and Calcium Carbonate Oral Suspension=A-Monos.pdf)
Analysis: Pipet 10 mL of Sample solution into a 250-mL DEFINITION
beaker, add 20 mL of water, then add, in the order named Alumina, Magnesia, and Calcium Carbonate Oral Suspension
and with continuous stirring, 25 mL of Edetate disodium contains NLT 90.0% and NMT 110.0% of the labeled amounts
titrant and 20 mL of acetic acidammonium acetate buffer of Al(OH)3, Mg(OH)2, and CaCO3.
TS, and heat near the boiling point for 5 min. Cool, add 50
mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M IDENTIFICATION
zinc sulfate VS until the color changes from green-violet to A. IDENTIFICATION TESTSGENERAL, Calcium 191
rose-pink. Perform a blank determination, substituting 10 Sample solution: To 5 g of Oral Suspension, add 25 mL of
mL of water for the Sample solution, and make any necessary 2 N sulfuric acid, stir, and allow to stand for 5 min. Add 25
correction. Each mL of 0.05 M Edetate disodium titrant is mL of alcohol, stir, and place in an ice bath for 30 min.
equivalent to 3.900 mg of Al(OH)3. Filter while cold. [NOTERetain the filtrate for Identification
Acceptance criteria: Equivalent of 90.0%110.0% of the test B.] Wash the precipitate with 50 mL of 0.75 N sulfuric
labeled amounts of Al(OH)3 acid, and discard the washings. Dissolve the precipitate in 3
MAGNESIUM HYDROXIDE N hydrochloric acid, and filter.
Sample solution: Prepare as directed in the Assay for Acceptance criteria: Meets the requirements
Aluminum Hydroxide. B. IDENTIFICATION TESTSGENERAL, Aluminum 191
Analysis: Pipet a volume of Sample solution, equivalent to Sample solution: To the filtrate obtained in Identification
40 mg of magnesium hydroxide, into a 400-mL beaker. Add test A, add 5 drops of methyl red TS, and heat to boiling.
200 mL of water and 20 mL of triethanolamine, and stir. Add 6 N ammonium hydroxide until the color of the
Add 10 mL of ammoniaammonium chloride buffer TS and solution changes to deep yellow, continue boiling for 2 min,
3 drops of an eriochrome black indicator solution (prepared and filter through hardened filter paper. [NOTERetain the
by dissolving 200 mg of eriochrome black TS in a mixture of filtrate for Identification test C.] Wash the precipitate with
15 mL of triethanolamine and 5 mL of dehydrated alcohol). 350 mL of a hot ammonium chloride solution (1 in 50),
Cool the solution to between 3 and 4 by immersion of the discarding the washings. Dissolve the precipitate so obtained
beaker in an ice bath, then remove, and titrate with 0.05 M in 3 N hydrochloric acid.
edetate disodium VS to a blue endpoint. Perform a blank Acceptance criteria: Meets the requirements.
determination, substituting 10 mL of water for the Sample C. IDENTIFICATION TESTSGENERAL, Aluminum Magnesium 191
solution, and make any necessary correction. Each mL of Sample solution: The filtrate obtained in Identification test B
0.05 M edetate disodium consumed is equivalent to 2.916 Acceptance criteria: Meets the requirements
mg of Mg(OH)2.
Acceptance criteria: Equivalent of 90.0%110.0% of the ASSAY
labeled amounts of Mg(OH)2 ALUMINUM HYDROXIDE
Edetate disodium titrant: 18.6 mg/mL of edetate disodium
PERFORMANCE TESTS in water
DISINTEGRATION 701 Standardization of titrant: Transfer 2 g of aluminum wire
Medium: Simulated gastric fluid TS being substituted for to a 1000-mL volumetric flask, and add 50 mL of a mixture
water in the test of hydrochloric acid and water (1:1). Swirl the flask to
Time: 10 min ensure contact of the aluminum and the acid, and allow the
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements reaction to proceed until all of the aluminum has dissolved.
for Weight Variation with respect to alumina and to magnesia Dilute with water to volume. Pipet 10 mL of this solution
into a 250-mL beaker and add, in the order named and
SPECIFIC TESTS with continuous stirring, 25.0 mL of Edetate disodium titrant
ACID-NEUTRALIZING CAPACITY 301: The acid consumed by and 20 mL of acetic acidammonium acetate buffer TS, and
the minimum single dose recommended in the labeling is boil gently for 5 min. Cool, and add 50 mL of alcohol and 2
NLT 5 mEq, and NLT the number of mEq calculated: mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a
bright rose-pink color. Perform a blank determination,
Result = 0.55(ANC1A) + 0.8(ANC2M) substituting 10 mL of water for the aluminum solution.
ANC1 = theoretical acid-neutralizing capacity of Al(OH)3, Calculate the molarity of the solution taken:
0.0385 mEq Result = W/ArV
A = quantity of Al(OH)3 in the speciment tested,
based on the labeled quantity (mg) W = weight of aluminum in the portion of solution
ANC2 = theoretical acid-neutralizing capacity of Mg(OH)2, taken (mg)
0.0343 mEq Ar = atomic weight of aluminum, 26.98
M = quantitiy of Mg(OH)2 in the specimen tested, V = volume of Edetate disodium titrant consumed
based on the labeled quantity (mg) (mL)
ADDITIONAL REQUIREMENTS Sample solution: Transfer the Oral Suspension, previously
PACKAGING AND STORAGE: Preserve in well-closed containers. well shaken in its original container, equivalent to 600 mg of
LABELING: Tablets prepared with the use of Dried Aluminum aluminum hydroxide, to a tared beaker and weigh. Add 20
Hydroxide Gel may be labeled to state the aluminum mL of water, stir, and slowly add 40 mL of 3 N hydrochloric
hydroxide content in terms of the equivalent amount of acid. Heat gently, if necessary, to aid solution, cool, and
dried aluminum hydroxide gel, on the basis that each mg of transfer to a 200-mL volumetric flask. Wash the beaker with
dried gel is equivalent to 0.765 mg of Al(OH)3.
water, adding the washings to the flask. Add water to CHLORIDE AND SULFATE, Sulfate 221
volume. Sample solution: Dissolve 5.0 g in 7 mL of 3 N
Analysis: Pipet 10 mL of Sample solution into a 250-mL hydrochloric acid, and gently heat. Cool, add water to
beaker, add 20 mL of water, then add, in the order named make 250 ml, and filter.
and with continuous stirring, 25.0 mL of Edetate disodium Acceptance criteria: A 20-mL portion of the Sample
titrant and 20 mL of acetic acidammonium acetate buffer solution shows no more sulfate than corresponds to 0.40
TS, and heat the solution near the boiling temperature for 5 mL of 0.020 N sulfuric acid (0.1%).
min. Cool, and add 50 mL of alcohol and 2 mL of dithizone
TS. Titrate with 0.05 M zinc sulfate VS until the color SPECIFIC TESTS
changes from green-violet to rose-pink. Perform a blank MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED
determination, substituting 10 mL of water for the Sample MICROORGANISMS 62
solution. Each mL of 0.05 M Edetate disodium titrant Acceptance criteria: Its total aerobic microbial count does
consumed is equivalent to 3.900 mg of Al(OH)3. not exceed 100 cfu/mL, and it meets the requirements of
Acceptance criteria: 90.0%110.0% the test for absence of Escherichia coli.
MAGNESIUM HYDROXIDE PH 791
Sample solution: Transfer Oral Suspension, previously well Acceptance criteria: Between 7.5 and 8.5
shaken in its original container, equivalent to 600 mg of ACID-NEUTRALIZING CAPACITY 301
aluminum hydroxide, to a tared beaker and weigh. Add 20 Acceptance criteria: The acid consumed by the minimum
mL of water, stir, and slowly add 40 mL of 3 N hydrochloric single dose recommended in the labeling is NLT 5 mEq, and
acid. Heat gently, if necessary, to aid solution, cool, and NLT the number of mEq calculated:
transfer to a 200-mL volumetric flask. Wash the beaker with
water, adding the washings to the flask. Add water to Result = 0.55(ANC1 A) + 0.8(ANC2 M) + 0.9(ANC3 C)
volume.
Analysis: Pipet a volume of Sample solution, equivalent to ANC1 = theoretical acid-neutralizing capacity of Al(OH)3,
40 mg of magnesium hydroxide, into a 400-mL beaker, and 0.0385 mEq
add 200 mL of water and 20 mL of trolamine, and mix. Add A = amount of Al(OH)3 in the specimen tested, based
50 mL of ammoniaammonium chloride buffer TS and 2 on the labeled quantity (mg)
drops of an eriochrome black indicator solution (prepared ANC2 = theoretical acid-neutralizing capacity of Mg(OH)2,
by dissolving 200 mg of eriochrome black T in a mixture of 0.0343 mEq
15 mL of trolamine and 5 mL of dehydrated alcohol, and M = amount of Mg(OH)2 in the specimen tested,
mixing). Cool the solution to between 3 and 4 by based on the labeled quantity (mg)
immersing the beaker in an ice bath, and titrate with 0.05 ANC3 = theoretical acid-neutralizing capacity of CaCO3,
M edetate disodium VS until the color changes to pure blue. 0.02 mEq
Perform a blank determination, substituting 10 mL of water C = amount of CaCO3 in the specimen tested, based
for the Sample solution. From the volume of 0.05 M edetate on the labeled quantity (mg)
disodium consumed, subtract the volume of 0.05 M edetate OTHER REQUIREMENTS
disodium consumed in the Calcium Carbonate. Each mL of ARSENIC, Method I 211
0.05 M edetate disodium is equivalent to 2.916 mg of Sample solution: Gel, equivalent to 0.5 g of Al(OH)3, in
Mg(OH)2. 20 mL of 7 N sulfuric acid
Acceptance criteria: 90.0%110.0% of the labeled amount Standard solution: Prepare as directed in the test for
of magnesium hydroxide [Mg(OH)2] Arsenic 211, except to prepare it to contain 5 g of
CALCIUM CARBONATE arsenic instead of 3 g.
Sample solution: Transfer the Oral Suspension, previously Acceptance criteria: NMT 10 ppm, based on the Al(OH)3
well shaken in its original container, equivalent to 600 mg of content
aluminum hydroxide, to a tared beaker and weigh. Add 20 HEAVY METALS 231
mL of water, stir, and slowly add 40 mL of 3 N hydrochloric Sample solution: Gel, equivalent to 0.24 g of Al(OH)3, in
acid. Heat gently, if necessary, to aid solution, cool, and 10 mL of 3 N hydrochloric acid with the aid of heat, filter if
transfer to a 200-mL volumetric flask. Wash the beaker with necessary, and dilute with water to 25 mL.
water, adding the washings to the flask. Add water to Acceptance criteria: NMT 83 ppm, based on the Al(OH)3
volume. content
Analysis: Pipet a volume of the Sample solution, equivalent ADDITIONAL REQUIREMENTS
to 50 mg of calcium carbonate, into a 400-mL beaker, and PACKAGING AND STORAGE: Preserve in tight containers, and
add 200 mL of water, 5 mL of sodium hydroxide solution (1 avoid freezing.
in 2), and 250 mg of hydroxy naphthol blue. Stir with a LABELING: Oral Suspension may be labeled to state the
magnetic stirrer, and titrate immediately with 0.05 M aluminum hydroxide content in terms of the equivalent
edetate disodium VS until the solution is distinctly blue. Each amount of dried aluminum hydroxide gel, on the basis that
mL of 0.05 M edetate disodium is equivalent to 5.004 mg of each mg of dried gel is equivalent to 0.765 mg of Al(OH)3.
CaCO3.
Acceptance criteria: NLT 90.0% and NMT 110.0% of the
labeled amount of calcium carbonate (CaCO3)
Alumina, Magnesia, and Calcium
IMPURITIES Carbonate Tablets
CHLORIDE AND SULFATE, Chloride 221 (Comment on this Monograph)id=m1713a=Alumina, Magnesia,
Sample solution: Dissolve 5.0 g 3 mL of nitric acid, add and Calcium Carbonate Tablets=A-Monos.pdf)
water to make 100 mL, and filter. (Current titlenot to change until February 1, 2010)
Acceptance criteria: A 10-mL portion of the Sample Monograph title changeto become official February 1, 2010
solution shows no more chloride than corresponds to 1.0 See Alumina, Magnesia, and Calcium Carbonate Chewable Tablets
mL of 0.020 N hydrochloric acid (0.14%).
DEFINITION and with continuous stirring, 25.0 mL of 0.05 M Edetate

Alumina, Magnesia, and Calcium Carbonate Chewable Tablets disodium titrant and 20 mL of acetic acidammonium
contain NLT 90.0% and NMT 110.0% of the labeled amounts acetate buffer TS, and heat the solution near the boiling
of aluminum hydroxide [Al(OH)3], magnesium hydroxide temperature for 5 min. Cool, add 50 mL of alcohol and 2
[Mg(OH)2], and calcium carbonate (CaCO3). mL of dithizone TS, and mix. Titrate with 0.05 M zinc sulfate
VS until the color changes from green-violet to rose-pink.
IDENTIFICATION Perform a blank determination, substituting 10 mL of water
A. PROCEDURE for the Sample solution, and make any necessary correction.
Sample solution: To 3 g of finely powdered Chewable Each mL of 0.05 M Edetate disodium titrant consumed is
Tablets, add 25 mL of water and 25 mL of 2 N sulfuric acid, equivalent to 3.900 mg of Al(OH)3.
stir, and heat on a steam bath for 10 min. Cool, add 50 mL Acceptance criteria: 90.0%110.0% of the labeled amount
of alcohol, and stir. of aluminum hydroxide [Al(OH)3]
Analysis 1: Place the mixture obtained above in an ice bath MAGNESIUM HYDROXIDE
for 30 min. Filter while cold, retaining the filtrate for Edetate disodium titrant and Sample solution: Prepare as
Identification test B. Wash the precipitate with 50 mL of 0.75 directed in the Assay for Aluminum Hydroxide.
N sulfuric acid, and discard the washings: the precipitate so Analysis: Pipet a volume of Sample solution, equivalent to
obtained, dissolved in 3 N hydrochloric acid and filtered, 40 mg of magnesium hydroxide, into a 400-mL beaker, add
meets the requirements for Identification TestsGeneral, 200 mL of water and 20 mL of trolamine, and mix. Add 50
Calcium 191. mL of ammoniaammonium chloride buffer TS and 2 drops
B. PROCEDURE of eriochrome black indicator solution (prepared by
Analysis 2: To the filtrate obtained in Identification test A dissolving 200 mg of eriochrome black T in a mixture of 15
add 5 drops of methyl red TS, and heat to boiling. Add 6 N mL of trolamine and 5 mL of dehydrated alcohol, and
ammonium hydroxide until the color of the solution mixing). Cool the solution to 34 by immersing the beaker
changes to deep yellow, continue boiling for 2 min, and in an ice bath, and titrate with 0.05 M edetate disodium VS
filter through hardened filter paper. [NOTERetain the until the color changes to pure blue. Perform a blank
filtrate for Identification test C.] Wash the precipitate with determination, substituting 10 mL of water for the Sample
350 mL of a hot ammonium chloride solution (1 in 50), solution, and make any necessary correction. From the
discarding the washings: the precipitate so obtained, volume of 0.05 M edetate disodium consumed, subtract the
dissolved in 3 N hydrochloric acid meets the requirements volume of 0.05 M edetate disodium consumed in the Assay
for Identification TestsGeneral, Aluminum 191. for Calcium Carbonate. Each mL of 0.05 M edetate disodium
C. PROCEDURE is equivalent to 2.916 mg of Mg(OH)2.
Analysis 3: The filtrate obtained in Identification test B Acceptance criteria: 90.0%110.0% of the labeled amount
meets the requirements for Identification TestsGeneral, of magnesium hydroxide [Mg(OH)2]
Magnesium 191. CALCIUM CARBONATE
Edetate disodium titrant and Sample solution: Prepare as
ASSAY directed in the Assay for Aluminum Hydroxide.
ALUMINUM HYDROXIDE Analysis: Pipet a volume of the Sample solution, equivalent
Edetate disodium titrant: 18.6 g of edetate disodium in to 50 mg of calcium carbonate, into a 400-mL beaker, and
water to make 1000 mL add 200 mL of water, 5 mL of sodium hydroxide solution (1
Standardization of titrant: Transfer 2 g of aluminum wire in 2), and 250 mg of hydroxy naphthol blue. Stir with a
to a 1000-mL volumetric flask, and add 50 mL of a mixture magnetic stirrer, and titrate immediately with 0.05 M
of hydrochloric acid and water (1:1). Swirl the flask to edetate disodium VS until the solution is distinctly blue. Each
ensure contact of the aluminum and the acid, and allow the mL of 0.05 M edetate disodium is equivalent to 5.004 mg of
reaction to proceed until all of the aluminum has dissolved. calcium carbonate (CaCO3).
Dilute with water to volume. Pipet 10 mL of this solution Acceptance criteria: 90.0%110.0% of the labeled amount
into a 250-mL beaker, add, in the order named and with of calcium carbonate (CaCO3)
continuous stirring, 25.0 mL of Edetate disodium titrant and
20 mL of acetic acidammonium acetate buffer TS, and boil PERFORMANCE TESTS
gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL DISINTEGRATION 701
of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a Time: 45 min
bright rose-pink color. Perform a blank determination, UNIFORMITY OF DOSAGE UNITS 905
substituting 10 mL of water for the aluminum solution, and Acceptance criteria: Meets the requirements for Weight
make any necessary correction. Calculate the molarity of the Variation with respect to alumina, to magnesia, and to
solution taken: calcium carbonate
Result = W/ArV SPECIFIC TESTS
ACID-NEUTRALIZING CAPACITY 301
W = weight of aluminum in the portion of solution Acceptance criteria: The acid consumed by the minimum
taken (mg) single dose recommended in the labeling is NLT 5 mEq, and
Ar = atomic weight of aluminum, 26.98 NLT the number of mEq calculated:
V = volume of Edetate disodium titrant consumed
(mL) Result = 0.55(ANC1 A) + 0.8(ANC2 M) + 0.9(ANC3 C)
Sample solution: Weigh and finely powder NLT 20
Chewable Tablets. Transfer a portion of the powder, ANC1 = theoretical acid-neutralizing capacity of Al(OH)3,
equivalent to 600 mg of aluminum hydroxide, to a beaker, 0.0385 mEq
add 20 mL of water, and slowly add 40 mL of 3 N A = amount of Al(OH)3 in the specimen tested, based
hydrochloric acid, with mixing. Heat the mixture to boiling, on the labeled quantity (mg)
cool, and filter into a 200-mL volumetric flask. Wash the ANC2 = theoretical acid-neutralizing capacity of Mg(OH)2,
beaker with water, adding the washings to the filter. Add 0.0343 mEq
water to volume. M = amount of Mg(OH)2 in the specimen tested,
Analysis: Pipet 10 mL of the Sample solution into a 250-mL based on the labeled quantity (mg)
beaker, add 20 mL of water, then add, in the order named
ANC3 = theoretical acid-neutralizing capacity of CaCO3, solution into a 250-mL beaker, add, in the order named and
0.02 mEq with continuous stirring, 25.0 mL of Edetate disodium titrant
C = amount of CaCO3 in the specimen tested, based and 20 mL of acetic acidammonium acetate buffer TS, and
on the labeled quantity (mg) boil gently for 5 min. Cool, and add 50 mL of alcohol, and
2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to
ADDITIONAL REQUIREMENTS a bright rose-pink color. Perform a blank determination,
PACKAGING AND STORAGE: Preserve in well-closed containers. substituting 10 mL of water for the aluminum solution, and
LABELING: Label the Chewable Tablets to indicate that they make any necessary correction. Calculate the molarity of the
are to be chewed before being swallowed. Chewable Tablets solution taken:
prepared using dried aluminum hydroxide gel may be
labeled to state the aluminum hydroxide content in terms of Result = W/ArV
the equivalent amount of dried aluminum hydroxide gel, on
the basis that each mg of dried gel is equivalent to 0.765 W = weight of aluminum in the portion of solution
mg of Al(OH)3. taken (mg)
Ar = atomic weight of aluminum, 26.98
(mL)
Alumina, Magnesia, and Calcium Sample solution: Weigh and finely powder NLT 20
Carbonate Chewable Tablets Chewable Tablets. Transfer a portion of the powder,
(Comment on this Monograph)id=m1713b=Alumina, Magnesia, equivalent to 600 mg of aluminum hydroxide, to a beaker,
and Calcium Carbonate Chewable Tablets=A-Monos.pdf) add 20 mL of water, and slowly add 40 mL of 3 N
(Monograph under this new titleto become official February hydrochloric acid, with mixing. Heat the mixture to boiling,
1, 2010) cool, and filter into a 200-mL volumetric flask. Wash the
Current monograph title is Alumina, Magnesia, and Calcium beaker with water, adding the washings to the filter. Add
Carbonate Tablets water to volume.
Analysis: Pipet 10 mL of the Sample solution into a 250-mL
DEFINITION beaker, add 20 mL of water, then add, in the order named
Alumina, Magnesia, and Calcium Carbonate Chewable Tablets and with continuous stirring, 25.0 mL of 0.05 M Edetate
contain NLT 90.0% and NMT 110.0% of the labeled amounts disodium titrant and 20 mL of acetic acidammonium
of aluminum hydroxide [Al(OH)3], magnesium hydroxide acetate buffer TS, and heat the solution near the boiling
[Mg(OH)2], and calcium carbonate (CaCO3). temperature for 5 min. Cool, add 50 mL of alcohol and 2
mL of dithizone TS, and mix. Titrate with 0.05 M zinc sulfate
IDENTIFICATION VS until the color changes from green-violet to rose-pink.
Sample solution: To 3 g of finely powdered Chewable Perform a blank determination, substituting 10 mL of water
Tablets, add 25 mL of water and 25 mL of 2 N sulfuric acid, for the Sample solution, and make any necessary correction.
stir, and heat on a steam bath for 10 min. Cool, add 50 mL of Each mL of 0.05 M Edetate disodium titrant consumed is
alcohol, and stir: the mixture so obtained meets the following equivalent to 3.900 mg of Al(OH)3.
requirements: Acceptance criteria: 90.0%110.0% of the labeled amount
A. PROCEDURE of aluminum hydroxide [Al(OH)3]
Analysis 1: Place the mixture obtained above in an ice bath MAGNESIUM HYDROXIDE
for 30 min. Filter while cold retaining the filtrate for Edetate disodium titrant and Sample solution: Prepare as
Identification test B. Wash the precipitate with 50 mL of 0.75 directed in the Assay for Aluminum Hydroxide.
N sulfuric acid, and discard the washings: the precipitate so Analysis: Pipet a volume of Sample solution, equivalent to
obtained, dissolved in 3 N hydrochloric acid and filtered, 40 mg of magnesium hydroxide, into a 400-mL beaker, add
meets the requirements for Identification TestsGeneral, 200 mL of water and 20 mL of trolamine, and mix. Add 50
Calcium 191. mL of ammoniaammonium chloride buffer TS and 2 drops
B. PROCEDURE of eriochrome black indicator solution (prepared by
Analysis 2: To the filtrate obtained in Identification test A dissolving 200 mg of eriochrome black T in a mixture of 15
add 5 drops of methyl red TS, and heat to boiling. Add 6 N mL of trolamine and 5 mL of dehydrated alcohol, and
ammonium hydroxide until the color of the solution mixing). Cool the solution to 3 to 4 by immersing the
changes to deep yellow, continue boiling for 2 min, and beaker in an ice bath, and titrate with 0.05 M edetate
filter through hardened filter paper. [NOTERetain the disodium VS until the color changes to pure blue. Perform a
filtrate for Identification test C.] Wash the precipitate with blank determination, substituting 10 mL of water for the
350 mL of a hot ammonium chloride solution (1 in 50), Sample solution, and make any necessary correction. From
discarding the washings: the precipitate so obtained, the volume of 0.05 M edetate disodium consumed, subtract
dissolved in 3 N hydrochloric acid meets the requirements the volume of 0.05 M edetate disodium consumed in the
for Identification TestsGeneral, Aluminum 191. Assay for Calcium Carbonate. Each mL of 0.05 M edetate
C. PROCEDURE disodium is equivalent to 2.916 mg of Mg(OH)2.
Analysis 3: The filtrate obtained in Identification test B Acceptance criteria: 90.0%110.0% of the labeled amount
meets the requirements for Identification TestsGeneral, of magnesium hydroxide [Mg(OH)2]
Magnesium 191. CALCIUM CARBONATE
Edetate disodium titrant and Sample solution: Prepare as
ASSAY directed in the Assay for Aluminum Hydroxide.
ALUMINUM HYDROXIDE Analysis: Pipet a volume of the Sample solution, equivalent
Edetate disodium titrant: 18.6 g of edetate disodium in to 50 mg of calcium carbonate, into a 400-mL beaker, and
water add 200 mL of water, 5 mL of sodium hydroxide solution (1
Standardization of titrant: Transfer 2 g of aluminum wire, in 2), and 250 mg of hydroxy naphthol blue. Stir with a
transfer to a 1000-mL volumetric flask, and add 50 mL of a magnetic stirrer, and titrate immediately with 0.05 M
mixture of hydrochloric acid and water (1:1). Swirl the flask edetate disodium VS until the solution is distinctly blue. Each
to ensure contact of the aluminum and the acid, and allow mL of 0.05 M edetate disodium is equivalent to 5.004 mg of
the reaction to proceed until all of the aluminum has calcium carbonate (CaCO3).
dissolved. Dilute with water to volume. Pipet 10 mL of this
Acceptance criteria: 90.0%110.0% of the labeled amount Acceptance criteria: Meets the requirements
of calcium carbonate (CaCO3) B. IDENTIFICATION TESTSGENERAL, Aluminum 191
Sample solution: To the filtrate obtained in Identification
PERFORMANCE TESTS test A, add 5 drops of methyl red TS, and heat to boiling.
DISINTEGRATION 701 Add 6 N ammonium hydroxide until the color of the
Time: 45 min solution changes to deep yellow, continue boiling for 2 min,
UNIFORMITY OF DOSAGE UNITS 905 and filter through hardened filter paper. [NOTERetain the
Acceptance criteria: Meets the requirements for Weight filtrate for Identification test C.] Wash the precipitate with
Variation with respect to alumina, to magnesia, and to 350 mL of a hot ammonium chloride solution (1 in 50),
calcium carbonate. discarding the washings. Dissolve the precipitate so obtained
in 3 N hydrochloric acid.
SPECIFIC TESTS Acceptance criteria: Meets the requirements
ACID-NEUTRALIZING CAPACITY 301 C. IDENTIFICATION TESTSGENERAL, Magnesium 191
Acceptance criteria: The acid consumed by the minimum Sample solution: The filtrate obtained in Identification test B
single dose recommended in the labeling is NLT 5 mEq, and Acceptance criteria: Meets the requirements
NLT the number of mEq calculated: D. INFRARED ABSORPTION 197S
Result = 0.55(ANC1 A) + 0.8(ANC2 M) + 0.9(ANC3 C) Sample solution: Transfer the equivalent to 60 mg of
simethicone from Chewable Tablets (weigh and finely
ANC1 = theoretical acid-neutralizing capacity of Al(OH)3, powder NLT 20 Chewable Tablets), to a suitable screw-
0.0385 mEq capped bottle.
A = amount of Al(OH)3 in the specimen tested, based Add 30.0 mL of chloroform and 60 mL of Dilute hydrochloric
on the labeled quantity (mg) acid (see the Assay for Dimethylpolysiloxane), and allow to
ANC2 = theoretical acid-neutralizing capacity of Mg(OH)2, stand, with frequent shaking, until the Chewable Tablets
0.0343 mEq are dissolved. Transfer the contents of the bottle to a
M = amount of Mg(OH)2 in the specimen tested, separator, shake, and allow the phases to separate. Remove
based on the labeled quantity (mg) 10 mL of the lower, organic layer to a screw-capped, 15-
ANC3 = theoretical acid-neutralizing capacity of CaCO3, mL test tube containing 0.5 g of anhydrous sodium sulfate.
0.02 mEq Close the tube with a screw-cap having an inert liner,
C = amount of CaCO3 in the specimen tested, based agitate vigorously, and centrifuge the mixture until a clear
on the labeled quantity (mg) supernatant is obtained. The supernatant so obtained is the
Sample solution.
ADDITIONAL REQUIREMENTS Analysis: Proceed as directed using a 0.5-mm cell.
LABELING: Label the Chewable Tablets to indicate that they ASSAY
are to be chewed before being swallowed. Chewable Tablets ALUMINUM HYDROXIDE
prepared using dried aluminum hydroxide gel may be Edetate disodium titrant: 18.6 mg/mL of edetate disodium
labeled to state the aluminum hydroxide content in terms of in water
the equivalent amount of dried aluminum hydroxide gel, on Standardization of titrant: Transfer 2 g of aluminum wire,
the basis that each mg of dried gel is equivalent to 0.765 transfer to a 1000-mL volumetric flask, and add 50 mL of a
mg of Al(OH)3. mixture of hydrochloric acid and water (1:1). Swirl the flask
to ensure contact of the aluminum and the acid, and allow
the reaction to proceed until all of the aluminum has
Alumina, Magnesia, Calcium Carbonate, solution into a 250-mL beaker and add, in the order named
and Simethicone Tablets and with continuous stirring, 25.0 mL of Edetate disodium
titrant and 20 mL of acetic acidammonium acetate buffer
(Comment on this Monograph)id=m1720=Alumina, Magnesia, TS, and boil gently for 5 min. Cool, and add 50 mL of
Calcium Carbonate, and Simethicone Tablets=A-Monos.pdf) alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc
(Current titlenot to change until February 1, 2010) sulfate VS to a bright rose-pink color. Perform a blank
Monograph title changeto become official February 1, 2010 determination, substituting 10 mL of water for the
See Alumina, Magnesia, Calcium Carbonate, and Simethicone aluminum solution, and make any necessary correction.
Chewable Tablets. Calculate the molarity of the solution taken:
DEFINITION
Alumina, Magnesia, Calcium Carbonate, and Simethicone Result = W/Ar V
Chewable Tablets contain NLT 90.0% and NMT 110.0% of the W = weight of aluminum in the portion of solution
labeled amounts of aluminum hydroxide [Al(OH)3], taken (mg)
magnesium hydroxide [Mg(OH)2], and calcium carbonate V = volume of Edetate disodium titrant consumed
(CaCO3), and an amount of polydimethylsiloxane ([(CH3)2 (mL)
SiO]n) that is NLT 85.0% and NMT 115.0% of the labeled Ar = atomic weight of aluminum, 26.98
amount of simethicone. Sample solution: Transfer the equivalent to 665 mg of
IDENTIFICATION aluminum hydroxide from a number of Chewable Tablets, to
A. IDENTIFICATION TESTSGENERAL, Calcium 191 a suitable beaker.
Sample solution: Cut a Chewable Tablet into pieces, add Add 15 mL of hydrochloric acid, and swirl to dissolve the
50 mL of 1 N sulfuric acid, stir until the pieces disintegrate, Chewable Tablets. Add 80 mL of water and filter into a
and heat on a steam bath for 10 min. Cool, add 50 mL of 200-mL volumetric flask. Wash the filter with water into the
alcohol, and stir. Place in an ice bath for 30 min. Filter while flask, and add water to volume.
cold. [NOTERetain the filtrate for Identification test B.] Wash Analysis: Pipet 20 mL of Sample solution into a 250-mL
the precipitate with 50 mL of 0.75 N sulfuric acid, and beaker, then add, in the order named and with continuous
discard the washings. Dissolve the precipitate so obtained in stirring, 25.0 mL of Edetate disodium titrant and 20 mL of
3 N hydrochloric acid and filter. acetic acidammonium acetate buffer TS, and heat the
solution near the boiling temperature for 5 min. Cool, add Acceptance criteria: 90.0%110.0%
50 mL of alcohol and 2 mL of dithizone TS. Titrate with CALCIUM CARBONATE
0.05 M zinc sulfate VS until the color changes from green- Sample solution: Transfer the equivalent to 665 mg of
violet to rose-pink. Perform a blank determination, aluminum hydroxide from a number of Chewable Tablets, to
substituting 20 mL of water for the Sample solution, and a suitable beaker.
make any necessary correction. Each mL of 0.05 M Edetate Add 15 mL of hydrochloric acid, and swirl to dissolve the
disodium titrant consumed is equivalent to 3.900 mg of Chewable Tablets. Add 80 mL of water and filter into a
Al(OH)3. 200-mL volumetric flask. Wash the filter with water into the
Acceptance criteria: 90.0%110.0% flask, and add water to volume.
MAGNESIUM HYDROXIDE Analysis: Pipet a volume of the Sample solution, equivalent
Lanthanum chloride solution: Transfer 17.6 g of to 50 mg of calcium carbonate, into a 400-mL beaker, and
lanthanum chloride to a 200-mL volumetric flask, add 100 add 200 mL of water, a volume of sodium hydroxide
mL of water, and carefully add 50 mL of hydrochloric acid. solution (1 in 2) equivalent to the volume of the Sample
Allow to cool, and dilute with water to volume. solution taken, and 250 mg of hydroxy naphthol blue. Stir
Dilute hydrochloric acid: Dilute 226 mL of hydrochloric with a magnetic stirrer, and titrate immediately with 0.05 M
acid with water to 1000 mL. edetate disodium VS until the solution is distinctly blue.
Potassium chloride solution: 30 mg/mL Perform a blank determination, substituting a volume of
Magnesium stock solution: Transfer 1.000 g of magnesium water equivalent to the volume of the Sample solution taken,
metal to a 1000-mL volumetric flask containing 10 mL of and make any necessary correction. Each mL of 0.05 M
water, slowly add 10 mL of hydrochloric acid, and swirl to edetate disodium is equivalent to 5.004 mg of CaCO3.
dissolve the metal. Dilute with water to volume. Acceptance criteria: 90.0%110.0%
Magnesium solution: 20 g/mL of magnesium (Mg) in POLYDIMETHYLSILOXANE
water, from Magnesium stock solution Dilute hydrochloric acid: 400 mL of hydrochloric acid with
Standard solutions: To three separate, 100-mL volumetric sufficient water to make 1000 mL
flasks each containing 5.0 mL of Lanthanum chloride solution, Standard solution: Transfer 60 mg of USP
add 1.0, 2.0, and 3.0 mL, respectively, of the Magnesium Polydimethylsiloxane RS to a separator, add 30.0 mL of
solution. Dilute each with water to volume. These solutions chloroform and 60 mL of Dilute hydrochloric acid, shake for
contain 0.1, 0.2, and 0.3 g/mL of magnesium (Mg), 30 s, and allow the phases to separate. Remove 10 mL of
respectively. the lower, organic layer to a screw-capped, 15-mL test tube
Sample stock solution: Transfer the equivalent to 250 mg containing 0.5 g of anhydrous sodium sulfate. Close the
of magnesium hydroxide (100 mg of magnesium) from a tube with a screw-cap having an inert liner, agitate
number of Chewable Tablets, to a 1000-mL volumetric flask. vigorously, and centrifuge the mixture until a clear
Add 500 mL of Dilute hydrochloric acid, and swirl to supernatant is obtained. The supernatant so obtained is the
disintegrate the Chewable Tablets. Add 100.0 mL of Standard solution.
Potassium chloride solution, and dilute with water to Sample solution: Transfer the equivalent to 60 mg of
volume. Transfer 10.0 mL of this solution to a 100-mL simethicone from Chewable Tablets (weigh and finely
volumetric flask, and dilute with water to volume. powder NLT 20 Chewable Tablets), to a suitable screw-
Sample solution: Transfer 2.0 mL of Sample stock solution capped bottle.
to a second 100-mL volumetric flask, add 5.0 mL of Add 30.0 mL of chloroform and 60 mL of Dilute hydrochloric
Lanthanum chloride solution, and dilute with water to acid, and allow to stand, with frequent shaking, until the
volume. Chewable Tablets are dissolved. Transfer the contents of the
Blank: Add 50 mL of Dilute hydrochloric acid and 10.0 mL of bottle to a separator, shake, and allow the phases to
Potassium chloride solution to a 100-mL volumetric flask, and separate. Remove 10 mL of the lower, organic layer to a
dilute with water to volume. Transfer 10.0 mL of this screw-capped, 15-mL test tube containing 0.5 g of
solution to a second 100-mL volumetric flask, and dilute to anhydrous sodium sulfate. Close the tube with a screw-cap
volume with water. Transfer 2.0 mL of this solution to a having an inert liner, agitate vigorously, and centrifuge the
third, 100-mL volumetric flask, add 5.0 mL of Lanthanum mixture until a clear supernatant is obtained. The
chloride solution, and dilute with water to volume. supernatant so obtained is the Sample solution.
Analysis: Concomitantly determine the absorbances of the Blank: Place 30.0 mL of chloroform and 60 mL of Dilute
Standard solutions and the Sample solution at the magnesium hydrochloric acid in a separator, shake for 30 s, and allow the
emission line at 285.2 nm, with a suitable atomic absorption phases to separate. Remove 10 mL of the lower, organic
spectrophotometer (see Spectrophotometry and Light- layer to a screw-capped, 15-mL test tube containing 0.5 g of
Scattering 851) equipped with a magnesium hollow- anhydrous sodium sulfate. Close the tube with a screw-cap
cathode lamp and an airacetylene flame, using the Blank to having an inert liner, agitate vigorously, and centrifuge the
set the instrument. Plot the absorbances of the Standard mixture until a clear supernatant is obtained. The
solutions versus concentration, in g/mL, of magnesium, and supernatant so obtained is the Blank.
draw the straight line best fitting the three plotted points. Analysis: Concomitantly determine the absorbances of the
From the graph so obtained, determine the concentration, Standard solution and the Sample solution in 0.5-mm cells at
C, in g/mL, of magnesium in the Sample solution. the wavelength of maximum absorbance at 7.9 m, with a
Calculate the percentage of Mg(OH)2 in each Tablet taken: suitable IR spectrophotometer, using the Blank to set the
instrument.
Result = (Mr/Ar) (500C/N) 100 Calculate the percentage of [(CH3)2 SiO]n in each Tablet
taken:
Mr = molecular weight of magnesium hydroxide,
58.34 Result = (AU/AS) (CS/CU) 100
Ar = atomic weight of magnesium, 24.305
C = nominal concentration of magnesium in the AU = absorbance of the Sample solution
Sample solution (g/mL) AS = absorbance of the Standard solution
N = number of Chewable Tablets taken to prepare CS = concentration of polydimethylsiloxane in the
the Sample solution Standard solution (mg/mL)
CU = nominal concentration of polydimethylsiloxane in Standard stock solution: Transfer 2.5420 g of sodium

the Sample solution (mg/mL) chloride, previously dried at 105 for 2 h, to a 1000-mL
Acceptance criteria: 85.0%115.0% volumetric flask, dissolve in and dilute with water to volume.
Transfer 10.0 mL of this solution to a 100-mL volumetric
PERFORMANCE TESTS flask, and dilute with water to volume. Transfer 10.0 mL of
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements this solution to a second 100-mL volumetric flask, and dilute
for Weight Variation with respect to aluminum hydroxide, to with water to volume.
magnesium hydroxide, and to calcium carbonate Standard solutions: To three separate, 100-mL volumetric
flasks, each containing 10.0 mL of Potassium chloride solution
SPECIFIC TESTS and 3.0 mL of Dilute hydrochloric acid, add 10.0, 20.0, and
MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED 30.0 mL, respectively, of the Standard stock solution. The
MICROORGANISMS 62: The total aerobic microbial count is resulting Standard solutions contain 1.0, 2.0, and 3.0 g/mL
NMT 200 cfu/g; the total combined molds and yeasts count of sodium, respectively.
is NMT 200 cfu/g; and the Chewable Tablets meet the Sample stock solution: Weigh 10 Chewable Tablets, and
requirements of the test for the absence of Salmonella determine the average weight, A, in mg. Cut 4 Chewable
species and Escherichia coli. Tablets into pieces, combine the pieces, and weigh them.
ACID-NEUTRALIZING CAPACITY 301 Transfer the combined pieces to a 500-mL volumetric flask,
Sample solution: Equivalent to 120 mEq of acid- add 150 mL of Dilute hydrochloric acid, and swirl gently to
neutralizing capacity from a counted number of Chewable dissolve the pieces. Dilute with water to volume.
Tablets, in 400 mL of water Sample solution: Transfer 10.0 mL of the Sample stock
Transfer the mixture to a 500-mL volumetric flask, and dilute solution to a 100-mL volumetric flask, add 10.0 mL of
with water to volume. Potassium chloride solution, and dilute with water to volume.
Analysis: Proceed as directed in the section Procedure for Blank solution: Combine 3.0 mL of Dilute hydrochloric acid
Powders, Effervescent Solids, Suspensions and Other Liquids, and 10.0 mL of Potassium chloride solution in a 100-mL
Nonchewable Chewable Tablets, Chewable Chewable Tablets, volumetric flask, and dilute with water to volume.
and Capsules using 75.0 mL of the Sample solution. Analysis: Concomitantly determine the absorbances of the
Acceptance criteria: The acid consumed by the minimum Standard solutions and the Sample solution at the sodium
single dose recommended in the labeling is NLT 5 mEq, and emission line at 589.0 nm with a suitable atomic absorption
NLT the number of mEq calculated: spectrophotometer (see Spectrophotometry and Light-
Result = 0.55(ANC1 A) + 0.8(ANC2 M) + 0.9(ANC3 C) Scattering 851) equipped with a sodium hollow-cathode
lamp and an airacetylene flame, using the Blank. Plot the
ANC1 = theoretical acid-neutralizing capacity of Al(OH)3, absorbances of the Standard solutions versus concentration,
0.0385 mEq in g/mL, of sodium, and draw the straight line best fitting
A = quantity of Al(OH)3 in the specimen tested, the three plotted points. From the graph so obtained,
based on the labeled quantity (mg) determine the concentration, C, in g/mL, of sodium in the
ANC2 = theoretical acid-neutralizing capacity of Mg(OH)2, Sample solution.
0.0343 mEq Calculate the quantity of Na, in mg, in each Chewable
M = quantity of Mg(OH)2 in the specimen tested, Tablet taken:
based on the labeled quantity (mg)
ANC3 = theoretical acid-neutralizing capacity of CaCO3, Result = (A/W) C F
0.02 mEq A = average weight of each tablet (mg)
C = quantity of CaCO3 in the specimen tested, based W = weight of the portion of Chewable Tablets taken
on the labeled quantity (mg) to prepare the Sample solution (mg)
DEFOAMING ACTIVITY C = concentration of sodium in the Sample solution
Foaming solution: 10 mg/mL of octoxynol 9 in 0.3 N (g/mL)
hydrochloric acid F = correction factor, 5
Sample: Equivalent to 20 mg of simethicone from an Acceptance criteria: Chewable Tablets contain NMT 5
amount of Chewable Tablets, cut into small pieces and mg/Tablet of sodium, except when labeled as containing
mixed more than 5 mg/Tablet of sodium; then they contain NMT
[NOTEWeigh NLT 10 Chewable Tablets.] 110% of the labeled amount.
Analysis: [NOTEFor each test, use a clean, 250-mL
cylindrical glass jar.] Transfer the Sample to a clean, 250-mL ADDITIONAL REQUIREMENTS
cylindrical glass jar, fitted with a 50-mm cap, containing 100 PACKAGING AND STORAGE: Preserve in well-closed containers.
mL of Foaming solution that has been warmed to 37. After LABELING: The labeling indicates that the Chewable Tablets
effervescence ceases, cap the jar, and clamp it in an upright are to be chewed before swallowing. Label the Chewable
position on a wrist-action shaker. Using a radius of 13.3 Tablets to state the sodium content, if it is greater than 5
0.4 cm (measured from the center of shaft to the center of mg/Chewable Tablet.
the bottle), shake for 10 s through an arc of 10 at a USP REFERENCE STANDARDS 11
frequency of 300 30 strokes/min. Record the time required USP Polydimethylsiloxane RS
for the foam to collapse. The time, in seconds, for foam
collapse is determined at the instant the first portion of
foam-free liquid surface appears, measured from the end of
the shaking period. Alumina, Magnesia, Calcium Carbonate,
Acceptance criteria: The defoaming activity time does not and Simethicone Chewable Tablets
exceed 45 s. (Comment on this Monograph)id=m1720=Alumina, Magnesia,
SODIUM CONTENT Calcium Carbonate, and Simethicone Chewable Tablets=A-
Potassium chloride solution: 30 mg/mL of potassium Monos.pdf)
chloride (Monograph under this new titleto become official February
Dilute hydrochloric acid: Dilute 226 mL of hydrochloric 1, 2010)
acid with sufficient water to make 1000 mL.
(Current monograph title is Alumina, Magnesia, Calcium Calculate the molarity of the solution taken:
Carbonate, and Simethicone Tablets.)
Result = W/Ar V
DEFINITION
Alumina, Magnesia, Calcium Carbonate, and Simethicone W = weight of aluminum in the portion of solution
Chewable Tablets contain NLT 90.0% and NMT 110.0% of the taken (mg)
labeled amounts of aluminum hydroxide [Al(OH)3], V = volume of Edetate disodium titrant consumed
magnesium hydroxide [Mg(OH)2], and calcium carbonate (mL)
(CaCO3), and an amount of polydimethylsiloxane ([(CH3)2 Ar = atomic weight of aluminum, 26.98
SiO]n) that is NLT 85.0% and NMT 115.0% of the labeled Sample solution: Transfer the equivalent to 665 mg of
amount of simethicone. aluminum hydroxide from a counted number of Chewable
Tablets, to a suitable beaker.
IDENTIFICATION Add 15 mL of hydrochloric acid, and swirl to dissolve the
A. IDENTIFICATION TESTSGENERAL, Calcium 191 Chewable Tablets. Add 80 mL of water and filter into a
Sample solution: Cut a Chewable Tablet into pieces, add 200-mL volumetric flask. Wash the filter with water into the
50 mL of 1 N sulfuric acid, stir until the pieces disintegrate, flask, and add water to volume.
and heat on a steam bath for 10 min. Cool, add 50 mL of Analysis: Pipet 20 mL of Sample solution into a 250-mL
alcohol, and stir. Place in an ice bath for 30 min. Filter while beaker, then add, in the order named and with continuous
cold. [NOTERetain the filtrate for Identification test B.] Wash stirring, 25.0 mL of Edetate disodium titrant and 20 mL of
the precipitate with 50 mL of 0.75 N sulfuric acid, and acetic acidammonium acetate buffer TS, and heat the
discard the washings. Dissolve the precipitate so obtained in solution near the boiling temperature for 5 min. Cool, add
3 N hydrochloric acid, and filter. 50 mL of alcohol and 2 mL of dithizone TS. Titrate with
Acceptance criteria: Meets the requirements 0.05 M zinc sulfate VS until the color changes from green-
B. IDENTIFICATION TESTSGENERAL, Aluminum 191 violet to rose-pink. Perform a blank determination,
Sample solution: To the filtrate obtained in Identification substituting 20 mL of water for the Sample solution, and
test A, add 5 drops of methyl red TS, and heat to boiling. make any necessary correction. Each mL of 0.05 M Edetate
Add 6 N ammonium hydroxide until the color of the disodium titrant consumed is equivalent to 3.900 mg of
solution changes to deep yellow, continue boiling for 2 min, Al(OH)3.
and filter through hardened filter paper. [NOTERetain the Acceptance criteria: 90.0%110.0%
filtrate for Identification test C.] Wash the precipitate with MAGNESIUM HYDROXIDE
350 mL of a hot ammonium chloride solution (1 in 50), Lanthanum chloride solution: Transfer 17.6 g of
discarding the washings. Dissolve the precipitate so obtained lanthanum chloride to a 200-mL volumetric flask, add 100
in 3 N hydrochloric acid. mL of water, and carefully add 50 mL of hydrochloric acid.
Acceptance criteria: Meets the requirements Allow to cool, and dilute with water to volume.
C. IDENTIFICATION TESTSGENERAL, Magnesium 191 Dilute hydrochloric acid: Dilute 226 mL of hydrochloric
Sample solution: The filtrate obtained in Identification test B acid to 1000 mL with water.
Acceptance criteria: Meets the requirements Potassium chloride solution: 30 mg/mL
D. INFRARED ABSORPTION 197S Magnesium stock solution: Transfer 1.000 g of magnesium
Sample solution: Transfer the equivalent to 60 mg of metal to a 1000-mL volumetric flask containing 10 mL of
simethicone from Chewable Tablets (weigh and finely water, slowly add 10 mL of hydrochloric acid, and swirl to
powder NLT 20 Chewable Tablets), to a suitable screw- dissolve the metal. Dilute with water to volume.
capped bottle, add 30.0 mL of chloroform and 60 mL of Magnesium solution: 20 g/mL of magnesium (Mg) in
Dilute hydrochloric acid (see the Assay for water, from Magnesium stock solution
Dimethylpolysiloxane), and allow to stand, with frequent Standard solutions: To three separate, 100-mL volumetric
shaking, until the Chewable Tablets are dissolved. Transfer flasks each containing 5.0 mL of Lanthanum chloride solution,
the contents of the bottle to a separator, shake, and allow add 1.0, 2.0, and 3.0 mL, respectively, of the Magnesium
the phases to separate. Remove 10 mL of the lower, organic solution. Dilute each with water to volume. These solutions
layer to a screw-capped, 15-mL test tube containing 0.5 g of contain 0.1, 0.2, and 0.3 g/mL of Mg, respectively.
anhydrous sodium sulfate. Close the tube with a screw-cap Sample stock solution: Transfer the equivalent to 250 mg
having an inert liner, agitate vigorously, and centrifuge the of magnesium hydroxide (100 mg of magnesium) from a
mixture until a clear supernatant is obtained. The number of Chewable Tablets, to a 1000-mL volumetric flask.
supernatant so obtained is the Sample solution. Add 500 mL of Dilute hydrochloric acid, and swirl to dissolve
Analysis: Proceed as directed using a 0.5-mm cell. the Chewable Tablets. Add 100.0 mL of Potassium chloride
solution, and dilute with water to volume. Transfer 10.0 mL
ASSAY of this solution to a 100-mL volumetric flask, and dilute
ALUMINUM HYDROXIDE with water to volume.
Edetate disodium titrant: 18.6 mg/mL of edetate disodium Sample solution: Transfer 2.0 mL of Sample stock solution
in water to a second 100-mL volumetric flask, add 5.0 mL of
Standardization of titrant: Transfer 2 g of aluminum wire, Lanthanum chloride solution, and dilute with water to
transfer to a 1000-mL volumetric flask, and add 50 mL of a volume.
mixture of hydrochloric acid and water (1:1). Swirl the flask Blank: Add 50 mL of Dilute hydrochloric acid and 10.0 mL of
to ensure contact of the aluminum and the acid, and allow Potassium chloride solution to a 100-mL volumetric flask, and
the reaction to proceed until all of the aluminum has dilute with water to volume. Transfer 10.0 mL of this
dissolved. Dilute with water to volume. Pipet 10 mL of this solution to a second 100-mL volumetric flask, and dilute
solution into a 250-mL beaker and add, in the order named with water to volume. Transfer 2.0 mL of this solution to a
and with continuous stirring, 25.0 mL of Edetate disodium third 100-mL volumetric flask, add 5.0 mL of Lanthanum
titrant and 20 mL of acetic acidammonium acetate buffer chloride solution, and dilute with water to volume.
TS, and boil gently for 5 min. Cool, and add 50 mL of Analysis: Concomitantly determine the absorbances of the
alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc Standard solutions and the Sample solution at the magnesium
sulfate VS to a bright rose-pink color. Perform a blank emission line at 285.2 nm, with a suitable atomic absorption
determination, substituting 10 mL of water for the spectrophotometer (see Spectrophotometry and Light-
aluminum solution, and make any necessary correction.
Scattering 851) equipped with a magnesium hollow- that has an inert liner, agitate vigorously, and centrifuge the
cathode lamp and an airacetylene flame, using the Blank to mixture until a clear supernatant is obtained. The
set the instrument. Plot the absorbances of the Standard supernatant so obtained is the Blank.
solutions versus concentration, in g/mL, of magnesium, and Analysis: Concomitantly determine the absorbances of the
draw the straight line best fitting the three plotted points. Standard solution and the Sample solution in 0.5-mm cells at
From the graph so obtained, determine the concentration, the wavelength of maximum absorbance at 7.9 m, with a
C, in g/mL, of magnesium in the Sample solution. suitable IR spectrophotometer, using the Blank to set the
Calculate the percentage of Mg(OH)2 in each Tablet taken: instrument.
Calculate the percentage of [(CH3)2 SiO]n in each Tablet
Result = (Mr/Ar) (500C/N) 100 taken:
Mr = molecular weight of magnesium hydroxide, Result = (AU/AS) (CS/CU) 100
58.34
Ar = atomic weight of magnesium, 24.305 AU = absorbance of the Sample solution
C = nominal concentration of magnesium in the AS = absorbance of the Standard solution
Sample solution (g/mL) CS = concentration of polydimethylsiloxane in the
N = number of Chewable Tablets taken to prepare Standard solution (mg/mL)
the Sample solution CU = nominal concentration of polydimethylsiloxane in
Acceptance criteria: 90.0%110.0% the Sample solution (mg/mL)
CALCIUM CARBONATE Acceptance criteria: 85.0%115.0%
Sample solution: Transfer the equivalent to 665 mg of
aluminum hydroxide from a counted number of Chewable PERFORMANCE TESTS
Tablets, to a suitable beaker. UNIFORMITY OF DOSAGE UNITS 905 Meet the requirements
Add 15 mL of hydrochloric acid, and swirl to dissolve the for Weight Variation with respect to aluminum hydroxide, to
Chewable Tablets. Add 80 mL of water and filter into a magnesium hydroxide, and to calcium carbonate.
200-mL volumetric flask. Wash the filter with water into the
flask, and add water to volume. SPECIFIC TESTS
Analysis: Pipet a volume of the Sample solution, equivalent MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED
to 50 mg of calcium carbonate, into a 400-mL beaker, and MICROORGANISMS 62: The total aerobic microbial count is
add 200 mL of water, a volume of sodium hydroxide NMT 200 cfu/g; the total combined molds and yeasts count
solution (1 in 2) equivalent to the volume of the Sample is NMT 200 cfu/g; and the Chewable Tablets meet the
solution taken, and 250 mg of hydroxy naphthol blue. Stir requirements of the test for the absence of Salmonella
with a magnetic stirrer, and titrate immediately with 0.05 M species and Escherichia coli.
edetate disodium VS until the solution is distinctly blue. ACID-NEUTRALIZING CAPACITY 301
Perform a blank determination, substituting a volume of Sample solution: Equivalent to 120 mEq of acid-
water equivalent to the volume of the Sample solution taken, neutralizing capacity from a counted number of Chewable
and make any necessary correction. Each mL of 0.05 M Tablets, in 400 mL of water. Transfer the mixture to a 500-
edetate disodium is equivalent to 5.004 mg of calcium mL volumetric flask, and dilute with water to volume.
carbonate (CaCO3). Analysis: Proceed as directed in the section Procedure for
Acceptance criteria: 90.0%110.0% Powders, Effervescent Solids, Suspensions and Other Liquids,
POLYDIMETHYLSILOXANE Nonchewable Chewable Tablets, Chewable Chewable Tablets,
Dilute hydrochloric acid: Dilute 400 mL of hydrochloric and Capsules using 75.0 mL of the Sample solution.
acid with sufficient water to make 1000 mL. Acceptance criteria: The acid consumed by the minimum
Standard solution: Transfer 60 mg of USP single dose recommended in the labeling is NLT 5 mEq, and
Polydimethylsiloxane RS to a separator, add 30.0 mL of NLT the number of mEq calculated:
chloroform and 60 mL of Dilute hydrochloric acid, shake for Result = 0.55(ANC1 A) + 0.8(ANC2 M) + 0.9(ANC3 C)
30 s, and allow the phases to separate. Remove 10 mL of
the lower, organic layer to a screw-capped, 15-mL test tube ANC1 = theoretical acid-neutralizing capacity of Al(OH)3,
containing 0.5 g of anhydrous sodium sulfate. Close the 0.0385 mEq
tube with a screw-cap having an inert liner, agitate A = quantity of Al(OH)3 in the specimen tested,
vigorously, and centrifuge the mixture until a clear based on the labeled quantity (mg)
supernatant is obtained. The supernatant so obtained is the ANC2 = theoretical acid-neutralizing capacity of Mg(OH)2,
Standard solution. 0.0343 mEq
Sample solution: Transfer the equivalent to 60 mg of M = quantity of Mg(OH)2 in the specimen tested,
simethicone from Chewable Tablets (weigh and finely based on the labeled quantity (mg)
powder NLT 20 Chewable Tablets), to a suitable screw- ANC3 = theoretical acid-neutralizing capacity of CaCO3,
capped bottle. 0.02 mEq
Add 30.0 mL of chloroform and 60 mL of Dilute C = quantity of CaCO3 in the specimen tested, based
hydrochloric acid, and allow to stand, with frequent shaking, on the labeled quantity (mg)
until the Chewable Tablets are dissolved. Transfer the DEFOAMING ACTIVITY
contents of the bottle to a separator, shake, and allow the Foaming solution: 10 mg/mL of octoxynol 9 in 0.3 N
phases to separate. Remove 10 mL of the lower, organic hydrochloric acid
layer to a screw-capped, 15-mL test tube containing 0.5 g Sample: Equivalent to 20 mg of simethicone from an
of anhydrous sodium sulfate. Close the tube with a screw- amount of Chewable Tablets, cut into small pieces and
cap having an inert liner, agitate vigorously, and centrifuge mixed
the mixture until a clear supernatant is obtained. The [NOTEWeigh NLT 10 Chewable Tablets.]
supernatant so obtained is the Sample solution. Analysis: [NOTEFor each test, use a clean, 250-mL
Blank: Place 30.0 mL of chloroform and 60 mL of Dilute cylindrical glass jar.] Transfer the Sample to a clean, 250-mL
hydrochloric acid in a separator, shake for 30 s, and allow the cylindrical glass jar, fitted with a 50-mm cap, containing
phases to separate. Remove 10 mL of the lower, organic 100 mL of Foaming solution that has been warmed to 37.
layer to a screw-capped, 15-mL test tube containing 0.5 g of After effervescence ceases, cap the jar, and clamp it in an
anhydrous sodium sulfate. Close the tube with a screw-cap
upright position on a wrist-action shaker. Using a radius of Tablets to state the sodium content, if it is greater than 5
13.3 0.4 cm (measured from the center of the shaft to mg/Chewable Tablet.
the center of the bottle), shake for 10 s through an arc of USP REFERENCE STANDARDS 11
10 at a frequency of 300 30 strokes/min. Record the USP Polydimethylsiloxane RS
time required for the foam to collapse. The time, in
seconds, for foam collapse is determined at the instant the
first portion of foam-free liquid surface appears, measured
from the end of the shaking period. Alumina, Magnesia, and Simethicone
Acceptance criteria: The defoaming activity time does not Oral Suspension
exceed 45 s. (Comment on this Monograph)id=m1750=Alumina, Magnesia,
SODIUM CONTENT and Simethicone Oral Suspension=A-Monos.pdf)
Potassium chloride solution: 30 mg/mL of potassium
chloride DEFINITION
Dilute hydrochloric acid: Dilute 226 mL of hydrochloric Alumina, Magnesia, and Simethicone Oral Suspension contains
acid with sufficient water to make 1000 mL. the equivalent of NLT 90.0% and NMT 115.0% of the labeled
Standard stock solution: Transfer 2.5420 g of sodium amounts of aluminum hydroxide [Al(OH)3] and magnesium
chloride, previously dried at 105 for 2 h, to a 1000-mL hydroxide [Mg(OH)2], and an amount of polydimethylsiloxane
volumetric flask, and dissolve in and dilute with water to [(CH3)2 SiO]n that is NLT 85.0% and NMT 115.0% of the
volume. Transfer 10.0 mL of this solution to a 100-mL labeled amount of simethicone.
volumetric flask, and dilute with water to volume. Transfer
10.0 mL of this solution to a second 100-mL volumetric IDENTIFICATION
flask, and dilute with water to volume. A. INFRARED ABSORPTION 197S
Standard solutions: To three separate 100-mL volumetric Sample solution: Transfer an amount of Oral Suspension
flasks, each containing 10.0 mL of Potassium chloride solution equivalent to 50 mg of simethicone to a suitable round,
and 3.0 mL of Dilute hydrochloric acid, add 10.0, 20.0, and narrow-mouth, screw-capped, 120-mL bottle, add 40 mL of
30.0 mL, respectively, of the Standard stock solution. The 0.1 N sodium hydroxide, and swirl to disperse. Add 25.0 mL
resulting Standard solutions contain 1.0, 2.0, and 3.0 g/mL of toluene, close the bottle securely with a cap having an
of sodium (Na), respectively. inert liner, and shake for 15 min on a reciprocating shaker
Sample stock solution: Weigh 10 Chewable Tablets, and (e.g., about 200 oscillations per minute and a stroke of 38
determine the average weight, A, in mg. Cut 4 Chewable 2 mm). Transfer the mixture to a 125-mL separator. Remove
Tablets into pieces, combine the pieces, and weigh them. about 5 mL of the upper, organic layer to a screw-capped,
Transfer the combined pieces to a 500-mL volumetric flask, 15-mL test tube containing 0.5 g of anhydrous sodium
add 150 mL of Dilute hydrochloric acid, and swirl gently to sulfate. Close the tube with a screw-cap having an inert
dissolve the pieces. Dilute with water to volume. liner, agitate vigorously, and centrifuge the mixture until a
Sample solution: Transfer 10.0 mL of the Sample stock clear supernatant is obtained. The clear supernatant is the
solution to a 100-mL volumetric flask, add 10.0 mL of Sample solution.
Potassium chloride solution, and dilute with water to volume. Analysis: Proceed as directed using a 0.5-mm cell.
Blank solution: Combine 3.0 mL of Dilute hydrochloric acid B. IDENTIFICATION TESTSGENERAL, Magnesium 191
and 10.0 mL of Potassium chloride solution in a 100-mL Sample solution: Add 5 g of sample to 10 mL of 3 N
volumetric flask, and dilute with water to volume. hydrochloric acid, then add 5 drops of methyl red TS, heat
Analysis: Concomitantly determine the absorbances of the to boiling, add 6 N ammonium hydroxide until the color of
Standard solutions and the Sample solution at the sodium the solution just changes to deep yellow, then continue
emission line at 589.0 nm with a suitable atomic absorption boiling for 2 min and filter. The filtrate so obtained is the
spectrophotometer (see Spectrophotometry and Light- Sample solution.
Scattering 851) equipped with a sodium hollowcathode Acceptance criteria: Meets the requirements
lamp and an airacetylene flame, using the Blank solution as C. IDENTIFICATION TESTSGENERAL, Aluminum 191
the blank. Plot the absorbances of the Standard solutions Sample solution: Wash the precipitate from Identification
versus concentration, in g/mL, of sodium, and draw the test B with hot ammonium chloride solution (1 in 50), and
straight line best fitting the three plotted points. From the dissolve the precipitate in hydrochloric acid. Divide this
graph so obtained, determine the concentration, C, in solution into two portions.
g/mL, of sodium in the Sample solution. Analysis 1: Add, dropwise, 6 N ammonium hydroxide to
Calculate the quantity, in mg, of Na in each Chewable one portion of the Sample solution.
Tablet taken: Acceptance criteria 1: Yields a gelatinous white precipitate,
which does not dissolve in an excess of 6 N ammonium
Result = (A/W) C F hydroxide.
Analysis 2: Add, dropwise, 1 N sodium hydroxide to the
A = average weight of each Tablet (mg) second portion of the Sample solution.
W = weight of the portion of Chewable Tablets taken Acceptance criteria 2: Yields a gelatinous white precipitate,
to prepare the Sample solution (mg) which dissolves in an excess of 1 N sodium hydroxide,
C = concentration of sodium in the Sample solution leaving some turbidity.
(g/mL)
F = correction factor, 5 ASSAY
Acceptance criteria: Chewable Tablets contain NMT 5 ALUMINUM HYDROXIDE
mg/Tablet of sodium, except when labeled as containing Edetate disodium titrant: Prepare a solution with a
more than 5 mg/Tablet of sodium; then they contain NMT concentration of 18.6 mg/mL of edetate disodium in water
110% of the labeled amount. and standardize as follows. Weigh 2 g of aluminum wire,
transfer to a 1000-mL volumetric flask, and add 50 mL of a
ADDITIONAL REQUIREMENTS mixture of hydrochloric acid and water (1:1). Swirl the flask
PACKAGING AND STORAGE: Preserve in well-closed containers. to ensure contact of the aluminum and the acid, and allow
LABELING: The labeling indicates that the Chewable Tablets the reaction to proceed until all of the aluminum has
are to be chewed before swallowing. Label the Chewable
dissolved. Dilute with water to volume. Pipet 10 mL of this of toluene, close the bottle securely with a cap having an
solution into a 250-mL beaker and add, in the order named inert liner, and shake for 15 min on a reciprocating shaker
and with continuous stirring, 25.0 mL of Edetate disodium (e.g., about 200 oscillations per minute and a stroke of 38
titrant and 20 mL of acetic acidammonium acetate buffer 2 mm). Transfer the mixture to a 125-mL separator. Remove
TS, and boil gently for 5 min. Cool, and add 50 mL of about 5 mL of the upper, organic layer to a screw-capped,
alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc 15-mL test tube containing 0.5 g of anhydrous sodium
sulfate VS to a bright rose-pink color. Perform a blank sulfate. Close the tube with a screw-cap having an inert
determination, substituting 10 mL of water for the liner, agitate vigorously, and centrifuge the mixture until a
aluminum solution, and make any necessary correction. clear supernatant is obtained. The clear supernatant is the
Calculate the molarity of the solution taken by the formula: Sample solution.
Blank: Mix 10 mL of toluene with 0.5 g of anhydrous
W/ArV sodium sulfate and centrifuge to obtain a clear supernatant.
W = weight of aluminum in the portion of solution (See Spectrophotometry and Light-Scattering 851.)
taken (mg) Mode: IR spectrophotometry
Ar = atomic weight of aluminum, 26.98 Analytical wavelength: 7.9 m
V = volume of Edetate disodium titrant consumed Cell: 0.5 mm
(mL) Analysis
Sample solution: Transfer a measured amount of Oral Samples: Standard solution, Blank, and Sample solution
Suspension, previously well-shaken in its original container, Calculate the amount in mg of [(CH3)2 SiO]n in each mL
equivalent to 800 mg of aluminum hydroxide, to a suitable of the Oral Suspension taken:
beaker. Add 20 mL of water, stir, and slowly add 10 mL of
hydrochloric acid. Heat gently, if necessary, to aid solution, Result = (AU/AS) (W/V) 100
cool, and filter into a 200-mL volumetric flask. Wash the
filter with water into the flask, and add water to volume. AU = absorbance of the Sample solution
Analysis: Pipet 10 mL of Sample solution into a 250-mL AS = absorbance of the Standard solution
beaker, add 20 mL of water, then add, in the order named W = weight of USP Polydimethylsiloxane RS used in
and with continuous stirring, 25.0 mL of Edetate disodium the Standard solution (mg)
titrant and 20 mL of acetic acidammonium acetate buffer V = volume of Oral suspension used in the Sample
TS, and heat the solution near the boiling temperature for 5 solution (mL)
min. Cool, add 50 mL of alcohol and 2 mL of dithizone TS. Acceptance criteria: NLT 85.0% and NMT 115.0%
Titrate with 0.05 M zinc sulfate VS until the color changes
from green-violet to rose-pink. Perform a blank SPECIFIC TESTS
determination, substituting 10 mL of water for the Sample Microbial Enumeration Tests 61 and Tests for Specified
solution, and make any necessary correction. Each mL of Microorganisms 62
0.05 M Edetate disodium titrant consumed is equivalent to Acceptance criteria: The total aerobic microbial count is
3.900 mg of Al(OH)3. NMT 100 cfu/g and it meets the requirements of the test
Acceptance criteria: NLT 90.0% and NMT 115.0% for the absence of Escherichia coli.
MAGNESIUM HYDROXIDE ACID-NEUTRALIZING CAPACITY 301
Sample solution: Transfer an amount of Oral Suspension, Acceptance criteria: The acid consumed by the minimum
previously well-shaken in its original container, equivalent to single dose recommended in the labeling is NLT 5 mEq, and
800 mg of aluminum hydroxide, to a suitable beaker. Add NLT the number of mEq calculated:
20 mL of water, stir, and slowly add 10 mL of hydrochloric
acid. Heat gently, if necessary, to aid solution, cool, and 0.55(0.0385A) + 0.8(0.0343M)
filter into a 200-mL volumetric flask. Wash the filter with
water into the flask, and add water to volume. 0.0385 = theoretical acid-neutralizing capacity of Al(OH)3
Analysis: Pipet a volume of the Sample solution, equivalent (mEq)
to 40 mg of magnesium hydroxide, into a 400-mL beaker, A = quantity of Al(OH)3 in the specimen tested,
add 200 mL of water and 20 mL of triethanolamine, and based on the labeled quantity (mg)
stir. Add 10 mL of ammoniaammonium chloride buffer TS 0.0343 = theoretical acid-neutralizing capacity of Mg(OH)2
and 3 drops of an eriochrome black indicator solution (mEq)
(prepared by dissolving 200 mg of eriochrome black T in a M = quantity of Mg(OH)2 in the specimen tested,
mixture of 15 mL of triethanolamine and 5 mL of based on the labeled quantity (mg)
dehydrated alcohol, and mixing). Cool the solution to PH 791
between 3 and 4 by immersion of the beaker in an ice Acceptance criteria: 7.08.6
bath, then remove, and titrate with 0.05 M edetate SODIUM CONTENT
disodium VS to a blue endpoint. Perform a blank Potassium chloride solution: 38 mg/mL of potassium
determination, substituting water for the Sample solution, chloride
and make any necessary correction. Each mL of 0.05 M Sodium chloride stock solution: 25.42 g/mL of sodium
edetate disodium consumed is equivalent to 2.916 mg of chloride (previously dried at 105 for 2 h) in water
Mg(OH)2. [NOTEThe solution contains 10 g/mL of sodium.]
Acceptance criteria: NLT 90.0% and NMT 115.0% Standard solutions: On the day of use, transfer 4.0 mL of
POLYDIMETHYLSILOXANE 1 N hydrochloric acid and 10.0 mL of Potassium chloride
Standard solution: Prepare similarly to the Sample solution, solution to each of two 100-mL volumetric flasks. To the
except dissolve 50 mg of USP Polydimethylsiloxane RS in respective flasks, add 5.0 mL and 10.0 mL of Sodium chloride
25.0 mL of toluene, add 40 mL of 0.1 N sodium hydroxide, stock solution. Dilute with water to volume. The resulting
and add a volume of water equal to that of the specimen of Standard solutions contain 0.5 and 1.0 g/mL of sodium
Oral Suspension taken for the Sample solution. (Na), respectively.
Sample solution: Transfer an amount of Oral Suspension Sample solution: Transfer 5.0 mL of Oral Suspension,
equivalent to 50 mg of simethicone to a suitable round, previously well-shaken in its original container, to a 100-mL
narrow-mouth, screw-capped, 120-mL bottle, add 40 mL of volumetric flask, add 50 mL of 1 N hydrochloric acid, boil
0.1 N sodium hydroxide, and swirl to disperse. Add 25.0 mL for 15 mins, cool to room temperature, dilute with water to
volume and filter, discarding the first few mL of the filtrate. C. PROCEDURE
Transfer 5.0 mL of the filtrate to a 100-mL volumetric flask Sample: Wash the precipitate obtained in Identification test
containing 10.0 mL of Potassium chloride solution, and dilute B with hot ammonium chloride solution (1 in 50).
with water to volume. Analysis: Dissolve the precipitate in hydrochloric acid.
Blank solution: Combine 4.0 mL of 1 N hydrochloric acid Divide this solution into two portions.
and 10.0 mL of Potassium chloride solution in a 100-mL Acceptance criteria: The dropwise addition of 6 N
volumetric flask, and dilute with water to volume. ammonium hydroxide to one portion yields a gelatinous
Spectrometric conditions white precipitate, which does not dissolve in an excess of 6
(See Spectrophotometry and Light-Scattering 851.) N ammonium hydroxide. The dropwise addition of 1 N
Mode: Atomic absorption sodium hydroxide to the other portion yields a gelatinous
[NOTEUse an airacetylene flame.] white precipitate, which dissolves in an excess of 1 N
Analytical wavelength: 589.0 nm sodium hydroxide, leaving some turbidity.
Lamp: Sodium hollow-cathode lamp
Analysis: Standard solutions, Blank solution, and Sample ASSAY
solution ALUMINUM HYDROXIDE
Plot the absorbances of the Standard solutions versus Edetate disodium titrant: 18.6 mg/mL of edetate disodium
concentration, in g/mL, of sodium, and draw the straight Standardization of titrant: Transfer 2 g of aluminum wire
line between the plotted points. From the graph so to a 1000-mL volumetric flask, and add 50 mL of a mixture
obtained, determine the concentration, C, in g/mL, of of hydrochloric acid and water (1:1). Swirl the flask to
sodium in the Sample solution. ensure contact of the aluminum and the acid, and allow the
Calculate the quantity, in mg, of sodium in each mL of reaction to proceed until all of the aluminum has dissolved.
Oral Suspension taken: Dilute with water to volume. Pipet 10 mL of this solution
into a 250-mL beaker, add, in the order named and with
0.4 C continuous stirring, 25.0 mL of Edetate disodium titrant and
20 mL of acetic acidammonium acetate buffer TS, and boil
C = concentration of sodium in the Sample solution gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL
(g/mL) of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a
ADDITIONAL REQUIREMENTS substituting 10 mL of water for the aluminum solution, and
PACKAGING AND STORAGE: Preserve in tight containers, and make any necessary correction.
avoid freezing. Calculate the molarity of the solution taken:
LABELING: Oral Suspension may be labeled to state the
aluminum hydroxide content in terms of the equivalent Result = W/ArV
amount of dried aluminum hydroxide gel, on the basis that
each mg of dried gel is equivalent to 0.765 mg of Al(OH)3. W = weight of aluminum in the portion of solution
Label it to state the sodium content if it is greater than 1 mg taken (mg)
per mL. Ar = atomic weight of aluminum, 26.98
USP REFERENCE STANDARDS 11 V = volume of Edetate disodium titrant consumed
USP Polydimethylsiloxane RS (mL)
aluminum hydroxide from Chewable Tablets (finely powder
NLT 20 Chewable Tablets), to a 150-mL beaker.
Alumina, Magnesia, and Simethicone Add 20 mL of water, stir, and slowly add 30 mL of 3 N
Tablets hydrochloric acid. Heat gently, if necessary, to aid solution,
(Comment on this Monograph)id=m1753=Alumina, Magnesia, cool to room temperature, and filter into a 200-mL
and Simethicone Tablets=A-Monos.pdf) volumetric flask. Wash the filter with water into the flask,
(Current titlenot to change until February 1, 2010) and add water to volume.
Monograph title changeto become official February 1, 2010 Analysis: Pipet 10 mL of the Sample solution into a 250-mL
See Alumina, Magnesia, and Simethicone Chewable Tablets beaker.
Add 20 mL of water, then add, in the order named and
DEFINITION with continuous stirring, 25.0 mL of Edetate disodium titrant
Alumina, Magnesia, and Simethicone Chewable Tablets contain and 20 mL of acetic acidammonium acetate buffer TS,
the equivalent of NLT 90.0% and NMT 115.0% of the labeled and heat near the boiling temperature for 5 min. Cool, add
amounts of aluminum hydroxide [Al(OH)3] and magnesium 50 mL of alcohol and 2 mL of dithizone TS. Titrate with
hydroxide [Mg(OH)2], and an amount of polydimethylsiloxane 0.05 M zinc sulfate VS until the color changes from green-
[(CH3)2 SiO]n that is NLT 85.0% and NMT 115.0% of the violet to rose-pink. Perform a blank determination,
labeled amount of simethicone. substituting 10 mL of water for the Sample solution, and
making any necessary correction. Each mL of 0.05 M
IDENTIFICATION Edetate disodium titrant consumed is equivalent to 3.900
A. INFRARED ABSORPTION 197S mg of Al(OH)3.
Cell: 0.5 mm Acceptance criteria: 90.0%115.0%
Solution: Prepared as directed in the Assay for MAGNESIUM HYDROXIDE
Polydimethylsiloxane. Sample solution: Prepare as directed in the Assay for
B. IDENTIFICATION TESTSGENERAL, Magnesium 191 Aluminum Hydroxide.
Sample solution: Equivalent to 600 mg of magnesium Analysis: Pipet a volume of the Sample solution, equivalent
hydroxide from finely powdered Chewable Tablets to 40 mg of magnesium hydroxide, into a 400-mL beaker,
Add 25 mL of 3 N hydrochloric acid and 25 mL of water, add 200 mL of water and 20 mL of triethanolamine, and
and mix. Boil gently for 2 min. Allow to cool, and filter. stir. Add 10 mL of ammoniaammonium chloride buffer TS
Add 5 drops of methyl red TS, heat to boiling, and add 6 and 3 drops of an eriochrome black indicator solution
N ammonium hydroxide until the color of the solution just (prepared by dissolving 200 mg of eriochrome black T in a
turns to deep yellow. Continue boiling for 2 min, and filter: mixture of 15 mL of triethanolamine and 5 mL of
the filtrate so obtained meets the requirements.
dehydrated alcohol, and mixing). Cool the solution to 3 to Analysis: [NOTEFor each test, use a clean, unused, 250-mL
4 by immersion of the beaker in an ice bath, then remove, glass jar.] Transfer a quantity of finely powdered Chewable
and titrate with 0.05 M edetate disodium VS to a blue Tablets, passed completely through an 80-mesh sieve,
endpoint. Perform a blank determination, substituting water equivalent to 20 mg of simethicone, to a clean, unused,
for the Sample solution, and make any necessary correction. cylindrical 250-mL glass jar, fitted with a 50-mm cap,
Each mL of 0.05 M edetate disodium consumed is containing 100 mL of Foaming solution that has been
equivalent to 2.916 mg of Mg(OH)2. warmed to 37. Cap the jar, and clamp it in an upright
Acceptance criteria: 90.0%115.0% position on a wrist-action shaker. Using a radius of 13.3
POLYDIMETHYLSILOXANE 0.4 cm (measured from center of shaft to center of bottle),
Sample solution: Transfer the equivalent to 33 mg of shake for 10 s through an arc of 10 at a frequency of 300
simethicone from Chewable Tablets (finely powder NLT 20 30 strokes/min. Record the time required for the foam to
Chewable Tablets), to a suitable round, narrow-mouth, collapse. The time, in seconds, for foam collapse is
screw-capped, 120-mL bottle. determined at the instant the first portion of foam-free
Add 40 mL of 0.1 N sodium hydroxide, and swirl to liquid surface appears, measured from the end of the
disperse. Add 20.0 mL of toluene, close the bottle securely shaking period.
with a cap having an inert liner, and shake for 30 min on a Acceptance criteria: The defoaming activity time does not
reciprocating shaker (e.g., 200 oscillations per minute and exceed 45 s.
a stroke of 38 2 mm). Transfer the mixture to a 125-mL SODIUM CONTENT
separator, and allow to separate. Remove the upper, Potassium chloride solution: 38 mg/mL of potassium
organic layer to a screw-capped, centrifuge tube containing chloride
2 g of anhydrous sodium sulfate. Close the tube with a Sodium chloride stock solution: Dissolve sodium chloride,
screw-cap having an inert liner, agitate vigorously, and previously dried at 105 for 2 h and weighed, in water, and
centrifuge the mixture until a clear supernatant is obtained. dilute with water to obtain a solution containing 25.42
Standard solution: Similarly prepare a Standard solution, g/mL (10 g of sodium/mL).
using 33 mg of USP Polydimethylsiloxane RS. Standard solutions: On the day of use, transfer 4.0 mL of
Blank: Mix 10 mL of toluene with 0.5 g of anhydrous 1 N hydrochloric acid and 10.0 mL of Potassium chloride
sodium sulfate, and centrifuge to obtain a clear supernatant. solution to each of two 100-mL volumetric flasks. To the
Analysis respective flasks add 5.0 mL and 10.0 mL of Sodium chloride
Samples: Sample solution, Standard solution, and Blank stock solution. Dilute with water to volume. These solutions
Spectrometric conditions contain 0.5 g/mL and 1.0 g/mL of sodium, respectively.
(See Spectrophotometry and Light-Scattering 851.) Sample solution: Transfer the equivalent to the average
Mode: IR spectrophotometer weight of 1 Tablet from Chewable Tablets (weigh and finely
Analytical wavelength: Wavelength of maximum powder NLT 20 Chewable Tablets), to a 100-mL volumetric
absorbance at 7.9 m (1265.8 cm1) flask.
Cell: 0.5 mm Add 50 mL of 1 N hydrochloric acid, boil for 15 min, cool to
Analysis: Use Blank to set the instrument. room temperature, and dilute with water to volume. Filter,
Calculate the quantity in mg of [(CH3)2 SiO]n in the discarding the first few mL of the filtrate. Transfer 5.0 mL
portion of Chewable Tablets taken: of the filtrate to a 100-mL volumetric flask containing 10.0
mL of Potassium chloride solution, and dilute with water to
Result = (AU/AS) W volume.
Blank: 1 N hydrochloric acid, Potassium chloride solution,
AU = absorbance of the Sample solution and water (4:10:86)
AS = absorbance of the Standard solution Spectrometric conditions
W = weight in mg of USP Polydimethylsiloxade RS (See Spectrophotometry and Light-Scattering 851.)
used to prepare the Standard solution Mode: Atomic absorption spectrophotometer
Acceptance criteria: 85.0%115.0% of simethicone Analytical wavelength: 589.0 nm, sodium emission line
PERFORMANCE TESTS Flame: Air acetylene
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Analysis
for Weight Variation with respect to aluminum hydroxide and Samples: Standard solutions and Sample solution
to magnesium hydroxide Plot the absorbances of the Standard solutions versus
SPECIFIC TESTS concentration, in g/mL, of sodium, and draw the straight
ACID-NEUTRALIZING CAPACITY 301 line between the plotted points. From the graph so
Analysis: The acid consumed by the minimum single dose obtained, determine the concentration, C, in g/mL, of
recommended in the labeling is NLT 5 mEq, and NLT the sodium in the Sample solution.
number of mEq calculated: Calculate the quantity, in mg, of sodium per Tablet taken:
Result = 0.55(ANC1 A) + 0.8(ANC2 M) Result = 2C
ANC1 = theoretical acid-neutralizing capacity of Al(OH)3, ADDITIONAL REQUIREMENTS

0.0385 mEq PACKAGING AND STORAGE: Preserve in well-closed containers.
A = quantity of Al(OH)3 in the specimen tested, LABELING: Label the Chewable Tablets to indicate that they
based on the labeled quantity (mg) are to be chewed before being swallowed. Label the
ANC2 = theoretical acid-neutralizing capacity of Mg(OH)2, Chewable Tablets to state the sodium content if it is greater
0.0343 mEq than 5 mg/Tablet. The Chewable Tablets may be labeled to
M = quantity of Mg(OH)2 in the specimen tested, state the aluminum hydroxide content in terms of the
based on the labeled quantity (mg) equivalent amount of dried aluminum hydroxide gel, on the
DEFOAMING ACTIVITY basis that each mg of dried gel is equivalent to 0.765 mg of
Foaming solution: 500 g of FD&C Blue No. 1 and 1 g of Al(OH)3.
octoxynol 9 in 100 mL of 0.1 N hydrochloric acid
USP REFERENCE STANDARDS 11 Ar = atomic weight of aluminum, 26.98

USP Polydimethylsiloxane RS V = volume of Edetate disodium titrant consumed
(mL)
aluminum hydroxide from Chewable Tablets (finely powder
Alumina, Magnesia, and Simethicone NLT 20 Chewable Tablets), to a 150-mL beaker.
Chewable Tablets Add 20 mL of water, stir, and slowly add 30 mL of 3 N
(Comment on this Monograph)id=m1753=Alumina, Magnesia, hydrochloric acid. Heat gently, if necessary, to aid solution,
and Simethicone Chewable Tablets=A-Monos.pdf) cool to room temperature, and filter into a 200-mL
(Monograph under this new titleto become official February volumetric flask. Wash the filter with water into the flask,
1, 2010) and add water to volume.
(Current monograph title is Alumina, Magnesia, and Simethicone Analysis: Pipet 10 mL of the Sample solution into a 250-mL
Tablets.) beaker, add 20 mL of water, then add, in the order named
and with continuous stirring, 25.0 mL of Edetate disodium
DEFINITION titrant and 20 mL of acetic acidammonium acetate buffer
Alumina, Magnesia, and Simethicone Chewable Tablets contain TS, and heat near the boiling temperature for 5 min. Cool,
the equivalent of NLT 90.0% and NMT 115.0% of the labeled add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with
amounts of aluminum hydroxide [Al(OH)3] and magnesium 0.05 M zinc sulfate VS until the color changes from green-
hydroxide [Mg(OH)2], and an amount of polydimethylsiloxane violet to rose-pink. Perform a blank determination,
[(CH3)2 SiO]n that is NLT 85.0% and NMT 115.0% of the substituting 10 mL of water for the Sample solution, and
labeled amount of simethicone. making any necessary correction. Each mL of 0.05 M Edetate
disodium titrant consumed is equivalent to 3.900 mg of
IDENTIFICATION Al(OH)3.
A. INFRARED ABSORPTION 197S Acceptance criteria: 90.0%115.0%
Cell: 0.5 mm MAGNESIUM HYDROXIDE
Solution: Prepared as directed in the Assay for Sample solution: Prepare as directed in the Assay for
Polydimethylsiloxane. Aluminum Hydroxide.
B. IDENTIFICATION TESTSGENERAL, Magnesium 191 Analysis: Pipet a volume of the Sample solution, equivalent
Sample solution: Equivalent to 600 mg of magnesium to 40 mg of magnesium hydroxide, into a 400-mL beaker,
hydroxide from finely powdered Chewable Tablets add 200 mL of water and 20 mL of triethanolamine, and
Add 25 mL of 3 N hydrochloric acid and 25 mL of water, stir. Add 10 mL of ammoniaammonium chloride buffer TS
and mix. Boil gently for 2 min. Allow to cool, and filter. and 3 drops of an eriochrome black indicator solution
Add 5 drops of methyl red TS, heat to boiling, and add 6 (prepared by dissolving 200 mg of eriochrome black T in a
N ammonium hydroxide until the color of the solution just mixture of 15 mL of triethanolamine and 5 mL of
turns to deep yellow. Continue boiling for 2 min, and filter: dehydrated alcohol and mixing). Cool the solution to 34
the filtrate so obtained meets the requirements. by immersion of the beaker in an ice bath, then remove,
C. PROCEDURE and titrate with 0.05 M edetate disodium VS to a blue
Sample: Wash the precipitate obtained in Identification test endpoint. Perform a blank determination, substituting water
B with hot ammonium chloride solution (1 in 50). for the Sample solution, and make any necessary correction.
Analysis: Dissolve the precipitate in hydrochloric acid. Each mL of 0.05 M edetate disodium consumed is
Divide this solution into two portions. equivalent to 2.916 mg of Mg(OH)2.
Acceptance criteria: The dropwise addition of 6 N Acceptance criteria: 90.0%115.0%
ammonium hydroxide to one portion yields a gelatinous POLYDIMETHYLSILOXANE
white precipitate, which does not dissolve in an excess of 6 Sample solution: Transfer the equivalent to 33 mg of
N ammonium hydroxide. The dropwise addition of 1 N simethicone from Chewable Tablets (finely powder NLT 20
sodium hydroxide to the other portion yields a gelatinous Chewable Tablets), to a suitable round, narrow-mouth,
white precipitate, which dissolves in an excess of 1 N screw-capped, 120-mL bottle.
sodium hydroxide, leaving some turbidity. Add 40 mL of 0.1 N sodium hydroxide, and swirl to
disperse. Add 20.0 mL of toluene, close the bottle securely
ASSAY with a cap having an inert liner, and shake for 30 min on a
ALUMINUM HYDROXIDE reciprocating shaker (e.g., 200 oscillations/min and a stroke
Edetate disodium titrant: 18.6 mg/mL of edetate disodium of 38 2 mm). Transfer the mixture to a 125-mL separator,
Standardization of titrant: Transfer 2 g of aluminum wire and allow to separate. Remove the upper, organic layer to
to a 1000-mL volumetric flask, and add 50 mL of a mixture a screw-capped, centrifuge tube containing 2 g of
of hydrochloric acid and water (1:1). Swirl the flask to anhydrous sodium sulfate. Close the tube with a screw-cap
ensure contact of the aluminum and the acid, and allow the having an inert liner, agitate vigorously, and centrifuge the
reaction to proceed until all of the aluminum has dissolved. mixture until a clear supernatant is obtained.
Dilute with water to volume. Pipet 10 mL of this solution Standard solution: Similarly prepare a Standard solution,
into a 250-mL beaker, add, in the order named and with using 33 mg of USP Polydimethylsiloxane RS.
continuous stirring, 25.0 mL of Edetate disodium titrant and Blank: Mix 10 mL of toluene with 0.5 g of anhydrous
20 mL of acetic acidammonium acetate buffer TS, and boil sodium sulfate, and centrifuge to obtain a clear supernatant.
gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL Analysis
of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a Samples: Sample solution, Standard solution, and Blank
bright rose-pink color. Perform a blank determination, Spectrometric conditions
substituting 10 mL of water for the aluminum solution, and (See Spectrophotometry and Light-Scattering 851.)
make any necessary correction. Mode: IR spectrophotometer
Calculate the molarity of the solution taken: Analytical wavelength: Wavelength of maximum
Result = W/ArV absorbance at 7.9 m (1265.8 cm1)
W = weight of aluminum in the portion of solution

taken (mg)
Cell: 0.5 mm Add 50 mL of 1 N hydrochloric acid, boil for 15 min, cool to

Analysis: Use the Blank to set the instrument. room temperature, and dilute with water to volume. Filter,
Calculate the quantity in mg, of [(CH3)2 SiO]n in the discarding the first few mL of the filtrate. Transfer 5.0 mL
portion of Chewable Tablets taken: of the filtrate to a 100-mL volumetric flask containing 10.0
mL of Potassium chloride solution, and dilute with water to
Result = (AU/AS) W volume.
Blank: 1 N hydrochloric acid, Potassium chloride solution,
AU = absorbance of the Sample solution and water (4:10:86)
AS = absorbance of the Standard solution Spectrometric conditions
W = weight in mg of USP Polydimethylsiloxane RS in (See Spectrophotometry and Light-Scattering 851.)
Standard solution Mode: Atomic absorption spectrophotometer
Acceptance criteria: 85.0%115.0% of simethicone Analytical wavelength: 589.0 nm, sodium emission line
PERFORMANCE TESTS Flame: Air acetylene
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Analysis
for Weight Variation with respect to aluminum hydroxide and Samples: Standard solutions and Sample solution
to magnesium hydroxide Plot the absorbances of the Standard solutions versus
SPECIFIC TESTS concentration, in g/mL, of sodium, and draw the straight
ACID-NEUTRALIZING CAPACITY 301 line between the plotted points. From the graph so
Analysis: The acid consumed by the minimum single dose obtained, determine the concentration, C, in g/mL, of
recommended in the labeling is NLT 5 mEq, and NLT the sodium in the Sample solution.
number of mEq calculated: Calculate the quantity, in mg, of sodium per Tablet taken:
Result = 0.55(ANC1 A) + 0.8(ANC2 M) Result = 2C
ANC1 = theoretical acid-neutralizing capacity of Al(OH)3, ADDITIONAL REQUIREMENTS

0.0385 mEq PACKAGING AND STORAGE: Preserve in well-closed containers.
A = quantitly of Al(OH)3 in the specimen tested, LABELING: Label the Chewable Tablets to indicate that they
based on the labeled quantity (mg) are to be chewed before being swallowed. Label the
ANC2 = theoretical acid-neutralizing capacity of Mg(OH)2, Chewable Tablets to state the sodium content if it is greater
0.0343 mEq than 5 mg/Tablet. The Chewable Tablets may be labeled to
M = quantity of Mg(OH)2 in the specimen tested, state the aluminum hydroxide content in terms of the
based on the labeled quantity (mg) equivalent amount of dried aluminum hydroxide gel, on the
DEFOAMING ACTIVITY basis that each mg of dried gel is equivalent to 0.765 mg of
Foaming solution: 500 g of FD&C Blue No. 1 and 1 g of Al(OH)3.
octoxynol 9 in 100 mL of 0.1 N hydrochloric acid. USP REFERENCE STANDARDS 11
Analysis: [NOTEFor each test, use a clean, unused, 250-mL USP Polydimethylsiloxane RS
glass jar.] Transfer a quantity of finely powdered Chewable
Tablets, passed completely through an 80-mesh sieve,
equivalent to 20 mg of simethicone, to a clean, unused,
cylindrical 250-mL glass jar, fitted with a 50-mm cap, Alumina and Magnesium Carbonate
containing 100 mL of Foaming solution that has been
warmed to 37. Cap the jar, and clamp it in an upright
Oral Suspension
position on a wrist-action shaker. Using a radius of 13.3 (Comment on this Monograph)id=m1770=Alumina and
0.4 cm (measured from center of shaft to center of bottle), Magnesium Carbonate Oral Suspension=A-Monos.pdf)
shake for 10 s through an arc of 10 at a frequency of 300 DEFINITION
30 strokes/min. Record the time required for the foam to Alumina and Magnesium Carbonate Oral Suspension contains
collapse. The time, in seconds, for foam collapse is the equivalent of NLT 90.0% and NMT 110.0% of the labeled
determined at the instant the first portion of foam-free amounts of aluminum hydroxide [Al(OH)3] and magnesium
liquid surface appears, measured from the end of the carbonate (MgCO3).
shaking period.
Acceptance criteria: The defoaming activity time does not IDENTIFICATION
exceed 45 s. A. PROCEDURE: Place 1 g in a flask equipped with a stopper
SODIUM CONTENT and glass tubing, the tip of which is immersed in calcium
Potassium chloride solution: 38 mg/mL of potassium hydroxide TS in a test tube. Add 5 mL of 3 N hydrochloric
chloride acid to the flask, and immediately insert the stopper: gas
Sodium chloride stock solution: Dissolve sodium chloride, evolves in the flask and a precipitate is formed in the test
previously dried at 105 for 2 h and weighed, in water, and tube.
dilute with water to obtain a solution containing 25.42 B. IDENTIFICATION TESTSGENERAL, Magnesium 191
g/mL (10 g of sodium/mL). Sample solution: To a solution of 5 g in 10 mL of 3 N
Standard solutions: On the day of use, transfer 4.0 mL of hydrochloric acid, add 5 drops of methyl red TS, heat to
1 N hydrochloric acid and 10.0 mL of Potassium chloride boiling, add 6 N ammonium hydroxide until the color of the
solution to each of two 100-mL volumetric flasks. To the solution changes to deep yellow, then continue boiling for 2
respective flasks, add 5.0 mL and 10.0 mL of Sodium chloride min, and filter: the filtrate meets the requirements.
stock solution. Dilute with water to volume. These solutions C. IDENTIFICATION TESTSGENERAL, Aluminum 191
contain 0.5 g/mL and 1.0 g/mL of sodium, respectively. Sample solution: Wash the precipitate obtained in
Sample solution: Transfer the equivalent to the average Identification test B with a hot solution of ammonium
weight of 1 Tablet from Chewable Tablets (weigh and finely chloride (1 in 50), and dissolve the precipitate in
powder NLT 20 Chewable Tablets), to a 100-mL volumetric hydrochloric acid: the solution meets the requirements.
flask
ASSAY Lamp: Magnesium hollowcathode

ALUMINUM HYDROXIDE Flame: Airacetylene
Potassium chloride solution: 45 mg/mL of potassium Blank: Water
chloride solution Analysis
Aluminum stock solution: Transfer 1.000 g of aluminum Samples: Standard solutions and Sample solution
wire to a 1000-mL volumetric flask, and add 50 mL of 6 N Calculate the percentage of Mg(OH)2 in the portion of Oral
hydrochloric acid. Swirl to ensure contact of the aluminum Suspension taken:
and the acid, and allow the reaction to proceed until all of
the aluminum has dissolved. Dilute with water to volume. Result = (AU/AS) (CS/CU) (Mr/Ar) 100
Standard solutions: To separate 100-mL volumetric flasks,
each containing 10 mL of Potassium chloride solution, AU =
absorbance of the Sample solution
transfer 9.0, 10.0, and 11.0 mL, respectively, of Aluminum AS =
absorbance of the Standard solution
stock solution, and dilute with water to volume. These CS =
concentration of the Standard solution
Standard solutions contain 90.0, 100.0, and 110.0 g of CU nominal concentration, in g of magnesium
=
aluminum per mL, respectively. carbonate per mL, of the Sample solution,
Sample solution: Transfer a quantity of Oral Suspension, based on the labeled amount of magnesium
previously shaken in its original container, equivalent to 75 carbonate in the portion of Oral Suspension
mg of aluminum hydroxide, to a suitable beaker. Add 25 mL taken (mg) and the extent of dilution
of 6 N hydrochloric acid, and heat on a steam bath for 30 Mr = molecular weight of magnesium carbonate,
min, with occasional swirling. Cool, and transfer with the aid 84.31
of water to a 250-mL volumetric flask containing 25 mL of Ar = atomic weight of magnesium, 24.31
Potassium chloride solution. Dilute with water to volume and Acceptance criteria: 90.0%110.0%
filter.
Spectrometric conditions SPECIFIC TESTS
(See Spectrophotometry and Light-Scattering 851.) MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED
Mode: Atomic absorption spectrophotometer MICROORGANISMS 62: Its total aerobic microbial count
Analytical wavelength: Aluminum emission line at 309.3 does not exceed 100 cfu/mL, and it meets the requirements
nm of the test for absence of Escherichia coli, Salmonella species,
Lamp: Aluminum hollowcathode Staphylococcus aureus, and Pseudomonas aeruginosa.
Flame: Nitrous oxideacetylene PH 791: 7.59.5
Blank: Water ACID-NEUTRALIZING CAPACITY 301
Analysis Analysis: NLT 5 mEq of acid is consumed by the minimum
Samples: Standard solutions and Sample solution single dose recommended in the labeling, and NLT the
Calculate the quantity in mg, of Al(OH)3 in the portion of number of mEq calculated:
Oral Suspension taken: Result = 0.55(ANC1 A) + 0.8(ANC2 M)
Result = Mr/Ar 0.25 AU/RS ANC1 = theoretical acid-neutralizing capacity of Al(OH)3
Mr = the molecular weight of aluminum hydroxide, (mEq), 0.0385
78.00 A = quantity of Al(OH)3 in the specimen tested (mg),
Ar = the atomic weight of aluminum, 26.98 based on the labeled quantity
AU = absorbances from the Sample solution ANC2 = theoretical acid-neutralizing capacity of MgCO3
RS = average of the ratio of the absorbances of (mEq), 0.024
Standard solution to their respective M = quantity of MgCO3 in the specimen tested (mg),
concentration of Al in g/mL based on the labeled quantity
MAGNESIUM CARBONATE PACKAGING AND STORAGE: Preserve in tight containers, and
Lanthanum chloride solution: 5 mg/mL of lanthanum avoid freezing.
chloride in water
Magnesium stock solution: Transfer 1.000 g of magnesium
metal to a 1000-mL volumetric flask containing 50 mL of
water, and slowly add 10 mL of hydrochloric acid. Dilute Alumina and Magnesium Carbonate
with water to volume. Transfer 10.0 mL of this solution to a Tablets
100-mL volumetric flask, and dilute with water to volume.
Standard solutions: To separate 100-mL volumetric flasks, (Comment on this Monograph)id=m1780=Alumina and
each containing 10 mL of Lanthanum chloride solution, Magnesium Carbonate Tablets=A-Monos.pdf)
transfer 1.70 mL and 1.80 mL, respectively, of Magnesium DEFINITION
stock solution, and dilute with water to volume. These Alumina and Magnesium Carbonate Tablets contain the
Standard solutions contain 1.7 g/mL of magnesium and 1.8 equivalent of NLT 90.0% and NMT 110.0% of the labeled
g/mL of magnesium, respectively. amounts of aluminum hydroxide [Al(OH)3] and magnesium
Sample solution: Quantitatively dilute a measured volume carbonate (MgCO3).
of the Sample solution prepared as directed in the Assay for
Aluminum hydroxide with water to obtain a solution having a IDENTIFICATION
concentration of 6 g/mL of magnesium carbonate. A. PROCEDURE
Spectrometric conditions Analysis: Place 1 g of finely powdered Tablets in a flask
(See Spectrophotometry and Light-Scattering 851.) equipped with a stopper and glass tubing, the tip of which
Mode: Atomic absorption spectrophotometer is immersed in calcium hydroxide TS in a test tube. Add 5
Analytical wavelength: Magnesium emission line at 285.2
nm
mL of 3 N hydrochloric acid to the flask, and immediately L = label claim (mg/unit)

insert the stopper Acceptance criteria: 90.0%110.0% of the labeled
Acceptance criteria: Gas evolves in the flask and a amounts of Al(OH)3
precipitate is formed in the test tube. MAGNESIUM CARBONATE
B. IDENTIFICATION TESTSGENERAL, Magnesium 191 Lanthanum chloride solution: Transfer 17.6 g of
Sample solution: To a 7-g portion of finely powdered lanthanum chloride to a 1000-mL volumetric flask, add 500
Tablets, add 10 mL of 3 N hydrochloric acid and 5 drops of mL of water, and carefully add 50 mL of hydrochloric acid.
methyl red TS, heat to boiling, and add 6 N ammonium Mix, and allow to cool. Dilute with water to volume.
hydroxide until the color of the solution changes to deep Digestion fluid: Hydrochloric acid, nitric acid, and water
yellow. Continue boiling for 2 min, and filter: the filtrate (1:2:2)
meets the requirements. [NOTEUse promptly]
C. IDENTIFICATION TESTSGENERAL, Aluminum 191 Magnesium stock solution: Transfer 1.000 g of magnesium
Sample solution: Wash the precipitate obtained in metal to a 1000-mL volumetric flask containing 50 mL of
Identification test B with a hot solution of ammonium water, and slowly add 10 mL of hydrochloric acid. Dilute
chloride (1 in 50), and dissolve the precipitate in with water to volume. Transfer 5.0 mL of this solution to a
hydrochloric acid: the solution meets the requirements. 500-mL volumetric flask, and dilute with water to volume.
Standard solutions: To separate 100-mL volumetric flasks,
ASSAY transfer 4.0, 6.0, and 8.0 mL of Magnesium stock solution,
ALUMINUM HYDROXIDE respectively. To each flask, add 0.5 mL of Digestion fluid and
Potassium chloride solution: 38.1 mg/mL of potassium 10 mL of Lanthanum chloride solution, and dilute with water
chloride solution to volume. These Standard solutions contain 0.40, 0.60, and
Digestion fluid: Hydrochloric acid, nitric acid, and water 0.80 g/mL of magnesium, respectively.
(1:2:2) Sample solution: Transfer a measured volume of the filtrate
[NOTEUse promptly] used to prepare the Sample solution in the Assay for
Aluminum stock solution: Transfer 1.000 g of aluminum Aluminum hydroxide, equivalent to 0.4 mg of magnesium
wire to a 1000-mL volumetric flask, and add 50 mL of 6 N carbonate, to a 200-mL volumetric flask, add 20 mL of
hydrochloric acid. Swirl to ensure contact of the aluminum Lanthanum chloride solution, and dilute with water to
and the acid, and allow the reaction to proceed until all of volume.
the aluminum has dissolved. Dilute with water to volume. Spectrometric conditions
Standard solutions: To separate 100-mL volumetric flasks Mode: Atomic absorption spectrophotometer (see
transfer 3.0, 4.0, and 5.0 mL of Aluminum stock solution, Spectrophotometry and Light-Scattering 851)
respectively. To each flask add 10 mL of Potassium chloride Analytical wavelength: Magnesium emission line at 285.2
solution and 7.5 mL of Digestion fluid, dilute with water to nm
volume, and mix. These Standard solutions contain 30, 40, Lamp: Magnesium hollowcathode
and 50 g/mL of aluminum, respectively. Flame: Airacetylene
Sample solution: Transfer an equivalent to 30 mg of Blank: Water
aluminum hydroxide, from finely powdered Tablets (NLT Analysis
20), to a 100-mL volumetric flask, add 25-mL of Digestion Samples: Standard solutions and Sample solution
fluid, and heat on a steam bath for 30 min or on a hot plate Analysis: Plot the absorbances of the Standard solutions
until the volume is reduced by about one-half. Cool, and versus concentration, in g/mL, of magnesium, and draw
dilute with water to volume. Filter, discarding the first 20 the straight line best fitting the three plotted points. From
mL of the filtrate. Transfer 15.0 mL of the filtrate to a 50-mL the graph so obtained, determine the concentration, C, in
volumetric flask, add 5.0 mL of Potassium chloride solution, g/mL, of magnesium in each mL of the Sample solution.
and dilute with water to volume. Calculate the quantity in mg, of MgCO3 in the portion of
[NOTEReserve a portion of the filtrate for use in the Assay Tablets taken:
for Magnesium carbonate.]
Spectrometric conditions Result = (20C/V) (Mr/Ar)
(See Spectrophotometry and Light-Scattering 851.)
Mode: Atomic absorption spectrophotometer C = concentration of the Sample solution (g/mL)
Analytical wavelength: Aluminum emission line at 309.3 V = sample volume taken (mL)
nm Mr = molecular weight of magnesium carbonate,
Lamp: Aluminum hollow-cathode 84.31
Flame: Nitrous oxideacetylene Ar = atomic weight of magnesium, 24.31
Blank: Water Acceptance criteria: 90.0%110.0%
Analysis
Samples: Standard solutions and Sample solution PERFORMANCE TESTS
Analysis: Plot the absorbances of the Standard preparations DISINTEGRATION 701
versus concentration, in g/mL, of aluminum, and draw Medium: Simulated gastric fluid TS being substituted for
the straight line best fitting the three plotted points. From water in the test
the graph so obtained, determine the concentration, C, in Time: 10 min
g/mL, of aluminum in each mL of the Sample solution. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Calculate the percentage of Al(OH)3 in the portion of for Weight Variation with respect to aluminum hydroxide and
Tablets taken: to magnesium carbonate
Result = (C/3) (Mr/Ar) (100/L) SPECIFIC TESTS

ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is
C = concentration for the Sample solution (g/mL) consumed by the minimum single dose recommended in
Mr = molecular weight of aluminum hydroxide, 78.00 the labeling.
ADDITIONAL REQUIREMENTS and slowly add 30 mL 3 N hydrochloric acid. Heat gently, if

PACKAGING AND STORAGE: Preserve in tight containers. necessary, to aid solution, cool, and filter into a 200-mL
volumetric flask. Wash the filter with water into the flask,
and dilute with water to volume.
Alumina, Magnesium Carbonate, and beaker, add 20 mL of water, then add, in the order named
Magnesium Oxide Tablets and with continuous stirring, 25.0 mL of Edetate disodium
(Comment on this Monograph)id=m1795=Alumina, Magnesium titrant and 20 mL of acetic acidammonium acetate buffer
Carbonate, and Magnesium Oxide Tablets=A-Monos.pdf) TS, and heat near the boiling point for 5 min. Cool, add 50
mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M
DEFINITION zinc sulfate VS until the color changes from green-violet to
Alumina, Magnesium Carbonate, and Magnesium Oxide Tablets rose-pink. Perform a blank determination, substituting 10
contain the equivalent of NLT 90.0% and NMT 110.0% of the mL of water for the Sample solution, and make any necessary
labeled amounts of aluminum hydroxide [Al(OH)3] and correction. Each mL of 0.05M Edetate disodium titrant
magnesium carbonate (MgCO3), and NLT 85.0% and NMT consumed is equivalent to 3.900 mg of Al(OH)3.
115.0% of the labeled amount of magnesium oxide (MgO). Acceptance criteria: 90.0%110.0%
MAGNESIUM CARBONATE
IDENTIFICATION Sample solution: Finely powder NLT 20 Tablets. Transfer a
A. PROCEDURE portion of the powder, equivalent to 750 mg of magnesium
Place 3 g of finely powdered Tablets in a flask equipped carbonate, to a 250-mL conical flask fitted with a two-hole
with a stopper and glass tubing, the tip of which is stopper.
immersed in calcium hydroxide TS in a test tube. Add 5 mL Analysis: Fill the lower transverse section of a U-shaped
of 3 N hydrochloric acid to the flask, and immediately insert drying tube of about 15-mm internal diameter and 15-cm
the stopper. height with loosely packed glass wool. Place in one arm of
Acceptance criteria: Gas evolves in the flask and a the tube, about 5 g of anhydrous calcium chloride, and
precipitate is formed in the test tube. weigh the tube and the contents. Into the other arm of the
B. IDENTIFICATION TESTSGENERAL, Aluminum 191 tube, place 9.5 g to 10.5 g of soda lime, and again weigh.
Sample solution: Solution in the flask obtained in Insert stoppers in the open arms of the U-tube, and connect
Identification test A the side tube of the arm filled with soda lime to a calcium
Analysis: To the Sample solution add 5 drops of methyl red chloride drying tube, which in turn is connected to one of
TS, and heat to boiling. Add 6 N ammonium hydroxide until the holes in the stopper of the 250-mL conical flask. Attach
the color of the solution changes to deep yellow, continue a dropping funnel to the other hole in the stopper of the
boiling for 2 min, and filter through hardened filter paper. 250-mL conical flask. Add 100 mL of water and 10 mL of a
(Retain the filtrate for Identification test C.) Wash the mixture of hydrochloric acid and nitric acid (4:1) to the 250-
precipitate with 350 mL of a hot ammonium chloride mL conical flask through the dropping funnel, and close the
solution (1 in 50), discarding the washings. dropping funnel. Heat the 250-mL conical flask at 95 for 1
Acceptance criteria: The precipitate so obtained, dissolved h, and allow the evolved carbon dioxide to pass through the
in 3 N hydrochloric acid, meets the requirements of the tests U-tube. Replace the dropping funnel with a source of
for Aluminum. carbon dioxide-free air, and pass the carbon dioxide-free air
C. IDENTIFICATION TESTSGENERAL, Magnesium 191: The through the apparatus at a rate of about 75 mL/min for 30
filtrate obtained in Identification test B meets the min. Disconnect the U-tube, cool to room temperature,
requirements of the tests for Magnesium. remove the stoppers, and weigh. The increase in weight
corresponds to the quantity of carbon dioxide evolved.
ASSAY Calculate the quantity, in mg, of magnesium carbonate in
ALUMINUM HYDROXIDE each Tablet taken:
Standardization of titrant: Transfer 2 g of aluminum wire Result = (Mr1/Mr2) (I) (WA/WP)
to a 1000-mL volumetric flask, and add 50 mL of a mixture
of hydrochloric acid and water (1:1). Swirl the flask to Mr1 = molecular weight of magnesium carbonate,
ensure contact of the aluminum and the acid, and allow the 84.31
reaction to proceed until all of the aluminum has dissolved. Mr2 = molecular weight of carbon dioxide, 44.01
Dilute with water to volume. Pipet 10 mL of this solution I = quantity of carbon dioxide evolved from the
into a 250-mL beaker, add, in the order named and with portion of Tablets taken (mg)
continuous stirring, 25.0 mL of Edetate disodium titrant and WA = average weight of 1 Tablet (g)
20 mL of acetic acidammonium acetate buffer TS, and boil WP = weight of the portion of Tablets taken (g)
gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL Acceptance criteria: 90.0%110.0%
of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a MAGNESIUM OXIDE
bright rose-pink color. Perform a blank determination, Sample solution: Finely powder NLT 20 Tablets. Transfer a
substituting 10 mL of water for the aluminum solution, and portion of the powder, equivalent to 1000 mg of
make any necessary correction. magnesium carbonate and magnesium oxide combined, to
Calculate the molarity of the solution taken: a beaker, add 20 mL of water, and slowly add 40 mL of 3 N
hydrochloric acid, with mixing.
Result = W/ArV Analysis: Heat the mixture to boiling, cool, and filter into a
200-mL volumetric flask. Wash the beaker with water,
W = weight of aluminum in the portion of solution adding the washings to the filter. Add water to volume.
taken (mg) Transfer 20.0 mL of this solution to a 400-mL beaker, add
Ar = atomic weight of aluminum, 26.98 180 mL of water and 20 mL of triethanolamine, and stir.
V = volume of Edetate disodium titrant consumed Add 10 mL of ammoniaammonium chloride buffer TS and
(mL) 3 drops of an eriochrome black indicator solution (prepared
Sample solution: Finely powder NLT 20 Tablets. Transfer a by dissolving 200 mg of eriochrome black T in a mixture of
portion of the powder, equivalent to 1200 mg of aluminum 15 mL of triethanolamine and 5 mL of dehydrated alcohol
hydroxide, to a 150-mL beaker, add 20 mL of water, stir,
and mixing). Cool the solution to between 3 and 4 by Acceptance criteria: The filtrate meets the requirements.
immersion of the beaker in an ice bath, then remove, and C. PROCEDURE Transfer the filter paper and contents from
titrate with 0.05 M edetate disodium VS to a blue endpoint. Identification test B to a small platinum dish, ignite, cool in a
Perform a blank determination, substituting 20 mL of water desiccator, and weigh. Moisten the residue with water and
for the assay solution, and make any necessary correction. add 6 mL of hydrofluoric acid. Evaporate to dryness, ignite
Each mL of 0.05 M edetate disodium consumed is for 5 min, cool in a desiccator, and weigh: a loss of more
equivalent to 1.216 mg of Mg. than 10% in relation to the weight of the residue from the
Calculate the quantity, in mg, of magnesium equivalent in initial ignition indicates SiO2.
each Tablet taken:
ASSAY
Result = 10T (WA/WP) ALUMINUM HYDROXIDE
T = magnesium equivalent obtained in the titration in water
WA = average weight of 1 Tablet (g) Standardization of titrant: Transfer 2 g of aluminum wire
WP = weight of the portion of Tablets taken (g) to a 1000-mL volumetric flask, and add 50 mL of a mixture
Calculate the quantity, in mg, of magnesium oxide in each of hydrochloric acid and water (1:1). Swirl the flask to
Tablet taken: ensure contact of the aluminum and the acid, and allow the
reaction to proceed until all of the aluminum has dissolved.
Result = (Mr3/Ar1) (A 0.2883B) Dilute with water to volume. Pipet 10 mL of this solution
Mr3 = molecular weight of magnesium oxide, 40.30 continuous stirring, 25.0 mL of Edetate disodium titrant and
Ar1 = atomic weight of magnesium, 24.31 20 mL of acetic acidammonium acetate buffer TS, and boil
A = quantity of magnesium equivalent in each Tablet gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL
(mg) of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a
B = quantity of magnesium carbonate in each Tablet, bright rose-pink color. Perform a blank determination,
as determined in the Assay for magnesium substituting 10 mL of water for the aluminum solution, and
carbonate (mg) make any necessary correction.
Acceptance criteria: 85.0%115.0% Calculate the molarity of the solution taken:
SPECIFIC TESTS W/ArV
ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is
consumed by the minimum single dose recommended in W = weight of aluminum in the portion of solution
the labeling. taken (mg)
PERFORMANCE TESTS (mL)
DISINTEGRATION 701: Ar = atomic weight of aluminum, 26.98
Medium: Simulated gastric fluid TS being substituted for Sample solution: Transfer 10 g of well-shaken Oral
water in the test Suspension to a tared beaker, and weigh. Add 50 mL of
Time: 10 min water and 10 mL of hydrochloric acid, and digest on a
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements steam bath for 1 h. Cool, and filter into a 200-mL
for Weight Variation with respect to alumina, to magnesium volumetric flask, washing the filter with water into the flask.
carbonate, and to magnesium oxide Dilute with water to volume.
ADDITIONAL REQUIREMENTS Analysis: Pipet 20 mL of Sample solution into a 250-mL
PACKAGING AND STORAGE: Preserve in tight containers. beaker, add 20 mL of water, then add, in the order named
and with continuous stirring, 25.0 mL of Edetate disodium
TS, and heat near the boiling point for 5 min. Cool, and
Alumina and Magnesium Trisilicate add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with
Oral Suspension 0.05 M zinc sulfate VS until the color changes from green-
violet to rose-pink. Perform a blank determination,
(Comment on this Monograph)id=m1810=Alumina and substituting 20 mL of water for the Sample solution, and
Magnesium Trisilicate Oral Suspension=A-Monos.pdf) make any necessary correction. Each mL of 0.05 M Edetate
DEFINITION disodium titrant consumed is equivalent to 3.900 mg of
Alumina and Magnesium Trisilicate Oral Suspension contains the Al(OH)3.
equivalent of NLT 90.0% and NMT 110.0% of the labeled Acceptance criteria: Equivalent of 90.0%110.0% of the
amounts of aluminum hydroxide [Al(OH)3], and magnesium labeled amounts of Al(OH)3
trisilicate (Mg2Si3O8). MAGNESIUM TRISILICATE
Sample solution: Prepare as directed in the Assay for
IDENTIFICATION Aluminum hydroxide.
A. IDENTIFICATION TESTSGENERAL, Magnesium 191 Analysis: Pipet 20 mL of Sample solution into a 400-mL
Sample solution To a mixture of 5 mL in 10 mL of 3 N beaker, add 180 mL of water and 20 mL of triethanolamine,
hydrochloric acid add 5 drops of methyl red TS, heat to and stir. Add 10 mL of ammoniaammonium chloride buffer
boiling, add 6 N ammonium hydroxide until the color of the TS and 3 drops of an eriochrome black indicator solution
solution changes to deep yellow, then continue boiling for 2 (prepared by dissolving 200 mg of eriochrome black T in a
min, and filter. mixture of 15 mL of triethanolamine and 5 mL of
Acceptance criteria: The filtrate meets the requirements. dehydrated alcohol). Cool the solution to between 3 and 4
B. IDENTIFICATION TESTSGENERAL, Aluminum 191 by immersion of the beaker in an ice bath, then remove and
Sample solution: Wash the solids on the filter obtained in titrate with 0.05 M edetate disodium VS to a blue endpoint.
Identification A with hot ammonium chloride solution (1 in Perform a blank determination, substituting 20 mL of water
50), add 10 mL of 3 N hydrochloric acid, and filter. for the Sample solution, and make any necessary correction.
Each mL of 0.05 M edetate disodium consumed is W = weight of aluminum in the portion of solution
equivalent to 6.521 mg of Mg2Si3O8. taken
Acceptance criteria: Equivalent of 90.0%110.0% of the Ar = atomic weight of aluminum, 26.98
labeled amounts of Mg2Si3O8 V = volume of the Edetate disodium titrant consumed
Sample solution: Finely powder NLT 20 Tablets. Transfer a
SPECIFIC TESTS portion of the powder, equivalent to 600 mg of aluminum
ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is hydroxide, to a beaker, add 20 mL of water, stir, and slowly
consumed by the minimum single dose recommended in add 40 mL of 3 N hydrochloric acid. Heat gently, if
the labeling. necessary, to aid solution, cool, and transfer to a 200-mL
PH 791: 7.58.5 volumetric flask. Wash the beaker with water, adding the
washings to the flask, and add water to volume.
ADDITIONAL REQUIREMENTS Analysis: Pipet 10 mL of the Sample solution into a 250-mL
PACKAGING AND STORAGE: Preserve in tight containers. beaker, add 20 mL of water, then add, in the order named
and with continuous stirring, 25.0 mL of 0.05 M Edetate
disodium titrant and 20 mL of acetic acidammonium
Alumina and Magnesium Trisilicate acetate buffer TS, and heat the solution near the boiling
Tablets temperature for 5 min. Cool, add 50 mL of alcohol and 2
mL of dithizone TS, and mix. Titrate with 0.05 M zinc sulfate
(Comment on this Monograph)id=m1820=Alumina and VS until the color changes from green-violet to rose-pink.
Magnesium Trisilicate Tablets=A-Monos.pdf) Perform a blank determination, substituting 10 mL of water
for the Sample solution. Each mL of 0.05 M Edetate disodium
DEFINITION titrant consumed is equivalent to 3.900 mg of Al(OH)3.
Alumina and Magnesium Trisilicate Tablets contain NLT 90.0% Acceptance criteria: 90.0%110.0%
and NMT 110.0% of the labeled amounts of [Al(OH)3] and MAGNESIUM TRISILICATE
Mg2Si3O8. Potassium chloride solution: 50 mg/mL of potassium
IDENTIFICATION chloride in water
One powdered Tablet responds to the following tests. Magnesium standard solution: Transfer 1.000 g of
A. IDENTIFICATION TESTSGENERAL, Magnesium 191 magnesium metal to a 1000-mL volumetric flask containing
Sample solution: To a mixture of 5 mL in 10 mL of 3 N 50 mL of water, and slowly add 10 mL of hydrochloric acid.
hydrochloric acid, add 5 drops of methyl red TS, heat to Dilute with water to volume. Transfer 5.0 mL of this solution
boiling, add 6 N ammonium hydroxide until the color of to a 500-mL volumetric flask, and dilute with water to
the solution changes to deep yellow, continue boiling for 2 volume.
min, and filter. Standard solutions: Transfer 16.0, 18.0, and 20.0 mL of
Acceptance criteria: The filtrate meets the requirements. Magnesium standard solution to separate 100-mL volumetric
B. IDENTIFICATION TESTSGENERAL, Aluminum 191 flasks, add 2.0 mL of Potassium chloride solution to each
Sample solution: Wash the solids on the filter obtained in flask, and dilute with water to volume. These Standard
Identification Test A with hot ammonium chloride solution solutions contain 1.6, 1.8, and 2.0 g/mL of magnesium
(1 in 50), add 10 mL of 3 N hydrochloric acid, and filter. respectively.
Acceptance criteria: The filtrate meets the requirements. [NOTEPrepare these solutions on the day of use.]
C. PROCEDURE Sample solution: Finely powder NLT 20 Tablets. Transfer a
Sample: Transfer the filter paper and contents from portion of the powder, equivalent to 5 mg of magnesium
Identification Test B to a small platinum dish, ignite, cool in trisilicate, to a 100-mL volumetric flask, and add 10 mL of
a desiccator, and weigh. 18 N sulfuric acid. Heat on a steam bath for 30 min with
Analysis: Moisten the residue with water, and add 6 mL occasional swirling. Allow to cool, and dilute with water to
of hydrofluoric acid. Evaporate to dryness, ignite for 5 min, volume. Filter this solution, discarding the first 20 mL of the
cool in a desiccator, and weigh: a loss of more than 10% filtrate. Transfer 20.0 mL of the filtrate to a second 100-mL
in relation to the weight of the residue from the initial volumetric flask, add 2.0 mL of Potassium chloride solution,
ignition indicates SiO2. and dilute with water to volume.
Acceptance criteria: Meets the requirements Spectrometric conditions
(See Spectrophotometry and Light-Scattering 851.)
ASSAY Mode: Atomic absorption spectrophotometer
ALUMINUM HYDROXIDE Analytical wavelength: 285.2 nm
Edetate disodium titrant: 18.6 mg/mL of edetate disodium Lamp: Magnesium hollowcathode lamp
in water Flame: Nitrous oxideacetylene flame
Standardization of titrant: Transfer 2 g of aluminum wire Blank: Water
to a 1000-mL volumetric flask, and add 50 mL of a mixture Analysis
of hydrochloric acid and water (1:1). Swirl the flask to Samples: Standard solutions and Sample solution
ensure contact of the aluminum and the acid, and allow the Analysis: Plot the absorbances of the Standard solutions of
reaction to proceed until all of the aluminum has dissolved. magnesium, and draw the line best fitting the three plotted
Dilute with water to volume. Pipet 10 mL of this solution points in g/mL. From the graph so obtained, determine
into a 250-mL beaker, add, in the order named and with the concentration of magnesium in the Sample solution in
continuous stirring, 25.0 mL of Edetate disodium titrant and g/mL.
20 mL of acetic acidammonium acetate buffer TS, and boil Calculate the quantity, in mg, of Mg2Si3O8 in the portion of
gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL Tablets taken:
of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a
bright rose-pink color. Perform a blank determination, Result = 0.5 CU Mr/Ar
substituting 10 mL of water for the aluminum solution.
Calculate the molarity of the portion of solution taken: CU = concentration of the Sample solution (g/mL)
Mr = molecular weight of anhydrous magnesium
Result = W/ArV trisilicate, 260.86
USP 32 Official Monographs / Aluminum 123
Ar = twice the atomic weight of magnesium, 48.62 DEFINITION

Acceptance criteria: 90.0%110.0% Aluminum Acetate Topical Solution yields, from each 100 mL,
NLT 1.20 g and NMT 1.45 g of aluminum oxide (Al2O3), and
SPECIFIC TESTS NLT 4.24 g and NMT 5.12 g of acetic acid (C2H4O2),
ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is corresponding to NLT 4.8 g and NMT 5.8 g of aluminum
consumed by the minimum single dose recommended in acetate (C6H9AlO6). Aluminum Acetate Topical Solution may be
the labeling. stabilized by the addition of NMT 0.6% of Boric Acid.
[NOTETablets labeled for the temporary relief of heartburn
(acid indigestion) due to acid reflux are exempt from this
Aluminum Subacetate Topical Solution 545 mL
requirement.]
PH 791: NLT 4.5, determined on the foam layer obtained Glacial Acetic Acid 15 mL
in the Foam test, where Tablets are labeled for the Purified Water A sufficient quantity
temporary relief of heartburn (acid indigestion) due to acid
To make 1000 mL
reflux.
[NOTETake care that the electrodes do not touch the liquid
beneath the foam.] Add the Glacial Acetic Acid to the Aluminum Subacetate Topical
FOAM [Where Tablets are labeled for the temporary relief of Solution and sufficient water to make 1000 mL. Mix, and filter,
heartburn (acid indigestion) due to acid reflux] if necessary.
Sample solution: Finely powder a number of Tablets [NOTEDispense only clear Aluminum Acetate Topical Solution.]
equivalent to the minimum single dose recommended in the IDENTIFICATION
labeling, and transfer the powder to a 100-mL beaker IDENTIFICATION TESTSGENERAL, Aluminum AND Acetate 191
having an inside diameter of 45 mm. Add 5 mL of alcohol Acceptance criteria: Meets the requirements of the tests
and sufficient water to make 40 mL.
Analysis: Mix at 300 rpm for 60 s, using a magnetic stirrer ASSAY
and a 9.5- 38-mm polytef-coated stirring bar. Stop the ALUMINUM OXIDE
stirrer, and carefully add 10 mL of 0.5 N hydrochloric acid Edetate disodium titrant: 18.6 mg/mL of edetate disodium
down the side of the beaker. Stir for 30 s at 300 rpm. Allow in water
to stand for 10 min, and measure the thickness of the foam Standardization of Titrant: Transfer 2 g of aluminum wire
layer above the liquid in the beaker. to a 1000-mL volumetric flask, and add 50 mL of a mixture
Acceptance criteria: The thickness of the foam is NLT 10 of hydrochloric acid and water (1:1). Swirl the flask to
mm. ensure contact of the aluminum and the acid, and allow the
PERFORMANCE TESTS Dilute with water to volume. Pipet 10 mL of this solution
DISINTEGRATION 701 into a 250-mL beaker and add, in the order named and
Medium: Simulated gastric fluid TS being substituted for with continuous stirring, 25.0 mL of Edetate disodium titrant
water in the test and 20 mL of acetic acidammonium acetate buffer TS, and
[NOTETablets that must be chewed before swallowing are boil gently for 5 min. Cool, and add 50 mL of alcohol, and
exempt from this requirement.] 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to
Time: 10 min a bright rose-pink color. Perform a blank determination,
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements substituting 10 mL of water for the aluminum solution, and
for Weight Variation with respect to aluminum hydroxide and make any necessary correction.
to magnesium trisilicate. Calculate the molarity of the solution taken:
ADDITIONAL REQUIREMENTS W/ArV
LABELING: Tablets prepared with the use of Dried Aluminum W = weight of aluminum in the portion of solution
Hydroxide Gel may be labeled to state the aluminum taken (mg)
hydroxide content in terms of the equivalent amount of Ar = atomic weight of aluminum, 26.98
dried aluminum hydroxide gel, on the basis that each mg of V = volume of Edetate disodium titrant consumed
dried gel is equivalent to 0.765 mg of Al(OH)3. Tablets (mL)
intended for the temporary relief of heartburn (acid Sample: 25 mL of Aluminum Acetate Topical Solution
indigestion) due to acid reflux are so labeled. Tablets that Analysis: Pipet the Sample into a 250-mL volumetric flask,
must be chewed before swallowing are so labeled. add 5 mL of hydrochloric acid, and dilute with water to
volume. Pipet 25 mL of this solution into a 250-mL beaker,
and add, in the order named and with continuous stirring,
Aluminum Acetate Topical Solution 25.0 mL of Edetate disodium titrant and 20 mL of acetic
acidammonium acetate buffer TS, then heat the solution
(Comment on this Monograph)id=m1890=Aluminum Acetate near the boiling point for 5 min. Cool, and add 50 mL of
Topical Solution=A-Monos.pdf) alcohol and 2 mL of dithizone TS. Titrate the solution with
0.05 M zinc sulfate VS to a bright rose-pink color. Perform a
blank determination, substituting water for the Sample, and
make any necessary correction. Each mL of 0.05 M Edetate
disodium titrant is equivalent to 2.549 mg of Al2O3.
Acceptance criteria: NLT 1.20 g/100 mL and NMT 1.45
g/100 mL of aluminum oxide (Al2O3)
ACETIC ACID
C6H9AlO6 204.11 Sample: 20 mL Aluminum Acetate Topical Solution
Acetic acid, aluminum salt; Analysis: Pipet the Sample into a Kjeldahl flask containing a
Aluminum acetate [139-12-8]. mixture of 20 mL of phosphoric acid and 150 mL of water.
124 Aluminum / Official Monographs USP 32
Connect the flask to a condenser, the delivery tube from Standardization of titrant: Transfer 2 g of aluminum wire
which dips beneath the surface of 50.0 mL of 0.5 N sodium to a 1000-mL volumetric flask, and add 50 mL of a mixture
hydroxide VS contained in a receiving flask. Distill about 160 of hydrochloric acid and water (1:1). Swirl the flask to
mL, then remove the delivery tube from below the surface ensure contact of the aluminum and the acid, and allow the
of the liquid, allow the distilling flask to cool, add 50 mL of reaction to proceed until all of the aluminum has dissolved.
water, and distill an additional 40 to 45 mL into the Dilute with water to volume. Pipet 10 mL of this solution
receiving flask. Add phenolphthalein TS to the distillate, and into a 250-mL beaker and add, in the order named and
titrate the excess 0.5 N sodium hydroxide VS with 0.5 N with continuous stirring, 25.0 mL of Edetate disodium titrant
sulfuric acid VS. Each mL of 0.5 N sodium hydroxide is and 20 mL of acetic acidammonium acetate buffer TS, and
equivalent to 30.03 mg of C2H4O2. boil gently for 5 min. Cool, and add 50 mL of alcohol, and
Acceptance criteria: NLT 4.24 g/100 mL and NMT 5.12 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to
g/100 mL of C2H4O2 a bright rose-pink color. Perform a blank determination,
[NOTEThe results for both assays correspond to NLT 4.8 substituting 10 mL of water for the aluminum solution, and
g/100 mL and NMT 5.8 g/100 mL of C6H9AlO6.] make any necessary correction. Calculate the molarity of the
solution taken:
OTHER COMPONENTS
LIMIT OF BORIC ACID Result = W/Ar V
Sample: 25 mL Aluminum Acetate Topical Solution
Analysis: Pipet the Sample into 75 mL of water in a conical W = weight of aluminum (mg)
flask. Add 3 mL of phenolphthalein TS, then add 0.5 N Ar = atomic weight of aluminum, 26.98
sodium hydroxide VS from a buret until a faint pink color is V = volume of Edetate disodium titrant consumed
obtained. Heat to boiling, and again neutralize. Add 150 mL (mL)
of glycerin to the neutralized solution, and titrate with 0.5 N Sample solution: 20 mg/mL
sodium hydroxide VS. Perform a blank determination in a Analysis: Pipet 10 mL of the Sample solution into a 250-mL
similar manner. From the volume of 0.5 N sodium hydroxide beaker, and add, in the order named and with continuous
VS used after the addition of the glycerin, subtract the stirring, 25.0 mL of Edetate disodium titrant and 20 mL of
volume used in the blank. Each mL of 0.5 N sodium acetic acidammonium acetate buffer TS, and boil gently for
hydroxide is equivalent to 30.92 mg of H3BO3. 5 min. Cool, and add 50 mL of alcohol and 2 mL of
Acceptance criteria: NMT 0.6% dithizone TS. Titrate the solution with 0.05 M zinc sulfate VS
to a bright rose-pink color. Perform a blank determination,
IMPURITIES substituting 10 mL of water for the sample, and make any
Inorganic Impurities necessary correction. Each mL of 0.05 M Edetate disodium
HEAVY METALS 231 titrant is equivalent to 6.667 mg of AlCl3.
Sample solution: 2 mL of Sample diluted to 25 mL Acceptance criteria: 95.0%102.0%
Acceptance criteria: NMT 10 ppm
IMPURITIES
SPECIFIC TESTS Inorganic Impurities
PH 791 CHLORIDE AND SULFATE, Sulfate 221
Acceptance criteria: 3.64.4 Sample solution: 10 mg/mL
Analysis: Add 0.2 mL of barium chloride TS to 10 mL of
ADDITIONAL REQUIREMENTS the Sample solution.
PACKAGING AND STORAGE: Preserve in tight containers. Acceptance criteria: No turbidity is produced within 1
min.
IRON 241
Aluminum Chloride Sample solution: Dissolve 1.0 g of sample in 45 mL of
water, and add 2 mL of hydrochloric acid.
(Comment on this Monograph)id=m2040=Aluminum Acceptance criteria: NMT 10 ppm
Chloride=A-Monos.pdf) HEAVY METALS, Method I 231
AlCl3 6H2O 241.43 Sample solution: Dissolve 1 g of sample in 1 mL of 1 N
Aluminum chloride, hexahydrate; acetic acid, and sufficient water to make 25 mL.
Aluminum chloride hexahydrate [7784-13-6]. Acceptance criteria: NMT 20 ppm
Anhydrous 133.34
[7446-70-0]. SPECIFIC TESTS
WATER DETERMINATION, Method I 921
DEFINITION Acceptance criteria: Between 42.0% and 48.0%
Aluminum Chloride contains NLT 95.0% and NMT 102.0% of LIMIT OF ALKALIES AND ALKALINE EARTHS
AlCl3, calculated on the anhydrous basis. Sample solution: 66.7 mg/mL
Analysis: To 100 mL of boiling Sample solution, add a few
IDENTIFICATION drops of methyl red TS, then add 6 N ammonium hydroxide
IDENTIFICATION TESTSGENERAL, Aluminum 191 until the color of the solution just changes to a distinct
Sample solution: 100 mg/mL yellow. Add hot water to restore the volume to 150 mL, and
Acceptance criteria: Meets the requirements filter while hot. Evaporate 75 mL of the filtrate to dryness,
IDENTIFICATION TESTSGENERAL, Chloride 191 and ignite to a constant weight.
Sample solution: 100 mg/mL Acceptance criteria: The weight of the residue does not
Acceptance criteria: Meets the requirements exceed 2.5 mg (0.5%).
ASSAY
PROCEDURE
in water
ADDITIONAL REQUIREMENTS of hydrochloric acid and water (1:1). Swirl the flask to
PACKAGING AND STORAGE: Preserve in tight containers. ensure contact of the aluminum and the acid, and allow the
Dilute with water to volume. Pipet 10 mL of this solution
Aluminum Chlorohydrate with continuous stirring, 25.0 mL of Edetate disodium titrant
(Comment on this Monograph)id=m2050=Aluminum and 20 mL of acetic acidammonium acetate buffer TS, and
Chlorohydrate=A-Monos.pdf) boil gently for 5 min. Cool, and add 50 mL of alcohol and 2
AIy(OH)3y-zClz H2O mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a
Aluminum chlorohydroxide; bright rose-pink color. Perform a blank determination,
Aluminum hydroxychloride; substituting 10 mL of water for the aluminum solution, and
Dihydrate [12042-91-0]. make any necessary correction.
Anhydrous [1327-41-9]. Calculate the molarity of the solution taken:
Aluminum chlorohydroxide, dihydrate; Result = W/Ar V
Aluminum hydroxychloride, dihydrate;
Dihydrate 210.48 W = weight of aluminum (mg)
[12042-91-0]. Ar = atomic weight of aluminum, 26.98
Anhydrous 174.45 V = volume of Edetate disodium titrant consumed
[1327-41-9]. (mL)
DEFINITION Sample solution: Transfer 200 mg of Aluminum
Aluminum Chlorohydrate consists of complex basic aluminum Chlorohydrate to a 250-mL beaker, add 20 mL of water and
chloride that is polymeric and loosely hydrated and 5 mL of hydrochloric acid, boil on a hot plate for NLT 5
encompasses a range of aluminum-to-chloride atomic ratios min, and allow to cool.
between 1.91:1 and 2.10:1. It contains the equivalent of NLT Analysis: To the Sample solution, add 25.0 mL of Edetate
90.0% and NMT 110.0% of the labeled amount of anhydrous disodium titrant, and adjust with 2.5 N ammonium
aluminum chlorohydrate. hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20
mL of acetic acidammonium acetate buffer TS, 50 mL of
IDENTIFICATION alcohol, and 5 mL of dithizone TS. The pH of this solution
A. IDENTIFICATION TESTSGENERAL, Aluminum 191 should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until
Sample solution: 100 mg/mL the color changes from green-violet to rose-pink. Perform a
Acceptance criteria: Meets the requirements blank titration, and make any necessary correction. Each mL
B. IDENTIFICATION TESTSGENERAL, Chloride 191 of 0.1 M Edetate disodium titrant consumed is equivalent to
Sample solution: 100 mg/mL 2.698 mg of aluminum (Al).
Acceptance criteria: Meets the requirements Use the aluminum content obtained to calculate the
Aluminum/Chloride Atomic Ratio.
ASSAY ALUMINUM/CHLORIDE ATOMIC RATIO
PROCEDURE Analysis: Divide the percentage of aluminum found in the
Calculate the percentage of anhydrous aluminum test for Content of Aluminum by the percentage of chloride
chlorohydrate in the Aluminum Chlorohydrate taken: found in the test for Content of Chloride, and multiply by
35.453/26.98, in which 35.453 and 26.98 are the atomic
Result = Al({Ar1x + [Mr(3x -1)] + Ar2}/Ar1x) weights of chlorine and aluminum, respectively.
Acceptance criteria: The ratio is between 1.91:1 and
Al = percentage of aluminum as obtained in the test 2.10:1.
for Content of Aluminum
Ar1 = atomic weight of aluminum, 26.98 IMPURITIES
x = Aluminum/Chloride Atomic Ratio Inorganic Impurities
Mr = molecular weight of the hydroxide anion (OH), ARSENIC, Method I 211
17.01 Acceptance criteria: NMT 2 ppm
Ar2 = atomic weight of chlorine (Cl), 35.453 HEAVY METALS, Method I 231
Acceptance criteria: NLT 90.0% and NMT 110.0% Acceptance criteria: NMT 20 ppm
LIMIT OF IRON
OTHER COMPONENTS Standard solution: 2.0 mL of Standard Iron Solution,
CONTENT OF CHLORIDE prepared as directed for Iron 241
Sample: 700 mg of Aluminum Chlorohydrate Sample solution: 27 mg/mL
Analysis: Transfer the Sample to a 250-mL beaker, and add, Analysis: Pipet 5.0 mL of the Standard solution into a 50-
with stirring, 100 mL of water and 10 mL of diluted nitric mL beaker. Pipet 5.0 mL of the Sample solution to a second
acid. Titrate with 0.1 N silver nitrate VS using a glass 50-mL beaker. To each of the beakers, add 5 mL of 6 N
silversilver chloride electrode and a silver billet electrode nitric acid, cover with a watch glass, and boil on a hot
system, determining the endpoint potentiometrically. Each plate for 35 min. Allow to cool, add 5 mL of Ammonium
mL of 0.1 N silver nitrate is equivalent to 3.545 mg of Thiocyanate Solution, prepared as directed for Iron 241,
chloride (Cl). transfer to separate 50-mL color comparison tubes, and
Use the chloride content obtained to calculate the Aluminum dilute with water to volume.
/Chloride Atomic Ratio. Acceptance criteria: The color of the solution from the
CONTENT OF ALUMINUM Sample solution is not darker than that of the solution from
Edetate disodium titrant: 37.2 mg/mL of edetate disodium the Standard solution (150 ppm).
in water
Standardization of titrant: Transfer 2 g of aluminum wire
SPECIFIC TESTS ASSAY

PH 791 [NOTEConduct Procedure 1, Procedure 2, and Procedure 3.]
Sample solution: 15 in 100 (w/w) PROCEDURE 1: CONTENT OF CHLORIDE
Acceptance criteria: Between 3.0 and 5.0 Sample: 1.4 g of Aluminum Chlorohydrate Solution
Analysis: Transfer the Sample to a 250-mL beaker, and add
ADDITIONAL REQUIREMENTS 100 mL of water and 10 mL of diluted nitric acid with
PACKAGING AND STORAGE: Preserve in well-closed containers. stirring. Titrate with 0.1 N silver nitrate VS using a glass
LABELING: The label states the content of anhydrous silversilver chloride electrode and a silver billet electrode
aluminum chlorohydrate. system, determining the endpoint potentiometrically. Each
mL of 0.1 N silver nitrate is equivalent to 3.545 mg of
chloride (Cl).
Aluminum Chlorohydrate Solution Calculate the percentage of chloride (Cl) found and use the
chloride content thus obtained to calculate the Aluminum/
(Comment on this Monograph)id=m2052=Aluminum Chloride Atomic Ratio in Procedure 3.
Chlorohydrate Solution=A-Monos.pdf) PROCEDURE 2: CONTENT OF ALUMINUM
DEFINITION in water
Aluminum Chlorohydrate Solution consists of complex basic Standardization of titrant: Transfer 2 g of aluminum wire
aluminum chloride that is polymeric and encompasses a range to a 1000-mL volumetric flask, and add 50 mL of a mixture
of aluminum-to-chloride ratios between 1.91:1 and 2.10:1. of hydrochloric acid and water (1:1). Swirl the flask to
The following solvents may be used: water, propylene glycol, ensure contact of the aluminum and the acid, and allow the
dipropylene glycol, or alcohol. It contains the equivalent of reaction to proceed until all of the aluminum has dissolved.
NLT 90.0% and NMT 110.0% of the labeled concentration of Dilute with water to volume. Pipet 10 mL of this solution
anhydrous aluminum chlorohydrate. into a 250-mL beaker and add, in the order named and
IDENTIFICATION with continuous stirring, 25.0 mL of Edetate disodium titrant
A. IDENTIFICATION TESTSGENERAL, Aluminum and Chloride and 20 mL of acetic acidammonium acetate buffer TS. Boil
191 gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL
Sample solution: Amount equivalent to 100 mg/mL of of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a
anhydrous aluminum chlorohydrate bright rose-pink color. Perform a blank determination,
Acceptance criteria: Meets the requirements substituting 10 mL of water for the aluminum solution, and
B. PROPYLENE GLYCOL make any necessary correction. Calculate the molarity of the
[NOTEPerform this test where propylene glycol is stated on solution taken:
the label.]
Sample solution: 2 g of Aluminum Chlorohydrate Solution Result = W/Ar V
in 10 mL of isopropyl alcohol W = weight of aluminum in the portion of solution
[NOTEFilter the solution after preparation.] taken (mg)
Analysis: Evaporate the Sample solution to about 1 mL on a Ar = atomic weight of aluminum, 26.98
steam bath. V = volume of Edetate disodium titrant consumed
Acceptance criteria: The IR spectrum of a film of the (mL)
resulting solution on a silver chloride disk exhibits maxima Sample solution: Transfer 400 mg of Aluminum
only at the same wavelengths as that of a similar Chlorohydrate Solution to a 250-mL beaker. Add 20 mL of
preparation of a film of propylene glycol. water and 5 mL of hydrochloric acid, boil on a hot plate for
C. DIPROPYLENE GLYCOL NLT 5 min, and allow to cool.
[NOTEPerform this test where dipropylene glycol is stated Analysis: To the Sample solution, add 25.0 mL of Edetate
on the label.] disodium titrant and adjust with 2.5 N ammonium hydroxide
Sample solution: 2 g of Aluminum Chlorohydrate Solution or 1 N acetic acid to a pH of 4.7 0.1. Add 20 mL of acetic
in 10 mL of isopropyl alcohol acidammonium acetate buffer TS, 50 mL of alcohol, and 5
[NOTEFilter the solution after preparation.] mL of dithizone TS. The pH of this solution should be 4.7
Analysis: Evaporate the Sample solution to about 1 mL on a 0.1. Titrate with 0.1 M zinc sulfate VS until the color
steam bath. changes from green-violet to rose-pink. Perform a blank
Acceptance criteria: The IR spectrum of a film of the titration, and make any necessary correction. Each mL of 0.1
resulting solution on a silver chloride disk exhibits maxima M Edetate disodium titrant consumed is equivalent to 2.698
only at the same wavelengths as that of a similar mg of aluminum (Al).
preparation of a film of dipropylene glycol. Calculate the percentage of aluminum (Al) found and use
D. ALCOHOL the aluminum content thus obtained to calculate the
[NOTEPerform this test where alcohol is stated on the Aluminum/Chloride Atomic Ratio (below).
label.] PROCEDURE 3: ALUMINUM/CHLORIDE ATOMIC RATIO
Analysis: In a small beaker, mix 5 drops of Aluminum Analysis: Divide the percentage of aluminum found in
Chlorohydrate Solution with 1 mL of potassium Procedure 2: Content of Aluminum by the percentage of
permanganate solution (1 in 100), and 5 drops of 2 N chloride found in Procedure 1: Content of Chloride, and
sulfuric acid. Immediately cover the beaker with filter paper multiply by 35.453/26.98, in which 35.453 and 26.98 are
moistened with a freshly prepared solution of 0.1 g of the atomic weights of chlorine and aluminum, respectively.
sodium nitroferricyanide and 0.25 g of piperazine in 5 mL of Acceptance criteria: The ratio is between 1.91:1 and
water. 2.10:1.
Acceptance criteria: An intense blue color is produced on
the filter paper, the color fading after a few minutes.
PROCEDURE 4 IDENTIFICATION
Analysis: Calculate the percentage of anhydrous aluminum A. IDENTIFICATION TESTSGENERAL, Aluminum and Chloride
chlorohydrate in the portion of Aluminum Chlorohydrate 191
taken: Sample solution: 100 mg/mL
Acceptance criteria: Meets the requirements of the tests
Result = Al({Ar1X + [Mr(3X 1)] + Ar2}/Ar1X) B. INFRARED ABSORPTION 197F
Sample solution: Dissolve 0.5 g in 40 mL of water and,
Al% = percentage of aluminum as obtained in Procedure while mixing, adjust with 2.5 N sodium hydroxide to a pH
2: Content of Aluminum of 9.55 0.05. Filter the suspension of precipitate thus
Ar1 = atomic weight of aluminum, 26.98 obtained. Evaporate about 15 mL of the filtrate to about 1
X = aluminum/chloride atomic ratio as obtained in mL on a hot plate. Deposit the solution on a silver chloride
Procedure 3 disk.
Mr = molecular weight of the hydroxide anion (OH), Standard solution: A similar preparation of polyethylene
17.01 glycol
Ar2 = atomic weight of chlorine (Cl), 35.453
Acceptance criteria: NLT 90.0% and NMT 110.0% ASSAY
PROCEDURE
IMPURITIES Analysis
Inorganic Impurities Calculate the percentage of anhydrous aluminum
ARSENIC, Method I 211: NMT 2 ppm chlorohydrate in the Aluminum Chlorohydrex Polyethylene
HEAVY METALS, Method I 231: NMT 10 ppm Glycol:
LIMIT OF IRON
Standard solution: 2.0 mL of Standard Iron Solution, Result = Al({AR1X + [MR(3X -1)] + AR2}/AR1X)
prepared as directed under Iron 241
Sample solution: Transfer 5.3 g of Aluminum Chlorohydrate Al = percentage of aluminum as obtained in the test
Solution to a 100-mL volumetric flask, and dilute to volume for Content of Aluminum
with water. X = aluminum/chloride atomic ratio
Analysis: Pipet 5.0 mL of the Standard solution into a 50-mL AR1 = atomic weight of aluminum, 26.98
beaker. Pipet 5.0 mL of the Sample solution into a second MR = molecular weight of the hydroxide anion (OH),
50-mL beaker. To each of the beakers, add 5 mL of 6 N 17.01
nitric acid, cover with a watch glass, and boil on a hot plate AR2 = atomic weight of chlorine (Cl), 35.453
for 3 to 5 min. Allow to cool, add 5 mL of Ammonium Acceptance criteria: NLT 90.0% and NMT 110.0%
Thiocyanate Solution, prepared as directed under Iron 241,
transfer to separate 50-mL color comparison tubes, and OTHER COMPONENTS
dilute with water to volume. CONTENT OF CHLORIDE
Acceptance criteria: The color of the solution from the Sample: 700 mg of Aluminum Chlorohydrex Polyethylene
Sample solution is not darker than that of the solution from Glycol
the Standard solution (75 ppm). Analysis: Transfer the Sample to a 250-mL beaker and add
100 mL of water and 10 mL of diluted nitric acid with
SPECIFIC TESTS stirring. Titrate with 0.1 N silver nitrate VS using a glass
PH 791 silver-silver chloride electrode and a silver billet electrode
Sample solution: Dilute 3 g of the Aluminum system, determining the endpoint potentiometrically. Each
Chlorohydrate Solution with water to 10 mL. mL of 0.1 N silver nitrate is equivalent to 3.545 mg of
Acceptance criteria: 3.05.0 chloride (Cl). Use the chloride content thus obtained to
calculate the Aluminum/Chloride Atomic Ratio.
ADDITIONAL REQUIREMENTS CONTENT OF ALUMINUM
PACKAGING AND STORAGE: Preserve in well-closed containers. Edetate disodium titrant: 37.2 mg/mL of edetate disodium
LABELING: Label Solution to state the solvent used and the Standardization of titrant: Transfer 2 g of aluminum wire
claimed concentration of anhydrous aluminum chlorohydrate to a 1000-mL volumetric flask, and add 50 mL of a mixture
contained therein. of hydrochloric acid and water (1:1). Swirl the flask to
ensure contact of the aluminum and the acid, and allow the
Aluminum Chlorohydrex Polyethylene Dilute with water to volume. Pipet 10 mL of this solution
into a 250-mL beaker, and add, in the order named and
Glycol with continuous stirring, 25.0 mL of Edetate disodium titrant
(Comment on this Monograph)id=m2054=Aluminum and 20 mL of acetic acidammonium acetate buffer TS, and
Chlorohydrex Polyethylene Glycol=A-Monos.pdf) boil gently for 5 min. Cool, and add 50 mL of alcohol, and
Aly(OH)3y-zClz nH2O mH(OCH2CH2)nOH 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to
Aluminum chlorohydroxide polyethylene glycol complex; a bright rose-pink color. Perform a blank determination,
Aluminum hydroxychloride polyethylene glycol complex. substituting 10 mL of water for the aluminum solution, and
make any necessary correction. Calculate the molarity of the
DEFINITION solution taken:
Aluminum Chlorohydrex Polyethylene Glycol consists of
aluminum chlorohydrate in which some of the waters of Result = W/Ar V
hydration have been replaced by polyethylene glycol. It
encompasses a range of aluminum-to-chloride atomic ratios W = weight of aluminum in the portion of solution
between 1.91:1 and 2.10:1. It contains NLT 90.0% and NMT taken (mg)
110.0% of the labeled amount of anhydrous aluminum Ar = atomic weight of aluminum, 26.98
chlorohydrate.
V = volume of Edetate disodium titrant consumed DEFINITION

(mL) Aluminum Chlorohydrex Propylene Glycol is a complex of
Sample solution: Transfer 200 mg of Aluminum aluminum chlorohydrate and propylene glycol in which some
Chlorohydrex Polyethylene Glycol to a 250-mL beaker, add of the waters of hydration of the aluminum chlorohydrate
20 mL of water and 5 mL of hydrochloric acid, boil on a hot have been replaced by propylene glycol. It contains the
plate for NLT 5 min, and allow to cool. equivalent of NLT 90.0% and NMT 110.0% of the labeled
Analysis: To the Sample solution, add 25.0 mL of Edetate amount of anhydrous aluminum chlorohydrate.
disodium titrant, and adjust with 2.5 N ammonium
hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 IDENTIFICATION
mL of acetic acid-ammonium acetate buffer TS, 50 mL of A. IDENTIFICATION TESTSGENERAL, Aluminum and Chloride
alcohol, and 5 mL of dithizone TS. The pH of this solution 191
should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until Sample solution: 100 mg/mL
the color changes from green-violet to rose-pink. Perform a Acceptance criteria: Meets the requirements
blank titration, and make any necessary correction. Each mL B. INFRARED ABSORPTION 197F
of 0.1 M Edetate disodium titrant consumed is equivalent to Sample solution: Dissolve 0.5 g in 40 mL of water and,
2.698 mg of aluminum (Al). Use the aluminum content thus while mixing, adjust with 2.5 N sodium hydroxide to a pH
obtained to calculate the Aluminum/Chloride Atomic Ratio. of 9.55 0.05. Filter the suspension of precipitate thus
ALUMINUM/CHLORIDE ATOMIC RATIO obtained. Evaporate about 15 mL of the filtrate to about 1
Analysis: Divide the percentage of aluminum found in the mL on a hot plate. Deposit the solution on a silver chloride
test for Content of Aluminum by the percentage of chloride disk.
found in the test for Content of Chloride, and multiply by Standard solution: A similar preparation of propylene
35.453/26.98, in which 35.453 and 26.98 are the atomic glycol
weights of chlorine and aluminum, respectively.
Acceptance criteria: The ratio is between 1.91:1 and ASSAY
2.10:1. PROCEDURE
Analysis
IMPURITIES Calculate the percentage of anhydrous aluminum
Inorganic Impurities chlorohydrate in the Aluminum Chlorohydrex Propylene
ARSENIC, Method I 211 Glycol:
HEAVY METALS, Method I 231 Result = Al({Ar1X + [Mr(3X 1)] + Ar2}/Ar1X)
LIMIT OF IRON Al% = percentage of aluminum as obtained in the test
Standard solution: 2.0 mL of Standard Iron Solution for Procedure 2: Content of Aluminum (see
Prepared as directed under Iron 241. below)
Sample solution: 27 mg/mL Ar1 = atomic weight of aluminum, 26.98
Analysis: Pipet 5.0 mL of the Standard solution into a 50- X = Aluminum/Chloride Atomic Ratio
mL beaker. Pipet 5.0 mL of the Sample solution to a second Mr = molecular weight of the hydroxide anion (OH),
50-mL beaker. To each of the beakers, add 5 mL of 6 N 17.01
nitric acid, cover with a watch glass, and boil on a hot Ar2 = atomic weight of chlorine (Cl), 35.453
plate for 3 to 5 min. Allow to cool, add 5 mL of Ammonium Acceptance criteria: NLT 90.0% and NMT 110.0%
Thiocyanate Solution, prepared as directed under Iron 241, OTHER COMPONENTS
transfer to separate 50-mL color comparison tubes, and CONTENT OF CHLORIDE
dilute with water to volume. Sample: 700 mg of Aluminum Chlorohydrex Propylene
Acceptance criteria: The color of the solution from the Glycol
Sample solution is not darker than that of the solution from Analysis: Transfer the Sample to a 250-mL beaker and add
the Standard solution (150 ppm). 100 mL of water and 10 mL of diluted nitric acid with
SPECIFIC TESTS stirring. Titrate with 0.1 N silver nitrate VS using a glass
PH 791 silversilver chloride electrode and a silver billet electrode
Sample solution: 15 in 100 (w/w) system, determining the endpoint potentiometrically. Each
Acceptance criteria: Between 3.0 and 5.0 mL of 0.1 N silver nitrate is equivalent to 3.545 mg of
chloride (Cl). Use the chloride content thus obtained to
ADDITIONAL REQUIREMENTS calculate the Aluminum/Chloride Atomic Ratio.
PACKAGING AND STORAGE: Preserve in well-closed containers. CONTENT OF ALUMINUM
LABELING: The label states the content of anhydrous Edetate disodium titrant: 37.2 mg/mL of edetate disodium
aluminum chlorohydrate. in water
Standardization of titrant: Transfer 2 g of aluminum wire
of hydrochloric acid and water (1:1). Swirl the flask to
Aluminum Chlorohydrex Propylene ensure contact of the aluminum and the acid, and allow the
Glycol reaction to proceed until all of the aluminum has dissolved.
(Comment on this Monograph)id=m2057=Aluminum Dilute with water to volume. Pipet 10 mL of this solution
Chlorohydrex Propylene Glycol=A-Monos.pdf) into a 250-mL beaker and add, in the order named and
with continuous stirring, 25.0 mL of Edetate disodium titrant
Aly(OH)3y-zClz nH2O mC3H8O2 Al2(H2O)y-z(OH)6-n(Cl)n(C3H8O2)z and 20 mL of acetic acidammonium acetate buffer TS, and
Aluminum chlorohydroxide, hydrate:propylene glycol complex boil gently for 5 min. Cool, and add 50 mL of alcohol and 2
(1:1); mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a
Aluminum hydroxychloride, hydrate:propylene glycol complex bright rose-pink color. Perform a blank determination,
(1:1) [53026-85-0].
substituting 10 mL of water for the aluminum solution, and Aluminum Dichlorohydrate

make any necessary correction. (Comment on this Monograph)id=m2060=Aluminum
Calculate the molarity of the solution taken: Dichlorohydrate=A-Monos.pdf)
Result = W/Ar V Aly(OH)3y-zClz nH2O
Aluminum chlorohydroxide;
W = weight aluminum in the portion of solution taken Aluminum hydroxychloride.
(mg)
Ar = atomic weight of aluminum (Al), 26.98 DEFINITION
V = volume of Edetate disodium titrant consumed Aluminum Dichlorohydrate consists of complex basic aluminum
(mL) chloride that is polymeric and loosely hydrated and
Sample solution: Transfer 1.6 g of Aluminum Chlorohydrex encompasses a range of aluminum-to-chloride atomic ratios
Propylene Glycol to a 100-mL beaker, add 15 to 20 mL of between 0.90:1 and 1.25:1. It contains the equivalent of NLT
water and 5 to 6 mL of hydrochloric acid, boil on a hot 90.0% and NMT 110.0% of the labeled amount of anhydrous
plate for 15 to 20 min, and allow to cool. With the aid of aluminum dichlorohydrate.
water, transfer to a 100-mL volumetric flask and dilute to
volume. IDENTIFICATION
Analysis: Transfer 5.0 mL of the Sample solution to a 250- A. IDENTIFICATION TESTSGENERAL, Aluminum 191
mL beaker, add 10 to 15 mL of water and adjust with 1 N Sample solution: 100 mg/mL
sodium hydroxide to a pH of 1.5 0.5. Add 10.0 mL of Acceptance criteria: Meets the requirements
Edetate disodium titrant and heat to boiling. Cool the B. IDENTIFICATION TESTSGENERAL, Chloride 191
solution, and carefully introduce a magnetic stirring bar into Sample solution: 100 mg/mL
the beaker. Add 10 to 15 mL of acetic acidammonium Acceptance criteria: Meets the requirements
acetate buffer TS, 40 to 50 mL of alcohol, and, while ASSAY
stirring, adjust with glacial acetic acid to a pH of 4.6 0.1. PROCEDURE
Add 1 to 2 mL of dithizone TS and 40 to 50 mL of alcohol, Analysis
and titrate with 0.1 M zinc sulfate VS until the color changes Calculate the percentage of anhydrous aluminum
from green-violet to rose-pink. Perform a blank titration, and dichlorohydrate in the portion of Aluminum
make any necessary correction. Each mL of 0.1 M Edetate Dichlorohydrate taken:
disodium titrant consumed is equivalent to 2.698 mg of
aluminum (Al). Use the aluminum content thus obtained to Result = Al({Ar1X + [Mr(3X 1)] + Ar2}/Ar1X)
ALUMINUM/CHLORIDE ATOMIC RATIO Al = percentage of aluminum as obtained in the test
Analysis: Divide the percentage of aluminum found in the for Content of Aluminum
test for Content of Aluminum by the percentage of chloride Ar1 = atomic weight of aluminum, 26.98
found in the test for Content of Chloride, and multiply by X = Aluminum/Chloride Atomic Ratio, as determined
35.453/26.98, in which 35.453 and 26.98 are the atomic below
weights of chlorine and aluminum, respectively. Mr = molecular weight of the hydroxide anion (OH),
Acceptance criteria: The ratio is between 1.91:1 and 2.1:1. 17.01
IMPURITIES Acceptance criteria: NLT 90.0% and NMT 110.0%
ARSENIC, Method I 211 OTHER COMPONENTS
Acceptance criteria: NMT 2 ppm CONTENT OF CHLORIDE
HEAVY METALS, Method I 231 Sample: 700 mg of Aluminum Dichlorohydrate
Acceptance criteria: NMT 20 ppm Analysis: Transfer the Sample to a 250-mL beaker and add
LIMIT OF IRON 100 mL of water and 10 mL of diluted nitric acid with
Standard solution: 2.0 mL of Standard Iron Solution, stirring. Titrate with 0.1 N silver nitrate VS using a glass
prepared as directed under Iron 241 silversilver chloride electrode and a silver billet electrode
Sample solution: 27 mg/mL system, determining the endpoint potentiometrically. Each
Analysis: Pipet 5.0 mL of the Standard solution into a 50- mL of 0.1 N silver nitrate is equivalent to 3.545 mg of
mL beaker. Pipet 5.0 mL of the Sample solution to a second chloride (Cl). Use the chloride content thus obtained to
50-mL beaker. To each of the beakers add 5 mL of 6 N calculate the Aluminum/Chloride Atomic Ratio (below).
nitric acid, cover with a watch glass, and boil on a hot CONTENT OF ALUMINUM
plate for 3 to 5 min. Allow to cool, add 5 mL of Ammonium Edetate disodium titrant: 37.2 mg/mL of edetate disodium
Thiocyanate Solution, prepared as directed under Iron 241, in water
transfer to separate 50-mL color comparison tubes, and Standardization of titrant: Transfer 2 g of aluminum wire
dilute with water to volume. to a 1000-mL volumetric flask, and add 50 mL of a mixture
Acceptance criteria: The color of the solution from the of hydrochloric acid and water (1:1). Swirl the flask to
Sample solution is not darker than that of the solution from ensure contact of the aluminum and the acid, and allow the
the Standard solution (150 ppm). reaction to proceed until all of the aluminum has dissolved.
SPECIFIC TESTS into a 250-mL beaker and add, in the order named and
PH 791 with continuous stirring, 25.0 mL of Edetate disodium titrant
Sample solution: 15 in 100 (w/w) and 20 mL of acetic acidammonium acetate buffer TS, and
Acceptance criteria: Between 3.0 and 5.0 boil gently for 5 min. Cool, and add 50 mL of alcohol and 2
ADDITIONAL REQUIREMENTS mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a
PACKAGING AND STORAGE: Preserve in well-closed containers. bright rose-pink color. Perform a blank determination,
LABELING: The label states the content of anhydrous substituting 10 mL of water for the aluminum solution, and
aluminum chlorohydrate. make any necessary correction.
Calculate the molarity of the solution taken: Aluminum Dichlorohydrate Solution

(Comment on this Monograph)id=m2062=Aluminum
Result = W/Ar V Dichlorohydrate Solution=A-Monos.pdf)
W = weight of aluminum in the portion of solution DEFINITION
taken (mg) Aluminum Dichlorohydrate Solution consists of complex basic
Ar = atomic weight of aluminum, 26.98 aluminum chloride that is polymeric and encompasses a range
V = volume of Edetate disodium titrant consumed of aluminum-to-chloride atomic ratios between 0.90:1 and
(mL) 1.25:1. The following solvents may be used: water, propylene
Sample solution: Transfer 200 mg of Aluminum glycol, dipropylene glycol, or alcohol. It contains the
Dichlorohydrate to a 250-mL beaker, add 20 mL of water equivalent to NLT 90.0% and NMT 110.0% of the labeled
and 5 mL of hydrochloric acid, boil on a hot plate for NLT 5 amount of anhydrous aluminum dichlorohydrate.
min, and allow to cool.
Analysis: To the Sample solution, add 25.0 mL of Edetate IDENTIFICATION
disodium titrant and adjust with 2.5 N ammonium hydroxide A. IDENTIFICATION TESTSGENERAL, Aluminum 191:
or 1 N acetic acid to a pH of 4.7 0.1. Add 20 mL of acetic Sample solution: Amount equivalent to 100 mg/mL of
acidammonium acetate buffer TS, 50 mL of alcohol, and 5 anhydrous aluminum dichlorohydrate
mL of dithizone TS. The pH of this solution should be 4.7 Acceptance criteria: Meets the requirements
0.1. Titrate with 0.1 M zinc sulfate VS until the color B. IDENTIFICATION TESTSGENERAL, Chloride 191:
changes from green-violet to rose-pink. Perform a blank Sample solution: Amount equivalent to 100 mg/mL of
titration, and make any necessary correction. Each mL of 0.1 anhydrous aluminum dichlorohydrate
M Edetate disodium titrant consumed is equivalent to 2.698 Acceptance criteria: Meets the requirements
mg of aluminum (Al). Use the aluminum content thus C. INFRARED ABSORPTION 197F
obtained to calculate the Aluminum/Chloride Atomic Ratio [NOTEPerform this test where either propylene glycol or
(below). dipropylene glycol is stated on the label.]
ALUMINUM/CHLORIDE ATOMIC RATIO Standard solution: A similar preparation of propylene
Analysis: Divide the percentage of aluminum found in glycol or dipropylene glycol (where propylene glycol or
Content of Aluminum (above) by the percentage of chloride dipropylene glycol are indicated on the label)
found in Content of Chloride, and multiply by 35.453/26.98, Sample solution: Add 10 mL of isopropyl alcohol to 2 g of
in which 35.453 and 26.98 are the atomic weights of Aluminum Dichlorohydrate Solution, and filter. Evaporate
chlorine and aluminum, respectively. the filtrate to about 1 mL on a steam bath. Deposit this
Acceptance criteria: The ratio is between 0.90:1 and solution on a silver chloride disk.
1.25:1. D. IDENTIFICATION OF ALCOHOL
[NOTEPerform this test where alcohol is stated on the
IMPURITIES label.]
ARSENIC, Method I 211 Analysis: Mix 5 drops of Aluminum Dichlorohydrate
Acceptance criteria: NMT 2 ppm Solution in a small beaker with 1 mL of potassium
HEAVY METALS, Method I 231 permanganate solution (1 in 100) and 5 drops of 2 N
Acceptance criteria: NMT 20 ppm sulfuric acid, and cover the beaker immediately with filter
LIMIT OF IRON paper moistened with a freshly prepared solution of 0.1 g of
Standard solution: 2.0 mL of Standard Iron Solution, sodium nitroferricyanide and 0.25 g of piperazine in 5 mL of
prepared as directed in Iron 241 water.
Sample solution: 27 mg/mL Acceptance criteria: An intense blue color is produced on
Analysis: Pipet 5.0 mL of the Standard solution into a 50-mL the filter paper, the color becoming paler after a few
beaker. Pipet 5.0 mL of the Sample solution to a second 50- minutes.
mL beaker. To each of the beakers, add 5 mL of 6 N nitric
acid, cover with a watch glass, and boil on a hot plate for 3 ASSAY
to 5 min. Allow to cool. Add 5 mL of Ammonium PROCEDURE
Thiocyanate Solution (prepared as directed in Iron 241), Calculate the percentage of anhydrous aluminum
transfer to separate 50-mL color comparison tubes, and dichlorohydrate in the portion of Solution taken:
Acceptance criteria: The color of the solution from the Result = Al({Ar1x + [Mr(3x 1)] + Ar2}/Ar1x)
Sample solution is not darker than that of the solution from
the Standard solution (150 ppm). Al = percentage of aluminum as obtained in the test
for Content of Aluminum
SPECIFIC TESTS Ar1 = atomic weight of aluminum, 26.98
PH 791 x = Aluminum/Chloride Atomic Ratio
Sample solution: 15 in 100 (w/w) Mr = molecular weight of the hydroxide anion (OH),
Acceptance criteria: Between 3.0 and 5.0 17.0
LABELING: The label states the content of anhydrous
aluminum dichlorohydrate.
Acceptance criteria: NLT 90.0% and NMT 110.0% Acceptance criteria: NMT 2 ppm
HEAVY METALS, Method I 231
OTHER COMPONENTS Sample: Use a quantity of Aluminum Dichlorohydrate
CONTENT OF CHLORIDE Solution
Sample: 1.4 g of Aluminum Dichlorohydrate Solution Acceptance criteria: NMT 10 ppm
Analysis: Transfer the Sample to a 250-mL beaker, and add, LIMIT OF IRON
with stirring, 100 mL of water and 10 mL of diluted nitric Standard solution: 2.0 mL of Standard Iron Solution,
acid. Titrate with 0.1 N silver nitrate VS using a glass prepared as directed for Iron 241
silversilver chloride electrode and a silver billet electrode Sample solution: Transfer 5.3 g of Aluminum
system, determining the endpoint potentiometrically. Each Dichlorohydrate Solution to a 100-mL volumetric flask, and
mL of 0.1 N silver nitrate is equivalent to 3.545 mg of dilute to volume with water.
chloride (Cl). Analysis: Pipet 5.0 mL of the Standard solution into a 50-
Calculate the percentage of chloride (Cl) found and use the mL beaker. Pipet 5.0 mL of the Sample solution into a
chloride content thus obtained to calculate the Aluminum/ second 50-mL beaker. To each of the beakers, add 5 mL of
Chloride Atomic Ratio. 6 N nitric acid, cover with a watch glass, and boil on a hot
CONTENT OF ALUMINUM plate for 35 min. Allow to cool, add 5 mL of Ammonium
Edetate disodium titrant: 37.2 mg/mL of edetate disodium Thiocyanate Solution, prepared as directed for Iron 241,
in water transfer to separate 50-mL color comparison tubes, and
Standardization of titrant: Transfer 2 g of aluminum wire dilute with water to volume.
to a 1000-mL volumetric flask, and add 50 mL of a mixture Acceptance criteria: The color of the solution from the
of hydrochloric acid and water (1:1). Swirl the flask to Sample solution is not darker than that from the Standard
ensure contact of the aluminum and the acid, and allow the solution (75 ppm).
Dilute with water to volume. Pipet 10 mL of this solution SPECIFIC TESTS
into a 250-mL beaker and add, in the order named and PH 791
with continuous stirring, 25.0 mL of Edetate disodium titrant Sample solution: Dilute 3 g of Aluminum Dichlorohydrate
and 20 mL of acetic acidammonium acetate buffer TS, and Solution with water to 10 mL.
boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 Acceptance criteria: Between 3.0 and 5.0
mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a
bright rose-pink color. Perform a blank determination, ADDITIONAL REQUIREMENTS
substituting 10 mL of water for the aluminum solution, and PACKAGING AND STORAGE: Preserve in well-closed containers.
make any necessary correction. Calculate the molarity of the LABELING: Label Aluminum Dichlorohydrate Solution to state
solution taken: the solvent used and the claimed concentration of
anhydrous aluminum dichlorohydrate contained therein.
Result = W/Ar V
W = weight of aluminum (mg)

Ar = atomic weight of aluminium, 26.98 Aluminum Dichlorohydrex Polyethylene
V = volume of Edetate disodium titrant consumed Glycol
(mL) (Comment on this Monograph)id=m2064=Aluminum
Sample solution: Transfer 400 mg of Aluminum Dichlorohydrex Polyethylene Glycol=A-Monos.pdf)
Dichlorohydrate Solution to a 250-mL beaker, add 20 mL of
water and 5 mL of hydrochloric acid, boil on a hot plate for Aly(OH)3y-zClz nH2O mH(OCH2CH2)nOH.
NLT 5 min, and allow to cool. Aluminum chlorohydroxide polyethylene glycol complex;
Analysis: To the Sample solution, add 25.0 mL of Edetate Aluminum hydroxychloride polyethylene glycol complex.
disodium titrant, and adjust with 2.5 N ammonium DEFINITION
hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 Aluminum Dichlorohydrex Polyethylene Glycol consists of
mL of acetic acid-ammonium acetate buffer TS, 50 mL of aluminum dichlorohydrate, in which some of the waters of
alcohol, and 5 mL of dithizone TS. The pH of this solution hydration have been replaced by polyethylene glycol. It
should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until encompasses a range of aluminum-to-chloride atomic ratios
the color changes from green-violet to rose-pink. Perform a between 0.90:1 and 1.25:1. It contains NLT 90.0% and NMT
blank titration, and make any necessary correction. Each mL 110.0% of the labeled amount of anhydrous aluminum
of 0.1 M Edetate disodium titrant consumed is equivalent to dichlorohydrate.
2.698 mg of aluminum (Al).
Calculate the percentage of aluminum (Al) found and use IDENTIFICATION
the aluminum content obtained to calculate the Aluminum/ A. IDENTIFICATION TESTSGENERAL, Aluminum 191:
Chloride Atomic Ratio. Sample solution: 100 mg/mL
ALUMINUM/CHLORIDE ATOMIC RATIO Acceptance criteria: Meets the requirements of the test
Analysis: Divide the percentage of aluminum found in the B. IDENTIFICATION TESTSGENERAL, Chloride 191:
test for Content of Aluminum by the percentage of chloride Sample solution: 100 mg/mL
found in the test for Content of Chloride, and multiply by Acceptance criteria: Meets the requirements of the test
35.453/26.98, in which 35.453 and 26.98 are the atomic C. INFRARED ABSORPTION 197F
weights of chlorine and aluminum, respectively. Sample solution: Dissolve 0.5 g in 40 mL of water, and,
Acceptance criteria: The ratio is between 0.90:1 and while mixing, adjust with 2.5 N sodium hydroxide to a pH
1.25:1. of 9.55 0.05. Filter the suspension of precipitate obtained.
Evaporate about 15 mL of the filtrate to about 1 mL on a
IMPURITIES hot plate. Deposit the solution on a silver chloride disk.
Inorganic Impurities Standard solution: A similar preparation of polyethylene
ARSENIC, Method I 211 glycol
Sample: Use a weighed quantity of Aluminum
Dichlorohydrate Solution.

Analysis: Calculate the percentage of anhydrous aluminum test for Content of Aluminum by the percentage of chloride
dichlorohydrate in the Aluminum Dichlorohydrex found in the test for Content of Chloride, and multiply by
Polyethylene Glycol taken: 35.453/26.98, in which 35.453 and 26.98 are the atomic
weights of chlorine and aluminum, respectively.
Result = Al({Ar1X + [Mr(3X1)] + Ar2}/Ar1X) Acceptance criteria: Between 0.90:1 and 1.25:1.
Al = percentage of aluminum as obtained in the test IMPURITIES
for Content of Aluminum Inorganic Impurities
Ar1 = atomic weight of aluminum, 26.98 ARSENIC, Method I 211
X = aluminum/chloride atomic ratio (as determined Acceptance criteria: NMT 2 ppm
below) HEAVY METALS, Method I 231
Mr = molecular weight of hydroxide anion (OH), Acceptance criteria: NMT 20 ppm
17.01 LIMIT OF IRON
Ar2 = atomic weight of chlorine (Cl), 35.453 Standard solution: 2.0 mL of Standard Iron Solution,
Acceptance criteria: NLT 90.0% and NMT 110.0% prepared as directed for Iron 241
Sample solution: 27 mg/mL
OTHER COMPONENTS Analysis: Pipet 5.0 mL of the Standard solution into a 50-
CONTENT OF ALUMINUM mL beaker. Pipet 5.0 mL of the Sample solution to a second
Edetate disodium titrant: 37.2 mg/mL of edetate disodium 50-mL beaker. To each of the beakers, add 5 mL of 6 N
Standardization of titrant: Transfer 2 g of aluminum wire nitric acid, cover with a watch glass, and boil on a hot
to a 1000-mL volumetric flask, and add 50 mL of a mixture plate for 35 min. Allow to cool, add 5 mL of Ammonium
of hydrochloric acid and water (1:1). Swirl the flask to Thiocyanate Solution, prepared as directed for Iron 241,
ensure contact of the aluminum and the acid, and allow the transfer to separate 50-mL color comparison tubes, and
reaction to proceed until all of the aluminum has dissolved. dilute with water to volume.
Dilute with water to volume. Pipet 10 mL of this solution Acceptance criteria: The color of the solution from the
into a 250-mL beaker, add, in the order named and with Sample solution is not darker than that from the Standard
continuous stirring, 25.0 mL of Edetate disodium titrant and solution (150 ppm).
gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL SPECIFIC TESTS
of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a PH 791
bright rose-pink color. Perform a blank determination, Sample solution: 15 in 100 (w/w)
substituting 10 mL of water for the aluminum solution, and Acceptance criteria: Between 3.0 and 5.0
make any necessary correction.
Calculate the molarity of the solution taken: ADDITIONAL REQUIREMENTS
Result = W/Ar V LABELING: The label states the content of anhydrous
aluminum dichlorohydrate.
W = weight of aluminum in the portion of solution
taken (mg)
V = volume of Edetate disodium titrant consumed Aluminum Dichlorohydrex Propylene
(mL) Glycol
Sample solution: Transfer 200 mg of Aluminum (Comment on this Monograph)id=m2068=Aluminum
Dichlorohydrex Polyethylene Glycol to a 250-mL beaker, add Dichlorohydrex Propylene Glycol=A-Monos.pdf)
20 mL of water and 5 mL of hydrochloric acid, boil on a hot
plate for NLT 5 min, and allow to cool. Aly(OH)3y-zClz nH2O mC3H8O2
Analysis: To the Sample solution, add 25.0 mL of Edetate Aluminum chlorohydroxide propylene glycol complex;
disodium titrant, and adjust with 2.5 N ammonium Aluminum hydroxychloride propylene glycol complex.
hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 DEFINITION
mL of acetic acid-ammonium acetate buffer TS, 50 mL of Aluminum Dichlorohydrex Propylene Glycol consists of
alcohol, and 5 mL of dithizone TS. The pH of this solution aluminum dichlorohydrate in which some of the waters of
should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until hydration have been replaced by propylene glycol. It
the color changes from green-violet to rose-pink. Perform a encompasses a range of aluminum-to-chloride atomic ratios
blank titration, and make any necessary correction. Each mL between 0.90:1 and 1.25:1. It contains NLT 90.0% and NMT
of 0.1 M Edetate disodium titrant consumed is equivalent to 110.0% of the labeled amount of anhydrous aluminum
2.698 mg of aluminum (Al). Use the aluminum content dichlorohydrate.
obtained to calculate the Aluminum/Chloride Atomic Ratio.
CONTENT OF CHLORIDE IDENTIFICATION
Sample: 700 mg of Aluminum Dichlorohydrex Polyethylene A. IDENTIFICATION TESTSGENERAL, Aluminum and Chloride
Glycol 191
Analysis: Transfer the Sample to a 250-mL beaker and add, Sample solution: 100 mg/mL
with stirring, 100 mL of water and 10 mL of diluted nitric Acceptance criteria: Meets the requirements
acid. Titrate with 0.1 N silver nitrate VS using a glass B. PROCEDURE
silversilver chloride electrode and a silver billet electrode Sample solution: Dissolve 0.5 g in 40 mL of water and,
system, determining the endpoint potentiometrically. Each while mixing, adjust with 2.5 N sodium hydroxide to a pH
mL of 0.1 N silver nitrate is equivalent to 3.545 mg of of 9.55 0.05. Filter the suspension of precipitate thus
chloride (Cl). Use the chloride content thus obtained to obtained. Evaporate about 15 mL of the filtrate to about 1
mL on a hot plate. Deposit the solution on a silver chloride mL of dithizone TS. The pH of this solution should be 4.7
disk. 0.1. Titrate with 0.1 M zinc sulfate VS until the color
Standard solution: A similar preparation of propylene changes from green-violet to rose-pink. Perform a blank
glycol titration, and make any necessary correction. Each mL of 0.1
Acceptance criteria: The IR absorption spectrum of a film M Edetate disodium titrant consumed is equivalent to 2.698
of the Sample solution exhibits maxima only at the same mg of aluminum (Al). [NOTEUse the aluminum content
wavelengths as that of a film of the Standard solution. thus obtained to calculate the Aluminum/chloride atomic
ratio.]
Analysis test for Content of aluminum by the percentage of chloride
Calculate the percentage of anhydrous aluminum found in the test for Content of chloride, and multiply by
dichlorohydrate in the Aluminum Dichlorohydrex Propylene 35.453/26.98, in which 35.453 and 26.98 are the atomic
Glycol: weights of chlorine and aluminum, respectively.
Acceptance criteria: The ratio is between 0.90:1 and
Result = Al%({Ar1X + [Mr(3X 1)] + Ar2}/Ar1X) 1.25:1.
Al% = percentage of aluminum as obtained in the test IMPURITIES
for Procedure 2: Content of Aluminum Inorganic Impurities
Ar1 = atomic weight of aluminum, 26.98 ARSENIC, Method I 211
X = Aluminum/Chloride Atomic Ratio Acceptance criteria: NMT 2 ppm
Mr = molecular weight of the hydroxide ion (OH), HEAVY METALS, Method I 231
17.01 Acceptance criteria: NMT 20 ppm
Ar2 = atomic weight of chlorine (Cl), 35.453 LIMIT OF IRON 241
Acceptance criteria: NLT 90.0%NMT 110.0% Standard solution: Pipet 2.0 mL of Standard Iron Solution,
prepared as directed, into a 50-mL beaker
OTHER COMPONENTS Sample solution: 27 mg/mL
CONTENT OF CHLORIDE Analysis: Pipet 5.0 mL of the Sample solution to a second
Sample: 700 mg of Aluminum Dichlorohydrex Propylene 50-mL beaker. To each of the beakers add 5 mL of 6 N
Glycol nitric acid, cover with a watch glass, and boil on a hot
Analysis: Transfer the Sample to a 250-mL beaker and add plate for 3 to 5 min. Allow to cool, add 5 mL of Ammonium
100 mL of water and 10 mL of diluted nitric acid with Thiocyanate Solution, prepared as directed under Iron 241,
stirring. Titrate with 0.1 N silver nitrate VS using a glass transfer to separate 50-mL color comparison tubes, and
silversilver chloride electrode and a silver billet electrode dilute with water to volume.
system, determining the endpoint potentiometrically. Each Acceptance criteria: The color of the solution from the
mL of 0.1 N silver nitrate is equivalent to 3.545 mg of Sample solution is not darker than that of the solution from
chloride (Cl). [NOTEUse the results obtained to calculate the Standard solution (150 ppm).
the Aluminum/chloride atomic ratio.]
CONTENT OF ALUMINUM SPECIFIC TESTS
Edetate disodium titrant: Prepare a solution with a PH 791
concentration of 37.2 mg/mL of edetate disodium and Sample solution: 15 in 100 (w/w)
standardize as follows. Weigh 2 g of aluminum wire, transfer Acceptance criteria: Between 3.0 and 5.0
of hydrochloric acid and water (1:1). Swirl the flask to ADDITIONAL REQUIREMENTS
ensure contact of the aluminum and the acid, and allow the PACKAGING AND STORAGE: Preserve in well-closed containers.
reaction to proceed until all of the aluminum has dissolved. LABELING: The label states the content of anhydrous
Dilute with water to volume. Pipet 10 mL of this solution aluminum dichlorohydrate.
boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 Aluminum Hydroxide Gel
mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a (Comment on this Monograph)id=m2100=Aluminum Hydroxide
bright rose-pink color. Perform a blank determination, Gel=A-Monos.pdf)
substituting 10 mL of water for the aluminum solution, and Al(OH)3 78.00
make any necessary correction. Aluminum hydroxide [21645-51-2].
Calculate the molarity of the solution taken:
DEFINITION
Result = W/Ar V Aluminum Hydroxide Gel is a suspension of amorphous
aluminum hydroxide in which there is a partial substitution of
W = weight of aluminum in the portion of solution carbonate for hydroxide. It contains the equivalent of NLT
taken (mg) 90.0% and NMT 110.0% of the labeled amount of Al(OH)3. It
Ar = atomic weight of aluminum (Al), 26.98 may contain Peppermint Oil, Glycerin, Sorbitol, Sucrose,
V = volume of Edetate disodium titrant consumed Saccharin, or other suitable flavors, and it may contain suitable
(mL) antimicrobial agents.
Sample solution: Transfer 200 mg of Aluminum
Dichlorohydrex Propylene Glycol to a 250-mL beaker, add IDENTIFICATION
20 mL of water and 5 mL of hydrochloric acid, boil on a hot A. PROCEDURE
plate for NLT 5 min, and allow to cool. Sample: 1 g
Analysis: To the Sample solution, add 25.0 mL of Edetate Analysis: Place the Sample in a flask equipped with a
disodium titrant and adjust with 2.5 N ammonium hydroxide stopper and glass tubing, the tip of which is immersed in
or 1 N acetic acid to a pH of 4.7 0.1. Add 20 mL of acetic calcium hydroxide TS in a test tube. Add 5 mL of 3 N
acidammonium acetate buffer TS, 50 mL of alcohol, and 5
hydrochloric acid to the flask, and immediately insert the Analysis: Proceed as directed using a 20-mL portion of the
stopper. Sample solution.
Acceptance criteria: Gas evolves in the flask, and a Acceptance criteria: A 20-mL portion of the filtrate shows
precipitate is formed in the test tube. no more sulfate than corresponds to 0.20 mL of 0.020 N
B. IDENTIFICATION TESTSGENERAL, Aluminum 191 sulfuric acid [0.8%, based on the Al(OH)3 content].
Sample: The solution remaining in the flask from Procedure ARSENIC, Method I 211
A Sample solution: Amount of Aluminum Hydroxide Gel
Acceptance criteria: Meets the requirements equivalent to 25 mg/mL of Al(OH)3 in 7 N sulfuric acid
Standard solution: Prepare as directed in the test for
ASSAY Arsenic 211, except to prepare it to contain 5 g of
PROCEDURE arsenic instead of 3 g.
Edetate disodium titrant: Prepare a solution with a Acceptance criteria: NMT 10 ppm, based on the Al(OH)3
concentration of 18.6 mg/mL of edetate disodium and content
standardize as follows. Weigh 2 g of aluminum wire, transfer HEAVY METALS 231
to a 1000-mL volumetric flask, and add 50 mL of a mixture Sample solution: Dissolve an amount of Aluminum
of hydrochloric acid and water (1:1). Swirl the flask to Hydroxide Gel equivalent to 0.24 g of Al(OH)3 in 10 mL of
ensure contact of the aluminum and the acid, and allow the 3 N hydrochloric with the aid of heat, filter, if necessary,
reaction to proceed until all of the aluminum has dissolved. and dilute with water to 25 mL.
Dilute with water to volume. Pipet 10 mL of this solution Acceptance criteria: NMT 83 ppm, based on the Al(OH)3
into a 250-mL beaker and add, in the order named and content
and 20 mL of acetic acidammonium acetate buffer TS, and SPECIFIC TESTS
boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED
mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a MICROORGANISMS 62
bright rose-pink color. Perform a blank determination, Acceptance criteria: Its total aerobic microbial count does
substituting 10 mL of water for the aluminum solution, and not exceed 100 cfu/mL, and it meets the requirements of
make any necessary correction. the test for the absence of Escherichia coli.
Calculate the molarity of the solution taken: PH 791
Acceptance criteria: Between 5.5 and 8.0, determined
Result = W/Ar V potentiometrically
W = weight aluminum in the portion of solution taken Acceptance criteria: NLT 65.0% of the expected mEq
(mg) value, calculated from the results of the above Assay, is
Ar = atomic weight of aluminum (Al), 26.98 obtained. [NOTEAl(OH)3 has an expected acid-neutralizing
V = volume of Edetate disodium titrant consumed capacity value of 0.0385 mEq/mg.]
(mL)
Sample solution: Transfer an amount of Aluminum ADDITIONAL REQUIREMENTS
Hydroxide Gel equivalent to 1.5 g of Al(OH)3 to a beaker, PACKAGING AND STORAGE: Preserve in tight containers, and
add 15 mL of hydrochloric acid, and heat gently until avoid freezing.
solution is complete. Cool, transfer to a 500-mL volumetric
flask, and dilute to volume with water.
beaker and add, in the order named and with continuous Dried Aluminum Hydroxide Gel
stirring, 25.0 mL of Edetate disodium titrant and 20 mL of (Comment on this Monograph)id=m2110=Dried Aluminum
acetic acidammonium acetate buffer TS, then heat the Hydroxide Gel=A-Monos.pdf)
solution near the boiling point for 5 min. Cool, and add 50 Al(OH)3 78.00
mL of alcohol and 2 mL of dithizone TS. Titrate the solution Aluminum hydroxide [21645-51-2].
with 0.05 M zinc sulfate VS until the color changes from
green-violet to rose-pink. Perform a blank titration, DEFINITION
substituting 20 mL of water for the Sample solution, and Dried Aluminum Hydroxide Gel is an amorphous form of
make any necessary correction. Each mL of 0.05 M Edetate aluminum hydroxide in which there is a partial substitution of
disodium titrant consumed is equivalent to 3.9 mg of carbonate for hydroxide. It contains the equivalent of NLT
Al(OH)3. 76.5% of Al(OH)3, and it may contain varying quantities of
Acceptance criteria: NLT 90.0% and NMT 110.0% basic aluminum carbonate and bicarbonate.
IMPURITIES IDENTIFICATION
Inorganic Impurities A. INFRARED ABSORPTION 197K
CHLORIDE AND SULFATE, Chloride 221 B. IDENTIFICATION TESTSGENERAL, Aluminum 191
Sample: Amount of Aluminum Hydroxide Gel equivalent Sample: Dissolve 5 mg in 10 mL of 3 N hydrochloric acid,
to 0.6 g of Al(OH)3 with gentle warming.
Analysis: Transfer the Sample to a porcelain dish, and add Acceptance criteria: The solution responds to the tests.
0.1 mL of potassium chromate TS and 25 mL of water. Stir,
and add 0.10 N silver nitrate until a faint, persistent pink ASSAY
color is obtained. PROCEDURE
Acceptance criteria: NMT 8.0 mL of 0.10 N silver nitrate Edetate disodium titrant: Prepare a solution with a
is required [4.7%, based on the Al(OH)3 content]. concentration of 18.6 mg/mL of edetate disodium and
CHLORIDE AND SULFATE, Sulfate 221 standardize as follows. Weigh 2 g of aluminum wire, transfer
Sample solution: Add 5.0 mL of 3 N hydrochloric acid to to a 1000-mL volumetric flask, and add 50 mL of a mixture
an amount of Aluminum Hydroxide Gel equivalent to 0.3 g of hydrochloric acid and water (1:1). Swirl the flask to
of Al(OH)3 and heat to dissolve the specimen under test. ensure contact of the aluminum and the acid, and allow the
Cool, dilute with water to 250 mL, and filter if necessary.
reaction to proceed until all of the aluminum has dissolved. Acceptance criteria: NMT 60 ppm
into a 250-mL beaker and add, in the order named and SPECIFIC TESTS
with continuous stirring, 25.0 mL of Edetate disodium titrant PH 791
and 20 mL of acetic acidammonium acetate buffer TS, and Sample solution: 1 in 25
boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 Acceptance criteria: NMT 10.0
mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a ACID-NEUTRALIZING CAPACITY 301
bright rose-pink color. Perform a blank determination, Sample: 400 mg
substituting 10 mL of water for the aluminum solution, and Analysis: Test as directed for Powders under Test Preparation.
make any necessary correction. Acceptance criteria: NLT 25.0 mEq/g
Result = W/(Ar)(V) PACKAGING AND STORAGE: Preserve in tight containers.
W = weight aluminum in the portion of solution taken USP Dried Aluminum Hydroxide Gel RS
(mg)
Ar = atomic weight of aluminum (Al), 26.98
V = volume of Edetate disodium titrant consumed Dried Aluminum Hydroxide Gel
(mL)
Sample solution: Transfer an amount of Dried Aluminum Capsules
Hydroxide Gel equivalent to 2 g of Al(OH)3 to a beaker, add (Comment on this Monograph)id=m2120=Dried Aluminum
15 mL of hydrochloric acid, and heat gently until solution is Hydroxide Gel Capsules=A-Monos.pdf)
complete. Cool, transfer to a 500-mL volumetric flask, and
dilute to volume with water. DEFINITION
Analysis: Pipet 20 mL of the Sample solution into a 250-mL Dried Aluminum Hydroxide Gel Capsules contain NLT 90.0%
beaker and add, in the order named and with continuous and NMT 110.0% of the labeled amount of aluminum
stirring, 25.0 mL of Edetate disodium titrant and 20 mL of hydroxide [Al(OH)3].
acetic acidammonium acetate buffer TS, then heat the
solution near the boiling point for 5 min. Cool, and add 50 IDENTIFICATION
mL of alcohol and 2 mL of dithizone TS. Titrate the solution A. PROCEDURE
with 0.05 M zinc sulfate VS to a bright rose-pink color. Sample: Place a portion of Capsule contents, equivalent to
Perform a blank titration, substituting 20 mL of water for the about 500 mg of aluminum hydroxide.
Sample solution, and make any necessary correction. Each Analysis: Place in a flask equipped with a stopper and glass
mL of 0.05 M Edetate disodium titrant consumed is tubing, the tip of which is immersed in calcium hydroxide
equivalent to 3.9 mg of Al(OH)3. TS in a test tube. Add 10 mL of 3 N hydrochloric acid to the
Acceptance criteria: NLT 76.5% flask, and immediately insert the stopper: gas evolves in the
flask and a precipitate is formed in the test tube.
IMPURITIES Acceptance Criteria: Gas evolves in the flask and a
Inorganic Impurities precipitate is formed in the test tube.
CHLORIDE AND SULFATE, Chloride 221 B. IDENTIFICATION TESTSGENERAL, Aluminum 191: The
Sample solution: Dissolve 1.0 g in 30 mL of 2 N nitric solution remaining in the flask in Identification test A meets
acid, heat to boiling, add water to make 100 mL, and filter. the requirements.
Analysis: Proceed as directed using a 5.0-mL portion of
the Sample solution diluted with an equal volume of water. ASSAY
Acceptance criteria: A 5.0-mL portion of the Sample PROCEDURE
solution filtrate, diluted with an equal volume of water, Edetate disodium titrant: Prepare a solution with a
shows no more chloride than corresponds to 0.60 mL of concentration of 18.6 mg/mL of edetate disodium and
0.20 N hydrochloric acid (0.85%). standardize as follows. Weigh 2 g of aluminum wire, transfer
CHLORIDE AND SULFATE, Sulfate 221 to a 1000-mL volumetric flask, and add 50 mL of a mixture
Sample solution: Dissolve 330 mg in 15 mL of 3 N of hydrochloric acid and water (1:1). Swirl the flask to
hydrochloric acid, heat to boiling, add water to make 250 ensure contact of the aluminum and the acid, and allow the
mL, and filter. reaction to proceed until all of the aluminum has dissolved.
Analysis: Proceed as directed using a 25-mL portion of the Dilute with water to volume. Pipet 10 mL of this solution
Sample solution. into a 250-mL beaker and add, in the order named and
Acceptance criteria: A 25-mL portion of the filtrate shows with continuous stirring, 25.0 mL of Edetate disodium titrant
no more sulfate than corresponds to 0.20 mL of 0.020 N and 20 mL of acetic acidammonium acetate buffer TS, and
sulfuric acid (0.6%). boil gently for 5 min. Cool, and add 50 mL of alcohol and 2
ARSENIC, Method I 211 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a
Sample solution: Dissolve 1.5 g in 80 mL of 7 N sulfuric bright rose-pink color. Perform a blank determination,
acid, and dilute with water to 220 mL. substituting 10 mL of water for the aluminum solution, and
Analysis: Proceed as directed using a 55-mL portion of the make any necessary correction.
Sample solution, omitting the addition of 20 mL of 7 N Calculate the molarity of the solution taken:
sulfuric acid. Result = W/(Ar)(V)
HEAVY METALS 231 W = weight aluminum in the portion of solution taken
Sample solution: Dissolve 330 mg in 10 mL of 3 N (mg)
hydrochloric with the aid of heat, filter, if necessary, and Ar = atomic weight of aluminum (Al), 26.98
dilute with water to 25 mL.
V = volume of Edetate disodium titrant consumed add 50 mL of a mixture of hydrochloric acid and water
(mL) (1:1). Swirl the flask to ensure contact of the aluminum and
Sample solution: Weigh the contents of NLT 20 Capsules. the acid, and allow the reaction to proceed until all of the
Transfer a weighed portion of the powder, equivalent to 1.2 aluminum has dissolved. Dilute with water to volume. Pipet
g of aluminum hydroxide, to a beaker, add 15 mL of 10 mL of this solution into a 250-mL beaker, add, in the
hydrochloric acid, and heat until dissolved. Dilute with water order named and with continuous stirring, 25.0 mL of
to about 100 mL, and filter quantitatively into a 500-mL Edetate disodium titrant and 20 mL of acetic
volumetric flask, washing the filter with water, and dilute acidammonium acetate buffer TS, and boil gently for 5
with water to volume. min. Cool, and add 50 mL of alcohol and 2 mL of dithizone
Analysis: Pipet 20 mL of Sample solution into a 250-mL TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink
beaker, and add, in the order named and with continuous color. Perform a blank determination, substituting 10 mL of
stirring, 25.0 mL of Edetate disodium titrant and 20 mL of water for the aluminum solution, and make any necessary
acetic acidammonium acetate buffer TS, then heat the correction. Calculate the molarity of the solution taken:
solution near the boiling point for 5 min. Cool, and add 50
mL of alcohol and 2 mL of dithizone TS. Titrate the solution Result = W/26.98V
with 0.05 M zinc sulfate VS to a bright rose-pink color.
Perform a blank determination, substituting 20 mL of water W = weight of aluminum in the portion of solution
for the Sample solution, and make any necessary correction. taken (mg)
Each mL of 0.05 M Edetate disodium titrant is equivalent to V = volume of Edetate disodium titrant consumed
3.9 mg of Al(OH)3. (mL)
Acceptance criteria: 90.0%110.0% Sample solution: Weigh and finely powder NLT 20 Tablets.
Weigh a portion of the powder, equivalent to 1.2 g of
PERFORMANCE TESTS aluminum hydroxide, add 15 mL of hydrochloric acid, and
DISINTEGRATION 701: 10 min, simulated gastric fluid TS heat until dissolved. Dilute with water to about 100 mL, and
being substituted for water in the test filter quantitatively into a 500-mL volumetric flask, washing
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements the filter with water, and dilute with water to volume.
SPECIFIC TESTS beaker, and add, in the order named and with continuous
ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is stirring, 25.0 mL of Edetate disodium titrant and 20 mL of
consumed by the minimum single dose recommended in acetic acidammonium acetate buffer TS, then heat the
the labeling, and NLT 55.0% of the expected mEq value, solution near the boiling point for 5 min. Cool, and add 50
calculated from the labeled quantity of Al(OH)3, is obtained. mL of alcohol and 2 mL of dithizone TS. Titrate the solution
Each mg of Al(OH)3 has an expected acid-neutralizing with 0.05 M zinc sulfate VS to a bright rose-pink color.
capacity value of 0.0385 mEq. Perform a blank determination, substituting 20 mL of water
for the Sample solution, and make any necessary correction.
ADDITIONAL REQUIREMENTS Each mL of 0.05 M Edetate disodium titrant is equivalent to
PACKAGING AND STORAGE: Preserve in well-closed containers. 3.900 mg of Al(OH)3.
LABELING: Capsules may be labeled to state the aluminum Acceptance criteria: 90.0%110.0%
hydroxide content in terms of the equivalent amount of
dried aluminum hydroxide gel, on the basis that each mg of PERFORMANCE TESTS
dried gel is equivalent to 0.765 mg of Al(OH)3. DISINTEGRATION 701: 10 min, simulated gastric fluid TS
being substituted for water in the test
Dried Aluminum Hydroxide Gel Tablets SPECIFIC TESTS
(Comment on this Monograph)id=m2150=Dried Aluminum ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is
Hydroxide Gel Tablets=A-Monos.pdf) consumed by the minimum single dose recommended in
the labeling, and NLT 55.0% of the expected mEq value,
DEFINITION calculated from the labeled quantity of Al(OH)3, is obtained.
Dried Aluminum Hydroxide Gel Tablets contain NLT 90.0% and Each mg of Al(OH)3 has an expected acid-neutralizing
NMT 110.0% of the labeled amount of aluminum hydroxide capacity value of 0.0385 mEq.
[Al(OH)3].
IDENTIFICATION PACKAGING AND STORAGE: Preserve in well-closed containers.
A. PROCEDURE LABELING: Tablets may be labeled to state the aluminum
Sample: Finely ground Tablets, equivalent to 500 mg of hydroxide content in terms of the equivalent amount of
aluminum hydroxide dried aluminum hydroxide gel, on the basis that each mg of
Analysis: Place in a flask equipped with a stopper and glass dried gel is equivalent to 0.765 mg of Al(OH)3.
tubing, the tip of which is immersed in calcium hydroxide
TS in a test tube. Add 5 mL of 3 N hydrochloric acid to the
flask, and immediately insert the stopper.
Acceptance criteria: Gas evolves in the flask, and a Aluminum Phosphate Gel
precipitate is formed in the test tube. (Comment on this Monograph)id=m2270=Aluminum
B. IDENTIFICATION TESTSGENERAL, Aluminum 191: The Phosphate Gel=A-Monos.pdf)
solution remaining in the flask in Identification test A meets
the requirements. Phosphoric acid, aluminum salt (1:1);
Aluminum phosphate (1:1) [7784-30-7].
ASSAY
PROCEDURE DEFINITION
Edetate disodium titrant: 18.6 mg/mL of edetate disodium Aluminum Phosphate Gel is a water suspension containing NLT
in water 4.0% and NMT 5.0% (w/w) of aluminum phosphate (AlPO4).
Edetate disodium standardization: Weigh 2 g of It may contain sodium benzoate, benzoic acid, or other
aluminum wire, transfer to a 1000-mL volumetric flask, and
suitable agent, in an amount not exceeding 0.5%, as a Acceptance criteria: NMT 5 ppm
preservative.
SPECIFIC TESTS
IDENTIFICATION PH 791: 6.07.2
A. IDENTIFICATION TESTSGENERAL, Aluminum 191: A SOLUBLE PHOSPHATE: Filter 20 g and wash the residue with
solution of it in hydrochloric acid meets the requirements. 30 mL of water. Add to the filtrate 2 mL of nitric acid, heat
B. IDENTIFICATION TESTSGENERAL, Phosphate 191: A to 60, and add 20 mL of ammonium molybdate TS. Heat at
solution of it in 2 N nitric acid meets the requirements. 50 for 30 min, filter, wash the precipitate with dilute nitric
acid (1 in 36), then wash with potassium nitrate solution (1
ASSAY in 100) until the last portion of the filtrate is not acid to
PROCEDURE litmus paper. Dissolve the precipitate in 50.0 mL of 0.5 N
Sample solution: To 20 g of Gel, in a 100-mL volumetric sodium hydroxide VS, add phenolphthalein TS, and titrate
flask, add nitric acid to dissolve, dilute with water to the excess alkali with 0.5 N hydrochloric acid VS. Each mL of
volume. 0.5 N sodium hydroxide is equivalent to 2.065 mg of PO4.
Analysis: Transfer 10.0 mL of Sample solution to a 400-mL Acceptance criteria: Soluble phosphate, calculated as PO4,
beaker, dilute with water to 100 mL, heat to 60, add an NMT 0.30%
excess of ammonium molybdate TS, and maintain at 50 for
30 min. Filter, and wash the precipitate with dilute nitric ADDITIONAL REQUIREMENTS
acid (1 in 36), then with potassium nitrate solution (1 in PACKAGING AND STORAGE: Preserve in tight containers.
100) until the last portion of the filtrate is not acid to litmus
paper. Dissolve the precipitate in 50.0 mL of 0.5 N sodium
hydroxide VS, add phenolphthalein TS, and titrate the
excess sodium hydroxide with 0.5 N sulfuric acid VS. Each Aluminum Sesquichlorohydrate
mL of 0.5 N sodium hydroxide is equivalent to 2.651 mg of (Comment on this Monograph)id=m2285=Aluminum
AlPO4. Sesquichlorohydrate=A-Monos.pdf)
Acceptance criteria: 4.0%5.0% (w/w) Aly(OH)3y-zClz nH2O
IMPURITIES Aluminum chlorohydroxide;
Inorganic Impurities Aluminum hydroxychloride [11097-68-0].
CHLORIDE: DEFINITION
Sample Solution: Transfer 25 g to a beaker with the aid of Aluminum Sesquichlorohydrate consists of complex basic
50 mL of water, add 5 mL of nitric acid, then add, with aluminum chloride that is polymeric and loosely hydrated and
stirring, 30.0 mL of 0.1 N silver nitrate VS. Warm on a encompasses a range of aluminum-to-chloride atomic ratios
steam bath for 30 min, filter, and wash the precipitate with between 1.26:1 and 1.90:1. It contains NLT 90.0% and NMT
water acidified with nitric acid. 110.0% of the labeled amount of anhydrous aluminum
Analysis: To the filtrate add ferric ammonium sulfate TS, sesquichlorohydrate.
and titrate the excess silver nitrate with 0.1 N ammonium
thiocyanate VS. Each mL of 0.1 N silver nitrate is equivalent IDENTIFICATION
to 3.545 mg of Cl. IDENTIFICATION TESTSGENERAL, Aluminum and Chloride
Acceptance criteria: NMT 0.16% 191: A 100 mg/mL solution meets the requirements of
CHLORIDE AND SULFATE, Sulfate 221 the tests for Aluminum and for Chloride.
Sample solution: Add 10 mL of 3 N hydrochloric acid to
10 g of Gel, and heat to boiling. Cool, dilute with water to ASSAY
250 mL, and filter, if necessary. PROCEDURE
Acceptance criteria: A 10-mL portion of the solution Analysis: Calculate the percentage of anhydrous aluminum
shows no more sulfate than corresponds to 0.20 mL of sesquichlorohydrate in the Aluminum Sesquichlorohydrate:
0.020 N sulfuric acid: NMT 0.05%.
ARSENIC, Method I 211 Result = Al ({Ar1x + [Mr(3x -1)] + Ar2}/Ar1x)
Sample solution: Dissolve 5.0 g of Gel in the smallest
necessary volume of 3 N hydrochloric acid. Al = percentage of aluminum found in the test for
Acceptance criteria: NMT 0.6 ppm Content of aluminum
HEAVY METALS, Method I 231 Ar1 = atomic weight of aluminum, 26.98
[NOTEProceed as directed in the chapter, except to make x = aluminum/chloride atomic ratio found in the test
the following modifications.] for Aluminum/Chloride Atomic Ratio
Standard solution: Into a 50-mL color-comparison tube Mr = molecular weight of the hydroxide anion (OH),
pipet 4.0 mL of Standard Lead Solution, dilute with water to 17.01
25 mL, adjust with 6 N ammonium hydroxide to a pH Ar2 = atomic weight of chlorine (Cl), 35.453
between 1.9 and 2.1, and dilute with water to 40 mL. Acceptance criteria: 90.0%110.0%
Sample solution: Dissolve 8 g in 5 mL of 3 N hydrochloric
acid, warming if necessary, dilute with water to 25 mL, and OTHER COMPONENTS
adjust with 6 N ammonium hydroxide to a pH between 1.9 CONTENT OF CHLORIDE
and 2.1. Transfer to a 50-mL color-comparison tube, and Sample: 700 mg of Aluminum Sesquichlorohydrate to a
dilute with water to 40 mL. 250-mL beaker. Add 100 mL of water and 10 mL of diluted
Monitor solution: Into a 50-mL color-comparison tube nitric acid with stirring.
place 25 mL of the Sample solution, add 4.0 mL of Standard Analysis: Titrate with 0.1 N silver nitrate VS using a glass
Lead Solution, adjust with 1 N acetic acid or 6 N silver-silver chloride electrode and a silver billet electrode
ammonium hydroxide to a pH between 1.9 and 2.1, and system, determining the endpoint potentiometrically. Each
dilute with water to 40 mL. mL of 0.1 N silver nitrate is equivalent to 3.545 mg of
Analysis: Proceed as directed in the chapter, except to chloride (Cl). [NOTEUse the chloride content thus obtained
omit the addition of 2 mL of pH 3.5 Acetate Buffer. to calculate the Aluminum/chloride atomic ratio.]
CONTENT OF ALUMINUM SPECIFIC TESTS

Edetate disodium titrant: Prepare a solution with a PH 791: 3.05.0, in a solution [15 in 100 (w/w)]
concentration of 37.2 mg/mL of edetate disodium and
standarize as follows. Weigh 2 g of aluminum wire, transfer ADDITIONAL REQUIREMENTS
to a 1000-mL volumetric flask, and add 50 mL of a mixture PACKAGING AND STORAGE: Preserve in well-closed containers.
of hydrochloric acid and water (1:1). Swirl the flask to LABELING: The label states the content of anhydrous
ensure contact of the aluminum and the acid, and allow the aluminum sesquichlorohydrate.
into a 250-mL beaker, add, in the order named and with Aluminum Sesquichlorohydrate
continuous stirring, 25.0 mL of Edetate disodium titrant and
20 mL of acetic acidammonium acetate buffer TS, and boil Solution
gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL (Comment on this Monograph)id=m2286=Aluminum
of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a Sesquichlorohydrate Solution=A-Monos.pdf)
substituting 10 mL of water for the aluminum solution, and DEFINITION
make any necessary correction. Aluminum Sesquichlorohydrate Solution consists of complex
Calculate the molarity of the solution taken: basic aluminum chloride that is polymeric and encompasses a
range of aluminum-to-chloride atomic ratios between 1.26:1
W/Ar V and 1.90:1. The following solvents may be used: water,
propylene glycol, dipropylene glycol, or alcohol. It contains
W = weight of aluminum in the portion of Solution the equivalent of NLT 90.0% and NMT 110.0% of the labeled
taken (mg) concentration of anhydrous aluminum sesquichlorohydrate.
V = volume of Edetate disodium titrant consumed IDENTIFICATION
(mL) A. IDENTIFICATION TESTSGENERAL, Aluminum and Chloride
Sample solution: Transfer 200 mg of Aluminum 191: A solution containing the equivalent of 100 mg of
Sesquichlorohydrate to a 250-mL beaker, add 20 mL of anhydrous aluminum sesquichlorohydrate/mL meets the
water and 5 mL of hydrochloric acid, boil on a hot plate for requirements of the tests for Aluminum and for Chloride.
NLT 5 min, and allow to cool. B. IDENTIFICATION OF PROPYLENE GLYCOL (where stated on the
Analysis: To the Sample solution add 25.0 mL of Edetate label)
disodium titrant, and adjust with 2.5 N ammonium Sample solution: 200 mg/mL of Solution in isopropyl
hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 alcohol, and filter. Evaporate the filtrate to 1 mL on a steam
mL of acetic acidammonium acetate buffer TS, 50 mL of bath.
alcohol, and 5 mL of dithizone TS. The pH of this solution Acceptance criteria: The IR absorption spectrum of a film
should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until of this solution on a silver chloride disk exhibits maxima only
the color changes from green-violet to rose-pink. Perform a at the same wavelengths as that of a similar preparation of a
blank titration, and make any necessary correction. Each mL film of propylene glycol.
of 0.1 M Edetate disodium titrant consumed is equivalent to C. IDENTIFICATION OF DIPROPYLENE GLYCOL (where stated on
2.698 mg of aluminum (Al). [NOTEUse the aluminum the label)
content thus obtained to calculate the Aluminum/chloride Sample solution: 200 mg/mL of Solution in isopropyl
atomic ratio.] alcohol, and filter. Evaporate the filtrate to 1 mL on a steam
ALUMINUM/CHLORIDE ATOMIC RATIO: Divide the percentage bath.
of aluminum found in the test for Content of aluminum by Acceptance criteria: The IR absorption spectrum of a film
the percentage of chloride found in the test for Content of of this solution on a silver chloride disk exhibits maxima only
chloride, and multiply by 35.453/26.98, in which 35.453 and at the same wavelengths as that of a similar preparation of a
26.98 are the atomic weights of chlorine and aluminum, film of dipropylene glycol.
respectively. D. IDENTIFICATION OF ALCOHOL (where stated on the label)
Acceptance criteria: The ratio is between 1.26:1 and Sample solution: Mix 5 drops of Solution in a small beaker
1.90:1. with 1 mL of potassium permanganate solution (1 in 100)
and 5 drops of 2 N sulfuric acid, and cover the beaker
IMPURITIES immediately with filter paper moistened with a freshly
Inorganic Impurities prepared solution of 0.1 g of sodium nitroferricyanide and
ARSENIC, Method I 211: NMT 2 ppm 0.25 g of piperazine in 5 mL of water.
HEAVY METALS, Method I 231: NMT 20 ppm Acceptance criteria: An intense blue color is produced on
LIMIT OF IRON the filter paper, the color becoming paler after a few min.
Standard solution: Transfer 2.0 mL of Standard Iron
Solution, prepared as directed under Iron 241, to a 50-mL ASSAY
beaker. PROCEDURE
Sample solution: 27 mg/mL of Aluminum Analysis: Calculate the percentage of anhydrous aluminum
Sesquichlorohydrate in water. Transfer 5.0 mL of this sesquichlorohydrate in the Solution:
solution to a 50 mL beaker.
Analysis: To each of the beakers containing the Standard Result = Al({Ar1x + [Mr(3x -1)] + Ar2}/Ar1x)
solution and the Sample solution add 5 mL of 6 N nitric
acid, cover with a watch glass, and boil on a hot plate for Al = percentage of aluminum from the test for
3 to 5 min. Allow to cool, add 5 mL of Ammonium Content of aluminum
Thiocyanate Solution, prepared as directed under Iron Ar1 = atomic weight of aluminum, 26.98
241, transfer to separate 50-mL color comparison tubes, x = aluminum/chloride atomic ratio
and dilute with water to volume. Mr = molecular weight of the hydroxide anion, 17.01
Acceptance criteria: The color of the solution from the Ar2 = atomic weight of chlorine, 35.453
the Standard solution (150 ppm).
Acceptance criteria: 90.0%110.0% Acceptance criteria: NMT 10 ppm

LIMIT OF IRON
OTHER COMPONENTS Standard solution: Transfer 2.0 mL of Standard Iron
CONTENT OF CHLORIDE Solution, prepared as directed under Iron 241, to a 50-mL
Sample solution: 1.4 g of Aluminum Sesquichlorohydrate beaker.
Solution to a 250-mL beaker, add 100 mL of water and 10 Sample solution: 53 mg/mL of Aluminum
mL of diluted nitric acid with stirring. Sesquichlorohydrate Solution. Transfer 5.0 mL of this
Analysis: Titrate with 0.1 N silver nitrate VS using a glass solution to a 50-mL beaker.
silversilver chloride electrode and a silver billet electrode Analysis: To each of the beakers add 5 mL of 6 N nitric
system. Each mL of 0.1 N silver nitrate is equivalent to 3.545 acid, cover with a watch glass, and boil on a hot plate for
mg of chloride (Cl). [NOTEUse the chloride content thus 3 to 5 min. Allow to cool, add 5 mL of Ammonium
obtained to calculate the Aluminum/chloride atomic ratio.] Thiocyanate Solution, prepared as directed under Iron 241,
CONTENT OF ALUMINUM transfer to separate 50-mL color comparison tubes, and
Edetate disodium titrant: Prepare a solution with a dilute with water to volume.
concentration of 37.2 mg/mL of edetate disodium, and Acceptance criteria: The color of the solution from the
standardize as follows. Weigh 2 g of aluminum wire, transfer Sample solution is not darker than that of the solution from
to a 1000-mL volumetric flask, and add 50 mL of a mixture the Standard solution (75 ppm).
of hydrochloric acid and water (1:1). Swirl the flask to
ensure contact of the aluminum and the acid, and allow the SPECIFIC TESTS
reaction to proceed until all of the aluminum has dissolved. PH 791: 3.05.0, in a solution (3 in 10)
into a 250-mL beaker, add, in the order named and with ADDITIONAL REQUIREMENTS
continuous stirring, 25.0 mL of Edetate disodium titrant and PACKAGING AND STORAGE: Preserve in well-closed containers.
20 mL of acetic acidammonium acetate buffer TS, and boil LABELING: Label Solution to state the solvent used and the
gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL claimed concentration of anhydrous aluminum
of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a sesquichlorohydrate contained therein.
make any necessary correction. Aluminum Sesquichlorohydrex
Polyethylene Glycol
Result = W/ArV (Comment on this Monograph)id=m2290=Aluminum
Sesquichlorohydrex Polyethylene Glycol=A-Monos.pdf)
W = weight, of aluminum in the portion of solution
taken (mg) Aly(OH)3y-zClz nH2O mH(OCH2CH2)nOH
Ar = atomic weight of aluminum, 26.98 Aluminum chlorohydroxide polyethylene glycol complex;
V = volume of Edetate disodium titrant consumed Aluminum hydroxychloride polyethylene glycol complex.
(mL) DEFINITION
Sample solution: Transfer 400 mg of Aluminum Aluminum Sesquichlorohydrex Polyethylene Glycol consists of
Sesquichlorohydrate Solution, to a 250-mL beaker, add 20 aluminum sesquichlorohydrate in which some of the waters of
mL of water and 5 mL of hydrochloric acid, boil on a hot hydration have been replaced by polyethylene glycol. It
plate for NLT 5 min, and allow to cool. encompasses a range of aluminum-to-chloride atomic ratios
Analysis: To the Sample solution add 25.0 mL of Edetate between 1.26:1 and 1.90:1. It contains NLT 90.0% and NMT
disodium titrant, and adjust with 2.5 N ammonium 110.0% of the labeled amount of anhydrous aluminum
hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 sesquichlorohydrate.
mL of acetic acidammonium acetate buffer TS, 50 mL of
alcohol, and 5 mL of dithizone TS. The pH of this solution IDENTIFICATION
should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until A. IDENTIFICATION TESTSGENERAL, Aluminum and Chloride
the color changes from green-violet to rose-pink. Perform a 191: A 100 mg/mL solution meets the requirements.
blank titration, and make any necessary correction. Each mL B. INFRARED ABSORPTION 197F
of 0.1 M Edetate disodium titrant consumed is equivalent to Sample solution: Dissolve 0.5 g in about 40 mL of water,
2.698 mg of aluminum (Al). [NOTEUse the aluminum and while mixing, adjust with 2.5 N sodium hydroxide to a
content thus obtained to calculate the Aluminum/chloride pH of 9.55 0.05. Filter the suspension of precipitate thus
atomic ratio.] obtained. Evaporate 15 mL of the filtrate to 1 mL on a hot
ALUMINUM/CHLORIDE ATOMIC RATIO: Divide the percentage plate. Deposit this solution on a silver chloride disk.
of aluminum found in the test for Content of aluminum by Standard solution: A similar preparation of polyethylene
the percentage of chloride found in the test for Content of glycol
chloride, and multiply by 35.453/26.98, in which 35.453 and
26.98 are the atomic weights of chlorine and aluminum, ASSAY
respectively. PROCEDURE
Acceptance criteria: The ratio is between 1.26:1 and Analysis: Calculate the percentage of anhydrous aluminum
1.90:1. sesquichlorohydrex in the Aluminum Sesquichlorohydrex
Polyethylene Glycol:
IMPURITIES
Inorganic Impurities Result = Al%({Ar1X + [Mr(3X 1)] + Ar2}/Ar1X)
ARSENIC, Method I 211
Sample solution: Prepare as directed for Test Preparation Al% = percentage of aluminum found in the test for
in the chapter, using a weighed quantity of the Solution. Content of aluminum
Acceptance criteria: NMT 2 ppm Ar1 = atomic weight of aluminum, 26.98
HEAVY METALS, Method I 231 X = aluminum/chloride atomic ratio found in the test
Sample solution: Prepare as directed for Test Preparatiion for Aluminum/chloride atomic ratio
in the chapter, using a weighed quantity of the Solution.
Mr = molecular weight of the hydroxide anion (OH), Acceptance criteria: The ratio is between 1.26:1 and
17.01 1.90:1.
OTHER COMPONENTS ARSENIC, Method I 211: NMT 2 ppm
CONTENT OF CHLORIDE HEAVY METALS, Method I 231: NMT 20 ppm
Sample solution: 700 mg of Aluminum Sesquichlorohydrex LIMIT OF IRON
Polyethylene Glycol to a 250-mL beaker. Add 100 mL of Standard solution: Transfer 2.0 mL of Standard Iron
water and 10 mL of diluted nitric acid with stirring. Solution, and prepare as directed under Iron 241, to a 50-
Analysis: Titrate with 0.1 N silver nitrate VS using a glass mL beaker.
silversilver chloride electrode and a silver billet electrode Sample solution: 27 mg/mL of Aluminum
system, determining the endpoint potentiometrically. Each Sesquichlorohydrex Polyethylene Glycol in water. Transfer
mL of 0.1 N silver nitrate is equivalent to 3.545 mg of 5.0 mL of this solution to a 50-mL beaker.
chloride (Cl). [NOTEUse the chloride content thus obtained Analysis: To each of the beakers containing the Standard
to calculate the Aluminum/chloride atomic ratio.] solution and the Sample solution, add 5 mL of 6 N nitric
CONTENT OF ALUMINUM acid, cover with a watch glass, and boil on a hot plate for
Edetate disodium titrant: Prepare a solution with a 3 to 5 min. Allow to cool, add 5 mL of Ammonium
concentration of 37.2 mg/mL of edetate disodium in water, Thiocyanate Solution, prepared as directed in Iron 241,
and standardize as follows. Weigh 2 g of aluminum wire. transfer to separate 50-mL color comparison tubes, and
Transfer to a 1000-mL volumetric flask, and add 50 mL of a dilute with water to volume.
mixture of hydrochloric acid and water (1:1). Swirl the flask Acceptance criteria: The color of the solution from the
to ensure contact of the aluminum and the acid, and allow Sample solution is not darker than that of the solution from
the reaction to proceed until all of the aluminum has the Standard solution (150 ppm).
solution into a 250-mL beaker, add, in the order named and SPECIFIC TESTS
with continuous stirring, 25.0 mL of Edetate disodium titrant PH 791: 3.05.0, in a solution of 15 in 100 (w/w)
boil gently for 5 min. Cool, and add 50 mL of alcohol, and ADDITIONAL REQUIREMENTS
2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to PACKAGING AND STORAGE: Preserve in well-closed containers.
a bright rose-pink color. Perform a blank determination, LABELING: The label states the content of anhydrous
substituting 10 mL of water for the aluminum solution, and aluminum sesquichlorohydrate.
Result = W/(Ar)(V)
Aluminum Sesquichlorohydrex
Propylene Glycol
W = weight of aluminum in the portion of Solution (Comment on this Monograph)id=m2293=Aluminum
taken (mg) Sesquichlorohydrex Propylene Glycol=A-Monos.pdf)
Ar = atomic weight of aluminum, 26.98 Aly(OH)3yzClz nH2O mC3H8O2
V = volume of Edetate disodium titrant consumed Aluminum chlorohydroxide propylene glycol complex;
(mL) Aluminum hydroxychloride propylene glycol complex.
Sample solution: Transfer about 200 mg of Aluminum
Sesquichlorohydrex Polyethylene Glycol to a 250-mL DEFINITION
beaker, add 20 mL of water and 5 mL of hydrochloric acid, Aluminum Sesquichlorohydrex Propylene Glycol consists of
boil on a hot plate for NLT 5 min, and allow to cool. aluminum sesquichlorohydrate in which some of the waters of
Analysis: To the Sample solution, add 25.0 mL of Edetate hydration have been replaced by propylene glycol. It
disodium titrant, and adjust with 2.5 N ammonium encompasses a range of aluminum-to-chloride atomic ratios
hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 between 1.26:1 and 1.90:1. It contains NLT 90.0% and NMT
mL of acetic acidammonium acetate buffer TS, 50 mL of 110.0% of the labeled amount of anhydrous aluminum
alcohol, and 5 mL of dithizone TS. The pH of this solution sesquichlorohydrate.
should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until
the color changes from green-violet to rose-pink. Perform a IDENTIFICATION
blank titration, and make any necessary correction. Each A. IDENTIFICATION TESTSGENERAL, Aluminum and Chloride
mL of 0.1 M Edetate disodium titrant consumed is 191: A 100 mg/mL solution meets the requirements.
equivalent to 2.698 mg of aluminum (Al). [NOTEUse the B. INFRARED ABSORPTION 197F:
aluminum content thus obtained to calculate the Sample solution: Dissolve 0.5 g in about 40 mL of water,
Aluminum/chloride atomic ratio.] and while mixing adjust with 2.5 N sodium hydroxide to a
ALUMINUM/CHLORIDE ATOMIC RATIO: Divide the percentage pH of 9.55 0.05. Filter the suspension of precipitate thus
of aluminum found in the test for Content of aluminum by obtained. Evaporate 15 mL of the filtrate to 1 mL on a hot
the percentage of chloride found in the test for Content of plate. Deposit this solution on a silver chloride disk.
chloride, and multiply by 35.453/26.98, in which 35.453 and Standard solution: A similar preparation of propylene
26.98 are the atomic weights of chlorine and aluminum, glycol
respectively.
ASSAY the percentage of chloride found in the test for Content of

PROCEDURE chloride, and multiply by 35.453/26.98, in which 35.453 and
Analysis: Calculate the percentage of anhydrous aluminum 26.98 are the atomic weights of chlorine and aluminum,
sesquichlorohydrate in the Aluminum Sesquichlorohydrex respectively.
Propylene Glycol: Acceptance criteria: The ratio is between 1.26:1 and
1.90:1.
Result = Al({Ar1X + [Mr(3X -1)] + Ar2}/Ar1X)
IMPURITIES
Al = percentage of aluminum from the test for Inorganic Impurities
Content of aluminum ARSENIC, Method I 211: NMT 2 ppm
Ar1 = atomic weight of aluminum, 26.98 HEAVY METALS, Method I 231: NMT 20 ppm
X = aluminum/chloride atomic ratio found in the test LIMIT OF IRON
for Aluminum/chloride atomic ratio Standard solution: Transfer 2.0 mL of Standard Iron
Mr = molecular weight of the hydroxide anion (OH), Solution, prepared as directed under Iron 241, to a 50-mL
17.01 beaker.
Ar2 = atomic weight of chlorine (Cl), 35.453 Sample solution: 27 mg/mL of Aluminum
Acceptance criteria: 90.0%110.0% Sesquichlorohydrex Propylene Glycol. Transfer 5.0 mL of
this solution to a 50-mL beaker.
OTHER COMPONENTS Analysis: To each of the beakers containing the Standard
CONTENT OF CHLORIDE solution and the Sample solution add 5 mL of 6 N nitric
Sample solution: 700 mg of Aluminum Sesquichlorohydrex acid, cover with a watch glass, and boil on a hot plate for
Propylene Glycol to a 250-mL beaker. Add 100 mL of water 3 to 5 min. Allow to cool, add 5 mL of Ammonium
and 10 mL of diluted nitric acid with stirring. Thiocyanate Solution, prepared as directed under Iron
Analysis: Titrate with 0.1 N silver nitrate VS using a glass 241, transfer to separate 50-mL color comparison tubes,
silversilver chloride electrode and a silver billet electrode and dilute with water to volume.
system. Each mL of 0.1 N silver nitrate is equivalent to 3.545 Acceptance criteria: The color of the solution from the
mg of chloride (Cl). [NOTEUse the chloride content thus Sample solution is not darker than that of the solution from
obtained to calculate the Aluminum/chloride atomic ratio.] the Standard solution (150 ppm).
CONTENT OF ALUMINUM
Edetate disodium titrant: Prepare a solution with a SPECIFIC TESTS
concentration of 37.2 mg/mL of edetate disodium, and PH 791: 3.05.0, in a solution [15 in 100 (w/w)]
standardize as follows. Weigh 2 g of aluminum wire, transfer
to a 1000-mL volumetric flask, and add 50 mL of a mixture ADDITIONAL REQUIREMENTS
of hydrochloric acid and water (1:1). Swirl the flask to PACKAGING AND STORAGE: Preserve in well-closed containers.
ensure contact of the aluminum and the acid, and allow the LABELING: The label states the content of anhydrous
reaction to proceed until all of the aluminum has dissolved. aluminum sesquichlorohydrate.
continuous stirring, 25.0 mL of Edetate disodium titrant and Aluminum Subacetate Topical Solution
gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL (Comment on this Monograph)id=m2300=Aluminum
of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a Subacetate Topical Solution=A-Monos.pdf)
Result = W/ArV
W = weight of aluminum in the portion of Solution
taken (mg) C4H7AlO5 162.08
Ar = atomic weight of aluminum, 26.98 Aluminum, bis(acetato-O)hydroxy-;
V = volume of Edetate disodium titrant consumed Bis(acetato)hydroxyaluminum;
(mL) Basic aluminum acetate [142-03-0; 8000-61-1].
Sample solution: Transfer 200 mg of Aluminum
Sesquichlorohydrex Propylene Glycol to a 250-mL beaker, DEFINITION
add 20 mL of water and 5 mL of hydrochloric acid, boil on Aluminum Subacetate Topical Solution yields, from each 100
a hot plate for NLT 5 min, and allow to cool. mL, NLT 2.30 g and NMT 2.60 g of Al2O3, and NLT 5.43 g and
Analysis: To the 25 mL of Sample solution add 25.0 mL of NMT 6.13 g of C2H4O2. It may be stabilized by the addition of
Edetate disodium titrant, and adjust with 2.5 N ammonium NMT 0.9% of boric acid.
hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 Aluminum Subacetate Topical Solution may be prepared as
mL of acetic acidammonium acetate buffer TS, 50 mL of follows:
alcohol, and 5 mL of dithizone TS. The pH of this solution
should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until Aluminum Sulfate 145 g
the color changes from green-violet to rose-pink. Perform a Acetic Acid 160 mL
blank titration, and make any necessary correction. Each mL
of 0.1 M Edetate disodium titrant consumed is equivalent to Calcium Carbonate 70 g
2.698 mg of aluminum (AL). [NOTEUse the aluminum Purified Water A sufficient quantity
content thus obtained to calculate the Aluminum/chloride To make 1000 mL
atomic ratio.]
ALUMINUM/CHLORIDE ATOMIC RATIO: Divide the percentage
of aluminum found in the test for Content of aluminum by
Dissolve the aluminum sulfate in 600 mL of cold water, filter Acceptance criteria: 5.43 g6.13 g of C2H4O2
the solution, and add the calcium carbonate gradually, in
several portions, with constant stirring. Then slowly add the SPECIFIC TESTS
acetic acid, mix, and set the mixture aside for 24 h. Filter the PH 791: 3.84.6
product with the aid of vacuum if necessary, returning the first LIMIT OF BORIC ACID
portion of the filtrate to the funnel. Wash the magma on the Sample solution: Pipet 25 mL of Aluminum Subacetate
filter with small portions of cold water, until the total filtrate Topical Solution into 75 mL of water in a conical flask.
measures 1000 mL. Analysis: To 100 mL of Sample solution, add 3 mL of
phenolphthalein TS, then add 0.5 N sodium hydroxide VS
IDENTIFICATION from a buret until a faint pink color is obtained. Heat to
IDENTIFICATION TESTS GENERAL, Aluminum and Sulfate boiling, and again neutralize. Add 150 mL of glycerin to the
191: Meets the requirements neutralized solution, and titrate with 0.5 N sodium
hydroxide VS. Perform a blank determination in a similar
ASSAY manner. From the volume of 0.5 N sodium hydroxide VS
ALUMINUM OXIDE used after the addition of the glycerin, subtract the volume
Edetate disodium titrant: Prepare a solution with a used in the blank. Each mL of 0.5 N sodium hydroxide is
concentration of 18.6 mg/mL of edetate disodium and equivalent to 30.92 mg of H3BO3
standardize as follows. Weigh 2 g of aluminum wire, transfer Acceptance criteria: NMT 0.6%
of hydrochloric acid and water (1:1). Swirl the flask to ADDITIONAL REQUIREMENTS
ensure contact of the aluminum and the acid, and allow the PACKAGING AND STORAGE: Preserve in tight containers.
continuous stirring, 25.0 mL of Edetate disodium titrant and Aluminum Sulfate
20 mL of acetic acidammonium acetate buffer TS, and boil (Comment on this Monograph)id=m2350=Aluminum Sulfate=A-
gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL Monos.pdf)
of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a Al2(SO4)3 xH2O (anhydrous)
bright rose-pink color. Perform a blank determination, Sulfuric acid, aluminum salt (3:2), hydrate;
substituting 10 mL of water for the aluminum solution, and Aluminum sulfate (2:3) hydrate [17927-65-0].
make any necessary connection. Anhydrous 342.15
Calculate the molarity of the solution taken by the formula: [10043-01-3].
Result = W/Ar V DEFINITION
Aluminum Sulfate contains NLT 54.0% and NMT 59.0% of
W = weight in mg, of aluminum in the portion of Al2(SO4)3. It contains a varying amount of water of
Solution crystallization.
V = volume of Edetate disodium titrant consumed IDENTIFICATION
(mL) IDENTIFICATION TESTSGENERAL, Aluminum and Sulfate 191:
Sample solution: Pipet 20 mL of Topical Solution into a A 100 mg/mL solution meets the requirements of the tests
250-mL volumetric flask, add 5 mL of hydrochloric acid, and for Aluminum and for Sulfate.
Analysis: Pipet 25 mL of Sample solution into a 250-mL ASSAY
beaker, and add, in the order named and with continuous PROCEDURE
stirring, 25.0 mL of Edetate disodium titrant and 20 mL of Edetate disodium titrant: Prepare a solution with a
acetic acidammonium acetate buffer TS, then heat the concentration of 18.6 mg/mL of edetate disodium and
solution near the boiling point for 5 min. Cool, and add 50 standardize as follows. Weigh 2 g of aluminum wire, transfer
mL of alcohol and 2 mL of dithizone TS. Titrate the solution to a 1000-mL volumetric flask, and add 50 mL of a mixture
with 0.05 M zinc sulfate VS to a bright rose-pink color. of hydrochloric acid and water (1:1). Swirl the flask to
Perform a blank determination, substituting water for the ensure contact of the aluminum and the acid, and allow the
sample. Each mL of 0.05 M Edetate disodium titrant is reaction to proceed until all of the aluminum has dissolved.
equivalent to 2.549 mg of Al2O3 Dilute with water to volume. Pipet 10 mL of this solution
Acceptance criteria: 2.30 2.60 g of Al2O3 into a 250-mL beaker, add, in the order named and with
ACETIC ACID continuous stirring, 25.0 mL of Edetate disodium titrant and
Sample solution: Pipet 20 mL of Topical Solution into a 20 mL of acetic acidammonium acetate buffer TS, and boil
Kjeldahl flask containing a mixture of 20 mL of phosphoric gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL
acid and 150 mL of water. of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a
Analysis: Connect the flask to a condenser, the delivery bright rose-pink color. Perform a blank determination,
tube from which dips beneath the surface of 50.0 mL of 0.5 substituting 10 mL of water for the aluminum solution, and
N sodium hydroxide VS contained in a receiving flask. Distill make any necessary correction.
160 mL, then remove the delivery tube from below the Calculate the molarity of the solution taken:
surface of the liquid, allow the distilling flask to cool, add 50
mL of water, and distill an additional 40 to 45 mL into the Result = W/ArV
receiving flask. Add phenolphthalein TS to the distillate, and
titrate the excess 0.5 N sodium hydroxide VS with 0.5 N W = weight of aluminum in the portion of solution
sulfuric acid VS. Each mL of 0.5 N sodium hydroxide is taken (mg)
equivalent to 30.03 mg of C2H4O2 Ar = atomic weight of aluminum, 26.98
V = volume of Edetate disodium titrant consumed B. IDENTIFICATION TESTSGENERAL, Sulfate and Calcium 191:
(mL) Suspend 2 g of sample in 50 mL of, and filter. The filtrate
Sample solution: 30.0 mg/mL of Aluminum Sulfate meets the requirements.
beaker, and add, in the order named and with continuous ASSAY
stirring, 25.0 mL of Edetate disodium titrant and 20 mL of ALUMINUM SULFATE
acetic acidammonium acetate buffer TS, then heat the Sample solution: Transfer 10 g of Aluminum Sulfate and
solution near the boiling point for 5 min. Cool, and add 50 Calcium Acetate for Topical Solution to a 1000-mL
mL of alcohol and 2 mL of dithizone TS. Titrate the solution volumetric flask. Add 100 mL of 1.2 M hydrochloric acid and
with 0.05 M zinc sulfate VS to a bright rose-pink color. 250 mL of water. Heat on a steam bath or hot plate until
Perform a blank determination, substituting water for the dissolved. Cool, dilute with water to volume. [NOTERetain
sample, and make any necessary correction. Each mL of a portion of this Sample solution for the Assay for Calcium
0.05 M Edetate disodium titrant is equivalent to 8.554 mg of acetate.]
Al2(SO4)3. Analysis: Transfer a 5.0-mL aliquot of the Sample solution to
Acceptance criteria: 54.0%59.0% a 250-mL conical flask. Add 40.0 mL of 0.01 M edetate
disodium VS and 20 mL of acetic acidammonium acetate
IMPURITIES buffer TS. Add 50 mL of alcohol and 2 mL of dithizone TS.
Inorganic Impurities [NOTEFollow the given order of addition.] Titrate with 0.02
IRON 241 M zinc sulfate VS until the color changes from green-violet
Sample solution: To 20 mL of a solution (1 in 150) add to clear rose-pink. Perform a blank titration, substituting 5.0
0.3 mL of potassium ferrocyanide TS. mL of water for the Sample solution. Each mL of 0.01 M
Acceptance criteria: No blue color is produced edetate disodium is equivalent to 2.972 mg of Al2(SO4)3
immediately. 14H2O.
HEAVY METALS 231 Calculate the percentage of Al2(SO4)3 14H2O taken:
Sample solution: Dissolve 1.0 g in 2 mL of 1 N acetic
acid, and dilute with water to 25 mL. Result = [(1000)(100)F M(VB VU)]/5.0 MTW
D = dilution factor, 1000/5.0
SPECIFIC TESTS 100 = conversion factor to percentage
PH 791: NLT 2.9, in a solution (1 in 20) F = conversion factor (2.972 mg of sample/mL of
WATER DETERMINATION, Method I 921: 41.0%46.0% 0.01 M edetate disodium)
LIMIT OF ALKALIES AND ALKALINE EARTHS: To a boiling M = actual molarity of the titrant
solution of 1.0 g in 150 mL of water add a few drops of VB = blank titration volume (mL)
methyl red TS, and then add 6 N ammonium hydroxide just VU = sample titration volume (mL)
until the color of the solution changes to a distinct yellow. MT = theoretical molarity of the titrant, 0.02
Add hot water to restore the volume to 150 mL, and filter W = weight of the sample (mg)
while hot. Evaporate 75 mL of the filtrate to dryness, and CALCIUM ACETATE
ignite to constant weight. Analysis: Transfer a 5.0-mL aliquot of the Sample solution
Acceptance criteria: NMT 2 mg of residue remains (0.4%). retained from the Assay for aluminum sulfate to a 250-mL
LIMIT OF AMMONIUM SALTS: Heat 1 g with 10 mL of 1 N conical flask. [NOTEFollow the given order of addition.]
sodium hydroxide on a steam bath for 1 min. Add 1 to 2 mL of 50% triethanolamine to mask the
Acceptance criteria: The odor of ammonia is not aluminum, mix well. Add 100 mL of water, 15 mL of 1 N
perceptible. sodium hydroxide, and 300 mg of hydroxy naphthol blue.
Titrate the solution with 0.01 M edetate disodium VS. The
ADDITIONAL REQUIREMENTS indicator will change from purple to a clear blue color at the
PACKAGING AND STORAGE: Preserve in well-closed containers. endpoint. Each mL of 0.01 M edetate disodium is equivalent
to 1.762 mg of C4H6CaO4 H2O.
Calculate the percentage of C4H6CaO4 H2O taken:
Aluminum Sulfate and Calcium Acetate Result = [(1000)(100)VUF M]/5.0 MTW
for Topical Solution
(Comment on this Monograph)id=m1370=Aluminum Sulfate D =
dilution factor, 1000/5.0
and Calcium Acetate for Topical Solution=A-Monos.pdf) 100 =
conversion factor to percentage
VU =
sample titration volume (mL)
DEFINITION F =
conversion factor (1.762 mg of sample/mL of
Aluminum Sulfate and Calcium Acetate for Topical Solution 0.01 M edetate disodium)
contains NLT 90.0% and NMT 110.0% of the labeled amounts M = actual molarity of the titrant
of aluminum sulfate tetradecahydrate [Al2(SO4)3 14H2O] and MT = theoretical molarity of the titrant. 0.01
calcium acetate monohydrate (C4H6CaO4 H2O). W = weight of the sample (mg)
IDENTIFICATION
A. Place approximately 0.25 g of Aluminum Sulfate and SPECIFIC TESTS
Calcium Acetate for Topical Solution in a test tube. Add 10 PH 791: 4.04.8, in a solution (1 in 200)
mL of water and 0.25 g of calcium carbonate. Heat on a
steam bath for 10 min, and filter. Add 3 to 4 drops of ferric ADDITIONAL REQUIREMENTS
chloride TS to the filtrate. A reddish-brown color or PACKAGING AND STORAGE: Preserve in single-unit containers,
precipitate indicates acetate. [NOTEAfter the addition of the and protect from excessive heat.
ferric chloride TS, the solution may be heated for 1 min to
speed the reaction.]
Aluminum Sulfate and Calcium Acetate Aluminum Zirconium

Tablets for Topical Solution Octachlorohydrate
(Comment on this Monograph)id=m2353=Aluminum Sulfate (Comment on this Monograph)id=m2355=Aluminum Zirconium
and Calcium Acetate Tablets for Topical Solution=A-Monos.pdf) Octachlorohydrate=A-Monos.pdf)
DEFINITION AlyZr(OH)3y+4-xClx nH2O
Aluminum Sulfate and Calcium Acetate Tablets for Topical DEFINITION
Solution contain NLT 90.0% and NMT 110.0% of the labeled Aluminum Zirconium Octachlorohydrate is a polymeric, loosely
amounts of aluminum sulfate tetradecahydrate [Al2(SO4)3 hydrated complex of basic aluminum zirconium chloride that
14H2O] and calcium acetate monohydrate (C4H6CaO4 H2O). encompasses a range of aluminum-to-zirconium atomic ratios
IDENTIFICATION between 6.0:1 and 10.0:1, and a range of (aluminum plus
A. IDENTIFICATION TESTSGENERAL, Aluminum 191 zirconium)-to-chloride atomic ratios between 1.5:1 and 0.9:1.
Sample solution: 40 mg/mL of ground Tablet powder in It contains NLT 90.0% and NMT 110.0% of the labeled
water, and filter amount of anhydrous aluminum zirconium octachlorohydrate.
Analysis: Mix 2 mL of the Sample solution with 2 mL of IDENTIFICATION
water and 2 drops of 3 N hydrochloric acid. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution
Acceptance criteria: The solution responds to the containing the equivalent of 100 mg/mL of anhydrous
ammonium hydroxide test. aluminum zirconium octachlorohydrate meets the
[NOTERetain the remaining filtrate for Identification test B.] requirements of the test for Chloride.
B. IDENTIFICATION TESTSGENERAL, Sulfate and Calcium 191:
A portion of the filtrate retained from Identification test A ASSAY
responds to the tests for Sulfate and for Calcium. PROCEDURE
Analysis: Calculate the percentage of anhydrous aluminum
ASSAY zirconium octachlorohydrate in the Aluminum Zirconium
ALUMINUM SULFATE Octachlorohydrate:
Sample solution: Finely powder and mix not fewer than 20
Tablets. Weigh a portion of the powder, equivalent to 2.8 g Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z]
of aluminum sulfate, and transfer to a 1000-mL volumetric + Ar3(y + 1)/z}/Ar1y)
flask. Add 100 mL of 1.2 N hydrochloric acid and 100 mL of
water, and heat on a steam bath, with occasional swirling, Al% = percentage of aluminum found in the test for
to dissolve the powder. Allow the solution to cool, dilute Content of aluminum
with water to volume. Ar1 = atomic weight of aluminum, 26.98
[NOTERetain a portion of this Sample solution for the y = aluminum/zirconium atomic ratio found in the
Assay for Calcium Acetate.] test for Aluminum/zirconium atomic ratio
Analysis: Transfer 25.0 mL of the Sample solution to a 250- Ar2 = atomic weight of zirconium corrected for 2%
mL conical flask. Add 40.0 mL of 0.01 M edetate disodium hafnium content, 92.97
VS and 20 mL of acetic acidammonium acetate buffer TS, Mr = molecular weight of the hydroxide anion (OH),
and mix by swirling. Add 50 mL of alcohol and 2 mL of 17.01
dithizone TS, and titrate with 0.02 M zinc sulfate VS until z = (aluminum plus zirconium)/chloride atomic ratio
the color changes from green-violet to a clear rose-pink. found in the test for (Aluminum plus
Perform a blank determination, substituting 25 mL of water zirconium)/chloride atomic ratio
for the Sample solution, and make any necessary correction. Ar3 = atomic weight of chlorine (Cl), 35.453
Each mL of 0.01 M edetate disodium is equivalent to 2.972 Acceptance criteria: 90.0%110.0%
mg of Al2(SO4)3 14H2O.
Acceptance criteria: 90.0%110.0% OTHER COMPONENTS
CALCIUM ACETATE CONTENT OF CHLORIDE
Analysis: Transfer 20.0 mL of the Sample solution retained Sample: 250 mg of Aluminum Zirconium
from the Assay for Aluminum Sulfate to a 125-mL conical Octachlorohydrate to a 250-mL beaker
flask. With constant stirring, add in the order named, 0.5 mL Analysis: Add 100120 mL of water and 20 mL of diluted
of trolamine, 10 mL of ammoniaammonium chloride buffer nitric acid, and swirl to dissolve. Titrate with 0.05 N silver
TS, and 3 drops of a solution prepared by dissolving 500 nitrate VS using a calomel electrode and a silver billet
mg of eriochrome black T trituration in 10 mL of methanol, electrode system. Each mL of 0.05 N silver nitrate is
and titrate with 0.01 M edetate disodium VS to a violet equivalent to 1.773 mg of chloride (Cl). [NOTEUse the
endpoint. Each mL of 0.01 M edetate disodium is equivalent result obtained to calculate the (Aluminum plus
to 1.762 mg of C4H6CaO4 H2O. zirconium)/chloride atomic ratio.]
Acceptance criteria: 90.0%110.0% CONTENT OF ZIRCONIUM
Sample: 250 mg of Aluminum Zirconium
PERFORMANCE TESTS Octachlorohydrate to a 150-mL beaker
DISINTEGRATION 701: 10 min Analysis: Add 5 mL of water and 15 mL of hydrochloric
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements acid. Heat this solution to boiling, and continue boiling for
for Weight Variation 68 min. Add 3040 mL of water and 5 mL of hydrochloric
acid, and heat to boiling. Add 1 drop of xylenol orange TS,
SPECIFIC TESTS and, while still hot, titrate with 0.1 M edetate disodium VS
PH 791: Between 4.0 and 4.8, in a solution of 2 g of until the color of the solution changes from pink to yellow.
ground Tablet powder in 500 mL of water Perform a blank determination, and make any necessary
LOSS ON DRYING 731: Dry ground Tablet powder at 150 correction. Each mL of 0.1 M edetate disodium is equivalent
for 15 min: it loses NMT 18% of its weight. to 9.297 mg of zirconium (Zr). [NOTEUse the result
ADDITIONAL REQUIREMENTS obtained to calculate the Aluminum/zirconium atomic ratio
PACKAGING AND STORAGE: Preserve in tight containers, and and the (Aluminum plus zirconium)/chloride atomic ratio.]
avoid excessive heat.
CONTENT OF ALUMINUM Sample solution: 27 mg/mL of Aluminum Zirconium

Sample: 0.15 g of Aluminum Zirconium Octachlorohydrate Octachlorohydrate. Transfer 5.0 mL of this solution to a 50-
to a 150-mL beaker mL beaker.
Analysis: Add 5 mL of water and 15 mL of hydrochloric Analysis: To each of the beakers containing the Standard
acid. Heat this solution to boiling, and continue boiling for 5 solution and the Sample solution, add 5 mL of 6 N nitric
min. Add 40 mL of water and 15.0 mL of 0.1 M edetate acid, cover with a watch glass, and boil on a hot plate for
disodium VS. Heat the solution to boiling, and continue 35 min. Allow to cool. Add 5 mL of Ammonium
boiling for 5 min. Allow the solution to cool, add 1015 mL Thiocyanate Solution, prepared as directed under Iron 241,
of acetic acidammonium acetate buffer TS, and adjust with transfer to separate 50-mL color comparison tubes, and
ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of dilute with water to volume.
alcohol, and adjust with ammonium hydroxide to a pH of Acceptance criteria: The color of the solution from the
4.6 0.1. Add 510 drops of dithizone TS, and titrate with Sample solution is not darker than that of the solution from
0.1 M zinc sulfate VS until the first permanent purple-pink the Standard solution. (NMT 150 ppm)
color appears. Perform a blank determination, and make any HEAVY METALS, Method I 231
necessary correction. [NOTEProceed as directed, except use the following
Calculate the percentage of aluminum (Al) in the Aluminum modifications.]
Zirconium Octachlorohydrate: Sample solution: Prepare as directed in the chapter. If the
solution is not clear after dilution to 40 mL, heat for several
Result = 2.698[15.0 Me (zMz + Ze)]/W min between 60 and 80, and cool to room temperature.
If the solution remains cloudy, repeat from the beginning
Me = molarity of the edetate disodium VS with the following modification: add 3 mL of hydrochloric
z = volume of zinc sulfate VS consumed (mL) acid before adjustment of the pH with 6 N ammonium
Mz = molarity of the zinc sulfate VS hydroxide.
Ze = equivalent volume of edetate disodium VS Monitor solution: Prepare as directed, using the same
consumed (mL) by the zirconium moiety, modifications described for Sample solution, if necessary.
calculated as (Zr%/Me) (W/Ar), where Zr% is Analysis: To each of the three tubes containing the
the percentage of zirconium as determined in Standard solution, the Sample solution, and the Monitor
the test for Content of zirconium, and Ar is the solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently
atomic weight of zirconium corrected for 2% to between 60 and 80. To the Standard solution, add 6
hafnium content, 92.97 drops of sodium sulfide TS. To the Sample solution and the
W = quantity of aluminum zirconium Monitor solution, add 12 drops of sodium sulfide TS. Cool
octachlorohydrate taken (g) to room temperature, and dilute the contents of each tube
[NOTEUse the result obtained to calculate the Aluminum/ with water to 50 mL. Gently mix each tube by inverting
zirconium atomic ratio and the (Aluminum plus zirconium)/ twice. Allow to stand for 5 min, and view downward over a
chloride atomic ratio] white surface. The color of the solution from the Sample
ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage solution is not darker than that of the solution from the
of aluminum found in the test for Content of aluminum by Standard solution, and the color of the solution from the
the percentage of zirconium found in the test for Content of Monitor solution is the same as or darker than the color of
zirconium, and multiply by 92.97/26.98, in which 92.97 is the solution from the Standard solution. If the color of the
the atomic weight of zirconium corrected for 2% hafnium solution from the Monitor solution is lighter than the color
content, and 26.98 is the atomic weight of aluminum. of the solution from the Standard solution, repeat the
Acceptance criteria: 6.0:1 and 10.0:1. analysis with the following modification: after the heating
(ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO step, to the Monitor solution and the Sample solution add
Calculate the (aluminum plus zirconium)/chloride atomic 1.0 mL, instead of 12 drops, of sodium sulfide TS.
ratio: Acceptance criteria: NMT 20 ppm
Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3) ADDITIONAL REQUIREMENTS
Al% = percentage of aluminum, as determined in the LABELING: The label states the content of anhydrous
test for Content of aluminum aluminum zirconium octachlorohydrate.
Ar1 = atomic weight of aluminum, 26.98
Zr% = percentage of zirconium as determined in the
test for Content of zirconium
Ar2 = atomic weight of zirconium corrected for 2% Aluminum Zirconium
hafnium content, 92.97 Octachlorohydrate Solution
Cl% = percentage of chloride as determined in the test (Comment on this Monograph)id=m2356=Aluminum Zirconium
for Content of chloride Octachlorohydrate Solution=A-Monos.pdf)
Ar3 = atomic weight of chlorine, 35.453
Acceptance criteria: 1.5:10.9:1 DEFINITION
Aluminum Zirconium Octachlorohydrate Solution consists of
SPECIFIC TESTS complex basic aluminum chloride that is polymeric and
PH 791: 3.05.0, in a solution (15 in 100) encompasses a range of aluminum-to-zirconium atomic ratios
IMPURITIES between 6.0:1 and 10.0:1, and a range of (aluminum plus
Inorganic Impurities zirconium)-to-chloride atomic ratios between 1.5:1 and 0.9:1.
ARSENIC, Method I 211: NMT 2 ppm The following solvents may be used: water, propylene glycol,
LIMIT OF IRON or dipropylene glycol. It contains the equivalent of NLT 90.0%
Standard solution: Transfer 2.0 mL of Standard Iron and NMT 110.0% of the labeled concentration of anhydrous
Solution, prepared as directed under Iron 241, to a 50-mL aluminum zirconium octachlorohydrate.
beaker.
IDENTIFICATION CONTENT OF ALUMINUM

A. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution Sample: 0.3 g of Aluminum Zirconium Octachlorohydrate
containing the equivalent of 100 mg/mL of anhydrous Solution to a 150-mL beaker
aluminum zirconium octachlorohydrate meets the Analysis: Add 5 mL of water and 15 mL of hydrochloric
requirements of the test for Chloride. acid. Heat this solution to boiling, and continue boiling for 5
B. PROPYLENE GLYCOL (where stated on the label) min. Add 40 mL of water and 15.0 mL of 0.1 M edetate
Sample solution: 200 mg/mL in isopropyl alcohol, and disodium VS. Heat the solution to boiling, and continue
filter boiling for 5 min. Allow the solution to cool, add 1015 mL
Analysis: Evaporate the filtrate to 1 mL on a steam bath: of acetic acidammonium acetate buffer TS, and adjust with
the IR absorption spectrum of a film of this solution on a ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of
silver chloride disk exhibits maxima only at the same alcohol, and adjust with ammonium hydroxide to a pH of
wavelengths as that of a similar preparation of a film of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with
propylene glycol. 0.1 M zinc sulfate VS until the first permanent purple-pink
C. DIPROPYLENE GLYCOL (where stated on the label) color appears. Perform a blank determination, and make any
Sample solution: 200 mg/mL in isopropyl alcohol, and necessary correction.
filter Calculate the percentage of aluminum (Al) in the Aluminum
Analysis: Evaporate the filtrate to 1 mL on a steam bath: Zirconium Octachlorohydrate:
the IR absorption spectrum of a film of this solution on a
silver chloride disk exhibits maxima only at the same Result = 2.698[15.0 Me (zMz + Ze)]/W
wavelengths as that of a similar preparation of a film of
dipropylene glycol. Me = molarity of the edetate disodium VS
z = volume of zinc sulfate VS consumed (mL)
ASSAY Mz = molarity of the zinc sulfate VS
PROCEDURE Ze = equivalent volume of edetate disodium VS
Analysis: Calculate the percentage of anhydrous aluminum consumed (mL) by the zirconium moiety,
zirconium octachlorohydrate in the Solution taken: calculated as (Zr%/Me) (W/Ar2), where Zr% is
the percentage of zirconium as determined in
Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + the test for Content of zirconium, and Ar2 is the
1)/z}/Ar1y) atomic weight of zirconium corrected for 2%
hafnium content, 92.97
Al% = percentage of aluminum found in the test for W = quantity of Aluminum Zirconium
Content of aluminum Octachlorohydrate taken (g)
Ar1 = atomic weight of aluminum, 26.98 [NOTEUse the result obtained to calculate the Aluminum/
y = aluminum/zirconium atomic ratio found in the zirconium atomic ratio and the (Aluminum plus zirconium)/
test for Aluminum/zirconium atomic ratio chloride atomic ratio.]
Ar2 = atomic weight of zirconium corrected for 2% ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage
hafnium content, 92.97 of aluminum found in the test for Content of aluminum by
Mr = molecular weight of the hydroxide anion (OH), the percentage of zirconium found in the test for Content of
17.01 zirconium, and multiply by 92.97/26.98, in which 92.97 is
z = (aluminum plus zirconium)/chloride atomic ratio the atomic weight of zirconium corrected for 2% hafnium
found in the test for (Aluminum plus content, and 26.98 is the atomic weight of aluminum.
zirconium)/chloride atomic ratio Acceptance criteria: 6.0:1 and 10.0:1.
Ar3 = atomic weight of chlorine (Cl), 35.453 (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO
Acceptance criteria: 90.0%110.0% Calculate the (aluminum plus zirconium)/chloride atomic
ratio:
SPECIFIC TESTS
CONTENT OF CHLORIDE Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3)
Octachlorohydrate Solution to a 250-mL beaker Al% = percentage of aluminum from the test for
Analysis: Add 100120 mL of water and 20 mL of diluted Content of aluminum
nitric acid, and swirl to dissolve. Titrate with 0.05 N silver Ar1 = atomic weight of aluminum, 26.98
nitrate VS using a calomel electrode and a silver billet Zr% = percentage of zirconium as determined in the
electrode system. Each mL of 0.05 N silver nitrate is test for Content of zirconium
equivalent to 1.773 mg of chloride (Cl). [NOTEUse the Ar2 = atomic weight of zirconium corrected for 2%
result obtained to calculate the (Aluminum plus hafnium content, 92.97
zirconium)/chloride atomic ratio.] Cl% = percentage of chloride as determined in the test
CONTENT OF ZIRCONIUM for Content of chloride
Sample: 500 mg of Aluminum Zirconium Ar3 = atomic weight of chlorine, 35.453
Octachlorohydrate Solution to a 150-mL beaker Acceptance criteria: 1.5:10.9:1
Analysis: Add 5 mL of water and 15 mL of hydrochloric
acid. Heat this solution to boiling, and continue boiling for IMPURITIES
68 min. Add 3040 mL of water and 5 mL of hydrochloric Inorganic Impurities
acid, and heat to boiling. Add 1 drop of xylenol orange TS, ARSENIC, Method I 211: Prepare the Sample solution using a
and while still hot, titrate with 0.1 M edetate disodium VS weighed quantity of the Solution.
until the color of the solution changes from pink to yellow. Acceptance criteria: NMT 2 ppm
Perform a blank determination, and make any necessary LIMIT OF IRON
correction. Each mL of 0.1 M edetate disodium is equivalent Standard solution: Transfer 2.0 mL of Standard Iron
to 9.297 mg of zirconium (Zr). [NOTEUse the result Solution, prepared as directed under Iron 241, to a 50-mL
obtained to calculate the Aluminum/zirconium atomic ratio beaker.
and the (Aluminum plus zirconium)/chloride atomic ratio.]
Sample solution: 53 mg/mL of Aluminum Zirconium 10.0:1, and a range of (aluminum plus zirconium)-to-chloride
Octachlorohydrate Solution atomic ratios between 1.5:1 and 0.9:1. It contains NLT 90.0%
Transfer 5.0 mL of this solution to a 50-mL beaker. and NMT 110.0% of the labeled amount of anhydrous
Analysis: To each of the beakers containing the Standard aluminum zirconium octachlorohydrate.
solution and the Sample solution, add 5 mL of 6 N nitric
acid, cover with a watch glass, and boil on a hot plate for IDENTIFICATION
35 min. Allow to cool. Add 5 mL of Ammonium A. IDENTIFICATION TESTSGENERAL, Chloride 191: A 100
Thiocyanate Solution, prepared as directed under Iron 241, mg/mL solution meets the requirements.
transfer to separate 50-mL color comparison tubes, and B. PROCEDURE
dilute with water to volume. Sample solution: 25 mg/mL in water
Acceptance criteria: The color of the solution from the Analysis: Heat to boiling on a hot plate, and add 60 mg of
Sample solution is not darker than that of the solution from ninhydrin to 20 mL of Sample solution.
the Standard solution. (NMT 75 ppm). Acceptance criteria: A deep violet color immediately
HEAVY METALS, Method I 231 develops.
[NOTEProceed as directed, except use the following
modifications.] ASSAY
Sample solution: Prepare as directed in the chapter, using PROCEDURE
a weighed quantity of Solution. If the solution is not clear Analysis: Calculate the percentage of anhydrous aluminum
after dilution to 40 mL, heat for several min between 60 zirconium octachlorohydrate in the Aluminum Zirconium
and 80, and cool to room temperature. If the solution Octachlorohydrate Gly in the solution taken:
remains cloudy, repeat from the beginning with the Result = Al% ({Ar1y + Ar2 + Mr1[3y + 4 (y + 1)/z] + Ar3(y +
following modification: add 3 mL of hydrochloric acid 1)/z}/ Ar1y)
before adjustment of the pH with 6 N ammonium
hydroxide. Al% = percentage of aluminum from the test for
Monitor solution: Prepare as directed, using the same Content of Aluminum
modifications described for the Sample solution, if Ar1 = atomic weight of aluminum, 26.98
necessary. y = aluminum/zirconium atomic ratio from the test
Analysis: To each of the three tubes containing the for Aluminum/Zirconium Atomic Ratio
Standard solution, the Sample solution, and the Monitor Ar2 = atomic weight of zirconium corrected for 2%
solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently hafnium content, 92.97
to between 60 and 80. To the Standard solution, add 6 Mr1 = molecular weight of the hydroxide anion (OH),
drops of sodium sulfide TS. To the Sample solution and the 17.01
Monitor solution, add 12 drops of sodium sulfide TS. Cool z = (aluminum plus zirconium)/chloride atomic ratio
to room temperature, and dilute the contents of each tube from the test for (Aluminum plus
with water to 50 mL. Gently mix each tube by inverting Zirconium)/Chloride Atomic Ratio
twice. Allow to stand for 5 min, and view downward over a Ar3 = atomic weight of chlorine (Cl), 35.453
white surface. Acceptance criteria: 90.0%110.0%
The color of the solution from the Sample solution is not OTHER COMPONENTS
darker than that of the solution from the Standard CONTENT OF CHLORIDE
solution, and the color of the solution from the Monitor Sample: 250 mg of Aluminum Zirconium
solution is the same as or darker than the color of the Octachlorohydrate Gly to a 250-mL beaker
solution from the Standard solution. If the color of the Analysis: Add 100120 mL of water and 20 mL of diluted
solution from the Monitor solution is lighter than the color nitric acid, and swirl to dissolve. Titrate with 0.05 N silver
of the solution from the Standard solution, repeat the nitrate VS using a calomel electrode and a silver billet
analysis with the following modification: after the heating electrode system. Each mL of 0.05 N silver nitrate is
step, to the Monitor solution and the Sample solution add equivalent to 1.773 mg of chloride (Cl). Use the result
1.0 mL, instead of 12 drops, of sodium sulfide TS. obtained to calculate the (Aluminum plus Zirconium)/Chloride
Atomic Ratio.
SPECIFIC TESTS CONTENT OF ZIRCONIUM
PH 791: 3.05.0, in a solution (3 in 10) Sample: 250 mg of Aluminum Zirconium
ADDITIONAL REQUIREMENTS Octachlorohydrate Gly to a 150-mL beaker
PACKAGING AND STORAGE: Preserve in well-closed containers. Analysis: Add 5 mL of water and 15 mL of hydrochloric
LABELING: Label the Solution to state the solvent used and acid. Heat this solution to boiling, and continue boiling for
the claimed concentration of anhydrous aluminum zirconium 68 min. Add 3040 mL of water and 5 mL of hydrochloric
octachlorohydrate. acid, and heat to boiling. Add 1 drop of xylenol orange TS,
and, while still hot, titrate with 0.1 M edetate disodium VS
until the color of the solution changes from pink to yellow.
Perform a blank determination, and make any necessary
Aluminum Zirconium Octachlorohydrex correction. Each mL of 0.1 M edetate disodium is equivalent
Gly to 9.297 mg of zirconium (Zr). Use the result obtained to
calculate the Aluminum/Zirconium Atomic Ratio and the
(Comment on this Monograph)id=m2358=Aluminum Zirconium (Aluminum plus Zirconium)/Chloride Atomic Ratio.
Octachlorohydrex Gly=A-Monos.pdf) CONTENT OF ALUMINUM
DEFINITION Sample: 0.15 g of Aluminum Zirconium Octachlorohydrate
Aluminum Zirconium Octachlorohydrex Gly is a derivative of Gly to a 150-mL beaker
Aluminum Zirconium Octachlorohydrate in which some of the Analysis: Add 5 mL of water and 15 mL of hydrochloric
water molecules have been displaced by glycine, calcium acid. Heat this solution to boiling, and continue boiling for 5
glycinate, magnesium glycinate, potassium glycinate, sodium min. Add 40 mL of water and 15.0 mL of 0.1 M edetate
glycinate, or zinc glycinate. It encompasses a range of disodium VS. Heat the solution to boiling, and continue
aluminum-to-zirconium atomic ratios between 6.0:1 and boiling for 5 min. Allow the solution to cool, add 1015 mL
of acetic acidammonium acetate buffer TS, and adjust with HEAVY METALS, Method I 231
ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of [NOTEProceed as directed in the chapter, except to use the
alcohol, and adjust with ammonium hydroxide to a pH of following modifications.]
4.6 0.1. Add 510 drops of dithizone TS, and titrate with Sample solution: Prepare as directed in the chapter. If the
0.1 M zinc sulfate VS until the first permanent purple-pink solution is not clear after dilution to 40 mL, heat for several
color appears. Perform a blank determination, and make any min between 60 and 80, and cool to room temperature.
necessary correction. If the solution remains cloudy, repeat from the beginning
Calculate the percentage of aluminum (Al) in the Aluminum with the following modification: add 3 mL of hydrochloric
Zirconium Octachlorohydrate Gly: acid prior to adjustment of the pH with 6 N ammonium
hydroxide.
Result = 2.698[15.0 Me (zMz + Ze)]/W Monitor solution: Prepare as directed, using the same
modifications described for Sample solution, if necessary.
Me = molarity of the edetate disodium VS Analysis: To each of the three tubes containing the
z = volume of zinc sulfate VS consumed (mL) Standard solution, the Sample solution, and the Monitor
Mz = molarity of the zinc sulfate VS solution, add 2 mL of pH 3.5 Acetate Buffer, and heat
Ze = equivalent volume of edetate disodium VS gently to between 60 and 80. To the Standard solution
consumed (mL) by the zirconium moiety, add 6 drops of sodium sulfide TS. To the Sample solution
calculated as (Zr%/Me) (W/Ar2), where Zr% is and the Monitor solution add 12 drops of sodium sulfide TS.
the percentage of zirconium as determined in Cool to room temperature, and dilute the contents of each
the test for Content of zirconium, and Ar2 is the tube with water to 50 mL. Gently mix each tube by
atomic weight of zirconium corrected for 2% inverting twice. Allow to stand for 5 min, and view
hafnium content, 92.97 downward over a white surface.
W = quantity of Aluminum Zirconium Acceptance criteria: NMT 20 ppm
Octachlorohydrate taken (g) The color of the solution from the Sample solution is not
[NOTEUse the result obtained to calculate the Aluminum/ darker than that of the solution from the Standard
zirconium atomic ratio and the (Aluminum plus zirconium)/ solution, and the color of the solution from the Monitor
chloride atomic ratio.] solution is the same as or darker than the color of the
ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage solution from the Standard solution. If the color of the
of aluminum found in the test for Content of aluminum by solution from the Monitor solution is lighter than the color
the percentage of zirconium found in the test for Content of of the solution from the Standard solution, repeat the
zirconium, and multiply by 92.97/26.98, in which 92.97 is analysis with the following modification: after the heating
the atomic weight of zirconium corrected for 2% hafnium step, to the Monitor solution and the Sample solution add
content, and 26.98 is the atomic weight of aluminum: the 1.0 mL, instead of 12 drops, of sodium sulfide TS.
ratio is between 6.0:1 and 10.0:1.
(ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO SPECIFIC TESTS
Analysis: Calculate the (aluminum plus zirconium)/chloride PH 791: 3.05.0, in a solution (15 in 100)
atomic ratio:
Result = [(Al %/Ar1) + (Zr/Ar2)]/(Cl/Ar3) PACKAGING AND STORAGE: Preserve in well-closed containers.
LABELING: The label states the form of glycine used and the
Al % = percentage of aluminum from the test for claimed content of anhydrous aluminum zirconium
Content of aluminum octachlorohydrate.
Zr = percentage of zirconium from the test for Content
of zirconium
Cl = percentage of chloride from the test for Content
of chloride Aluminum Zirconium Octachlorohydrex
Ar1 = atomic weight of aluminum, 26.98 Gly Solution
Ar2 = atomic weight of zirconium corrected for 2% (Comment on this Monograph)id=m2359=Aluminum Zirconium
hafnium content, 92.97 Octachlorohydrex Gly Solution=A-Monos.pdf)
Aluminum Zirconium Octachlorohydrex Gly Solution is a
IMPURITIES solution of Aluminum Zirconium Octachlorohydrate in which
Inorganic Impurities some of the waters of hydration have been displaced by
ARSENIC, Method I 211: NMT 2 ppm glycine, calcium glycinate, magnesium glycinate, potassium
LIMIT OF IRON glycinate, sodium glycinate, or zinc glycinate. It encompasses
Standard solution: Transfer 2.0 mL of Standard Iron a range of aluminum-to-zirconium ratios between 6.0:1 and
Solution, prepared as directed under Iron 241, to a 50-mL 10.0:1, and a range of (aluminum plus zirconium)-to-chloride
beaker. atomic ratios between 1.5:1 and 0.9:1. The following solvents
Sample solution: 27 mg/mL of Aluminum Zirconium may be used: water, propylene glycol, or dipropylene glycol. It
Octachlorohydrate Gly. Transfer 5.0 mL of this solution to a contains the equivalent of NLT 90.0% and NMT 110.0% of
50-mL beaker. the labeled concentration of anhydrous aluminum zirconium
Analysis: To each of the beakers containing the Standard octachlorohydrate.
solution and the Sample solution add 5 mL of 6 N nitric
acid, cover with a watch glass, and boil on a hot plate for IDENTIFICATION
35 min. Allow to cool, add 5 mL of Ammonium A. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution
Thiocyanate Solution, prepared as directed under Iron 241, containing the equivalent of 100 mg/mL of anhydrous
transfer to separate 50-mL color comparison tubes, and aluminum zirconium octachlorohydrate meets the
dilute with water to volume. requirements.
Acceptance criteria: The color of the solution from the
the Standard solution (NMT 150 ppm).
B. PROPYLENE GLYCOL (where stated on the label) CONTENT OF ALUMINUM

Sample solution: 200 mg/mL in isopropyl alcohol, and Sample: 0.30 g of Aluminum Zirconium Octachlorohydrate
filter Gly Solution in a 150-mL beaker
Analysis: Evaporate the filtrate to 1 mL on a steam bath: Analysis: Add 5 mL of water and 15 mL of hydrochloric
the IR absorption spectrum of a film of this solution on a acid. Heat this solution to boiling, and continue boiling for 5
silver chloride disk exhibits maxima only at the same min. Add 40 mL of water and 15.0 mL of 0.1 M edetate
wavelengths as that of a similar preparation of a film of disodium VS. Heat the solution to boiling, and continue
propylene glycol. boiling for 5 min. Allow the solution to cool, add 1015 mL
C. DIPROPYLENE GLYCOL (where stated on the label) of acetic acidammonium acetate buffer TS, and adjust with
Sample solution: 200 mg/mL in isopropyl alcohol, and ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of
filter alcohol, and adjust with ammonium hydroxide to a pH of
Analysis: Evaporate the filtrate to 1 mL on a steam bath: 4.6 0.1. Add 510 drops of dithizone TS, and titrate with
the IR absorption spectrum of a film of this solution on a 0.1 M zinc sulfate VS until the first permanent purple-pink
silver chloride disk exhibits maxima only at the same color appears. Perform a blank determination, and make any
wavelengths as that of a similar preparation of a film of necessary correction.
dipropylene glycol. Calculate the percentage of aluminum (Al) in the Aluminum
D. GLYCINE Zirconium Octachlorohydrate Gly:
Sample solution: 50 mg/mL in isopropyl alcohol, and filter
Analysis: Heat to boiling on a hot plate, and add 60 mg of Result = 2.698[15.0 Me (zMz + Ze)]/W
ninhydrin for 20 mL of Sample solution.
Acceptance criteria: A deep violet color develops Me = molarity of the edetate disodium VS
immediately. z = volume of zinc sulfate VS consumed (mL)
Mz = molarity of the zinc sulfate VS
ASSAY Ze = equivalent volume of edetate disodium VS
PROCEDURE consumed (mL) by the zirconium moiety,
Analysis: Calculate the percentage of anhydrous aluminum calculated as (Zr%/Me) (W/Ar2), where Zr% is
zirconium octachlorohydrate gly in the Solution taken: the percentage of zirconium as determined in
the test for Content of zirconium, and Ar2 is the
Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + atomic weight of zirconium corrected for 2%
1)/z}/Ar1y) hafnium content, 92.97
W = quantity of Aluminum Zirconium
Al% = percentage of aluminum from the test for Octachlorohydrate taken (g)
Content of aluminum [NOTEUse the result obtained to calculate the Aluminum/
Ar1 = atomic weight of aluminum, 26.98 zirconium atomic ratio and the (Aluminum plus zirconium)/
y = aluminum/zirconium atomic ratio from the test chloride atomic ratio.]
for Aluminum/zirconium atomic ratio ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage
Ar2 = atomic weight of zirconium corrected for 2% of aluminum found in the test for Content of aluminum by
hafnium content, 92.97 the percentage of zirconium found in the test for Content of
Mr = molecular weight of the hydroxide anion (OH), zirconium, and multiply by 92.97/26.98, in which 92.97 is
17.01 the atomic weight of zirconium corrected for 2% hafnium
z = (aluminum plus zirconium)/chloride atomic ratio content, and 26.98 is the atomic weight of aluminum.
from the test for (Aluminum plus Acceptance criteria: 6.0:1 and 10.0:1.
zirconium)/chloride atomic ratio (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO
Ar3 = atomic weight of chlorine (Cl), 35.453 Analysis: Calculate:
Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3)
OTHER COMPONENTS
CONTENT OF CHLORIDE Al% = percentage of aluminum as determined in the
Sample: 500 mg of Aluminum Zirconium test for Content of aluminum
Octachlorohydrate Gly Solution in a 250-mL beaker Ar1 = atomic weight of aluminum, 26.98
Analysis: Add 100120 mL of water and 20 mL of diluted Zr% = percentage of zirconium as determined in the
nitric acid, and swirl to dissolve. Titrate with 0.05 N silver test for Content of zirconium
nitrate VS, using a calomel electrode and a silver billet Ar2 = atomic weight of zirconium, corrected for 2%
electrode system. Each mL of 0.05 N silver nitrate is hafnium content, 92.97
equivalent to 1.773 mg of chloride (Cl). [NOTEUse the Cl% = percentage of chloride as determined in the test
result obtained to calculate the (Aluminum plus for Content of chloride
zirconium)/chloride atomic ratio.] Ar3 = atomic weight of chlorine, 35.453
CONTENT OF ZIRCONIUM Acceptance criteria: 1.5:10.9:1
Octachlorohydrate Gly Solution in a 150-mL beaker IMPURITIES
Analysis: Add 5 mL of water and 15 mL of hydrochloric Inorganic Impurities
acid. Heat this solution to boiling, and continue boiling for ARSENIC, Method I 211: Prepare the Sample solution using a
68 min. Add 3040 mL of water and 5 mL of hydrochloric weighed quantity of the solution.
acid, and heat to boiling. Add 1 drop of xylenol orange TS, Acceptance criteria: NMT 2 ppm
and while still hot, titrate with 0.1 M edetate disodium VS LIMIT OF IRON
until the color of the solution changes from pink to yellow. Standard solution: Transfer 2.0 mL of Standard Iron
Perform a blank determination, and make any necessary Solution, prepared as directed under Iron 241, to a 50-mL
correction. Each mL of 0.1 M edetate disodium is equivalent beaker.
to 9.297 mg of zirconium (Zr). [NOTEUse the result Sample solution: 54 mg/mL of Aluminum Zirconium
obtained to calculate the Aluminum/zirconium atomic ratio Octachlorohydrate Gly Solution
and the (Aluminum plus zirconium)/chloride atomic ratio.] Transfer 5.0 mL of this solution to a 50-mL beaker.
Analysis: To each of the beakers containing the Standard amount of anhydrous aluminum zirconium
solution and the Sample solution add 5 mL of 6 N nitric pentachlorohydrate.
acid, cover with a watch glass, and boil on a hot plate for
35 min. Allow to cool. Add 5 mL of Ammonium IDENTIFICATION
Thiocyanate Solution, prepared as directed under Iron 241, IDENTIFICATION TESTSGENERAL, Chloride 191: A 100
transfer to separate 50-mL color comparison tubes, and mg/mL solution meets the requirements
Acceptance criteria: The color of the solution from the ASSAY
Sample solution is not darker than that of the solution from PROCEDURE
the Standard solution. (NMT 75 ppm) Analysis: Calculate the percentage of anhydrous aluminum
HEAVY METALS, Method I 231 zirconium pentachlorohydrate in the Aluminum Zirconium
[NOTEProceed as directed, except use the following Pentachlorohydrate:
modifications.] Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y +
Sample solution: Prepare as directed in the chapter, using 1)/z}/Ar1y)
an accurately weighed quantity of solution. If the solution
is not clear after dilution to 40 mL, heat for several min Al% = percentage of aluminum from the test for
between 60 and 80, and cool to room temperature. If Content of aluminum
the solution remains cloudy, repeat from the beginning Ar1 = atomic weight of aluminum, 26.98
with the following modification: add 3 mL of hydrochloric y = aluminum/zirconium atomic ratio from the test
acid before adjustment of the pH with 6 N ammonium for Aluminum/zirconium atomic ratio
hydroxide. Ar2 = atomic weight of zirconium corrected for 2%
Monitor solution: Prepare as directed, using the same hafnium content, 92.97
modifications described for Sample solution, if necessary. Mr = molecular weight of the hydroxide anion (OH),
Analysis: To each of the three tubes containing the 17.01
Standard solution, the Sample solution, and the Monitor z = (aluminum plus zirconium)/chloride atomic ratio
solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently from the test for (Aluminum plus
between 60 and 80. To the Standard solution, add 6 zirconium)/chloride atomic ratio
drops of sodium sulfide TS. To the Sample solution and the Ar3 = atomic weight of chlorine (Cl), 35.453
Monitor solution, add 12 drops of sodium sulfide TS. Cool Acceptance criteria: 90.0%110.0%
to room temperature, and dilute the contents of each tube
with water to 50 mL. Gently mix each tube by inverting OTHER COMPONENTS
twice. Allow to stand for 5 min, and view downward over a CONTENT OF CHLORIDE
white surface. Sample: 250 mg of Aluminum Zirconium
Acceptance criteria: NMT 10 ppm Pentachlorohydrate to a 250-mL beaker
The color of the solution from the Sample solution is not Analysis: Add 100120 mL of water and 20 mL of diluted
darker than that of the solution made from the Standard nitric acid, and swirl to dissolve. Titrate with 0.05 N silver
solution, and the color of the solution from the Monitor nitrate VS using a calomel electrode and a silver billet
solution is the same as or darker than the color of the electrode system. Each mL of 0.05 N silver nitrate is
solution from the Standard solution. If the color of the equivalent to 1.773 mg of chloride (Cl). [NOTEUse the
solution from the Monitor solution is lighter than the color result obtained to calculate the (Aluminum plus
of the solution made from the Standard solution, repeat zirconium)/chloride atomic ratio.]
the analysis with the following modification: after the CONTENT OF ZIRCONIUM
heating step, to the Monitor solution and the Sample Sample: 250 mg of Aluminum Zirconium
solution add 1.0 mL, instead of 12 drops, of sodium Pentachlorohydrate to a 150-mL beaker
sulfide TS. Analysis: Add 5 mL of water and 15 mL of hydrochloric
acid. Heat this solution to boiling, and continue boiling for
SPECIFIC TESTS 68 min. Add 3040 mL of water and 5 mL of hydrochloric
PH 791: 3.05.0, in a solution (3 in 10) acid, and heat to boiling. Add 1 drop of xylenol orange TS,
ADDITIONAL REQUIREMENTS and while still hot, titrate with 0.1 M edetate disodium VS
PACKAGING AND STORAGE: Preserve in well-closed containers. until the color of the solution changes from pink to yellow.
LABELING: Label the Solution to state the solvent and form Perform a blank determination, and make any necessary
of glycine used and the claimed concentration of anhydrous correction. Each mL of 0.1 M edetate disodium is equivalent
aluminum zirconium octachlorohydrate. to 9.297 mg of zirconium (Zr). [NOTEUse the result
obtained to calculate the Aluminum/zirconium atomic ratio
CONTENT OF ALUMINUM
Aluminum Zirconium Sample: 0.15 g of Aluminum Zirconium Pentachlorohydrate
Pentachlorohydrate to a 150-mL beaker
(Comment on this Monograph)id=m2360=Aluminum Zirconium acid. Heat this solution to boiling, and continue boiling for 5
Pentachlorohydrate=A-Monos.pdf) min. Add 40 mL of water and 15.0 mL of 0.1 M edetate
AlyZr(OH)3y+4-xClx nH2O disodium VS. Heat the solution to boiling, and continue
boiling for 5 min. Allow the solution to cool, add 1015 mL
DEFINITION of acetic acidammonium acetate buffer TS, and adjust with
Aluminum Zirconium Pentachlorohydrate is a polymeric, loosely ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of
hydrated complex of basic aluminum zirconium chloride that alcohol, and adjust with ammonium hydroxide to a pH of
encompasses a range of aluminum-to-zirconium atomic ratios 4.6 0.1. Add 510 drops of dithizone TS, and titrate with
between 6.0:1 and 10.0:1, and a range of (aluminum plus 0.1 M zinc sulfate VS until the first permanent purple-pink
zirconium)-to-chloride atomic ratios between 2.1:1 and 1.51:1.
It contains NLT 90.0% and NMT 110.0% of the labeled
color appears. Perform a blank determination, and make any min between 60 and 80, and cool to room temperature.
necessary correction. If the solution remains cloudy, repeat from the beginning
Calculate the percentage of aluminum (Al) in the Aluminum with the following modification: add 3 mL of hydrochloric
Zirconium Pentachlorohydrate: acid before adjustment of the pH with 6 N ammonium
hydroxide.
Result = 2.698[15.0 Me (zMz + Ze)]/W Monitor solution: Prepare as directed, using the same
modifications described for Sample solution, if necessary.
Me = molarity of the edetate disodium VS Analysis: To each of the three tubes containing the
z = volume of zinc sulfate VS consumed (mL) Standard solution, the Sample solution, and the Monitor
Mz = molarity of the zinc sulfate VS solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently
Ze = equivalent volume of edetate disodium VS to between 60 and 80. To the Standard solution, add 6
consumed (mL) by the zirconium moiety, drops of sodium sulfide TS. To the Sample solution and the
calculated as (Zr%/Me) (W/Ar2), where Zr% is Monitor solution, add 12 drops of sodium sulfide TS. Cool
the percentage of zirconium as determined in to room temperature, and dilute the contents of each tube
the test for Content of zirconium, and Ar2 is the with water to 50 mL. Gently mix each tube by inverting
atomic weight of zirconium corrected for 2% twice. Allow to stand for 5 min, and view downward over a
hafnium content, 92.97 white surface.
W = quantity of Aluminum Zirconium Acceptance criteria: NMT 20 ppm
Octachlorohydrate taken (g) The color of the solution from the Sample solution is not
[NOTEUse the result obtained to calculate the Aluminum/ darker than that of the solution made from the Standard
zirconium atomic ratio and the (Aluminum plus zirconium)/ solution, and the color of the solution from the Monitor
chloride atomic ratio.] solution is the same as or darker than the color of the
ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage solution made from the Standard solution. If the color of
of aluminum found in the test for Content of aluminum by the solution from the Monitor solution is lighter than the
the percentage of zirconium found in the test for Content of color of the solution made from the Standard solution,
zirconium, and multiply by 92.97/26.98, in which 92.97 is repeat the analysis with the following modification: after
the atomic weight of zirconium corrected for 2% hafnium the heating step, to the Monitor solution and the Sample
content, and 26.98 is the atomic weight of aluminum. solution add 1.0 mL, instead of 12 drops, of sodium
Acceptance criteria: 6.0:1 and 10.0:1. sulfide TS.
(ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO
Calculate the (aluminum plus zirconium)/chloride atomic SPECIFIC TESTS
ratio: PH 791: 3.05.0, in a solution (15 in 100)
Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3) ADDITIONAL REQUIREMENTS
Al% = percentage of aluminum as determined in the LABELING: The label states the content of anhydrous
test for Content of aluminum aluminum zirconium pentachlorohydrate.
Ar1 = atomic weight of aluminum, 26.98
Zr% = percentage of zirconium as determined in the
test for Content of zirconium
hafnium content, 92.97 Pentachlorohydrate Solution
Cl% = percentage of chloride as determined in the test (Comment on this Monograph)id=m2361=Aluminum Zirconium
for Content of chloride Pentachlorohydrate Solution=A-Monos.pdf)
Aluminum Zirconium Pentachlorohydrate Solution is a
IMPURITIES polymeric, loosely hydrated complex of basic aluminum
Inorganic Impurities zirconium chloride that encompasses a range of aluminum-to-
ARSENIC, Method I 211: NMT 2 ppm zirconium atomic ratios between 6.0:1 and 10.0:1, and a
LIMIT OF IRON range of (aluminum plus zirconium)-to-chloride atomic ratios
Standard solution: Transfer 2.0 mL of Standard Iron between 2.1:1 and 1.51:1. The following solvents may be
Solution, prepared as directed under Iron 241, to a 50-mL used: water, propylene glycol, or dipropylene glycol. It
beaker. contains the equivalent of NLT 90.0% and NMT 110.0% of
Sample solution: 27 mg/mL of Aluminum Zirconium the labeled concentration of anhydrous aluminum zirconium
Pentachlorohydrate. Transfer 5.0 mL of this solution to a pentachlorohydrate.
50-mL beaker.
Analysis: To each of the beakers containing the Standard IDENTIFICATION
solution and the Sample solution, add 5 mL of 6 N nitric A. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution
acid, cover with a watch glass, and boil on a hot plate for containing the equivalent amount of 100 mg/mL of
35 min. Allow to cool. Add 5 mL of Ammonium anhydrous aluminum zirconium pentachlorohydrate meets
Thiocyanate Solution, prepared as directed under Iron 241, the requirements.
transfer to separate 50-mL color comparison tubes, dilute B. PROPYLENE GLYCOL (where stated on the label)
with water to volume. Sample solution: 200 mg/mL in isopropyl alcohol, and
Acceptance criteria: The color of the solution from the filter
Sample solution is not darker than that of the solution from Analysis: Evaporate the filtrate to 1 mL on a steam bath:
the Standard solution. (NMT 150 ppm) the IR absorption spectrum of a film of this solution on a
HEAVY METALS, Method I 231 silver chloride disk exhibits maxima only at the same
[NOTEProceed as directed, except use the following wavelengths as that of a similar preparation of a film of
modifications.] propylene glycol.
Sample solution: Prepare as directed in the chapter. If the
C. DIPROPYLENE GLYCOL (where stated on the label) Calculate the percentage of aluminum (Al) in the Aluminum
Sample solution: 200 mg/mL in isopropyl alcohol, and Zirconium Pentachlorohydrate:
filter
Analysis: Evaporate the filtrate to 1 mL on a steam bath: Result = 2.698[15.0 Me (zMz + Ze)]/W
the IR absorption spectrum of a film of this solution on a
silver chloride disk exhibits maxima only at the same Me = molarity of the edetate disodium VS
wavelengths as that of a similar preparation of a film of z = volume of zinc sulfate VS consumed (mL)
dipropylene glycol. Mz = molarity of the zinc sulfate VS
Ze = equivalent volume of edetate disodium VS
ASSAY consumed (mL) by the zirconium moiety,
PROCEDURE calculated as (Zr%/Me) (W/Ar2), where Zr% is
Analysis: Calculate the percentage of anhydrous aluminum the percentage of zirconium as determined in
zirconium pentachlorohydrate in the Solution: the test for Content of zirconium, and Ar2 is the
atomic weight of zirconium corrected for 2%
Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] hafnium content, 92.97
+ Ar3(y + 1)/z}/Ar1y) W = quantity of aluminum zirconium
pentachlorohydrate taken (g)
Al% = percentage of aluminum from the test for [NOTEUse the result obtained to calculate the Aluminum/
Content of aluminum zirconium atomic ratio and the (Aluminum plus zirconium)/
Ar1 = atomic weight of aluminum, 26.98 chloride atomic ratio.]
y = aluminum/zirconium atomic ratio from the test ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage
for Aluminum/zirconium atomic ratio of aluminum found in the test for Content of Aluminum by
Ar2 = atomic weight of zirconium corrected for 2% the percentage of zirconium found in the test for Content of
hafnium content, 92.97 Zirconium, and multiply by 92.97/26.98, in which 92.97 is
Mr = molecular weight of the hydroxide anion the atomic weight of zirconium corrected for 2% hafnium
(OH),17.01 content, and 26.98 is the atomic weight of aluminum.
z = (aluminum plus zirconium)/chloride atomic ratio Acceptance criteria: 6.0:1 and 10.0:1.
found in the test for (Aluminum plus (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO
zirconium)/chloride atomic ratio Calculate the (Aluminum plus zirconium)/chloride atomic
Ar3 = atomic weight of chlorine (Cl), 35.453 ratio:
OTHER COMPONENTS
CONTENT OF CHLORIDE Al% = percentage of aluminum from the test for
Sample: 500 mg of Aluminum Zirconium Content of aluminum
Pentachlorohydrate Solution to a 250-mL beaker Ar1 = atomic weight of aluminum, 26.98
Analysis: Add 100120 mL of water and 20 mL of diluted Zr% = percentage of zirconium from the test for Content
nitric acid, and swirl to dissolve. Titrate with 0.05 N silver of zirconium
nitrate VS using a calomel electrode and a silver billet Ar2 = atomic weight of zirconium corrected for 2%
equivalent to 1.773 mg of chloride (Cl). [NOTEUse the Cl% = percentage of chloride as determined in the test
result obtained to calculate the (Aluminum plus for Content of chloride
zirconium)/chloride atomic ratio.] Ar3 = atomic weight of chlorine, 35.453
Pentachlorohydrate Solution to a 150-mL beaker IMPURITIES
acid. Heat this solution to boiling, and continue boiling for ARSENIC, Method I 211
68 min. Add 3040 mL of water and 5 mL of hydrochloric Sample solution: Use a weighed quantity of the Solution.
and, while still hot, titrate with 0.1 M edetate disodium VS LIMIT OF IRON
to 9.297 mg of zirconium (Zr). [NOTEUse the result Sample solution: 54 mg/mL of Aluminum Zirconium
obtained to calculate the Aluminum/zirconium atomic ratio Pentachlorohydrate Solution. Transfer 5.0 mL of this
and the (Aluminum plus zirconium)/chloride atomic ratio.] solution to a 50-mL beaker.
CONTENT OF ALUMINUM Analysis: To each of the beakers containing the Standard
Sample: 0.3 g of Aluminum Zirconium Pentachlorohydrate solution and the Sample solution, add 5 mL of 6 N nitric
Solution to a 150-mL beaker acid, cover with a watch glass, and boil on a hot plate for
Analysis: Add 5 mL of water and 15 mL of hydrochloric 35 min. Allow to cool. Add 5 mL of Ammonium
acid. Heat this solution to boiling, and continue boiling for 5 Thiocyanate Solution, prepared as directed under Iron 241,
min. Add 40 mL of water and 15.0 mL of 0.1 M edetate transfer to separate 50-mL color comparison tubes, and
disodium VS. Heat the solution to boiling, and continue dilute with water to volume.
boiling for 5 min. Allow the solution to cool, add 1015 mL Acceptance criteria: The color of the solution from the
of acetic acidammonium acetate buffer TS, and adjust with Sample solution is not darker than that of the solution from
ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of the Standard solution. (NMT 75 ppm)
alcohol, and adjust with ammonium hydroxide to a pH of HEAVY METALS, Method I 231
4.6 0.1. Add 510 drops of dithizone TS, and titrate with [NOTEProceed as directed, except use the following
0.1 M zinc sulfate VS until the first permanent purple-pink modifications.]
color appears. Perform a blank determination, and make any Sample solution: Prepare as directed in the chapter, using
necessary correction. a weighed quantity of Solution. If the solution is not clear
after dilution to 40 mL, heat for several min between 60 ASSAY

and 80, and cool to room temperature. If the solution PROCEDURE
remains cloudy, repeat from the beginning with the Analysis: Calculate the percentage of anhydrous aluminum
following modification: add 3 mL of hydrochloric acid zirconium pentachlorohydrate in the Aluminum Zirconium
before adjustment of the pH with 6 N ammonium Pentachlorohydrex Gly:
hydroxide.
Monitor solution: Prepare as directed, using the same Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y +
modifications described for Sample solution, if necessary. 1)/z}/Ar1y)
Analysis: To each of the three tubes containing the
Standard solution, the Sample solution, and the Monitor Al% = percentage of aluminum from the test for
solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently Content of aluminum
to between 60 and 80. To the Standard solution, add 6 Ar1 = atomic weight of aluminum, 26.98
drops of sodium sulfide TS. To the Sample solution and the y = aluminum/zirconium atomic ratio from the test
Monitor solution, add 12 drops of sodium sulfide TS. Cool for Aluminum/zirconium atomic ratio
to room temperature, and dilute the contents of each tube Ar2 = atomic weight of zirconium corrected for 2%
with water to 50 mL. Gently mix each tube by inverting hafnium content, 92.97
twice. Allow to stand for 5 min, and view downward over a Mr = molecular weight of the hydroxide anion (OH),
white surface. 17.01
Acceptance criteria: NMT 10 ppm z = (aluminum plus zirconium)/chloride atomic ratio
The color of the solution from the Sample solution is not from the test for (Aluminum plus
darker than that of the solution made from the Standard zirconium)/chloride atomic ratio
solution, and the color of the solution from the Monitor Ar3 = atomic weight of chlorine (Cl), 35.453
solution is the same as or darker than the color of the Acceptance criteria: 90.0%110.0%
solution made from the Standard solution. If the color of
the solution from the Monitor solution is lighter than the OTHER COMPONENTS
color of the solution made from the Standard solution, CONTENT OF CHLORIDE
repeat the analysis with the following modification: after Sample: 250 mg of Aluminum Zirconium
the heating step, to the Monitor solution and the Sample Pentachlorohydrex Gly to a 250-mL beaker
solution add 1.0 mL, instead of 12 drops, of sodium Analysis: Add 100120 mL of water and 20 mL of diluted
sulfide TS. nitric acid, and swirl to dissolve. Titrate with 0.05 N silver
nitrate VS using a calomel electrode and a silver billet
SPECIFIC TESTS electrode system. Each mL of 0.05 N silver nitrate is
pH 791: 3.05.0, in a solution (3 in 10) equivalent to 1.773 mg of chloride (Cl). [NOTEUse the
result obtained to calculate the (Aluminum plus
ADDITIONAL REQUIREMENTS zirconium)/chloride atomic ratio.]
PACKAGING AND STORAGE: Preserve in well-closed containers. CONTENT OF ZIRCONIUM
LABELING: Label the Solution to state the solvent used and Sample: 250 mg of Aluminum Zirconium
the claimed concentration of anhydrous aluminum zirconium Pentachlorohydrex Gly to a 150-mL beaker
pentachlorohydrate. Analysis: Add 5 mL of water and 15 mL of hydrochloric
68 min. Add 3040 mL of water and 5 mL of hydrochloric
Aluminum Zirconium and while still hot, titrate with 0.1 M edetate disodium VS
Pentachlorohydrex Gly until the color of the solution changes from pink to yellow.
(Comment on this Monograph)id=m2362=Aluminum Zirconium Perform a blank determination, and make any necessary
Pentachlorohydrex Gly=A-Monos.pdf) correction. Each mL of 0.1 M edetate disodium is equivalent
to 9.297 mg of zirconium (Zr). [NOTEUse the result
DEFINITION obtained to calculate the Aluminum/zirconium atomic ratio
Aluminum Zirconium Pentachlorohydrex Gly is a derivative of and the (Aluminum plus zirconium)/chloride atomic ratio.]
Aluminum Zirconium Pentachlorohydrate in which some of the CONTENT OF ALUMINUM
water molecules have been displaced by glycine, calcium Sample: 0.15 g of Aluminum Zirconium Pentachlorohydrex
glycinate, magnesium glycinate, potassium glycinate, sodium Gly to a 150-mL beaker
glycinate, or zinc glycinate. It encompasses a range of Analysis: Add 5 mL of water and 15 mL of hydrochloric
aluminum-to-zirconium atomic ratios between 6.0:1 and acid. Heat the solution to boiling, and continue boiling for 5
10.0:1, and a range of (aluminum plus zirconium)-to-chloride min. Add 40 mL of water and 15.0 mL of 0.1 M edetate
atomic ratios between 2.1:1 and 1.51:1. It contains NLT disodium VS. Heat the solution to boiling, and continue
90.0% and NMT 110.0% of the labeled amount of anhydrous boiling for 5 min. Allow the solution to cool, add 1015 mL
aluminum zirconium pentachlorohydrate. of acetic acidammonium acetate buffer TS, and adjust with
ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of
IDENTIFICATION alcohol, and adjust with ammonium hydroxide to a pH of
A. IDENTIFICATION TESTSGENERAL, Chloride 191: A 100 4.6 0.1. Add 510 drops of dithizone TS, and titrate with
mg/mL solution meets the requirements. 0.1 M zinc sulfate VS until the first permanent purple-pink
B. PROCEDURE color appears. Perform a blank determination, and make any
Sample solution: 25 mg/mL necessary correction.
Analysis: Heat to boiling on a hot plate, and add 60 mg of
ninhydrin to 20 mL of Sample solution.
Acceptance criteria: A deep violet color develops
immediately.
Calculate the percentage of aluminum (Al) in the Aluminum the following modification: add 3 mL of hydrochloric acid
Zirconium Pentachlorohydrex Gly: before adjustment of the pH with 6 N ammonium
hydroxide.
Result = 2.698[15.0 Me (zMz + Ze)]/W Monitor solution: Prepare as directed, using the
modifications described for the Sample solution, if
Me = molarity of the edetate disodium VS necessary.
z = volume of zinc sulfate VS consumed (mL) Analysis: To each of the three tubes containing the
Mz = molarity of the zinc sulfate VS Standard solution, the Sample solution, and the Monitor
Ze = equivalent volume of edetate disodium VS solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently
consumed (mL) by the zirconium moiety, to between 60 and 80. To the Standard solution, add 6
calculated as (Zr%/Me) (W/Ar2), where Zr% is drops of sodium sulfide TS. To the Sample solution and the
the percentage of zirconium as determined in Monitor solution, add 12 drops of sodium sulfide TS. Cool
the test for Content of zirconium, and Ar2 is the to room temperature, and dilute the contents of each tube
atomic weight of zirconium corrected for 2% with water to 50 mL. Gently mix each tube by inverting
hafnium content, 92.97 twice. Allow to stand for 5 min, and view downward over a
W = quantity of Aluminum Zirconium white surface.
Pentachlorohydrate taken (g) Acceptance criteria: NMT 20 ppm
[NOTEUse the result obtained to calculate the Aluminum/ The color of the solution from the Sample solution is not
zirconium atomic ratio and the (Aluminum plus zirconium)/ darker than that of the solution made from the Standard
chloride atomic ratio.] solution, and the color of the solution from the Monitor
ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage solution is the same as or darker than the color of the
of aluminum found in the test for Content of aluminum by solution made from the Standard solution. If the color of
the percentage of zirconium found in the test for Content of the solution from the Monitor solution is lighter than the
zirconium, and multiply by 92.97/26.98, in which 92.97 is color of the solution made from the Standard solution,
the atomic weight of zirconium corrected for 2% hafnium repeat the analysis with the following modification: after
content, and 26.98 is the atomic weight of aluminum. the heating step, to the Monitor solution and the Sample
Acceptance criteria: 6.0:1 and 10.0:1. solution add 1.0 mL, instead of 12 drops, of sodium
(ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO sulfide TS.
Calculate the (aluminum plus zirconium)/chloride ratio:
SPECIFIC TESTS
Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3) PH 791: 3.05.0, in a solution (15 in 100)
Al% = percentage of aluminum as determined in the ADDITIONAL REQUIREMENTS
test for Content of aluminum PACKAGING AND STORAGE: Preserve in well-closed containers.
Ar1 = atomic weight of aluminum, 26.98 LABELING: The label states the form of glycine used and the
Zr% = percentage of zirconium as determined in the claimed content of anhydrous aluminum zirconium
test for Content of zirconium pentachlorohydrate.
Ar2 = atomic weight of zirconium corrected for 2%
Cl% = percentage of chloride as determined in the test
for Content of chloride Aluminum Zirconium
Ar3 = atomic weight of chlorine, 35.453 Pentachlorohydrex Gly Solution
Acceptance criteria: 2.1:11.51:1 (Comment on this Monograph)id=m2363=Aluminum Zirconium
Pentachlorohydrex Gly Solution=A-Monos.pdf)
IMPURITIES
Inorganic Impurities DEFINITION
ARSENIC, Method I 211: NMT 2 ppm Aluminum Zirconium Pentachlorohydrex Gly Solution is a
LIMIT OF IRON solution of Aluminum Zirconium Pentachlorohydrate in which
Standard solution: Transfer 2.0 mL of Standard Iron some of the waters of hydration have been displaced by
Solution, prepared as directed under Iron 241, to a 50-mL glycine, calcium glycinate, magnesium glycinate, potassium
beaker. glycinate, sodium glycinate, or zinc glycinate. It encompasses
Sample solution: 27 mg/mL of Aluminum Zirconium a range of aluminum-to-zirconium ratios between 6.0:1 and
Pentachlorohydrex Gly. Transfer 5.0 mL of this solution to a 10.0:1, and a range of (aluminum plus zirconium)-to-chloride
50-mL beaker. atomic ratios between 2.1:1 and 1.51:1. The following
Analysis: To each of the beakers containing the Standard solvents may be used: water, propylene glycol, or dipropylene
solution and the Sample solution, add 5 mL of 6 N nitric glycol. It contains the equivalent of NLT 90.0% and NMT
acid, cover with a watch glass, and boil on a hot plate for 110.0% of the labeled concentration of anhydrous aluminum
35 min. Allow to cool. Add 5 mL of Ammonium zirconium pentachlorohydrate.
Thiocyanate Solution, prepared as directed under Iron 241,
transfer to separate 50-mL color comparison tubes, and IDENTIFICATION
dilute with water to volume. A. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution
Acceptance criteria: The color of the solution from the equivalent to 100 mg/mL meets the requirements.
Sample solution is not darker than that of the solution from B. PROPYLENE GLYCOL (where stated on the label)
the Standard solution. (NMT 150 ppm) Sample solution: 200 mg/mL in isopropyl alcohol, and
HEAVY METALS, Method I 231 filter
[NOTEProceed as directed in the chapter, except use the Analysis: Evaporate the filtrate to 1 mL on a steam bath:
following modifications.] the IR absorption spectrum of a film of this solution on a
Sample solution: Prepare as directed in the chapter. If the silver chloride disk exhibits maxima only at the same
solution is not clear after dilution to 40 mL, heat for several wavelengths as that of a similar preparation of a film of
min at 6080, and cool to room temperature. If the propylene glycol.
solution remains cloudy, repeat from the beginning with
C. DIPROPYLENE GLYCOL (where stated on the label) of acetic acidammonium acetate buffer TS, and adjust with
Sample solution: 200 mg/mL in isopropyl alcohol, and ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of
filter alcohol, and adjust with ammonium hydroxide to a pH of
Analysis: Evaporate the filtrate to 1 mL on a steam bath: 4.6 0.1. Add 510 drops of dithizone TS, and titrate with
the IR absorption spectrum of a film of this solution on a 0.1 M zinc sulfate VS until the first permanent purple-pink
silver chloride disk exhibits maxima only at the same color appears. Perform a blank determination, and make any
wavelengths as that of a similar preparation of a film of necessary correction.
dipropylene glycol. Calculate the percentage of aluminum (Al) in the Aluminum
D. GLYCINE Zirconium Pentachlorohydrate:
Sample solution: 1 g of Solution in a 50-mL beaker
Add 20 mL of water, and swirl to dissolve. Result = 2.698[15.0 Me (zMz + Ze)]/W
Analysis: Heat to boiling on a hot plate, and add 60 mg of
ninhydrin for 20 mL of Sample solution. Me = molarity of the edetate disodium VS
Acceptance criteria: A deep violet color develops z = volume of zinc sulfate VS consumed (mL)
immediately. Mz = molarity of the zinc sulfate VS
Ze = equivalent volume of edetate disodium VS
ASSAY consumed (mL) by the zirconium moiety,
PROCEDURE calculated as (Zr%/Me) (W/Ar2), where Zr% is
Analysis: Calculate the percentage of anhydrous aluminum the percentage of zirconium as determined in
zirconium pentachlorohydrate in the Solution taken: the test for Content of zirconium, and Ar2 is the
atomic weight of zirconium corrected for 2%
Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + hafnium content, 92.97
1)/z}/Ar1y) W = quantity of Aluminum Zirconium
Pentachlorohydrate taken (g)
Al% = percentage of aluminum from the test for [NOTEUse the result to calculate the Aluminum/zirconium
Content of aluminum atomic ratio and the (Aluminum plus zirconium)/chloride
Ar1 = atomic weight of aluminum, 26.98 atomic ratio.]
y = aluminum/zirconium atomic ratio from the test ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage
for Aluminum/zirconium atomic ratio of aluminum found in the test for Content of aluminum by
Ar2 = atomic weight of zirconium corrected for 2% the percentage of zirconium found in the test for Content of
hafnium content, 92.97 zirconium, and multiply by 92.97/26.98, in which 92.97 is
Mr = molecular weight of the hydroxide anion (OH), the atomic weight of zirconium corrected for 2% hafnium
17.01 content, and 26.98 is the atomic weight of aluminum.
z = (aluminum plus zirconium)/chloride atomic ratio Acceptance criteria: 6.0:1 and 10.0:1.
from the test for (Aluminum plus (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO
zirconium)/chloride atomic ratio Calculate the (aluminum plus zirconium)/chloride atomic
Ar3 = atomic weight of chlorine (Cl), 35.453 ratio:
OTHER COMPONENTS
CONTENT OF CHLORIDE Al% = percentage of aluminum from the test for
Sample: 500 mg of Aluminum Zirconium Content of aluminum
Pentachlorohydrate Gly Solution to a 250-mL beaker Ar1 = atomic weight of aluminum, 26.98
Analysis: Add 100120 mL of water and 20 mL of diluted Zr% = percentage of zirconium from the test for Content
nitric acid, and swirl to dissolve. Titrate with 0.05 N silver of zirconium
nitrate VS using a calomel electrode and a silver billet Ar2 = atomic weight of zirconium, corrected for 2%
equivalent to 1.773 mg of chloride (Cl). [NOTEUse the Cl% = percentage of chloride from the test for Content
result to calculate the (Aluminum plus zirconium)/chloride of chloride
atomic ratio.] Ar3 = atomic weight of chlorine, 35.453
Pentachlorohydratex Gly Solution to a 150-mL beaker IMPURITIES
acid. Heat this solution to boiling, and continue boiling for ARSENIC, Method I 211
68 min. Add 3040 mL of water and 5 mL of hydrochloric Sample solution: Use a weighed quantity of the Solution
to 9.297 mg of zirconium (Zr). [NOTEUse the result to Sample solution: 54 mg/mL of Aluminum Zirconium
calculate the Aluminum/zirconium atomic ratio and the Pentachlorohydrate Gly Solution. Transfer 5.0 mL of this
(Aluminum plus zirconium)/chloride atomic ratio.] solution to a 50-mL beaker.
CONTENT OF ALUMINUM Analysis: To each of the beakers containing the Standard
Sample: 0.30 g of Aluminum Zirconium Pentachlorohydrate solution and the Sample solution, add 5 mL of 6 N nitric
Gly Solution to a 150-mL beaker acid, cover with a watch glass, and boil on a hot plate for
Analysis: Add 5 mL of water and 15 mL of hydrochloric 35 min. Allow to cool. Add 5 mL of Ammonium
acid. Heat the solution to boiling, and continue boiling for 5 Thiocyanate Solution, prepared as directed under Iron 241,
min. Add 40 mL of water and 15.0 mL of 0.1 M edetate transfer to separate 50-mL color comparison tubes, and
disodium VS. Heat the solution to boiling, and continue dilute with water to volume.
Acceptance criteria: The color of the solution from the ASSAY

Sample solution is not darker than that of the solution from PROCEDURE
the Standard solution. (NMT 75 ppm) Analysis: Calculate the percentage of anhydrous aluminum
HEAVY METALS, Method I 231 zirconium tetrachlorohydrate in the Aluminum Zirconium
[NOTEProceed as directed, except use the following Tetrachlorohydrate:
modifications.]
Sample solution: Prepare as directed in the chapter, using Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y +
a weighed quantity of Solution. If the solution is not clear 1)/z}/ Ar3y)
after dilution to 40 mL, heat for several min between 60
and 80, and cool to room temperature. If the solution Al% = percentage of aluminum from the test for
remains cloudy, repeat from the beginning with the Content of aluminum
following modification: add 3 mL of hydrochloric acid Ar1 = atomic weight of aluminum, 26.98
before adjustment of the pH with 6 N ammonium y = aluminum/zirconium atomic ratio from the test
hydroxide. for Aluminum/zirconium atomic ratio
Monitor solution: Prepare as directed, using the same Ar2 = atomic weight of zirconium corrected for 2%
modifications described for Sample solution if necessary. hafnium content, 92.97
Analysis: To each of the three tubes containing the Mr = molecular weight of the hydroxide anion (OH),
Standard solution, the Sample solution, and the Monitor 17.01
solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently z = (aluminum plus zirconium)/chloride atomic ratio
to between 60 and 80. To the Standard solution, add 6 from the test for (Aluminum plus
drops of sodium sulfide TS. To the Sample solution and the zirconium)/chloride atomic ratio
Monitor solution, add 12 drops of sodium sulfide TS. Cool Ar3 = atomic weight of chlorine (Cl), 35.453
to room temperature, and dilute the contents of each tube Acceptance criteria: 90.0%110.0%
with water to 50 mL. Gently mix each tube by inverting
twice. Allow to stand for 5 min, and view downward over a OTHER COMPONENTS
white surface. CONTENT OF CHLORIDE
Acceptance criteria: NMT 10 ppm Sample: 250 mg of Aluminum Zirconium
The color of the solution from the Sample solution is not Tetrachlorohydrate to a 250-mL beaker
darker than that of the solution made from the Standard Analysis: Add 100120 mL of water and 20 mL of diluted
solution, and the color of the solution made from the nitric acid, and swirl to dissolve. Titrate with 0.05 N silver
Monitor solution is the same as or darker than the color of nitrate VS using a calomel electrode and a silver billet
the solution from the Standard solution. If the color of the electrode system. Each mL of 0.05 N silver nitrate is
solution from the Monitor solution is lighter than the color equivalent to 1.773 mg of chloride (Cl). [NOTEUse the
of the solution made from the Standard solution, repeat result obtained to calculate the (Aluminum plus
the analysis with the following modification: after the zirconium)/chloride atomic ratio.]
heating step, to the Monitor solution and the Sample CONTENT OF ZIRCONIUM
solution add 1.0 mL, instead of 12 drops, of sodium Sample: 250 mg of Aluminum Zirconium
sulfide TS. Tetrachlorohydrate to a 150-mL beaker
SPECIFIC TESTS acid. Heat this solution to boiling, and continue boiling for 6
PH 791: 3.05.0, in a solution (3 in 10) to 8 min. Add 30 40 mL of water and 5 mL of hydrochloric
ADDITIONAL REQUIREMENTS and while still hot, titrate with 0.1 M edetate disodium VS
PACKAGING AND STORAGE: Preserve in well-closed containers. until the color of the solution changes from pink to yellow.
LABELING: Label the Solution to state the solvent and form Perform a blank determination, and make any necessary
of glycine used and the claimed concentration of anhydrous correction. Each mL of 0.1 M edetate disodium is equivalent
aluminum zirconium pentachlorohydrate. to 9.297 mg of zirconium (Zr). [NOTEUse the result
obtained to calculate the Aluminum/zirconium atomic ratio
CONTENT OF ALUMINUM
Aluminum Zirconium Sample: 0.15 g of Aluminum Zirconium Tetrachlorohydrate
Tetrachlorohydrate to a 150-mL beaker
(Comment on this Monograph)id=m2365=Aluminum Zirconium Analysis: Add 5 mL of water and 15 mL of hydrochloric
Tetrachlorohydrate=A-Monos.pdf) acid. Heat this solution to boiling, and continue boiling for 5
min. Add 40 mL of water and 15.0 mL of 0.1 M edetate
AlyZr(OH)3y+4-xClx nH2O disodium VS. Heat the solution to boiling, and continue
DEFINITION boiling for 5 min. Allow the solution to cool. Add 1015 mL
Aluminum Zirconium Tetrachlorohydrate is a polymeric, loosely of acetic acidammonium acetate buffer TS, and adjust with
hydrated complex of basic aluminum zirconium chloride that ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of
encompasses a range of aluminum-to-zirconium atomic ratios alcohol, and adjust with ammonium hydroxide to a pH of
between 2.0:1 and 5.99:1, and a range of (aluminum plus 4.6 0.1. Add 510 drops of dithizone TS, and titrate with
zirconium)-to-chloride atomic ratios between 1.5:1 and 0.9:1. 0.1 M zinc sulfate VS until the first permanent purple-pink
It contains NLT 90.0% and NMT 110.0% of the labeled color appears. Perform a blank determination, and make any
amount of anhydrous aluminum zirconium tetrachlorohydrate. necessary correction.
Calculate the percentage of aluminum (Al) in the Aluminum
IDENTIFICATION Zirconium Tetrachlorohydrate:
IDENTIFICATION TESTSGENERAL, Chloride 191: A solution
containing the equivalent of 100 mg/mL of anhydrous Result = 2.698[15.0 Me (zMz + Ze)]/W
aluminum zirconium tetrachloriohydrate meets the
requirements. Me = molarity of the edetate disodium VS
z = volume of zinc sulfate VS consumed (mL) Analysis: To each of the three tubes containing the
Mz = molarity of the zinc sulfate VS Standard solution, the Sample solution, and the Monitor
Ze = equivalent volume of edetate disodium VS solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently
consumed (mL) by the zirconium moiety, to between 60 and 80. To the Standard solution, add 6
calculated as (Zr%/Me) (W/Ar2), where Zr% is drops of sodium sulfide TS. To the Sample solution and the
the percentage of zirconium as determined in Monitor solution, add 12 drops of sodium sulfide TS. Cool
the test for Content of zirconium, and Ar2 is the to room temperature, and dilute the contents of each tube
atomic weight of zirconium corrected for 2% with water to 50 mL. Gently mix each tube by inverting
hafnium content, 92.97 twice. Allow to stand for 5 min, and view downward over a
W = quantity of aluminum zirconium white surface.
tetrachlorohydrate taken (g) Acceptance criteria: NMT 20 ppm
[NOTEUse the result obtained to calculate the Aluminum/ The color of the solution from the Sample solution is not
zirconium atomic ratio and the (Aluminum plus zirconium)/ darker than that of the solution made from the Standard
chloride atomic ratio.] solution, and the color of the solution from the Monitor
ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage solution is the same as or darker than the color of the
of aluminum found in the test for Content of aluminum by solution made from the Standard solution. If the color of
the percentage of zirconium found in the test for Content of the solution from the Monitor solution is lighter than the
zirconium, and multiply by 92.97/26.98, in which 92.97 is color of the solution made from the Standard solution,
the atomic weight of zirconium corrected for 2% hafnium repeat the analysis with the following modification: after
content, and 26.98 is the atomic weight of aluminum. the heating step, to the Monitor solution and the Sample
Acceptance criteria: 2.0:1 and 5.99:1 solution add 1.0 mL, instead of 12 drops, of sodium
(ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO sulfide TS.
Calculate the (aluminum plus zirconium)/chloride atomic
ratio: SPECIFIC TESTS
PH 791: 3.0 5.0, in a solution (15 in 100)
Al% = percentage of aluminum from the test for PACKAGING AND STORAGE: Preserve in well-closed containers.
Content of aluminum LABELING: The label states the content of anhydrous
Ar1 = atomic weight of aluminum, 26.98 aluminum zirconium tetrachlorohydrate.
Zr% = percentage of zirconium from the test fof Content
of zirconium
Cl% = percentage of chloride from the test for Content Tetrachlorohydrate Solution
of chloride (Comment on this Monograph)id=m2366=Aluminum Zirconium
Ar3 = atomic weight of chlorine, 35.453 Tetrachlorohydrate Solution=A-Monos.pdf)
Acceptance criteria: 1.5:10.9:1
DEFINITION
IMPURITIES Aluminum Zirconium Tetrachlorohydrate Solution is a
Inorganic Impurities polymeric, loosely hydrated complex of basic aluminum
ARSENIC, Method I 211: NMT 2 ppm zirconium chloride that encompasses a range of aluminum-to-
LIMIT OF IRON zirconium atomic ratios between 2.0:1 and 5.99:1, and a
Standard solution: Transfer 2.0 mL of Standard Iron range of (aluminum plus zirconium)-to-chloride atomic ratios
Solution, prepared as directed under Iron 241, to a 50-mL between 1.5:1 and 0.9:1. The following solvents may be used:
beaker. water, propylene glycol, or dipropylene glycol. It contains the
Sample solution: 27 mg/mL of Aluminum Zirconium equivalent of NLT 90.0% and NMT 110.0% of the labeled
Tetrachlorohydrate. Transfer 5.0 mL of this solution to a 50- concentration of anhydrous aluminum zirconium
mL beaker. tetrachlorohydrate.
Analysis: To each of the beakers containing the Standard
solution and the Sample solution, add 5 mL of 6 N nitric IDENTIFICATION
acid, cover with a watch glass, and boil on a hot plate for A. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution
35 min. Allow to cool. Add 5 mL of Ammonium containing the equivalent of 100 mg/mL of anhydrous
Thiocyanate Solution, prepared as directed under Iron 241, aluminum zirconium tetrachlorohydrate meets the
transfer to separate 50-mL color comparison tubes, and requirements.
dilute with water to volume. B. PROPYLENE GLYCOL (where stated on the label)
Acceptance criteria: The color of the solution from the Sample solution: 200 mg/mL in isopropyl alcohol, and
Sample solution is not darker than that of the solution from filter
the Standard solution. (NMT 150 ppm) Analysis: Evaporate the filtrate to 1 mL on a steam bath:
HEAVY METALS, Method I 231 the IR absorption spectrum of a film of this solution on a
[NOTEProceed as directed, except use the following silver chloride disk exhibits maxima only at the same
modifications.] wavelengths as that of a similar preparation of a film of
Sample solution: Prepare as directed in the chapter. If the propylene glycol.
solution is not clear after dilution to 40 mL, heat for several C. DIPROPYLENE GLYCOL (where stated on the label)
min between 60 and 80, and cool to room temperature. Sample solution: 200 mg/mL in isopropyl alcohol, and
If the solution remains cloudy, repeat from the beginning filter
with the following modification: add 3 mL of hydrochloric Analysis: Evaporate the filtrate to 1 mL on a steam bath:
acid before adjustment of the pH with 6 N ammonium the IR absorption spectrum of a film of this solution on a
hydroxide. silver chloride disk exhibits maxima only at the same
Monitor solution: Prepare as directed, using the same wavelengths as that of a similar preparation of a film of
modifications described for the Sample solution, if dipropylene glycol.
necessary.
ASSAY Ze = equivalent volume of edetate disodium VS

PROCEDURE consumed (mL) by the zirconium moiety,
Analysis: Calculate the percentage of anhydrous aluminum calculated as (Zr%/Me) (W/Ar2), where Zr% is
zirconium tetrachlorohydrate in the Solution taken: the percentage of zirconium as determined in
the test for Content of zirconium, and Ar2 is the
Result = Al%({Ar1y + Ar2 + Mr[3y + 4(y + 1)/z] + Ar3(y + atomic weight of zirconium corrected for 2%
1)/z}/ Ar1y) hafnium content, 92.97
W = quantity of Aluminum Zirconium
Al% = percentage of aluminum from the test for Tetrachlorohydrate taken (g)
Content of aluminum [NOTEUse the result obtained to calculate the Aluminum/
Ar1 = atomic weight of aluminum, 26.98 zirconium atomic ratio and the (Aluminum plus zirconium)/
y = aluminum/zirconium atomic ratio from the test chloride atomic ratio.]
for Aluminum/zirconium atomic ratio ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage
Ar2 = atomic weight of zirconium corrected for 2% of aluminum found in the test for Content of aluminum by
hafnium content, 92.97 the percentage of zirconium found in the test for Content of
Mr = molecular weight of the hydroxide anion (OH), zirconium, and multiply by 92.97/26.98, in which 92.97 is
17.01 the atomic weight of zirconium corrected for 2% hafnium
z = (aluminum plus zirconium)/chloride atomic ratio content, and 26.98 is the atomic weight of aluminum.
found in the test for (Aluminum plus Acceptance criteria: 2.0:1 and 5.99:1
zirconium)/chloride atomic ratio (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO
Ar3 = atomic weight of chlorine (Cl), 35.453 Calculate the (aluminum plus zirconium)/chloride atomic
Acceptance criteria: 90.0%110.0% ratio:
OTHER COMPONENTS Result = [(Al%/Ar1) + (Zr%/Ar2)] / (Cl%/Ar3)
CONTENT OF CHLORIDE
Sample: 500 mg of Aluminum Zirconium Al% = percentage of aluminum from the test for
Tetrachlorohydrate Solution to a 250-mL beaker Content of aluminum
Analysis: Add 100120 mL of water and 20 mL of diluted Ar1 = atomic weight of aluminum, 26.98
nitric acid, and swirl to dissolve. Titrate with 0.05 N silver Zr% = percentage of zirconium from the test for Content
nitrate VS using a calomel electrode and a silver billet of zirconium
electrode system. Each mL of 0.05 N silver nitrate is Ar2 = atomic weight of zirconium corrected for 2%
equivalent to 1.773 mg of chloride (Cl). [NOTEUse the hafnium content, 92.97
result obtained to calculate the (Aluminum plus Cl% = percentage of chloride from the test for Content
zirconium)/chloride atomic ratio.] of chloride
CONTENT OF ZIRCONIUM Ar3 = atomic weight of chlorine, 35.453
Sample: 500 mg of Aluminum Zirconium Acceptance criteria: 1.5:10.9:1
Tetrachlorohydrate Solution to a 150-mL beaker
Analysis: Add 5 mL of water and 15 mL of hydrochloric IMPURITIES
acid. Heat the solution to boiling, and continue boiling for Inorganic Impurities
68 min. Add 3040 mL of water and 5 mL of hydrochloric ARSENIC, Method I 211
acid, and heat to boiling. Add 1 drop of xylenol orange TS, Sample solution: Use a weighed quantity of the solution.
and while still hot, titrate with 0.1 M edetate disodium VS Acceptance criteria: NMT 2 ppm
until the color of the solution changes from pink to yellow. LIMIT OF IRON
Perform a blank determination. Each mL of 0.1 M edetate Standard solution: Transfer 2.0 mL of Standard Iron
disodium is equivalent to 9.297 mg of zirconium (Zr). Solution, prepared as directed under Iron 241, to a 50-mL
[NOTEUse the result obtained to calculate the beaker.
Aluminum/zirconium atomic ratio and the (Aluminum plus Sample solution: 54 mg/mL of Aluminum Zirconium
zirconium)/chloride atomic ratio.] Tetrachlorohydrate Solution. Transfer 5.0 mL of this solution
CONTENT OF ALUMINUM to a 50-mL beaker.
Sample: 0.30 g of Aluminum Zirconium Tetrachlorohydrate Analysis: To each of the beakers containing the Standard
Solution to a 150-mL beaker solution and the Sample solution add 5 mL of 6 N nitric
Analysis: Add 5 mL of water and 15 mL of hydrochloric acid, cover with a watch glass, and boil on a hot plate for
acid. Heat this solution to boiling, and continue boiling for 5 35 min. Allow to cool. Add 5 mL of Ammonium
min. Add 40 mL of water and 15.0 mL of 0.1 M edetate Thiocyanate Solution, prepared as directed under Iron 241,
disodium VS. Heat the solution to boiling, and continue transfer to separate 50-mL color comparison tubes, and
boiling for 5 min. Allow the solution to cool, add 1015 mL dilute with water to volume.
of acetic acidammonium acetate buffer TS, and adjust with Acceptance criteria: The color of the solution from the
ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of Sample solution is not darker than that of the solution from
alcohol, and adjust with ammonium hydroxide to a pH of the Standard solution. (NMT 75 ppm).
4.6 0.1. Add 510 drops of dithizone TS, and titrate with HEAVY METALS, Method I 231
0.1 M zinc sulfate VS until the first permanent purple-pink [NOTEProceed as directed in the chapter, except use the
color appears. Perform a blank determination. following modifications.]
Calculate the percentage of aluminum (Al) in the Aluminum Sample solution: Prepare as directed in the chapter, using
Zirconium Tetrachlorohydrate: a weighed quantity of solution. If the solution is not clear
after dilution to 40 mL, heat for several min between 60
Result = 2.698[15.0 Me (zMz + Ze)]/W and 80, and cool to room temperature. If the solution
remains cloudy, repeat from the beginning with the
Me = molarity of edetate disodium VS following modification: add 3 mL of hydrochloric acid
z = volume of zinc sulfate VS consumed (mL)
Mz = molarity of zinc sulfate VS
before adjustment of the pH with 6 N ammonium ASSAY

hydroxide. PROCEDURE
Monitor solution: Prepare as directed, using the same Analysis: Calculate the percentage of anhydrous aluminum
modifications described for the Sample solution, if zirconium tetrachlorohydrate in the Aluminum Zirconium
necessary. Tetrachlorohydrex Gly:
Analysis: To each of the three tubes containing the
Standard solution, the Sample solution, and the Monitor Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y +
solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently 1)/z}/Ar1y)
to between 60 and 80. To the Standard solution, add 6
drops of sodium sulfide TS. To the Sample solution and the Al% = percentage of aluminum found in the test for
Monitor solution, add 12 drops of sodium sulfide TS. Cool Content of aluminum
to room temperature, and dilute the contents of each tube Ar1 = atomic weight of aluminum, 26.98
with water to 50 mL. Gently mix each tube by inverting y = aluminum/zirconium atomic ratio found in the
twice. Allow to stand for 5 min, and view downward over a test for Aluminum/zirconium atomic ratio
white surface. Ar2 = atomic weight of zirconium corrected for 2%
Acceptance criteria: NMT 10 ppm hafnium content, 92.97
The color of the solution from the Sample solution is not Mr = molecular weight of the hydroxide anion (OH),
darker than that of the solution made from the Standard 17.01
solution, and the color of the solution made from the z = (aluminum plus zirconium)/chloride atomic ratio
Monitor solution is the same as or darker than the color of found in the test for (Aluminum plus
the solution from the Standard solution. If the color of the zirconium)/chloride atomic ratio
solution from the Monitor solution is lighter than the color Ar3 = atomic weight of chlorine (Cl), 35.453
of the solution made from the Standard solution, repeat Acceptance criteria: 90.0%110.0%
the analysis with the following modification: after the
heating step, to the Monitor solution and the Sample OTHER COMPONENTS
solution add 1.0 mL, instead of 12 drops, of sodium CONTENT OF CHLORIDE
sulfide TS. Sample: 250 mg of Aluminum Zirconium
Tetrachlorohydrex Gly to a 250-mL beaker
SPECIFIC TESTS Analysis: Add 100120 mL of water and 20 mL of diluted
PH 791: 3.05.0, in a solution (3 in 10) nitric acid, and swirl to dissolve. Titrate with 0.05 N silver
nitrate VS using a calomel electrode and a silver billet
ADDITIONAL REQUIREMENTS electrode system. Each mL of 0.05 N silver nitrate is
PACKAGING AND STORAGE: Preserve in well-closed containers. equivalent to 1.773 mg of chloride (Cl). [NOTEUse the
LABELING: Label the Solution to state the solvent used and result obtained to calculate the (Aluminum plus
the claimed concentration of anhydrous aluminum zirconium zirconium)/chloride atomic ratio.]
tetrachlorohydrate. CONTENT OF ZIRCONIUM
Tetrachlorohydrex Gly to a 150-mL beaker
Aluminum Zirconium acid. Heat this solution to boiling, and continue boiling for
Tetrachlorohydrex Gly 68 min. Add 3040 mL of water and 5 mL of hydrochloric
(Comment on this Monograph)id=m2368=Aluminum Zirconium acid, and heat to boiling. Add 1 drop of xylenol orange TS,
Tetrachlorohydrex Gly=A-Monos.pdf) and while still hot, titrate with 0.1 M edetate disodium VS
until the color of the solution changes from pink to yellow.
DEFINITION Perform a blank determination, and make any necessary
Aluminum Zirconium Tetrachlorohydrex Gly is a derivative of correction. Each mL of 0.1 M edetate disodium is equivalent
Aluminum Zirconium Tetrachlorohydrate in which some of the to 9.297 mg of zirconium (Zr). [NOTEUse the result
water molecules have been displaced by glycine, calcium obtained to calculate the Aluminum/zirconium atomic ratio
glycinate, magnesium glycinate, potassium glycinate, sodium and the (Aluminum plus zirconium)/chloride atomic ratio).]
glycinate, or zinc glycinate. It encompasses a range of CONTENT OF ALUMINUM
aluminum-to-zirconium atomic ratios between 2.0:1 and Sample: 0.15 g of Aluminum Zirconium Tetrachlorohydrex
5.99:1, and a range of (aluminum plus zirconium)-to-chloride Gly to a 150-mL beaker
atomic ratios between 1.5:1 and 0.9:1. It contains NLT 90.0% Analysis: Add 5 mL of water and 15 mL of hydrochloric
and NMT 110.0% of the labeled amount of anhydrous acid. Heat this solution to boiling, and continue boiling for 5
aluminum zirconium tetrachlorohydrate. min. Add 40 mL of water and 15.0 mL of 0.1 M edetate
disodium VS. Heat the solution to boiling, and continue
IDENTIFICATION boiling for 5 min. Allow the solution to cool, add 1015 mL
A. IDENTIFICATION TESTSGENERAL, Chloride 191: A 100 of acetic acidammonium acetate buffer TS, and adjust with
mg/mL solution meets the requirements of the test for ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of
Chloride. alcohol, and adjust with ammonium hydroxide to a pH of
B. PROCEDURE 4.6 0.1. Add 510 drops of dithizone TS, and titrate with
Sample solution: 25 mg/mL in water 0.1 M zinc sulfate VS until the first permanent purple-pink
Analysis: Heat 20 ml of the Sample solution to boiling on a color appears. Perform a blank determination, and make any
hot plate, and add 60 mg of ninhydrin. necessary correction.
Acceptance criteria: A deep violet color develops
immediately.
Calculate the percentage of aluminum (Al) in the Aluminum several min between 60 and 80, and cool to room
Zirconium Tetrachlorohydrex Gly: temperature. If the solution remains cloudy, repeat from
the beginning with the following modification: add 3 mL
Result = 2.698[15.0 Me (zMz + Ze)]/W of hydrochloric acid before adjustment of the pH with 6 N
ammonium hydroxide.
Me = molarity of the edetate disodium VS Monitor solution: Prepare as directed, using the
z = volume of zinc sulfate VS consumed (mL) modifications described for the Sample solution, if
Mz = molarity of the zinc sulfate VS necessary.
Ze = equivalent volume of edetate disodium VS Analysis
consumed (mL) by the zirconium moiety, Samples: Sample solution and Monitor solution
calculated as (Zr%/Me) (W/Ar2), where Zr% is To each of the three tubes containing the samples, add 2
the percentage of zirconium as determined in mL of pH 3.5 Acetate Buffer, and heat gently to between
the test for Content of zirconium, and Ar2 is the 60 and 80. To the Standard solution, add 6 drops of
atomic weight of zirconium corrected for 2% sodium sulfide TS. To the Sample solution and the
hafnium content, 92.97 Monitor solution, add 12 drops of sodium sulfide TS. Cool
W = quantity of aluminum zirconium to room temperature, and dilute the contents of each
tetrachlorohydrate taken (g) tube with water to 50 mL. Gently mix each tube by
[NOTEUse the result obtained to calculate the Aluminum/ inverting twice. Allow to stand for 5 min, and view
zirconium atomic ratio and the (Aluminum plus zirconium)/ downward over a white surface.
chloride atomic ratio).] Acceptance criteria: NMT 20 ppm
ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage The color of the solution from the Sample solution is not
of aluminum found in the test for Content of aluminum by darker than that of the solution from the Standard
the percentage of zirconium found in the test for Content of solution, and the color of the solution from the Monitor
zirconium, and multiply by 92.97/26.98, in which 92.97 is solution is the same as or darker than the color of the
the atomic weight of zirconium corrected for 2% hafnium solution from the Standard solution. If the color of the
content, and 26.98 is the atomic weight of aluminum. solution from the Monitor solution is lighter than the color
Acceptance criteria: 2.0:1 and 5.99:1 of the solution from the Standard solution, repeat the
(ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO analysis with the following modification: after the heating
Calculate the (aluminum plus zirconium)/chloride atomic step, to the Monitor solution and the Sample solution add
ratio: 1.0 mL, instead of 12 drops, of sodium sulfide TS.
Result = [(Al%/Ar1) + (Zr%/Ar2)/(Cl%/Ar3) SPECIFIC TESTS
Al% = percentage of aluminum as determined in the
test for Content of aluminum ADDITIONAL REQUIREMENTS
Ar1 = atomic weight of aluminum, 26.98 PACKAGING AND STORAGE: Preserve in well-closed containers.
Zr% = percentage of zirconium as determined in the LABELING: The label states the form of glycine used and the
test for Content of zirconium claimed content of anhydrous aluminum zirconium
Ar2 = atomic weight of zirconium corrected for 2% tetrachlorohydrate.
for Content of chloride
Ar3 = atomic weight of chlorine, 35.453 Aluminum Zirconium
Acceptance criteria: 1.5:10.9:1 Tetrachlorohydrex Gly Solution
(Comment on this Monograph)id=m2369=Aluminum Zirconium
IMPURITIES Tetrachlorohydrex Gly Solution=A-Monos.pdf)
ARSENIC, Method I 211: NMT 2 ppm DEFINITION
LIMIT OF IRON Aluminum Zirconium Tetrachlorohydrex Gly Solution is a
Standard solution: Transfer 2.0 mL of Standard iron solution of Aluminum Zirconium Tetrachlorohydrate in which
solution, prepared as directed under Iron 241, to a 50-mL some of the waters of hydration have been displaced by
beaker. glycine, calcium glycinate, magnesium glycinate, potassium
Sample solution: 27 mg/mL of Aluminum Zirconium glycinate, sodium glycinate, or zinc glycinate. It encompasses
Tetrachlorohydrex Gly in water. Transfer 5.0 mL of this a range of aluminum-to-zirconium ratios between 2.0:1 and
solution to a 50-mL beaker. 5.99:1, and a range of (aluminum plus zirconium)-to-chloride
Analysis atomic ratios between 1.5:1 and 0.9:1. The following solvents
Samples: Standard solution and Sample solution may be used: water, propylene glycol, or dipropylene glycol. It
To each of the Samples, add 5 mL of 6 N nitric acid, cover contains the equivalent of NLT 90.0% and NMT 110.0% of
with a watch glass, and boil on a hot plate for 35 min. the labeled concentration of anhydrous aluminum zirconium
Allow to cool. Add 5 mL of Ammonium Thiocyanate tetrachlorohydrate.
Solution, prepared as directed under Iron 241, transfer
to separate 50-mL color comparison tubes, and dilute IDENTIFICATION
with water to volume. A. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution
Acceptance criteria: NMT 150 ppm equivalent to 100 mg/mL of anhydrous aluminum zirconium
The color of the solution from the Sample solution is not tetrachlorohydrate meets the requirements of the test for
darker than that of the solution from the Standard Chloride.
solution. B. PROPYLENE GLYCOL (where stated on the label)
HEAVY METALS, Method I 231 Sample solution: 200 mg/mL in isopropyl alcohol, and
[NOTEProceed as directed, except use the following filter
modifications.] Analysis: Evaporate the filtrate to 1 mL on a steam bath:
Sample solution: Prepare as directed in the chapter. If the IR absorption spectrum of a film of this solution on a
the solution is not clear after dilution to 40 mL, heat for
silver chloride disk exhibits maxima only at the same of acetic acidammonium acetate buffer TS, and adjust with
wavelengths as that of a similar preparation of a film of ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of
propylene glycol. alcohol, and adjust with ammonium hydroxide to a pH of
C. DIPROPYLENE GLYCOL (where stated on the label) 4.6 0.1. Add 510 drops of dithizone TS, and titrate with
Sample solution: 200 mg/mL in isopropyl alcohol, and 0.1 M zinc sulfate VS until the first permanent purple-pink
filter color appears. Perform a blank determination, and make any
Analysis: Evaporate the filtrate to 1 mL on a steam bath: necessary correction.
the IR absorption spectrum of a film of this solution on a Calculate the percentage of aluminum (Al) in the aluminum
silver chloride disk exhibits maxima only at the same zirconium tetrachlorohydrate:
wavelengths as that of a similar preparation of a film of
dipropylene glycol. Result = 2.698[15.0 Me (zMz + Ze)]/W
D. GLYCINE
Sample solution: 50 mg/mL in isopropyl alcohol, and filter Me = molarity of the edetate disodium VS
Analysis: Heat to boiling on a hot plate, and add 60 mg of z = volume of zinc sulfate VS consumed (mL)
ninhydrin for 20 mL of Sample solution. Mz = molarity of the zinc sulfate VS
Acceptance criteria: A deep violet color develops Ze = equivalent volume of edetate disodium VS
immediately. consumed (mL) by the zirconium moiety,
calculated as (Zr%/Me) (W/Ar2), where Zr% is
ASSAY the percentage of zirconium as determined in
PROCEDURE the test for Content of zirconium, and Ar2 is the
Analysis: Calculate the percentage of anhydrous aluminum atomic weight of zirconium corrected for 2%
zirconium tetrachlorohydrate in the Solution taken: hafnium content, 92.97
W = quantity of aluminum zirconium
Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + tetrachlorohydrate taken (g)
1)/z}/Ar1y) [NOTEUse the result obtained to calculate the Aluminum/
zirconium atomic ratio and the (Aluminum plus zirconium)/
Al% = percentage of aluminum found in the test for chloride atomic ratio.]
Content of aluminum ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage
Ar1 = atomic weight of aluminum, 26.98 of aluminum found in the test for Content of aluminum by
y = aluminum/zirconium atomic ratio found in the the percentage of zirconium found in the test for Content of
test for Aluminum/zirconium atomic ratio zirconium, and multiply by 92.97/26.98, in which 92.97 is
Ar2 = atomic weight of zirconium corrected for 2% the atomic weight of zirconium corrected for 2% hafnium
hafnium content, 92.97 content, and 26.98 is the atomic weight of aluminum: the
Mr = molecular weight of the hydroxide anion (OH), ratio is between 2.0:1 and 5.99:1.
17.01 (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO
z = (aluminum plus zirconium)/chloride atomic ratio Calculate the (aluminum plus zirconium)/chloride atomic
found in the test for (Aluminum plus ratio:
zirconium)/chloride atomic ratio
Ar3 = atomic weight of chlorine (Cl), 35.453 Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3)
OTHER COMPONENTS test for Content of aluminum
CONTENT OF CHLORIDE Ar1 = atomic weight of aluminum, 26.98
Sample: 500 mg of Solution to a 250-mL beaker Zr% = percentage of zirconium as determined in the
Analysis: Add 100120 mL of water and 20 mL of diluted test for Content of zirconium
nitric acid, and swirl to dissolve. Titrate with 0.05 N silver Ar2 = atomic weight of zirconium corrected for 2%
nitrate VS using a calomel electrode and a silver billet hafnium content, 92.97
electrode system. Each mL of 0.05 N silver nitrate is Cl% = percentage of chloride as determined in the test
equivalent to 1.773 mg of chloride (Cl). [NOTEUse the for Content of chloride
result obtained to calculate the (Aluminum plus Ar3 = atomic weight of chlorine, 35.453
zirconium)/chloride atomic ratio.] Acceptance criteria: 1.5:10.9:1
CONTENT OF ZIRCONIUM
Sample: 500 mg of Solution to a 150-mL beaker IMPURITIES
acid. Heat this solution to boiling, and continue boiling for ARSENIC, Method I 211: Prepare the Sample solution using a
68 min. Add 3040 mL of water and 5 mL of hydrochloric weighed quantity of the Solution.
to 9.297 mg of zirconium (Zr).[NOTEUse the result Sample solution: 54 mg/mL of Solution in water. Transfer
obtained to calculate the Aluminum/zirconium atomic ratio 5.0 mL of this solution to a 50-mL beaker.
and the (Aluminum plus zirconium)/chloride atomic ratio.] Analysis
CONTENT OF ALUMINUM Samples: Standard solution and Sample solution
Sample: 0.3 g of Solution to a 150-mL beaker To each of the two samples, add 5 mL of 6 N nitric acid,
Analysis: Add 5 mL of water and 15 mL of hydrochloric cover with a watch glass, and boil on a hot plate for 35
acid. Heat this solution to boiling, and continue boiling for 5 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate
min. Add 40 mL of water and 15.0 mL of 0.1 M edetate Solution, prepared as directed under Iron 241, transfer
disodium VS. Heat the solution to boiling, and continue to separate 50-mL color comparison tubes, and dilute
boiling for 5 min. Allow the solution to cool, add 1015 mL with water to volume.
Acceptance criteria: NMT 75 ppm ASSAY

The color of the solution from the Sample solution is not PROCEDURE
darker than that of the solution from the Standard Analysis: Calculate the percentage of anhydrous aluminum
solution. zirconium trichlorohydrate in the Aluminum Zirconium
HEAVY METALS, Method I 231 Trichlorohydrate:
[NOTEProceed as directed, except use the following
modifications.] Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] +
Sample solution: Prepare as directed in the chapter, using Ar3(y + 1)/z}/Ar1y)
a weighed quantity of Solution. If the solution is not clear
after dilution to 40 mL, heat for several min between 60 Al% = percentage of aluminum found in the test for
and 80, and cool to room temperature. If the solution Content of aluminum
remains cloudy, repeat from the beginning with the Ar1 = atomic weight of aluminum, 26.98
following modification: add 3 mL of hydrochloric acid y = aluminum/zirconium atomic ratio found in the
before adjustment of the pH with 6 N ammonium test for Aluminum/airconium atomic ratio
hydroxide. Ar2 = atomic weight of zirconium corrected for 2%
Monitor solution: Prepare as directed, using the same hafnium content, 92.97
modifications described for Sample solution if necessary. Mr = molecular weight of the hydroxide anion (OH),
Analysis 17.01
Samples: Sample solution and Monitor solution z = (aluminum plus zirconium)/chloride atomic ratio
To each of the three samples, add 2 mL of pH 3.5 Acetate found in the test for (Aluminum plus
Buffer, and heat gently to between 60 and 80. To the zirconium)/chloride atomic ratio
Standard solution, add 6 drops of sodium sulfide TS. To Ar3 = atomic weight of chlorine (Cl), 35.453
the Sample solution and the Monitor solution, add 12 Acceptance criteria: 90.0%110.0%
drops of sodium sulfide TS. Cool to room temperature,
and dilute the contents of each tube with water to 50 OTHER COMPONENTS
mL. Gently mix each tube by inverting twice. Allow to CONTENT OF CHLORIDE
stand for 5 min, and view downward over a white Sample: 250 mg of Aluminum Zirconium Trichlorohydrate
surface. to a 250-mL beaker
Acceptance criteria: NMT 10 ppm. Analysis: Add 100120 mL of water and 20 mL of diluted
The color of the solution from the Sample solution is not nitric acid, and swirl to dissolve. Titrate with 0.05 N silver
darker than that of the solution from the Standard nitrate VS using a calomel electrode and a silver billet
solution, and the color of the solution from the Monitor electrode system. Each mL of 0.05 N silver nitrate is
solution is the same as or darker than the color of the equivalent to 1.773 mg of chloride (Cl). [NOTEUse the
solution from the Standard solution. If the color of the result obtained to calculate the (Aluminum plus
solution from the Monitor solution is lighter than the color zirconium)/chloride atomic ratio.]
of the solution from the Standard solution, repeat the CONTENT OF ZIRCONIUM
analysis with the following modification: after the heating Sample: 250 mg of Aluminum Zirconium Trichlorohydrate
step, to the Monitor solution and the Sample solution add to a 150-mL beaker
1.0 mL, instead of 12 drops, of sodium sulfide TS. Analysis: Add 5 mL of water and 15 mL of hydrochloric
SPECIFIC TESTS 68 min. Add 3040 mL of water and 5 mL of hydrochloric
PH 791: 3.05.0, in a solution (3 in 10) acid, and heat to boiling. Add 1 drop of xylenol orange TS,
and while still hot, titrate with 0.1 M edetate disodium VS
ADDITIONAL REQUIREMENTS until the color of the solution changes from pink to yellow.
PACKAGING AND STORAGE: Preserve in well-closed containers. Perform a blank determination, and make any necessary
LABELING: Label the Solution to state the solvent and form correction. Each mL of 0.1 M edetate disodium is equivalent
of glycine used and the claimed concentration of anhydrous to 9.297 mg of zirconium (Zr). [NOTEUse the result
aluminum zirconium tetrachlorohydrate. obtained to calculate the Aluminum/zirconium atomic ratio
CONTENT OF ALUMINUM
Sample: 0.15 g of Aluminum Zirconium Trichlorohydrate to
Aluminum Zirconium Trichlorohydrate a 150-mL beaker
(Comment on this Monograph)id=m2370=Aluminum Zirconium Analysis: Add 5 mL of water and 15 mL of hydrochloric
Trichlorohydrate=A-Monos.pdf) acid. Heat this solution to boiling, and continue boiling for 5
AlyZr(OH)3y+4-xClx nH2O min. Add 40 mL of water and 15.0 mL of 0.1 M edetate
disodium VS. Heat the solution to boiling, and continue
DEFINITION boiling for 5 min. Allow the solution to cool, add 1015 mL
Aluminum Zirconium Trichlorohydrate is a polymeric, loosely of acetic acidammonium acetate buffer TS, and adjust with
hydrated complex of basic aluminum zirconium chloride that ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of
encompasses a range of aluminum-to-zirconium atomic ratios alcohol, and adjust with ammonium hydroxide to a pH of
between 2.0:1 and 5.99:1, and a range of (aluminum plus 4.6 0.1. Add 510 drops of dithizone TS, and titrate with
zirconium)-to-chloride atomic ratios between 2.1:1 and 1.51:1. 0.1 M zinc sulfate VS until the first permanent purple-pink
It contains NLT 90.0% and NMT 110.0% of the labeled color appears. Perform a blank determination, and make any
amount of anhydrous aluminum zirconium trichlorohydrate. necessary correction.
Calculate the percentage of aluminum (Al) in the Aluminum
IDENTIFICATION Zirconium Trichlorohydrate:
IDENTIFICATION TESTSGENERAL, Chloride 191: A solution
containing the equivalent of 100 mg/mL of anhydrous Result = 2.698[15.0 Me (zMz + Ze)]/W
aluminum zirconium trichlorohydrate meets the
requirements of the test for Chloride. Me = molarity of the edetate disodium VS
z = volume of zinc sulfate VS consumed (mL) Monitor solution: Prepare as directed, using the same
Mz = molarity of the zinc sulfate VS modifications described for the Sample solution, if
Ze = equivalent volume of edetate disodium VS necessary.
consumed (mL) by the zirconium moiety, Analysis
calculated as (Zr%/Me) (W/Ar), where Zr% is Samples: Standard solution and Monitor solution
the percentage of zirconium as determined in To each of the three tubes containing the Samples, add 2
the test for Content of zirconium, and Ar is the mL of pH 3.5 Acetate Buffer, and heat gently to between
atomic weight of zirconium corrected for 2% 60 and 80. To the Standard solution, add 6 drops of
hafnium content, 92.7 sodium sulfide TS. To the Sample solution and the
W = quantity of Aluminum Zirconium Trichlorohydrate Monitor solution, add 12 drops of sodium sulfide TS. Cool
taken (g) to room temperature, and dilute the contents of each
[NOTEUse the result obtained to calculate the Aluminum/ tube with water to 50 mL. Gently mix each tube by
zirconium atomic ratio and the (Aluminum plus zirconium)/ inverting twice. Allow to stand for 5 min, and view
chloride atomic ratio.] downward over a white surface.
ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage Acceptance criteria: NMT 20 ppm.
of aluminum found in the test for Content of aluminum by The color of the solution from the Sample solution is not
the percentage of zirconium found in the test for Content of darker than that of the solution from the Standard
zirconium, and multiply by 92.97/26.98, in which 92.97 is solution, and the color of the solution from the Monitor
the atomic weight of zirconium corrected for 2% hafnium solution is the same as or darker than the color of the
content, and 26.98 is the atomic weight of aluminum. solution from the Standard solution. If the color of the
Acceptance criteria: 2.0:1 and 5.99:1 solution from the Monitor solution is lighter than the color
(ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO of the solution from the Standard solution, repeat the
Calculate the (aluminum plus zirconium)/chloride atomic analysis with the following modification: after the heating
ratio: step, to the Monitor solution and the Sample solution add
1.0 mL, instead of 12 drops, of sodium sulfide TS.
Result = [(Al%/Ar1) + (Zr%/Ar2)] /(Cl%/Ar3)
SPECIFIC TESTS
Al% = percentage of aluminum as determined in the PH 791: 3.05.0, in a solution (15 in 100)
test for Content of aluminum
Ar1 = atomic weight of aluminum, 26.98 ADDITIONAL REQUIREMENTS
Zr% = percentage of zirconium as determined in the PACKAGING AND STORAGE: Preserve in well-closed containers.
test for Content of zirconium LABELING: The label states the content of anhydrous
Ar2 = atomic weight of zirconium corrected for 2% aluminum zirconium trichlorohydrate.
for Content of chloride Aluminum Zirconium Trichlorohydrate
Acceptance criteria: 2.1:11.51:1 Solution
IMPURITIES Trichlorohydrate Solution=A-Monos.pdf)
LIMIT OF IRON Aluminum Zirconium Trichlorohydrate Solution is a polymeric,
Standard solution: Transfer 2.0 mL of Standard Iron loosely hydrated complex of basic aluminum zirconium
Solution, prepared as directed under Iron 241, to a 50-mL chloride that encompasses a range of aluminum-to-zirconium
beaker. atomic ratios between 2.0:1 and 5.99:1, and a range of
Sample solution: 27 mg/mL of Aluminum Zirconium (aluminum plus zirconium)-to-chloride atomic ratios between
Trichlorohydrate in water. Transfer 5.0 mL of this solution 2.1:1 and 1.51:1. The following solvents may be used: water,
to a 50-mL beaker. propylene glycol, or dipropylene glycol. It contains the
Analysis equivalent of NLT 90.0% and NMT 110.0% of the labeled
Samples: Standard soultion and Sample solution concentration of anhydrous aluminum zirconium
To each of the beakers containing the Samples, add 5 mL trichlorohydrate.
of 6 N nitric acid, cover with a watch glass, and boil on a
hot plate for 35 min. Allow to cool. Add 5 mL of IDENTIFICATION
Ammonium Thiocyanate Solution, prepared as directed A. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution
under Iron 241, transfer to separate 50-mL color containing the equivalent of about 100 mg/mL of anhydrous
comparison tubes, and dilute with water to volume. aluminum zirconium trichlorohydrate meets the
Acceptance criteria: NMT 150 ppm requirements of the test for Chloride.
The color of the solution from the Sample solution is not B. PROPYLENE GLYCOL (where stated on the label)
darker than that of the solution from the Standard Sample solution: 200 mg/mL in isopropyl alcohol, and
solution. filter
HEAVY METALS, Method I 231 Analysis: Evaporate the filtrate to 1 mL on a steam bath:
[NOTEProceed as directed, except use the following the IR absorption spectrum of a film of this solution on a
modifications.] silver chloride disk exhibits maxima only at the same
Sample solution: Prepare as directed in the chapter. If the wavelengths as that of a similar preparation of a film of
solution is not clear after dilution to 40 mL, heat for several propylene glycol.
min between 60 and 80, and cool to room temperature. C. DIPROPYLENE GLYCOL (where stated on the label)
If the solution remains cloudy, repeat from the beginning Sample solution: 200 mg/mL in isopropyl alcohol, and
with the following modification: add 3 mL of hydrochloric filter
acid before adjustment of the pH with 6 N ammonium Analysis: Evaporate the filtrate to 1 mL on a steam bath:
hydroxide. the IR absorption spectrum of a film of this solution on a
silver chloride disk exhibits maxima only at the same Ze = equivalent volume of edetate disodium VS
wavelengths as that of a similar preparation of a film of consumed (mL) by the zirconium moiety,
dipropylene glycol. calculated as (Zr%/Me) (W/Ar), where Zr% is
ASSAY the test for Content of zirconium, and Ar is the
PROCEDURE atomic weight of zirconium corrected for 2%
Analysis: Calculate the percentage of anhydrous aluminum hafnium content, 92.97
zirconium trichlorohydrate in the Solution taken: W = quantity of aluminum zirconium trichlorohydrate
taken (g)
Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + [NOTEUse the result obtained to calculate the Aluminum/
1)/z}/Ar1y) zirconium atomic ratio and the (Aluminum plus zirconium)/
chloride atomic ratio.]
Al% = percentage of aluminum found in the test for ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage
Content of aluminum of aluminum found in the test for Content of aluminum by
Ar1 = atomic weight of aluminum, 26.98 the percentage of zirconium found in the test for Content of
y = aluminum/zirconium atomic ratio found in the zirconium, and multiply by 92.97/26.98, in which 92.97 is
test for Aluminum/zirconium atomic ratio the atomic weight of zirconium corrected for 2% hafnium
Ar2 = atomic weight of zirconium corrected for 2% content, and 26.98 is the atomic weight of aluminum.
hafnium content, 92.97 Acceptance criteria 2.0:1 and 5.99:1
Mr = molecular weight of the hydroxide anion (OH), (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO
17.01 Calculate the (aluminum plus zirconium)/chloride atomic
z = (aluminum plus zirconium)/chloride atomic ratio ratio:
found in the test for (Aluminum plus
zirconium)/chloride atomic ratio Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3)
Acceptance criteria: 90.0%110.0% Al% = percentage of aluminum as determined in the
OTHER COMPONENTS Ar1 = atomic weight of aluminum, 26.98
CONTENT OF CHLORIDE Zr% = percentage of zirconium as determined in the
Sample: 500 mg of Solution to a 250-mL beaker test for Content of zirconium
Analysis: Add 100120 mL of water and 20 mL of diluted Ar2 = atomic weight of zirconium corrected for 2%
nitric acid, and swirl to dissolve. Titrate with 0.05 N silver hafnium content, 92.97
nitrate VS using a calomel electrode and a silver billet Cl% = percentage of chloride as determined in the test
electrode system. Each mL of 0.05 N silver nitrate is for Content of chloride
equivalent to 1.773 mg of chloride (Cl). [NOTEUse the Ar3 = atomic weight of chlorine, 35.453
result obtained to calculate the (Aluminum plus Acceptance criteria: 21:11.51:1
zirconium)/chloride atomic ratio.]
CONTENT OF ZIRCONIUM IMPURITIES
Sample: 500 mg of Solution to a 150-mL beaker Inorganic Impurities
Analysis: Add 5 mL of water and 15 mL of hydrochloric ARSENIC, Method I 211: Prepare the Sample solution using a
acid. Heat this solution to boiling, and continue boiling for quantity of the Solution.
68 min. Add 3040 mL of water and 5 mL of hydrochloric Acceptance criteria: NMT 2 ppm
acid, and heat to boiling. Add 1 drop of xylenol orange TS, LIMIT OF IRON
and while still hot, titrate with 0.1 M edetate disodium VS Standard solution: Transfer 2.0 mL of Standard iron
until the color of the solution changes from pink to yellow. solution, prepared as directed under Iron 241, to a 50-mL
Perform a blank determination, and make any necessary beaker.
correction. Each mL of 0.1 M edetate disodium is equivalent Sample solution: 54 mg/mL of Solution in water. Transfer
to 9.297 mg of zirconium (Zr). [NOTEUse the result 5.0 mL of this solution to a 50-mL beaker.
obtained to calculate the Aluminum/zirconium atomic ratio Analysis
and the (Aluminum plus zirconium)/chloride atomic ratio.] Samples: Standard solution and Sample solution
CONTENT OF ALUMINUM To each of the Samples, add 5 mL of 6 N nitric acid, cover
Sample: 0.3 g of Solution to a 150-mL beaker with a watch glass, and boil on a hot plate for 35 min.
Analysis: Add 5 mL of water and 15 mL of hydrochloric Allow to cool. Add 5 mL of Ammonium Thiocyanate
acid. Heat this solution to boiling, and continue boiling for 5 Solution, prepared as directed under Iron 241, transfer
min. Add 40 mL of water and 15.0 mL of 0.1 M edetate to separate 50-mL color comparison tubes, and dilute
disodium VS. Heat the solution to boiling, and continue with water to volume.
boiling for 5 min. Allow the solution to cool, add 1015 mL Acceptance criteria: NMT 75 ppm
of acetic acidammonium acetate buffer TS, and adjust with The color of the solution from the Sample solution is not
ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of darker than that of the solution from the Standard
alcohol, and adjust with ammonium hydroxide to a pH of solution.
4.6 0.1. Add 510 drops of dithizone TS, and titrate with HEAVY METALS, Method I 231
0.1 M zinc sulfate VS until the first permanent purple-pink [NOTEProceed as directed, except use the following
color appears. Perform a blank determination, and make any modifications.]
necessary correction. Sample solution: Prepare as directed in the chapter, using
Calculate the percentage of aluminum (Al) in the aluminum a weighed quantity of Solution. If the solution is not clear
zirconium trichlorohydrate: after dilution to 40 mL, heat for several min between 60
and 80, and cool to room temperature. If the solution
Result = 2.698[15.0 Me (zMz + Ze)]/W remains cloudy, repeat from the beginning with the
following modification: add 3 mL of hydrochloric acid
Me = molarity of the edetate disodium VS before adjustment of the pH with 6 N ammonium
z = volume of zinc sulfate VS consumed (mL) hydroxide.
Mz = molarity of the zinc sulfate VS
Monitor solution: Prepare as directed, using the same ASSAY

modifications described for the Sample solution, if PROCEDURE
necessary. Analysis: Calculate the percentage of anhydrous aluminum
Analysis zirconium trichlorohydrate in the Aluminum Zirconium
Samples: Standard solution, Sample solution, and Monitor Trichlorohydrate Gly:
solution
To each of the three Samples, add 2 mL of pH 3.5 Acetate Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] +
Buffer, and heat gently to between 60 and 80. To the Ar3(y + 1)/z}/Ar1y)
Standard solution, add 6 drops of sodium sulfide TS. To
the Sample solution and the Monitor solution, add 12 Al% = percentage of aluminum found in the test for
drops of sodium sulfide TS. Cool to room temperature, Content of aluminum
and dilute the contents of each tube with water to 50 Ar1 = atomic weight of aluminum, 26.98
mL. Gently mix each tube by inverting twice. Allow to y = aluminum/zirconium atomic ratio found in the
stand for 5 min, and view downward over a white test for Aluminum/zirconium atomic ratio
surface. Ar2 = atomic weight of zirconium corrected for 2%
Acceptance criteria: NMT 10 ppm. hafnium content, 92.97
The color of the solution from the Sample solution is not Mr = molecular weight of the hydroxide anion (OH),
darker than that of the solution from the Standard 17.01
solution, and the color of the solution from the Monitor z = (aluminum plus zirconium)/chloride atomic ratio
solution is the same as or darker than the color of the found in the test for (Aluminum plus
solution from the Standard solution. If the color of the zirconium)/chloride atomic ratio
solution from the Monitor solution is lighter than the color Ar3 = atomic weight of chlorine (Cl), 35.453
of the solution from the Standard solution, repeat the Acceptance criteria: 90.0%110.0%
analysis with the following modification: after the heating
step, to the Monitor solution and the Sample solution add OTHER COMPONENTS
1.0 mL, instead of 12 drops, of sodium sulfide TS CONTENT OF CHLORIDE
Sample: 250 mg of Aluminum Zirconium Trichlorohydrate
SPECIFIC TESTS Gly to a 250-mL beaker
PH 791: 3.05.0, in a solution (3 in 10) Analysis: Add 100120 mL of water and 20 mL of diluted
nitric acid, and swirl to dissolve. Titrate with 0.05 N silver
ADDITIONAL REQUIREMENTS nitrate VS using a calomel electrode and a silver billet
PACKAGING AND STORAGE: Preserve in well-closed containers. electrode system. Each mL of 0.05 N silver nitrate is
LABELING: Label the Solution to state the solvent used and equivalent to 1.773 mg of chloride (Cl). [NOTEUse the
the claimed concentration of anhydrous aluminum zirconium result obtained to calculate the (Aluminum plus
trichlorohydrate. zirconium)/chloride atomic ratio.]
CONTENT OF ZIRCONIUM
Sample: 250 mg of Aluminum Zirconium Trichlorohydrate
Gly to a 150-mL beaker
Aluminum Zirconium Trichlorohydrex Analysis: Add 5 mL of water and 15 mL of hydrochloric
Gly acid. Heat this solution to boiling, and continue boiling for
(Comment on this Monograph)id=m2373=Aluminum Zirconium 68 min. Add 3040 mL of water and 5 mL of hydrochloric
Trichlorohydrex Gly=A-Monos.pdf) acid, and heat to boiling. Add 1 drop of xylenol orange TS,
and while still hot, titrate with 0.1 M edetate disodium VS
DEFINITION until the color of the solution changes from pink to yellow.
Aluminum Zirconium Trichlorohydrex Gly is a derivative of Perform a blank determination, and make any necessary
Aluminum Zirconium Trichlorohydrate in which some of the correction. Each mL of 0.1 M edetate disodium is equivalent
water molecules have been displaced by glycine, calcium to 9.297 mg of zirconium (Zr). [NOTEUse the result
glycinate, magnesium glycinate, potassium glycinate, sodium obtained to calculate the Aluminum/zirconium atomic ratio
glycinate, or zinc glycinate. It encompasses a range of and the (Aluminum plus zirconium)/chloride atomic ratio.]
aluminum-to-zirconium atomic ratios between 2.0:1 and CONTENT OF ALUMINUM
5.99:1, and a range of (aluminum plus zirconium)-to-chloride Sample: 0.15 g of Aluminum Zirconium Trichlorohydrate
atomic ratios between 2.1:1 and 1.51:1. It contains NLT Gly to a 150-mL beaker
90.0% and NMT 110.0% of the labeled amount of anhydrous Analysis: Add 5 mL of water and 15 mL of hydrochloric
aluminum zirconium trichlorohydrate. acid. Heat this solution to boiling, and continue boiling for 5
min. Add 40 mL of water and 15.0 mL of 0.1 M edetate
IDENTIFICATION disodium VS. Heat the solution to boiling, and continue
A. IDENTIFICATION TESTSGENERAL, Chloride 191: A 100 boiling for 5 min. Allow the solution to cool, add 1015 mL
mg/mL solution meets the requirements of the test for of acetic acidammonium acetate buffer TS, and adjust with
Chloride. ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of
B. PROCEDURE alcohol, and adjust with ammonium hydroxide to a pH of
Sample solution: 25 mg/mL in water 4.6 0.1. Add 510 drops of dithizone TS, and titrate with
Analysis: Heat to boiling on a hot plate, and add 60 mg of 0.1 M zinc sulfate VS until the first permanent purple-pink
ninhydrin to 20 mL of Sample solution. color appears. Perform a blank determination, and make any
Acceptance criteria: A deep violet color develops necessary correction.
immediately.
Calculate the percentage of aluminum (Al) in the Aluminum min between 60 and 80, and cool to room temperature.
Zirconium Trichlorohydrate Gly: If the solution remains cloudy, repeat from the beginning
with the following modification: add 3 mL of hydrochloric
Result = 2.698[15.0 Me (zMz + Ze)]/W acid before adjustment of the pH with 6 N ammonium
hydroxide.
Me = molarity of the edetate disodium VS Monitor solution: Prepare as directed, using the same
z = volume of zinc sulfate VS consumed (mL) modifications described for Sample solution, if necessary.
Mz = molarity of the zinc sulfate VS Analysis
Ze = equivalent volume of edetate disodium VS Samples: Standard solution, Sample solution, and Monitor
consumed (mL) by the zirconium moiety, solution
calculated as (Zr%/Me) (W/Ar), where Zr% To each of the three Samples, add 2 mL of pH 3.5 Acetate
equals the percentage of zirconium as Buffer, and heat gently to between 60 and 80. To the
determined in the test for Content of zirconium, Standard solution, add 6 drops of sodium sulfide TS. To
and Ar equals the atomic weight of zirconium the Sample solution and the Monitor solution, add 12
corrected for 2% hafnium content, 92.97 drops of sodium sulfide TS. Cool to room temperature,
W = quantity of aluminum zirconium trichlorohydrate and dilute the contents of each tube with water to 50
taken (g) mL. Gently mix each tube by inverting twice. Allow to
[NOTEUse the result obtained to calculate the Aluminum/ stand for 5 min, and view downward over a white
zirconium atomic ratio and the (Aluminum plus zirconium)/ surface.
chloride atomic ratio.] Acceptance criteria: NMT 20 ppm.
ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage The color of the solution from the Sample solution is not
of aluminum found in the test for Content of aluminum by darker than that of the solution from the Standard
the percentage of zirconium found in the test for Content of solution, and the color of the solution from the Monitor
zirconium, and multiply by 92.97/26.98, in which 92.97 is solution is the same as or darker than the color of the
the atomic weight of zirconium corrected for 2% hafnium solution from the Standard solution. If the color of the
content, and 26.98 is the atomic weight of aluminum. solution from the Monitor solution is lighter than the color
Acceptance criteria: 2.0:1 and 5.99:1 of the solution from the Standard solution, repeat the
(ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO analysis with the following modification: after the heating
Calculate the (aluminum plus zirconium)/chloride atomic step, to the Monitor solution and the Sample solution add
ratio: 1.0 mL, instead of 12 drops, of sodium sulfide TS.
Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3) SPECIFIC TESTS
test for Content of aluminum ADDITIONAL REQUIREMENTS
Ar1 = atomic weight of aluminum, 26.98 PACKAGING AND STORAGE: Preserve in well-closed containers.
Zr% = percentage of zirconium as determined in the LABELING: The label states the form of glycine used and the
test for Content of zirconium claimed content of anhydrous aluminum zirconium
Ar2 = atomic weight of zirconium corrected for 2% trichlorohydrate.
for Content of chloride
Ar3 = atomic weight of chlorine, 35.453 Aluminum Zirconium Trichlorohydrex
Acceptance criteria: 2.1:11.51:1 Gly Solution
IMPURITIES Trichlorohydrex Gly Solution=A-Monos.pdf)
LIMIT OF IRON Aluminum Zirconium Trichlorohydrex Gly Solution is a solution
Standard solution: Transfer 2.0 mL of Standard Iron of Aluminum Zirconium Trichlorohydrate in which some of the
Solution, prepared as directed under Iron 241, to a 50-mL waters of hydration have been displaced by glycine, calcium
beaker. glycinate, magnesium glycinate, potassium glycinate, sodium
Sample solution: 27 mg/mL of Aluminum Zirconium glycinate, or zinc glycinate. It encompasses a range of
Trichlorohydrate Gly in water. Transfer 5.0 mL of this aluminum-to-zirconium ratios between 2.0:1 and 5.99:1, and a
solution to a 50-mL beaker. range of (aluminum plus zirconium)-to-chloride atomic ratios
Analysis between 2.1:1 and 1.51:1. The following solvents may be
Samples: Standard solution and Sample solution used: water, propylene glycol, or dipropylene glycol. It
To each of the Samples, add 5 mL of 6 N nitric acid, cover contains the equivalent NLT 90.0% and NMT 110.0% of the
with a watch glass, and boil on a hot plate for 35 min. labeled concentration of anhydrous aluminum zirconium
Allow to cool. Add 5 mL of Ammonium Thiocyanate trichlorohydrate.
Solution, prepared as directed under Iron 241, transfer
to separate 50-mL color comparison tubes, and dilute IDENTIFICATION
with water to volume. A. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution
Acceptance criteria: NMT 150 ppm containing the equivalent of 100 mg/mL of anhydrous
The color of the solution from the Sample solution is not aluminum zirconium trichlorohydrate meets the
darker than that of the solution from the Standard requirements of the test for Chloride.
solution. B. PROPYLENE GLYCOL (where stated on the label)
HEAVY METALS, Method I 231 Sample solution: 200 mg/mL in isopropyl alcohol, and
[NOTEProceed as directed, except use the following filter
modifications.] Analysis: Evaporate the filtrate to 1 mL on a steam bath:
Sample solution: Prepare as directed in the chapter. If the the IR absorption spectrum of a film of this solution on a
silver chloride disk exhibits maxima only at the same of acetic acidammonium acetate buffer TS, and adjust with
wavelengths as that of a similar preparation of a film of ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of
propylene glycol. alcohol, and adjust with ammonium hydroxide to a pH of
C. DIPROPYLENE GLYCOL (where stated on the label) 4.6 0.1. Add 510 drops of dithizone TS, and titrate with
Sample solution: 200 mg/mL in isopropyl alcohol, and 0.1 M zinc sulfate VS until the first permanent purple-pink
filter color appears. Perform a blank determination, and make any
Analysis: Evaporate the filtrate to 1 mL on a steam bath: necessary correction.
the IR absorption spectrum of a film of this solution on a Calculate the percentage of aluminum (Al) in the Solution:
silver chloride disk exhibits maxima only at the same
wavelengths as that of a similar preparation of a film of Result = 2.698[15.0 Me (zMz + Ze)]/W
dipropylene glycol.
D. GLYCINE Me = molarity of the edetate disodium VS
Sample solution: 50 mg/mL in isopropyl alcohol, and filter z = volume of zinc sulfate VS consumed (mL)
Analysis: Heat to boiling on a hot plate, and add 60 mg of Mz = molarity of the zinc sulfate VS
ninhydrin for 20 mL of Sample solution Ze = equivalent volume of edetate disodium VS
Acceptance criteria: A deep violet color develops consumed (mL) by the zirconium moiety,
immediately. calculated as (Zr%/Me) (W/Ar), where Zr% is
ASSAY the test for Content of zirconium, and Ar is the
PROCEDURE atomic weight of zirconium corrected for 2%
Analysis: Calculate the percentage of anhydrous aluminum hafnium content, 92.97
zirconium trichlorohydrate in the Solution taken: W = quantity of aluminum zirconium trichlorohydrate
taken (g)
Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + [NOTEUse the result obtained to calculate the Aluminum/
1)/z}/Ar1y) zirconium atomic ratio and the (Aluminum plus zirconium)/
chloride atomic ratio.]
Al% = percentage of aluminum found in the test for ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage
Content of aluminum of aluminum found in the test for Content of aluminum by
Ar1 = atomic weight of aluminum, 26.98 the percentage of zirconium found in the test for Content of
y = aluminum/zirconium atomic ratio found in the zirconium, and multiply by 92.97/26.98, in which 92.97 is
test for Aluminum/zirconium atomic ratio the atomic weight of zirconium corrected for 2% hafnium
Ar2 = atomic weight of zirconium corrected for 2% content, and 26.98 is the atomic weight of aluminum.
hafnium content, 92.97 Acceptance criteria: 2.0:1 and 5.99:1
Mr = molecular weight of the hydroxide anion (OH), (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO
17.01 Calculate the (aluminum plus zirconium)/chloride atomic
z = (aluminum plus zirconium)/chloride atomic ratio ratio:
found in the test for (Aluminum plus
zirconium)/chloride atomic ratio Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3)
Acceptance criteria: 90.0%110.0% Al% = percentage of aluminum as determined in the
OTHER COMPONENTS Ar1 = atomic weight of aluminum, 26.98
CONTENT OF CHLORIDE Zr% = percentage of zirconium as determined in the
Sample: 500 mg of Solution to a 250-mL beaker test for Content of zirconium
Analysis: Add 100120 mL of water and 20 mL of diluted Ar2 = atomic weight of zirconium corrected for 2%
nitric acid, and swirl to dissolve. Titrate with 0.05 N silver hafnium content, 92.97
nitrate VS using a calomel electrode and a silver billet Cl% = percentage of chloride as determined in the test
electrode system. Each mL of 0.05 N silver nitrate is for Content of chloride
equivalent to 1.773 mg of chloride (Cl). [NOTEUse the Ar3 = atomic weight of chlorine, 35.453
result obtained to calculate the (Aluminum plus Acceptance criteria: 2.1:11.51:1
zirconium)/chloride atomic ratio.]
CONTENT OF ZIRCONIUM IMPURITIES
Sample: 500 mg of Solution to a 150-mL beaker Inorganic Impurities
Analysis: Add 5 mL of water and 15 mL of hydrochloric ARSENIC, Method I 211: Prepare the Sample solution using a
acid. Heat this solution to boiling, and continue boiling for weighed quantity of the Solution.
68 min. Add 3040 mL of water and 5 mL of hydrochloric Acceptance criteria: NMT 2 ppm
acid, and heat to boiling. Add 1 drop of xylenol orange TS, LIMIT OF IRON
and, while still hot, titrate with 0.1 M edetate disodium VS Standard solution: Transfer 2.0 mL of Standard iron
until the color of the solution changes from pink to yellow. solution, prepared as directed under Iron 241, to a 50-mL
Perform a blank determination, and make any necessary beaker.
correction. Each mL of 0.1 M edetate disodium is equivalent Sample solution: 54 mg/mL of Solution in water. Transfer
to 9.297 mg of zirconium (Zr). [NOTEUse the result 5.0 mL of this solution to a 50-mL beaker.
obtained to calculate the Aluminum/zirconium atomic ratio Analysis
and the (Aluminum plus zirconium)/chloride atomic ratio.] Samples: Standard solution and Sample solution
CONTENT OF ALUMINUM To each of the Samples, add 5 mL of 6 N nitric acid, cover
Sample: 0.3 g of Solution to a 150-mL beaker with a watch glass, and boil on a hot plate for 35 min.
Analysis: Add 5 mL of water and 15 mL of hydrochloric Allow to cool. Add 5 mL of Ammonium Thiocyanate
acid. Heat this solution to boiling, and continue boiling for 5 Solution, prepared as directed under Iron 241, transfer
min. Add 40 mL of water and 15.0 mL of 0.1 M edetate to separate 50-mL color comparison tubes, and dilute
disodium VS. Heat the solution to boiling, and continue with water to volume.
Acceptance criteria: NMT 75 ppm IDENTIFICATION

The color of the solution from the Sample solution is not INFRARED ABSORPTION 197S
darker than that of the solution from the Standard Cell: 1 mm
solution. Sample solution: 50 mg in 10 mL of 0.1 N hydrochloric
HEAVY METALS, Method I 231 acid, and filter. Transfer the filtrate to a suitable separator,
[NOTEProceed as directed in the chapter, except use the add 1 mL of 5 N sodium hydroxide, and extract with 5 mL
following modifications.] of methylene chloride.
Sample solution: Prepare as directed in the chapter, using
a quantity of Solution. If the solution is not clear after ASSAY
dilution to 40 mL, heat for several min between 60 and PROCEDURE
80, and cool to room temperature. If the solution remains Sample: Dissolve 120 mg of Amantadine Hydrochloride in a
cloudy, repeat from the beginning with the following mixture of 30 mL glacial acetic acid and 10 mL of mercuric
modification: add 3 mL of hydrochloric acid before acetate TS.
adjustment of the pH with 6 N ammonium hydroxide. Analysis: Titrate with 0.1 N perchloric acid VS, using
Monitor solution: Prepare as directed, using the same suitable electrodes. Perform a blank determination (see
modifications described for Sample solution, if necessary. Titrimetry 541). Each mL of 0.1 N perchloric acid is
Analysis equivalent to 18.77 mg of C10H17N HCl.
Samples: Standard solution, Sample soution, and Monitor Acceptance criteria: 98.5%101.5%
solution
To each of the three Samples, add 2 mL of pH 3.5 Acetate IMPURITIES
Buffer, and heat gently to between 60 and 80. To the Inorganic Impurities
Standard solution, add 6 drops of sodium sulfide TS. To HEAVY METALS, Method I 231: NMT 10 ppm
the Sample solution and the Monitor solution, add 12 drops [NOTEUse 1 mL of 1 N acetic acid in the Sample solution.]
of sodium sulfide TS. Cool to room temperature, and Organic Impurities
dilute the contents of each tube with water to 50 mL. PROCEDURE
Gently mix each tube by inverting twice. Allow to stand Internal standard solution: 50 mg/mL of adamantane in
for 5 min, and view downward over a white surface. dichloromethane
Acceptance criteria: NMT 10 ppm. Sample solution: 1.0 g of Amantadine Hydrochloride in a
The color of the solution from the Sample solution is not separator. Add 20 mL of 5.0 N sodium hydroxide and 18
darker than that of the solution from the Standard solution, mL of dichloromethane, and shake for 10 min. Remove the
and the color of the solution from the Monitor solution is water layer, dry the organic layer by swirling with
the same as or darker than the color of the solution from anhydrous sodium sulfate, and allow to stand for a few min
the Standard solution. If the color of the solution from the to ensure that all remaining water has been removed.
Monitor solution is lighter than the color of the solution Filter, collect the filtrate in a 20-mL volumetric flask, add
from the Standard solution, repeat the analysis with the 0.2 mL of Internal standard solution, and dilute with
following modification: after the heating step, to the dichloromethane to volume.
Monitor solution and the Sample solution add 1.0 mL, Standard solution: 10 mg of USP Amantadine
instead of 12 drops, of sodium sulfide TS. Hydrochloride RS in a separator. Add 20 mL of 5.0 N
sodium hydroxide and 18 mL of dichloromethane, and
SPECIFIC TESTS shake for 10 min. Remove the water layer, dry the organic
PH 791: 3.05.0, in a solution (3 in 10) layer by swirling with anhydrous sodium sulfate, and allow
to stand for a few min to ensure that all remaining water
ADDITIONAL REQUIREMENTS has been removed. Filter, collect the filtrate in a 20-mL
PACKAGING AND STORAGE: Preserve in well-closed containers. volumetric flask, add 0.2 mL of Internal standard solution,
LABELING: Label Solution to state the solvent and form of and dilute with dichloromethane to volume.
glycine used and the claimed concentration of anhydrous Chromatographic system
aluminum zirconium trichlorohydrate. (See Chromatography 621, System Suitability.)
Mode: GC
Detector: Flame ionization
Column: 0.53-mm 30-m fused-silica column coated
Amantadine Hydrochloride with 1.0-m G27 stationary phase
(Comment on this Monograph)id=m2380=Amantadine Temperature: See Temperature Program Table. Increase
Hydrochloride=A-Monos.pdf) column temperature linearly at a rate of 10/min.
Temperature Program Table

Time (min) Temperature
Injection port 220
Detector 300
Column 0 70
C10H17N HCl 187.71 5 70
Tricyclo[3.3.1.1.3,7]decan-1-amine, hydrochloride;
1-Adamantanamine hydrochloride [665-66-7]. 23 250
40 250
DEFINITION
Amantadine Hydrochloride contains NLT 98.5% and NMT
101.5% of C10H17N HCl.
USP 32 Official Monographs / Amantadine 169
Carrier gas: Helium Standard stock solution: 2 mg/mL of USP Amantadine

Flow rate: 4 mL/min Hydrochloride RS in water
Injection size: 2 L Standard solution: Pipet 25.0 mL of Standard stock solution
Injection type: Split flow is 200 mL/min with a split flow into a 250-mL separator, and add 25 mL of 2.0 N sodium
ratio of 50:1. hydroxide and 50.0 mL of Internal standard solution. Shake
System suitability for 60 min, and collect the hexane layer (Standard solution).
Sample: Standard solution Sample solution: Transfer NLT 20 Capsules to a 200-mL
[NOTEThe relative retention times for adamantane and volumetric flask. Add 40 mL of 0.1 N hydrochloric acid, and
amantadine are about 0.7 and 1.0, respectively.] heat gently to achieve complete dissolution. Cool, and dilute
Suitability requirements with water to volume. Pipet 5.0 mL of the solution into a
Resolution: NLT 20 between adamantane and 250-mL separator, and add 40 mL of 1.0 N sodium
amantadine hydroxide and 50.0 mL of Internal standard solution. Shake
Relative standard deviation: NMT 5.0% determined for 60 min, and collect the hexane layer (Sample solution).
from the peak response ratios of amantadine to Chromatographic system
adamantane (See Chromatography 621, System Suitability.)
Analysis Mode: GC
Samples: Sample solution and Standard solution Detector: Flame ionization
Calculate the percentage of each impurity in the portion Column: 2-mm 1.22-m; glass column packed with 10%
of C10H17N HCl taken: phase G1 on 100- to 120-mesh support S1A
Temperature
Result = (RU/RS) (CS/CU) 100 Column: 115
Injection port: 250
RU = peak response ratio of each impurity to Detector block: 250
adamantane from the Sample solution Injection size: 1 L
RS = peak response ratio of amantadine to System suitability
adamantane from the Standard solution Sample: Standard solution
CS = concentration of USP Amantadine Hydrochloride Suitability requirements
RS in the Standard solution (mg/mL) Resolution: NLT 2 between napthalene and amantadine
CU = concentration of the Sample solution (mg/mL) Tailing factor: NMT 2.0 for the analyte peak
Acceptance criteria Relative standard deviation: NMT 2.0%
Individual impurities: NMT 0.3% Analysis
Total impurities: NMT 1.0% Samples: Standard solution and Sample solution
Calculate the percentage C10H17N HCl taken:
SPECIFIC TESTS
PH 791: 3.05.5, in a solution (1 in 5) Result = (RU/RS) (CS/CU) 100
CLARITY AND COLOR OF SOLUTION
Sample: Dissolve 2 g in 10 mL of water. RU = peak response ratios from the Sample solution
Acceptance criteria: Solution is clear and nearly colorless. RS = peak response ratios from the Standard solution
CS = concentration of USP Amantadine Hydrochloride
ADDITIONAL REQUIREMENTS RS in the Standard solution (mg/mL)
PACKAGING AND STORAGE: Preserve in well-closed containers. CU = nominal concentration of the Sample solution
USP Amantadine Hydrochloride RS Acceptance criteria: 95.0%105.0%
PERFORMANCE TESTS
Amantadine Hydrochloride Capsules DISSOLUTION, Procedure for a Pooled Sample 711
(Comment on this Monograph)id=m2390=Amantadine Apparatus 1: 100 rpm
Hydrochloride Capsules=A-Monos.pdf) Time: 45 min
Internal standard solution: 0.054 mg/mL of naphthalene
DEFINITION in hexane
Amantadine Hydrochloride Capsules contain NLT 95.0% and Standard stock solution: 0.1 mg/mL of USP Amantadine
NMT 105.0% of the labeled amount of amantadine Hydrochloride RS in water
hydrochloride (C10H17N HCl). Standard solution: Pipet 15.0 mL of Standard stock solution
IDENTIFICATION into a 50-mL screw-capped test tube, add 5.0 mL of 5 N
INFRARED ABSORPTION 197S sodium hydroxide and 10.0 mL of Internal standard solution,
Cell: 1 mm and shake for 60 min. Collect the hexane layer.
Sample solution: Place the contents of Capsules, equivalent Sample solution: Filter 15.0 mL of the solution under test,
to 200 mg of amantadine hydrochloride, in a vessel, dissolve and place into a 50-mL screw-capped test tube. Pipet 5.0
in 0.1 N hydrochloric acid, and filter. Transfer the filtrate to mL of 5 N sodium hydroxide and 10.0 mL of the Internal
a separator, add 1 mL of 5 N sodium hydroxide, and extract standard solution into the test tube, and shake for 60 min.
with 5 mL of methylene chloride. Filter the extract through Collect the hexane layer (Sample solution).
anhydrous sodium sulfate, and rinse the anhydrous sodium Chromatographic system: Proceed as directed in the
sulfate with 2 mL of methylene chloride. Assay.
Injection size: 2.5 L
ASSAY Analysis
PROCEDURE Sample: Standard solution and Sample solution
Internal standard solution: 0.4 mg/mL of naphthalene in Tolerances: NLT 75% (Q) of the labeled amount of
hexane C10H17N HCl is dissolved in 45 min.
170 Amantadine / Official Monographs USP 32
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements RS = peak response ratios from the Standard solution
CS = concentration of USP Amantadine Hydrochloride
PACKAGING AND STORAGE: Preserve in tight containers. CU = nominal concentration of amantadine
USP REFERENCE STANDARDS 11 hydrochloride in the Sample solution (mg/mL)
USP Amantadine Hydrochloride RS Acceptance criteria: 95.0%105.0%
Amantadine Hydrochloride Oral USP REFERENCE STANDARDS 11
Solution USP Amantadine Hydrochloride RS
(Comment on this Monograph)id=m2420=Amantadine
Hydrochloride Oral Solution=A-Monos.pdf)
DEFINITION Amcinonide
Amantadine Hydrochloride Oral Solution contains NLT 95.0% (Comment on this Monograph)id=m2550=Amcinonide=A-
and NMT 105.0% of the labeled amount of amantadine Monos.pdf)
hydrochloride (C10H17N HCl).
IDENTIFICATION
INFRARED ABSORPTION 197S
Cell: 1 mm
Sample solution: Place a volume of Oral Solution,
equivalent to about 200 mg of amantadine hydrochloride,
in a vessel, dissolve in 0.1 N hydrochloric acid, and filter.
Transfer the filtrate to a separator, add 10 mL of 0.5 N C28H35FO7 502.57
sodium hydroxide, and extract with 5 mL of methylene Pregna-1,4-diene-3,20-dione, 21-(acetyloxy)-16,17-
chloride. Filter the extract through anhydrous sodium [cyclopentylidenebis(oxy)]-9-fluoro-11-hydroxy-, (11,16)-;
sulfate, and rinse the anhydrous sodium sulfate with 2 mL of 9-Fluoro-11,16,17,21-tetrahydroxypregna-1,4-diene-3,20-
methylene chloride. dione cyclic 16,17-acetal with cyclopentanone, 21-acetate
[51022-69-6].
ASSAY
Internal standard solution: 0.4 mg/mL of naphthalene in Amcinonide contains NLT 97.0% and NMT 102.0% of
hexane C28H35FO7, calculated on the dried basis.
Standard stock solution: 2 mg/mL of USP Amantadine
Hydrochloride RS in water IDENTIFICATION
Standard solution: Pipet 25.0 mL of Standard stock solution A. INFRARED ABSORPTION 197K
into a 250-mL separator, add 25 mL of 2.0 N sodium B. ULTRAVIOLET ABSORPTION 197U
hydroxide and 50.0 mL of Internal standard solution. Shake Analytical wavelength: 238 nm
for 60 min, and collect the hexane layer. Solution: 40 g/mL in methanol
Sample solution: Pipet 5.0 mL of the Oral Solution into a Acceptance criteria: Absorptivities do not differ by more
250-mL conical flask, and add 45 mL of 1.0 N sodium than 3.0%, calculated on the dried basis
hydroxide and 50.0 mL of Internal standard solution. Shake
for 60 min, and collect the hexane layer. ASSAY
(See Chromatography 621, System Suitability.) Solution A: Acetonitrile and water (7:13)
Mode: GC Solution B: Acetonitrile and water (7:3)
Detector: Flame ionization System suitability solution: 12.5 g/mL of butylparaben
Column: 2-mm 1.22-m glass column packed with 10% and 20 g/mL of USP Amcinonide RS in Solution B
phase G1 on 100- to 120-mesh support S1A Standard solution: 0.02 mg/mL of USP Amcinonide RS in
Temperature Solution B
Column: 115 Sample stock solution: 0.2 mg/mL of Amcinonide in
Injection port: 250 Solution B [NOTESonicate for 5 min.]
Detector block: 250 Sample solution: 0.02 mg/mL of Sample stock solution in
Injection size 1 L Solution B
System suitability Mobile phase: See the gradient table below.
Suitability requirements Time Solution A Solution B
Resolution: NLT 2 between napthalene and amantadine (min) (%) (%)
Tailing factor: NMT 2.0 for the analyte peak
0 100 0
Analysis 2.5 100 0
Samples: Standard solution and Sample solution 10 0 100
Calculate the percentage of C10H17N HCl in the portion of
Oral Solution taken: Chromatographic system
RU = peak response ratios from the Sample solution
USP 32 Official Monographs / Amcinonide 171
Mode: LC Application volume: 25 L

Detector: UV 254 nm Developing solvent: Ether
Flow rate: 2 mL/min Samples: Standard solution and Sample solution
Injection size: 10 L Develop the chromatogram until the solvent front has
System suitability moved about 12 cm above the line of application.
Samples: System suitability solution and Standard solution Acceptance criteria: The intensity and the RF value of the
[NOTEThe relative retention times for butylparaben and principal spot of the Sample solution are similar to those of
amcinonide are 0.78 and 1.0, respectively.] the Standard solution.
Resolution: NLT 8.0 between butylparaben and ASSAY
amcinonide, System suitability solution PROCEDURE
Tailing factor: NMT 1.5, Standard solution Solution A: Acetonitrile and water (7:13)
Relative standard deviation: NMT 2.0%, Standard Solution B: Acetonitrile and water (7:3)
solution System suitability solution: 12.5 g/mL of butylparaben
Analysis and 20 g/mL of USP Amcinonide RS in Solution B
Samples: Standard solution and Sample solution Standard solution: 0.02 mg/mL of USP Amcinonide RS in
Calculate the percentage of C28H35FO7 in the portion of Solution B
Amcinonide taken: Sample solution: Transfer a quantity of Cream, equivalent
to 10 mg of Amcinonide, to a 50-mL volumetric flask. Add 5
Result = (rU/rS) (CS/CU) 100 mL of Solution B and 15 mL of acetonitrile, and heat over a
steam bath until dissolved. Add 20 mL of Solution B while
rU = peak responses from the Sample solution hot, cool to room temperature, dilute with Solution B to
rS = peak responses from the Standard solution volume, and refrigerate for 30 min. Vigorously shake the
CS = concentration of USP Amcinonide RSin the solution to disperse the mixture, and filter while cold.
Standard solution (mg/mL) Transfer 5 mL of this solution to a 50-mL volumetric flask,
CU = concentration of the Sample solution (mg/mL) and dilute with Solution B to volume.
Acceptance criteria: 97.0%102.0% Mobile phase: See the gradient table below.
IMPURITIES
Time Solution A Solution B
(min) (%) (%)
0 100 0
SPECIFIC TESTS 2.5 100 0
OPTICAL ROTATION, Specific Rotation 781S: +89.4 to
+94.0 10 0 100
Sample solution: 10 mg/mL, in chloroform
LOSS ON DRYING 731: Dry it at 105 for 4 h: it loses NMT Chromatographic system
1.0% of its weight. (See Chromatography 621, System Suitability.)
Mode: LC
ADDITIONAL REQUIREMENTS Detector: UV 254 nm
PACKAGING AND STORAGE: Preserve in well-closed containers. Column: 4.6-mm 25-cm; packing L1
USP REFERENCE STANDARDS 11 Flow rate: 2 mL/min
USP Amcinonide RS Injection size: 10 L
System suitability
[NOTEThe relative retention times for butylparaben and
Amcinonide Cream amcinonide are about 0.78 and 1.0, respectively.]
(Comment on this Monograph)id=m2555=Amcinonide Suitability requirements
Cream=A-Monos.pdf) Resolution: NLT 8.0 between butylparaben and
amcinonide, System suitability solution
DEFINITION Tailing factor: NMT 1.5, Standard solution
Amcinonide Cream is Amcinonide in a suitable cream base. It Relative standard deviation: NMT 2.0%, Standard
contains NLT 90.0% and NMT 115.0% of the labeled amount solution
of amcinonide (C28H35FO7). Analysis
IDENTIFICATION Calculate the percentage of C28H35FO7 in the portion of
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Cream taken:
Standard solution: 100 g/mL of USP Amcinonide RS in Result = (rU/rS) (CS/CU) 100
chloroform
Sample solution: Place 2 g of the Cream in a 150-mL rU = peak responses from the Sample solution
beaker, add 50 mL of chloroform and 15 g of anhydrous rS = peak responses from the Standard solution
sodium sulfate, and stir with a glass rod to dissolve the CS = concentration of USP Amcinonide RS in the
specimen. Filter the solution, and clarify the filtrate, if Standard solution (mg/mL)
necessary, by the further addition of anhydrous sodium CU = nominal concentration of amcinonide in the
sulfate and a second filtration. Evaporate the filtrate to Sample solution (mg/mL)
dryness, and dissolve the residue in chloroform to obtain a
solution containing 100 g/mL of amcinonide.
172 Amcinonide / Official Monographs USP 32
Acceptance criteria: 90.0%115.0% quantitatively with the same solvent mixture to obtain a
solution having a concentration of 0.2 mg/mL of
PERFORMANCE TESTS amcinonide. Cool to room temperature, dilute with
MINIMUM FILL 755: Meets the requirements acetonitrile to volume, and filter.
Sample solution: Dilute Sample stock solution with Solution
SPECIFIC TESTS B (5 in 50).
MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED Mobile phase: See the gradient table below.
MICROORGANISMS 62: It meets the requirements of the
tests for absence of Staphylococcus aureus and Pseudomonas
aeruginosa. Time Solution A Solution B
PH 791: 3.55.2 (min) (%) (%)
0 100 0
PACKAGING AND STORAGE: Preserve in tight containers. 2.5 100 0
USP REFERENCE STANDARDS 11 10 0 100
USP Amcinonide RS
Mode: LC
Amcinonide Ointment Detector: UV 240 nm
(Comment on this Monograph)id=m2560=Amcinonide Column: 4.6-mm 25-cm; packing L1
Ointment=A-Monos.pdf) Flow rate: 2 mL/min
DEFINITION System suitability
Amcinonide Ointment is Amcinonide in a suitable ointment Samples: System suitability solution and Standard solution
base. It contains NLT 90.0% and NMT 115.0% of the labeled [NOTEThe relative retention times for butylparaben and
amount of amcinonide (C28H35FO7). amcinonide are 0.78 and 1.0, respectively.]
IDENTIFICATION Resolution: NLT 8.0 between butylparaben and
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 amcinonide, System suitability solution
Adsorbent: 0.25-mm layer of chromatographic silica gel Tailing factor: NMT 1.5, Standard solution
Standard solution: 100 g/mL of USP Amcinonide RS in Relative standard deviation: NMT 2.0%, Standard
chloroform solution
Sample solution: Place 2 g of the Ointment in a 150-mL Analysis
beaker, add 50 mL of chloroform and 15 g of anhydrous Samples: Standard solution and Sample solution
sodium sulfate, and stir with a glass rod to dissolve the Calculate the percentage of C28H35FO7 in the portion of
specimen. Filter the solution, and clarify the filtrate, if Ointment taken:
necessary, by the further addition of anhydrous sodium
sulfate and a second filtration. Evaporate the filtrate to Result = (rU/rS) (CS/CU) 100
dryness, and dissolve the residue in chloroform to obtain a
solution containing 100 g/mL of amcinonide. rU = peak response from the Sample solution
Application volume: 25 L rS = peak response from the Standard solution
Developing solvent: Ether CS = concentration of USP Amcinonide RS in the
Analysis Standard solution (mg/mL)
Samples: Standard solution and Sample solution CU = nominal concentration of amcinonide in the
Develop the chromatogram until the solvent front has Sample solution (mg/mL)
moved about 12 cm above the line of application. Acceptance criteria: 90.0%115.0%
Acceptance criteria: The intensity and the RF value of the
principal spot of the Sample solution are similar to those of PERFORMANCE TESTS
the Standard solution. MINIMUM FILL 755: Meets the requirements
ASSAY SPECIFIC TESTS
PROCEDURE MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED
Solution A: Acetonitrile and water (7:13) MICROORGANISMS 62: Meets the requirements of the tests
Solution B: Acetonitrile and water (7:3) for absence of Staphylococcus aureus and Pseudomonas
System suitability solution: 12.5 g/mL of butylparaben aeruginosa
and 20 g/mL of USP Amcinonide RS in Solution B
Standard solution: 0.02 mg/mL of USP Amcinonide RS in ADDITIONAL REQUIREMENTS
Solution B PACKAGING AND STORAGE: Preserve in tight containers.
Sample stock solution: Dissolve Ointment in a suitable USP REFERENCE STANDARDS 11
volume of a mixture of acetonitrile and chloroform (4:1) by USP Amcinonide RS
heating in a hot water bath, cooling, and adjusting
USP 32 Official Monographs / Amifostine 173
Amifostine Acceptance criteria: 78.0%82.0%

(Comment on this Monograph)id=m2600=Amifostine=A- IMPURITIES
Monos.pdf) Inorganic Impurities
Organic Impurities
PROCEDURE
Mobile phase: 1.0 mL of nonafluorobutane sulfonic acid
in 1200 mL of HPLC grade water, add 400 L of
trifluoroacetic acid, and adjust with triethylamine to a pH
of 2.5. Prepare a mixture of this solution and acetonitrile
C5H15N2O3PS 3H2O 268.27 (68:32).
Ethanethiol, 2-[(3-aminopropyl)amino]-, dihydrogen phosphate Blank: Water
(ester), trihydrate; Standard thiol solution: 0.124 mg/mL of USP Amifostine
S-[2-(3-Aminopropyl)amino]ethyl]dihydrogen phosphorothioate, Thiol RS in water
trihydrate [112901-68-5]. [NOTEInject immediately after preparation, or refrigerate
until use. The solution is stable for 48 h if maintained at
DEFINITION about 5.]
Amifostine contains NLT 78.0% and NMT 82.0% of System suitability solution: 5 mg/mL of USP Amifostine
C5H15N2O3PS, calculated on the as-is basis. RS in Standard thiol solution [NOTEInject immediately after
preparation, or refrigerate until use. The solution is stable
IDENTIFICATION for 12 h if maintained at about 5.]
A. INFRARED ABSORPTION 197K Sample solution: 50 mg/mL of Amifostine in water
B. The retention time of the major peak of the Sample [NOTEInject immediately after preparation, or refrigerate
solution corresponds to that of the Standard solution, as until use. The solution is stable for 48 h if maintained at
obtained in the Assay. about 5.]
ASSAY (See Chromatography 621, System Suitability.)
PROCEDURE Mode: LC
Mobile phase: 1.0 mL of nonafluorobutane sulfonic acid in Detector: UV 220 nm
1200 mL of HPLC grade water. Prepare a mixture of this Column: 4.6-mm 25-cm; packing L1
solution and acetonitrile (90:10). Temperature: 30
Standard solution: 3 mg/mL of USP Amifostine RS in water [NOTEThe temperature of the solutions to be injected is
[NOTEInject immediately after preparation, or refrigerate maintained at 28.]
until use. The solution is stable for 48 h if maintained at Flow rate: 1 mL/min
about 5.] Injection size: 10 L
Sample solution: 3 mg/mL of Amifostine in water [NOTE System suitability
Inject immediately after preparation, or refrigerate until use. Samples: Standard thiol solution and System suitability
The solution is stable for 48 h if maintained at about 5.] solution
Chromatographic system Suitability requirements
(See Chromatography 621, System Suitability.) Resolution: NLT 2.0 between amifostine and amifostine
Mode: LC thiol, System suitability solution
Detector: UV 220 nm Tailing factor: NMT 4.0, System suitability solution and
Column: 4.6-mm 25-cm; 5-m packing L1 Standard solution
Temperature: 30 Capacity factor: More than 0.5, System suitability
[NOTEThe temperature of the solutions to be injected is solution and Standard solution
maintained at 28.] Column efficiency: NLT 2300 theoretical plates
Flow rate: 1 mL/min calculated for the amifostine thiol peak, System suitability
Injection size: 10 L solution and Standard solution
System suitability Relative standard deviation: NMT 4.0%, System
Samples: Standard solution and Sample solution suitability solution and Standard solution
Suitability requirements Analysis
Column efficiency: NLT 7500 theoretical plates, Standard Samples: Blank, Standard thiol solution, and Sample
solution and Sample solution solution
Tailing factor: NMT 2, Standard solution and Sample [NOTEMeasure the responses of all the peaks, excluding
solution the peaks corresponding to those from the Blank.]
Relative standard deviation: NMT 2.0%, Standard Calculate the percentage of amifostine thiol in the portion
solution and Sample solution of Amifostine taken:
Analysis
Samples: Standard solution and Sample solution Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
Calculate the percentage of C5H15N2O3PS in the portion of
Amifostine taken: rU = peak response of amifostine thiol from the
Sample solution
Result = (rU/rS) (CS/CU) 100 rS = peak response of amifostine thiol from the
Standard thiol solution
rU = peak responses from the Sample solution CS = concentration of amifostine thiol
rS = peak responses from the Standard solution dihydrochloride in the Standard thiol solution
CS = concentration of USP Amifostine RS in the (mg/mL)
Standard solution (mg/mL) CU = concentration of Amifostine in the Sample
CU = concentration of Amifostine in the Sample solution (mg/mL)
solution (mg/mL)
174 Amifostine / Official Monographs USP 32
Mr1 = molecular weight of amifostine thiol, 134.24 Flow rate: 1 mL/min

Mr2 =molecular weight of amifostine thiol Injection size: 10 L
dihydrochloride, 207.17 System suitability
Calculate the percentage of each of the other impurities in Samples: Standard solution and Sample solution
the portion of Amifostine taken: Suitability requirements
Column efficiency: NLT 7500 theoretical plates, Standard
Result = (ri/rU) 100 solution and Sample solution
Tailing factor: NMT 2, Standard solution and Sample
ri = peak response of each impurity from the Sample solution
solution Relative standard deviation: NMT 2.0%, Standard
rU = peak response of amifostine from the Sample solution and Sample solution
solution Analysis
Acceptance criteria Samples: Standard solution and Sample solution
Any individual impurity, excluding amifostine thiol: Calculate the percentage C5H15N2O3PS in the portion of
NMT 0.1% Amifostine for Injection taken:
Total impurities including amifostine thiol: NMT 0.3%
SPECIFIC TESTS
X-RAY DIFFRACTION 941: Its X-ray diffraction pattern rU = peak responses from the Sample solution
conforms to that of USP Amifostine RS, similarly determined. rS = peak responses from the Standard solution
PH 791: 6.57.5, in a solution (5 in 100) CS = concentration of USP Amifostine RS in the
WATER DETERMINATION, Method Ic 921 Standard solution (mg/mL)
Sample solution: To 100.0 mg of Amifostine, contained in CU = nominal concentration of the Sample solution
a stoppered centrifuge tube, add 10.0 mL of the solution of (mg/mL)
N-ethylmaleimide in methanol (4 in 100), and sonicate for Acceptance criteria: 90.0%110.0%
15 min. Shake to disperse, and sonicate for an additional 15
min. Use 1.0 mL of the supernatant. PERFORMANCE TESTS
Acceptance criteria: 19.2%21.2% UNIFORMITY OF DOSAGE UNITS 905: Meets the
requirements
PACKAGING AND STORAGE: Preserve in tight, light-resistant IMPURITIES
containers, and store in a refrigerator. Organic Impurities
USP REFERENCE STANDARDS 11 PROCEDURE
USP Amifostine RS Mobile phase: 1.0 mL of nonafluorobutane sulfonic acid
USP Amifostine Thiol RS in 1200 mL of HPLC grade water, add 400 L of
trifluoroacetic acid, and adjust with triethylamine to a pH
of 2.5. Prepare a degassed mixture of this solution and
acetonitrile (68:32).
Amifostine for Injection Blank: Water
(Comment on this Monograph)id=m2603=Amifostine for Standard disulfide solution: 0.186 mg/mL of USP
Injection=A-Monos.pdf) Amifostine Disulfide RS [NOTEInject immediately after
preparation, or refrigerate until use. The solution is stable
DEFINITION for 48 h if maintained at about 5.]
Amifostine for Injection is a sterile, crystalline substance suitable Standard thiol solution: 0.802 mg/mL of USP Amifostine
for parenteral use. It contains NLT 90.0% and NMT 110.0% of Thiol RS [NOTEInject immediately after preparation, or
the labeled amount of amifostine (C5H15N2O3PS). refrigerate until use. The solution is stable for 48 h if
maintained at about 5.]
ASSAY System suitability solution: 5 mg/mL of USP Amifostine
PROCEDURE RS in Standard thiol solution [NOTEInject immediately after
Mobile phase: 1.0 mL of nonafluorobutane sulfonic acid in preparation, or refrigerate until use. The solution is stable
1200 mL of HPLC grade water. Prepare a degassed mixture for 12 h if maintained at about 5.]
of this solution and acetonitrile (90:10). Sample solution: 50 mg/mL of amifostine, from
Standard solution: 3 mg/mL of USP Amifostine RS [NOTE Amifostine for Injection [NOTEInject immediately after
Inject immediately after preparation, or refrigerate until use. preparation, or refrigerate until use. The solution is stable
The solution is stable for 48 h if maintained at about 5.] for 48 h if maintained at about 5.]
Sample stock solution: Equivalent to 10 mg/mL of Chromatographic system
amifostine, from Amifostine for Injection (See Chromatography 621, System Suitability.)
Sample solution: Transfer 6.0 mL of Sample stock solution Mode LC
to a 25-mL volumetric flask, add 6.5 mL of water, and dilute Detector: UV, 220 nm and 247 nm
with methanol to volume. Column: 4.6-mm 25-cm; packing L1
Chromatographic system Temperature: 30
(See Chromatography 621, System Suitability.) [NOTEThe temperature of the solutions to be injected is
Mode: LC maintained at 28.]
Detector: UV 220 nm
Temperature: 30
[NOTEThe temperature of the solutions to be injected is
maintained at 28.]
USP 32 Official Monographs / Amikacin 175
Flow rate: 1 mL/min Acceptance criteria: NMT 0.1% of any individual impurity
Injection size: 10 L except amifostine thiol is found.
System suitability
Samples: Standard disulfide solution, Standard thiol SPECIFIC TESTS
solution, and System suitability solution CONSTITUTED SOLUTION: At the time of use, it meets the
Suitability requirements requirements for Injections 1, Constituted Solutions. When
Capacity factor, k: More than 0.5 for amifostine thiol; constituted with 0.9% Sodium Chloride Injection, the solution
more than 2.2 for amifostine disulfide, Standard disulfide must completely dissolve in 45 s.
solution, Standard thiol solution, and System suitability X-RAY DIFFRACTION 941: Its X-ray diffraction pattern
solution conforms to that of USP Amifostine RS, similarly determined.
Column efficiency: NLT 2300 theoretical plates, for STERILITY TESTS 71: It meets the requirements when tested
amifostine thiol; not more than 2000 for amifostine, for as directed for Test for Sterility of the Product to Be Examined,
amifostine disulfide Membrane Filtration.
Tailing factor: NMT 4.0 for amifostine thiol; NMT 4.5 PH 791: 6.57.5, in a solution constituted as directed in
for amifostine disulfide, Standard disulfide solution, the labeling
Standard thiol solution, and System suitability solution WATER DETERMINATION, Method Ic 921
Relative standard deviation: NMT 4.0% for amifostine Sample solution: To 100.0 mg of Amifostine for Injection,
thiol; NMT 4.0% for amifostine disulfide, System contained in a stoppered centrifuge tube, add 10.0 mL of
suitability solution, Standard disulfide solution, andStandard the solution of N-ethylmaleimide in methanol (4 in 100),
thiol solution and sonicate for 15 min. Shake to disperse, and sonicate for
Analysis an additional 15 min. Use 1.0 mL of the supernatant.
Samples: Blank, Standard disulfide solution, Standard thiol Acceptance criteria: 18.0%22.0%
solution and Sample solution PARTICULATE MATTER IN INJECTIONS 788: Meets the
[NOTEMeasure the responses of all the peaks, excluding requirements for small-volume injections
the peaks corresponding to those from the Blank.] BACTERIAL ENDOTOXINS TEST 85: Contains NMT 0.2 USP
Calculate the percentage of amifostine thiol in the portion Endotoxin Unit/mg of amifostine
of Amifostine for Injection taken: OTHER REQUIREMENTS: Meets the requirements for Labeling
under Injections 1. It also meets the requirements for
Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 Identification under Amifostine.
rU = amifostine thiol peak responses at 220 nm, from ADDITIONAL REQUIREMENTS
the Sample solution PACKAGING AND STORAGE: Preserve in tight Containers for
rS = amifostine thiol peak responses at 220 nm, from Sterile Solids as described under Injections 1, and store at
the Standard thiol solution controlled room temperature.
CS = concentration of amifostine thiol USP REFERENCE STANDARDS 11
dihydrochloride in the Standard thiol solution USP Amifostine RS
(mg/mL) USP Amifostine Disulfide RS
CU = concentration of the Sample solution (mg/mL) USP Amifostine Thiol RS
Mr1 = molecular weight of amifostine thiol, 134.24 USP Endotoxin RS
Mr2 = molecular weight of amifostine thiol
dihydrochloride, 207.17
Calculate the percentage of amifostine disulfide in the
portion of Amifostine for Injection taken: Amikacin
(Comment on this Monograph)id=m2610=Amikacin=A-
Result = (rU/rS) (CS/CU) (Mr3/Mr4) 100 Monos.pdf)
rU = peak responses at 247 nm, from the Sample
solution
rS = peak responses at 247 nm, from the Standard
disulfide solution
CS = concentration of USP Amifostine Disulfide RS in
the Standard disulfide solution (mg/ml)
CU = concentration for the Sample solution (mg/mL)
Mr3 = molecular weight of amifostine disulfide, 266.47 C22H43N5O13 585.60
Mr4 = molecular weight of amifostine disulfide D-Streptamine, O-3-amino-3-deoxy--D-glucopyranosyl-(16)-
tetrahydrochloride, 412.31 O-[6-amino-6-deoxy--D-glucopyranosyl(14)]-N1-(4-
Acceptance criteria: NMT 2.0% of total impurities, amino-2-hydroxy-1-oxobutyl)-2-deoxy-, (S)-;
including amifostine thiol and amifostine disulfide O-3-Amino-3-deoxy--D-glucopyranosyl(14)-O-[6-amino-6-
Calculate the percentage of each of the other impurities in deoxy--D-glucopyranosyl(16)]-N3-(4-amino-L-2-
the portion of Amifostine for Injection taken: hydroxybutyryl)-2-deoxy-L-streptamine [37517-28-5].
Result = (ri/rA) 100 DEFINITION
Amikacin has a potency of NLT 900 g of C22H43N5O13 per mg,
ri = peak response for each impurity from the Sample calculated on the anhydrous basis.
solution
rA = peak response for amifostine from the Sample
solution
176 Amikacin / Official Monographs USP 32
IDENTIFICATION Acceptance criteria: NLT 900 g

A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
Standard solution: 6 mg/mL IMPURITIES
Sample solution: 6 mg/mL Inorganic Impurities
Solution A: Sample solution and the Standard solution (1:1) RESIDUE ON IGNITION 281: NMT 1.0%, the charred residue
Application volume: 3 L being moistened with 2 mL of nitric acid and 5 drops of
Developing solvent system: Methanol, chloroform, and sulfuric acid
ammonium hydroxide (12:5:7)
Spray reagent: 10 mg/mL of ninhydrin in a mixture of SPECIFIC TESTS
butyl alcohol and pyridine (100:1) OPTICAL ROTATION, Specific Rotation 781S
Analysis Sample solution: 20 mg/mL
Samples: Standard solution, Sample solution, and Solution A Acceptance criteria: +97 to +105
Proceed as directed in the chapter, except to develop the CRYSTALLINITY 695: Meets the requirements
chromatogram by continuous flow for 5.5 h. Remove the PH 791: 9.511.5, in a solution containing 10 mg/mL
plate from the chamber, allow the solvent to evaporate, WATER DETERMINATION, Method I 921: NMT 8.5%
and heat the plate at 110 for 15 min. Spray the plate ADDITIONAL REQUIREMENTS
with Spray reagent, and immediately locate the spots. PACKAGING AND STORAGE: Preserve in tight containers.
Acceptance criteria: Amikacin appears as a pink spot, and USP REFERENCE STANDARDS 11
the spots obtained from the Sample solution and Solution A USP Amikacin RS
correspond in distance from the origin to that obtained USP Kanamycin Sulfate RS
from the Standard solution.
B. The retention time of the peak for amikacin of the
Sample solution corresponds to that of the Standard solution,
as obtained in the Assay. Amikacin Sulfate
ASSAY (Comment on this Monograph)id=m2630=Amikacin Sulfate=A-
PROCEDURE Monos.pdf)
Mobile phase: 0.115 N sodium hydroxide
System suitability solution: 0.02 mg/mL of USP Amikacin
RS and 0.008 mg/mL of USP Kanamycin Sulfate RS
Standard solution: 0.02 mg/mL of USP Amikacin RS
Sample solution: 0.02 mg/mL of Amikacin
Mode: LC C22H43N5O13 2H2SO4 781.76
Detector: Electrochemical detector, a gold working D-Streptamine, O-3-amino-3-deoxy--D-glucopyranosyl-(16)-
electrode, and a pH silversilver chloride reference O-[6-amino-6-deoxy--D-glucopyranosyl-(14)]-N1-(4-
electrode amino-2-hydroxy-1-oxobutyl)-2-deoxy-, (S)-, sulfate (1:2)
[NOTEThe electrochemical detector is used in the (salt);
integrated amperometric mode with a range of 300 nC, O-3-Amino-3-deoxy--D-glucopyranosyl-(14)-O-[6-amino-6-
an output of 1 V full scale, a rise time of 0.5 s, positive deoxy--D-glucopyranosyl-(16)]-N3-(4-amino-L-2-
polarity, potential E = 0.04 V; t1 = 200 ms; E2 = 0.8 V; t2 hydroxybutyryl)-2-deoxy-L-streptamine sulfate (1:2)
= 190 ms; E3 = 0.8 V; and t3 = 190 ms.] [39831-55-5; 3517-28-5].
Guard column: Packing L47
Analytical column: 4-mm 25-cm; packing L47 DEFINITION
Flow rate: 0.5 mL/min Amikacin Sulfate having a molar ratio of amikacin to H2SO4 of
Injection size: 20 L 1:2 contains the equivalent of NLT 674 g and NMT 786 g
System suitability of amikacin (C22H43N5O13) per mg, calculated on the dried
Samples: System suitability solution and Standard solution basis; Amikacin Sulfate having a molar ratio of amikacin to
[NOTEThe relative retention times of kanamycin and H2SO4 of 1:1.8 contains the equivalent of NLT 691 g and
amikacin are 0.8 and 1.0, respectively.] NMT 806 g of amikacin (C22H43N5O13) per mg, calculated on
Suitability requirements the dried basis.
Resolution: NLT 3 between kanamycin and amikacin,
System suitability solution IDENTIFICATION
Tailing factor: NMT 2, Standard solution A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
Relative standard deviation: NMT 3%, Standard solution Standard solution: 6 mg/mL in water
Analysis Sample solution: 6 mg/mL in water
Samples: Standard solution and Sample solution Solution A: Sample solution and the Standard solution (1:1)
Calculate the quantity, in g, of C22H43N5O13 in each mg of Application volume: 3 L
Amikacin taken: Developing solvent system: Methanol, ammonium
hydroxide, and chloroform (12:7:5)
Result = (rU/rS) (CS/CU) E Spray reagent: 10 mg/mL of ninhydrin in a mixture of
butyl alcohol and pyridine (100:1)
rU = amikacin peak areas from the Sample solution Analysis
rS = amikacin peak areas from the Standard solution Samples: Standard solution, Sample solution, and Solution A
CS = concentration of USP Amikacin RS in the Proceed as directed in the chapter, except to develop the
Standard solution (mg/mL) chromatogram by continuous flow for 5.5 h. Remove the
CU = concentration for the Sample solution (mg/mL) plate from the chamber, allow the solvent to evaporate,
E = designated amikacin content of USP Amikacin RS
(g/mg)
USP 32 Official Monographs / Amikacin 177
and heat the plate at 110 for 15 min. Spray the plate SPECIFIC TESTS
with Spray reagent, and immediately locate the spots. OPTICAL ROTATION, Specific Rotation 781: +76 to +84
Acceptance criteria: Amikacin appears as a pink spot, and Sample solution: 20 mg/mL, in water
the spots of the Sample solution and Solution A correspond in CRYSTALLINITY 695: Meets the requirements
distance from the origin to that of the Standard solution. PH 791: 2.04.0 (1:2 salt), or 6.07.3 (1:1.8 salt), in a
B. The retention time of the peak for amikacin of the solution containing 10 mg/mL
Sample solution corresponds to that of the Standard solution, LOSS ON DRYING 731: Dry 100 mg, in a vacuum at a
as obtained in the Assay. pressure not exceeding 5 mm of mercury at 110 for 3 h: it
loses NMT 13.0% of its weight.
ASSAY
PROCEDURE ADDITIONAL REQUIREMENTS
Mobile phase: 0.115 N sodium hydroxide PACKAGING AND STORAGE: Preserve in tight containers.
System suitability solution: 0.02 mg/mL of USP Amikacin LABELING: Label it to indicate whether its molar ratio of
RS and 0.008 mg/mL of USP Kanamycin Sulfate RS in water amikacin to H2SO4 is 1:2 or 1:1.8.
Standard solution: 0.02 mg/mL of USP Amikacin RS in USP REFERENCE STANDARDS 11
water USP Amikacin RS
Sample solution: Equivalent to 0.02 mg/mL of amikacin, USP Kanamycin Sulfate RS
from Amikacin Sulfate, in water
Mode: LC Amikacin Sulfate Injection
Detector: Electrochemical detector, a gold working (Comment on this Monograph)id=m2640=Amikacin Sulfate
electrode, and a pH silver-silver chloride reference electrode Injection=A-Monos.pdf)
[NOTEThe electrochemical detector is used in the
integrated amperometric mode with a range of 300 nC, an DEFINITION
output of 1 V full scale, a rise time of 0.5 s, positive Amikacin Sulfate Injection is a sterile solution of Amikacin
polarity, potential E = 0.04 V; t1 = 200 ms; E2 = 0.8 V; t2 Sulfate in Water for Injection, or of Amikacin in Water for
= 190 ms; E3 = 0.8 V; and t3 = 190 ms.] Injection prepared with the aid of Sulfuric Acid. It contains
Column NLT 90.0% and NMT 120.0% of the labeled amount of
Guard column: Packing L47 amikacin (C22H43N5O13).
Analytical column: 4-mm 25-cm; packing L47
Flow rate: 0.5 mL/min IDENTIFICATION
Injection size: 20 L A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
System suitability Standard solution: 6 mg/mL
Samples: System suitability solution and Standard solution Sample solution: 6 mg/mL
[NOTEThe relative retention times for kanamycin and Solution A: Sample solution and the Standard solution (1:1)
amikacin are 0.8 and 1.0, respectively.] Application volume: 3 L
Suitability requirements Developing solvent system: Methanol, chloroform, and
Resolution: NLT 3 between kanamycin and amikacin, ammonium hydroxide (12:5:7)
System suitability solution Spray reagent: 10 mg/mL of ninhydrin in a mixture of
Tailing factor: NMT 2, Standard solution butyl alcohol and pyridine (100:1)
Relative standard deviation: NMT 3%, Standard solution Analysis
Analysis Samples: Standard solution, Sample solution, and Solution A
Samples: Standard solution and Sample solution Proceed as directed in the chapter, except to develop the
Calculate the quantity, in g, of C22H43N5O13 in each mg of chromatogram by continuous flow for 5.5 h. Remove the
Amikacin Sulfate taken: plate from the chamber, allow the solvent to evaporate,
and heat the plate at 110 for 15 min. Spray the plate
Result = (rU/rS) (CS/CU) E with Spray reagent, and immediately locate the spots.
Acceptance criteria: Amikacin appears as a pink spot, and
rU = amikacin peak areas from the Sample solution the spots obtained from the Sample solution and the Solution
rS = amikacin peak areas from the Standard solution A correspond in distance from the origin to that obtained
CS = concentration of USP Amikacin RS in the from the Standard solution.
Standard solution (mg/mL) B. The retention time of the peak for amikacin of the
CU = concentration for the Sample solution Sample solution corresponds to that of the Standard solution,
(mg/mL) as obtained in the Assay.
E = designated amikacin content of USP Amikacin RS
(g/mg) ASSAY
Acceptance criteria: Amikacin Sulfate having a molar ratio PROCEDURE
of amikacin to H2SO4 of 1:2 contains the equivalent of NLT Mobile phase: 0.115 N sodium hydroxide
674 g and NMT 786 g of C22H43N5O13 per mg; Amikacin System suitability solution: 0.02 mg/mL of USP Amikacin
Sulfate having a molar ratio of amikacin to H2SO4 of 1:1.8 RS and 0.008 mg/mL of USP Kanamycin Sulfate RS
contains the equivalent of NLT 691 g and NMT 806 g of Standard solution: 0.02 mg/mL of USP Amikacin RS
C22H43N5O13 per mg Sample solution: 0.02 mg/mL of amikacin, from the
Injection
IMPURITIES Chromatographic system
Inorganic Impurities (See Chromatography 621, System Suitability.)
RESIDUE ON IGNITION 281: NMT 1.0%, the charred residue
being moistened with 2 mL of nitric acid and 5 drops of
sulfuric acid
178 Amikacin / Official Monographs USP 32
Mode: LC Amiloride Hydrochloride

Detector: Electrochemical detector, a gold working (Comment on this Monograph)id=m2650=Amiloride
electrode, and a pH silversilver chloride reference Hydrochloride=A-Monos.pdf)
electrode
[NOTEThe electrochemical detector is used in the
integrated amperometric mode with a range of 300 nC,
an output of 1 V full scale, a rise time of 0.5 s, positive
polarity, potential E = 0.04 V; t1 = 200 ms; E2 = 0.8 V; t2
= 190 ms; E3 = 0.8 V; and t3 = 190 ms.]
Column
Guard column: Packing L47
Analytical column: 4-mm 25-cm; packing L47 C6H8ClN7O HCl 2H2O 302.12
Flow rate: 0.5 mL/min Pyrazinecarboxamide, 3,5-diamino-N-(aminoiminomethyl)-6-
Injection size: 20 L chloro-, monohydrochloride dihydrate;
System suitability N-Amidino-3,5-diamino-6-chloropyrazinecarboxamide
Samples: System suitability solution and Standard solution monohydrochloride dihydrate [17440-83-4].
[NOTEThe relative retention times of kanamycin and
amikacin are 0.8 and 1.0, respectively.] DEFINITION
Suitability requirements Amiloride Hydrochloride contains NLT 98.0% and NMT 101.0%
Resolution: NLT 3 between kanamycin and amikacin, of C6H8ClN7O HCl, calculated on the dried basis.
System suitability solution
Tailing factor: NMT 2, Standard solution IDENTIFICATION
Relative standard deviation: NMT 3%, Standard solution A. INFRARED ABSORPTION 197M
Analysis B. ULTRAVIOLET ABSORPTION 197U
Samples: Standard solution and Sample solution Sample solution: 600 g/mL of water, diluted
Calculate the percentage of C22H43N5O13 in each mL of the quantitatively and stepwise with 0.1 N hydrochloric acid to a
Injection taken: concentration of about 9.6 g/mL
C. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets
Result = (rU/rS) (CS/CU) E F100 the requirements
rU = amikacin peak areas from the Sample solution ASSAY

rS = amikacin peak areas from the Standard solution PROCEDURE
CS = concentration of USP Amikacin RS in the Sample: 450 mg
Standard solution (mg/mL) Analysis: Dissolve in 100 mL of glacial acetic acid, add 10
CU = nominal concentration of amikacin in the Sample mL of mercuric acetate TS and 15 mL of dioxane. Add
solution crystal violet TS. Titrate with 0.1 N perchloric acid VS to a
E = designated amikacin content of USP Amikacin RS blue endpoint. Perform a blank determination (See Titrimetry
(g/mg) 541). Each mL of 0.1 N perchloric acid is equivalent to
F = unit conversion factor, 0.001 mg/g 26.61 mg of C6H8ClN7O HCl.
Acceptance criteria: 90.0%120.0% Acceptance criteria: 98.0%101.0%
SPECIFIC TESTS IMPURITIES
PH 791: 3.55.5 Inorganic Impurities
PARTICULATE MATTER IN INJECTIONS 788: Meets the RESIDUE ON IGNITION 281: NMT 0.1%
requirements for small-volume injections HEAVY METALS, Method II 231: NMT 20 ppm
BACTERIAL ENDOTOXINS TEST 85: NMT 0.33 USP Endotoxin Organic Impurities
Unit/mg of amikacin PROCEDURE
OTHER REQUIREMENTS: Meets the requirements under Standard solutions: Prepare a series of solutions, A, B, C,
Injections 1 D, E, and F, of USP Amiloride Hydrochloride RS in a mixture
of methanol and chloroform (4:1) having concentrations of
ADDITIONAL REQUIREMENTS 4000, 40, 20, 8, 4, and 2 g/mL, respectively.
PACKAGING AND STORAGE: Preserve in single-dose or in Sample solution: 4 mg/mL of Amiloride Hydrochloride in
multiple-dose containers, preferably of Type I or Type III methanol and chloroform (4:1)
glass. Chromatographic system
USP REFERENCE STANDARDS 11 (See Chromatography 621, Thin-Layer Chormatography.
USP Amikacin RS
USP Endotoxin RS
USP Kanamycin Sulfate RS
USP 32 Official Monographs / Amiloride 179
Mode: TLC Standard solution: 0.2 mg/mL of USP Amiloride

Adsorbent: 0.25-mm layer of chromatographic silica gel Hydrochloride RS in methanol
previously washed with methanol Chromatographic system
Application volume: 5 L (See Chromatography 621, Thin-Layer Chromatography)
Developing solvent system: Tetrahydrofuran and 3 N Mode: TLC
ammonium hydroxide (15:2) Adsorbent: 0.25-mm layer of chromatographic silica gel
Analysis mixture
Samples: Standard solutions A, B, C, D, E, and F and the Developing solvent system: Tetrahydrofuran and 3 N
Sample solution ammonium hydroxide (22:3)
Proceed as directed under General Chapter. Dry the spots Application volume: 10 L
with a stream of nitrogen, and develop the Analysis
chromatograms in the solvent system, until the solvent Samples: Sample solution and Standard solution
front has moved about three-fourths of the length of the Remove the plate from the developing chamber, air-dry,
plate. Remove the plate from the developing chamber, and examine under short-wavelength UV light.
mark the solvent front, allow to air-dry, and examine the Acceptance criteria: The RF value of the principal spot of
plate under long-wavelength UV light: the RF value of the the Sample solution corresponds to that of the Standard
principal spot of the Sample solution corresponds to that solution.
of Standard solution A. Estimate the levels of any
additional spots observed in the chromatogram of the ASSAY
Sample solution by comparison with the principal spots in PROCEDURE
the chromatograms of Standard solutions B, C, D, E, and Mobile phase: Methanol, water, and Buffer solution
F. (25:71:4)
Acceptance criteria: The sum of the intensities of any Solution A: 136 g of monobasic potassium phosphate in
additional spots observed is NMT that of the principal spot 800 mL of water. Adjust by the addition of phosphoric acid,
obtained from Standard solution B (equivalent to 1%). with mixing, to a pH of 3.0. Dilute with water to 1000 mL.
Standard stock solution: 1.0 mg/mL of amiloride
SPECIFIC TESTS hydrochloride, from USP Amiloride Hydrochloride RS in
ACIDITY methanol
Sample: 1.0 g Standard solution: 5.0 mL of the Standard stock solution to
Analysis: Dissolve in 100 mL of a mixture of methanol and a 50-mL volumetric flask. Add 10 mL of methanol, 2.0 mL
water (1:1). Titrate with 0.10 N sodium hydroxide. of 0.1 N hydrochloric acid, and dilute with water to volume.
Acceptance criteria: NMT 0.30 mL is required (0.1% as The concentration of USP Amiloride Hydrochloride RS in the
HCl) Standard solution is about 0.1 mg/mL.
LOSS ON DRYING Sample solution: Transfer an equivalent to 5 mg of
(See Thermal Analysis 891) amiloride hydrochloride, from finely powdered Tablets (NLT
[NOTEThe quantity taken for the determination may be 20), to a 50-mL volumetric flask containing 15.0 mL of
adjusted, if necessary, for instrument sensitivity.] methanol and 2.0 mL of 0.1 N hydrochloric acid. Sonicate
Sample: 10 mg of Amiloride Hydrochloride for 10 min, dilute with water to volume, sonicate for an
Analysis: Determine the percentage of volatile substances additional 10 min, and filter.
by thermogravimetric analysis on an appropriately calibrated Chromatographic system
instrument, using the Sample. Heat the specimen at the rate (See Chromatography 621, System Suitability.)
of 10/min between ambient temperature and 225 in an Mode: LC
atmosphere of nitrogen at a flow rate of 40 mL/min. From Detector: UV 286 nm
the thermogram determine the accumulated loss in weight Column: 3.9-mm 30-cm; packing L1
between ambient temperature and about 200 on the Flow rate: 1 mL/min
plateau. Injection size: 10 L
Acceptance criteria: It loses NLT 11.0% and NMT 13.0% System suitability
of its weight. Sample: Standard solution (five replicate injections)
ADDITIONAL REQUIREMENTS Tailing factor: NMT 2.0
PACKAGING AND STORAGE: Preserve in well-closed containers. Relative standard deviation: NMT 2.0%
USP REFERENCE STANDARDS 11 Analysis
USP Amiloride Hydrochloride RS Samples: Standard solution and Sample solution
Calculate the percentage of C6H8ClN7O HCl in the portion
of Tablets taken:
Amiloride Hydrochloride Tablets Result = (rU/rS) (CS/CU) 100
(Comment on this Monograph)id=m2660=Amiloride
Hydrochloride Tablets=A-Monos.pdf) rU = peak response from the Sample solution
DEFINITION CS = concentration of USP Amiloride Hydrochloride RS
Amiloride Hydrochloride Tablets contain NLT 90.0% and NMT (mg/mL)
110.0% of the labeled amount of C6H8ClN7O HCl. CU = nominal concentration of amiloride
hydrochloride in the Sample solution (mg/mL)
IDENTIFICATION
A. The retention time of the major peak of the Sample
solution corresponds to that of the Standard solution, as
B. Thin-Layer Chromatography 201
Sample solution: Equivalent to 0.2 mg/mL of amiloride
hydrochloride in methanol, from finely ground Tablets.
Filter.
180 Amiloride / Official Monographs USP 32
Acceptance criteria: 90.0%110.0% IDENTIFICATION

A. The retention times of the major peaks of the Sample
PERFORMANCE TESTS solution correspond to those of the Standard solution, as
DISSOLUTION 711 obtained in the Assay.
Medium: 0.1 N hydrochloric acid; 900 mL B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
Apparatus 2: 50 rpm Standard solution A: 0.2 mg/mL of USP Amiloride
Time: 30 min Hydrochloride RS in methanol
Detector: UV 363 nm Standard solution B: 2 mg/mL of USP Hydrochlorothiazide
Sample solutions: Sample per Dissolution 711. Dilute with RS in methanol
Medium as necessary. Sample solution: Equivalent to 0.2 mg/mL of amiloride
Standard solution: USP Amiloride Hydrochloride RS of hydrochloride, from ground Tablets in methanol and filter
known concentration in Medium Chromatographic system
[NOTEAn amount of methanol not to exceed 2% of the (See Chromatography 621, Thin-Layer Chromatography.)
total volume of the Standard solution may be used to Mode: TLC
dissolve the amiloride hydrochloride.] Adsorbent: 0.25-mm layer of chromatographic silica gel
Analysis mixture
Samples: Sample solution and Standard solution Application volume: 10 L
Tolerances: NLT 80% (Q) of the labeled amount of Developing solvent system: Tetrahydrofuran and 3 N
C6H8ClN7O HCl is dissolved. ammonium hydroxide (22:3)
UNIFORMITY OF DOSAGE UNITS 905 Analysis
Procedure for content uniformity Samples: Standard solution A, Standard solution B, and
Sample solution: Transfer 1 finely powdered Tablet to a Sample solution
100-mL volumetric flask, add 60 mL of 0.1 N hydrochloric Develop the chromatogram until the solvent has moved
acid, and shake by mechanical means for 30 min. Dilute about three-fourths of the length of the plate. Remove the
with 0.1 N hydrochloric acid to volume and centrifuge a plate from the developing chamber, air-dry, and examine
portion of the mixture. Dilute an accurately measured under short-wavelength UV light.
portion of the clear supernatant quantitatively to obtain a Acceptance criteria: The RF values of the amiloride
solution containing 10 g/mL of amiloride hydrochloride. hydrochloride and hydrochlorothiazide spots of the Sample
Standard solution: 10 g/mL of USP Amiloride solution correspond to those of the corresponding Standard
Hydrochloride RS in the same medium solutions.
Analytical wavelength: 363 nm ASSAY
Blank: 0.1 N hydrochloric acid PROCEDURE
Analysis Solution A: 136 g of monobasic potassium phosphate in
Samples: Sample solution and Standard solution 800 mL of water, and adjust with phosphoric acid to a pH
Calculate the percentage of C6H8ClN7O HCl in the Tablet of 3.0. Dilute with water to 1000 mL.
taken: Mobile phase: Methanol, water, and Solution A (25:71:4)
Standard stock solution: 1.0 mg/mL of USP Amiloride
Result = (AU/AS) (CS/CU) F (100/L) Hydrochloride RS in methanol
Standard solution: 0.1 mg/mL of USP Amiloride
AU = absorbance of the Sample solution Hydrochloride RS and 1 mg/mL USP Hydrochlorothiazide RS,
AS = absorbance of the Standard solution prepared by transfering 10.0 mL of Standard stock solution
CS = concentration of USP Amiloride Hydrochloride RS to a 100-mL volumetric flask containing 100 mg of USP
(g/mL) Hydrochlorothiazide RS and 20.0 mL of methanol. Add 4.0
CU = concentration of the Sample solution (unit/mL) mL of 1 N hydrochloric acid, and dilute with water to
(unit=Tablet) volume.
F = conversion factor (g to mg) Sample solution: Transfer an equivalent to 5 mg of
L = labeled quantity (mg) of amiloride hydrochloride amiloride hydrochloridefrom powdered Tablets (NLT 20) to
in the Tablet a 50-mL volumetric flask. Add 15.0 mL of methanol, and 2.0
Acceptance criteria: Meet the requriements mL of 1 N hydrochloric acid. Sonicate for 10 min, dilute
with water to volume, sonicate for an additional 10 min,
ADDITIONAL REQUIREMENTS and filter.
PACKAGING AND STORAGE: Preserve in well-closed containers. Chromatographic system
USP Amiloride Hydrochloride RS Mode: LC
Detector: UV 286 nm
Amiloride Hydrochloride and Flow rate: 1 mL/min
Hydrochlorothiazide Tablets Injection size: 10 L
System suitability
(Comment on this Monograph)id=m2665=Amiloride Sample: Standard solution
Hydrochloride and Hydrochlorothiazide Tablets=A-Monos.pdf) [NOTEThe relative retention times for hydrochlorothiazide
and amiloride hydrochloride are about 0.7 and 1.0,
DEFINITION respectively.]
Amiloride Hydrochloride and Hydrochlorothiazide Tablets Suitability requirements
contain NLT 90.0% and NMT 110.0% of the labeled amounts Resolution: NLT 2.0 between hydrochlorothiazide and
of amiloride hydrochloride (C6H8ClN7O HCl) and amiloride hydrochloride
hydrochlorothiazide (C7H8ClN3O4S2).
USP 32 Official Monographs / Amiloride 181
Relative standard deviation: NMT 2.0% AS = absorbance of the Amiloride standard solution
Analysis LC = tablet label claim of amiloride (mg)
Samples: Standard solution and Sample solution Correction for the interference of amiloride is made using
Calculate the percentage of C6H8ClN7O HCl in the portion the following equation:
of Tablets taken:
rU = peak response for amiloride hydrochloride from
the Sample solution AUC = corrected absorbance of Sample solution A, 270
rS = peak response for amiloride hydrochloride from nm
the Standard solution AU270 = absorbance of Sample solution B, 270 nm
CS = concentration of USP Amiloride Hydrochloride AU363 = absorbance of Sample solution A, 363 nm
RS, corrected for loss in weight in the Standard
solution (mg/mL)
CU = nominal concentration of amiloide hydrochloride
Calculate the percentage of C7H8ClN3O4S2 in the portion of ASAmiloride= absorbance of the Amiloride standard solution
Tablets taken: Calculate the amount of C7H8ClN3O4S2 dissolved, in
percentage:
rU = peak response of hydrochlorothiazide from the
Sample solution
rS = peak response of hydrochlorothiazide from the
Standard solution
CS = concentration of USP Hydrochlorothiazide RS in AUC = corrected absorbance of Sample solution A, 270
the Standard solution (mg/mL) nm
CU = nominal concentration of hydrochlorothiazide in CS = concentration of the Hydrochlorothiazide standard
the Sample solution (mg/mL) solution (mg/mL)
Acceptance criteria: 90.0%110.0% of the labeled 900 = volume of Medium (mL)
amounts of C6H8ClN7O HCl and C7H8ClN3O4S2 (25/5) = dilution factor of Sample solution B
100 = conversion factor to percentage
PERFORMANCE TESTS AS = absorbance of the Hydrochlorothiazide standard
DISSOLUTION 711 solution
Medium: 0.1 N hydrochloric acid; 900 mL LC = tablet label claim of hydrochlorothiazide (mg)
Apparatus 2: 50 rpm Tolerances: NLT 80% (Q) of the labeled amount of
Time: 30 min C6H8ClN7O HCl and NLT 75% (Q) of the labeled amount of
Amiloride standard solution: 60 mg of USP Amiloride C7H8ClN3O4S2 is dissolved.
Hydrochloride RS (equivalent to 52 mg of anhydrous UNIFORMITY OF DOSAGE UNITS, Content Uniformity 905:
amiloride hydrochloride) in a 200-mL volumetric flask. Meet the requirements with respect to amiloride
Dissolve in and dilute with methanol to volume. Transfer 2.0 hydrochloride and hydrochlorothiazide
mL of this solution to a 100-mL volumetric flask, and dilute
with Medium to volume. IMPURITIES
Hydrochlorothiazide standard solution: Transfer 100 mg Organic Impurities
of USP Hydrochlorothiazide RS to a 100-mL volumetric flask. PROCEDURE
Dissolve in and dilute with methanol to volume. Transfer 5.0 Solution A, Mobile phase, and Sample solution: Proceed
mL of this solution to a 100-mL volumetric flask, and dilute as directed in the Assay.
with Medium to volume. Transfer 10.0 mL of the resulting Standard solution: 10 g/mL of USP Benzothiadiazine
solution to a 50-mL volumetric flask, and dilute with Medium Related Compound A RS in the Mobile phase
to volume. Chromatographic system
Sample solution A: Pass a portion of the solution under (See Chromatography 621, System Suitability.)
test through a 0.45-m glass fiber filter. Mode: LC
Sample solution B: Transfer 5.0 mL of Sample solution A to Detector: UV 286 nm
a 25-mL volumetric flask, and dilute with Medium to volume. Column: 3.9-mm 30-cm; packing L1
Detector: UV 363 nm for amiloride hydrochloride, 270 nm Flow rate: 1 mL/min
for hydrochlorothiazide Injection size: 10 L
Blank: Medium System suitability
Analysis Sample: Standard solution
Samples: Amiloride standard solution, Hydrochlorothiazide [NOTEThe relative retention times for
standard solution, Sample solution A, and Sample solution B hydrochlorothiazide and amiloride hydrochloride are
Calculate the percentage of C6H8ClN7O HCl dissolved: about 0.7 and 1.0, respectively.]
Resolution: NLT 2.0 between hydrochlorothiazide and
amiloride hydrochloride
Analysis
Samples: Sample solution and Standard solution
AU = absorbance of Sample solution A Calculate the percentage of benzothiadiazine related
CS = concentration of the Amiloride standard solution compound A in the portion of Tablets taken:
(mg/mL)
900 = volume of Medium (mL) Result = (rU/rS) (CS/CU) F 100
100 = conversion factor to percentage
182 Amiloride / Official Monographs USP 32
rU = peak response of benzothiadiazine related Carrier gas: Helium

compound A from the Sample solution Flow rate: 6 mL/min
rS = peak response of benzothiadiazine related Injection size: 1 L
compound A from the Standard solution System suitability
CS = concentration of USP Benzothiadiazine Related Sample: Standard solution
Compound A RS in the Standard solution Suitability requirements
(g/mL) Resolution: NLT 1.0 between the amiloxate peak and any
CU = nominal concentration of benzothiadiazine in other peak
the Sample solution (mg/mL) Relative standard deviation: NMT 2.0%
F = unit conversion factor, 0.001 mg/g Analysis
Acceptance criteria: NMT 1.0% Samples: Standard solution and Sample solution
ADDITIONAL REQUIREMENTS Amiloxate taken:
USP REFERENCE STANDARDS 11 Result = (rU/rS) (CS/CU) 100
USP Amiloride Hydrochloride RS
USP Benzothiadiazine Related Compound A RS rU = peak response from the Sample solution
USP Hydrochlorothiazide RS rS = peak response from the Standard solution
CS = concentration of USP Amiloxate RS in the
Amiloxate Acceptance criteria: 95.0%105.0%
(Comment on this Monograph)id=m42600=Amiloxate=A-
Monos.pdf) IMPURITIES
Organic Impurities
PROCEDURE
Sample solution: Use the Sample solution as obtained in
the Assay.
Assay.
Injection size: 1 L
System suitability
C15H20O3 248.32 Sample: Use the Standard solution prepared as directed in
4-Methoxycinnamic acid, isoamyl ester; the Assay.
2-propenoic acid, 3-(4-methoxyphenyl)-3-methylbutyl ester Analysis
[71617-10-2]. Sample: Sample solution
DEFINITION of Amiloxate taken:
Amiloxate contains NLT 95.0% and NMT 105.0% of C15H20O3.
Result = (ri/rT) 100
IDENTIFICATION
A. INFRARED ABSORPTION 197F ri = peak response for each impurity
B. ULTRAVIOLET ABSORPTION 197U rT = sum of the responses of all the peaks
Sample solution: 5.0 g/mL in alcohol Acceptance criteria
Acceptance criteria: Absorptivities, calculated on the as-is Individual impurities: NMT 0.1%
basis, do not differ by more than 3.0%. Total impurities: NMT 2.0%
PROCEDURE SPECIFIC GRAVITY 841: 1.0371.041
Standard solution: 20 mg/mL of USP Amiloxate RS in tert- REFRACTIVE INDEX 831: 1.5561.560 at 20
butyl methyl ether ACIDITY
Sample solution: 20 mg/mL of Amiloxate in acetone Sample solution: Transfer 50 mL of alcohol to a suitable
Chromatographic system container. Add 1 mL of phenolphthalein TS and sufficient
(See Chromatography 621, System Suitability.) 0.1 N sodium hydroxide to obtain a persistent pink color.
Mode: GC Transfer 50 mL of this solution to a suitable container, and
Detector: Flame ionization add 5.0 mL of Amiloxate.
Column: 0.32-mm 25-m; coated with a 0.1-m film of Analysis: Titrate with 0.1 N sodium hydroxide.
G1 Acceptance criteria: NMT 0.2 mL of titrant/mL of
Temperature: See the temperature program table below. Amiloxate is required for neutralization.
Time ADDITIONAL REQUIREMENTS

(min) Temperature PACKAGING AND STORAGE: Preserve in tight containers.
Injection port 240 USP Amiloxate RS
Detector 260
Column 0 60
22.5 240
32.5 240
USP 32 Official Monographs / Aminobenzoate 183
Aminobenzoate Potassium Concomitantly determine the absorbances of the

(Comment on this Monograph)id=m2780=Aminobenzoate solutions.
Potassium=A-Monos.pdf) Acceptance criteria: The absorbance of the solution
obtained from the Sample solution does not exceed that of
DEFINITION the solution obtained from the Standard solution,
Aminobenzoate Potassium contains NLT 98.5% and NMT corresponding to NMT 0.002% of volatile diazotizable
101.0% of C7H6KNO2, calculated on the dried basis. substances, as p-toluidine.
IDENTIFICATION SPECIFIC TESTS

A. ULTRAVIOLET ABSORPTION 197U PH 791: 8.09.0, in a solution (1 in 20)
Solution: 10 g/mL in 0.001 N sodium hydroxide LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT
B. Dissolve 400 mg in 10 mL of water, add 1 mL of 3 N 1.0% of its weight.
hydrochloric acid, filter, and wash the precipitate with two
5-mL portions of cold water. Recrystallize from alcohol the ADDITIONAL REQUIREMENTS
precipitate so obtained, and dry at 110 for 1 h. The p- PACKAGING AND STORAGE: Preserve in well-closed containers.
aminobenzoic acid so obtained melts between 186 and USP REFERENCE STANDARDS 11
189. USP Aminobenzoate Potassium RS
C. A solution (1 in 100) meets the requirements of the
flame test under Identification TestsGeneral 191,
Potassium. Aminobenzoate Potassium Capsules
ASSAY (Comment on this Monograph)id=m2783=Aminobenzoate
PROCEDURE Potassium Capsules=A-Monos.pdf)
Sample: 500 mg
Analysis: Add 25 mL of water and 25 mL of 3 N DEFINITION
hydrochloric acid, mix and cool in an ice bath. Titrate with Aminobenzoate Potassium Capsules contain NLT 90.0% and
0.1 M sodium nitrite VS, using a calomel-platinum electrode NMT 110.0% of the labeled amount of aminobenzoate
system. Each mL of 0.1 M sodium nitrite is equivalent to potassium (C7H6KNO2).
17.52 mg of C7H6KNO2. IDENTIFICATION
Acceptance criteria: 98.5%101.0% Dissolve 1 g of the Capsule contents in 25 mL of water, add
IMPURITIES 5 mL of 3 N hydrochloric acid, and wash the precipitate
Inorganic Impurities with two 5-mL portions of cold water. Recrystallize from
CHLORIDE AND SULFATE, Chloride 221: A 1.4-g portion alcohol the precipitate so obtained, and dry at 110 for 1 h:
shows no more chloride than corresponds to 0.4 mL of the p-aminobenzoic acid so obtained melts between 186
0.020 N hydrochloric acid (0.02%). and 189.
CHLORIDE AND SULFATE, Sulfate 221: A 1.4-g portion shows ASSAY
no more sulfate than corresponds to 0.3 mL of 0.020 N PROCEDURE
sulfuric acid (0.02%). Standard solution: 5 g/mL of USP Aminobenzoate
HEAVY METALS, Method II 231: NMT 20 ppm Potassium RS
Organic Impurities Sample solution: Remove as completely as possible, and
PROCEDURE: VOLATILE DIAZOTIZABLE SUBSTANCES combine, the contents of NLT 20 Capsules. Transfer a
Standard solution: Dissolve 10 mg of p-toluidine in 5 mL portion of the combined contents, equivalent to 100 mg of
of methanol in a 100-mL volumetric flask, and add water aminobenzoate potassium, to a 200-mL volumetric flask,
to volume. Transfer 1 mL to a 100-mL volumetric flask, and add 150 mL of water, shake by mechanical means for 30
dilute with water to volume. min, dilute with water to volume, and filter. Pipet 2 mL of
Sample solution: Transfer 5.0 g of Aminobenzoate the filtrate into a 200-mL volumetric flask, and dilute with
Potassium to a suitable flask, and add a volume of 1.25 N water to volume. Concomitantly determine the absorbances
sodium hydroxide that is just sufficient to dissolve the of the Standard solution and Sample solution.
sample and to render the solution just alkaline to Spectrometric conditions
phenolphthalein TS. Dilute with water to 50 mL, and Mode: UV
steam-distill the solution, collecting 95 mL of the distillate Analytical wavelength: 270 nm
in a 100-mL volumetric flask. Add water to volume. Blank: Water
Spectrometric conditions Analysis
Mode: UV-Vis Samples: Standard solution and Sample solution
Analytical wavelength: 405 nm Calculate the percentage of C7H6KNO2 in the portion of
Analysis Capsule contents taken:
Transfer 20.0-mL portions of the Samples to separate 100- Result = (AU/AS) (CS/CU) F 100
mL beakers, and transfer 20.0 mL of water to a third
100-mL beaker to provide the blank. Treat each as AU = absorbance of the Sample solution
follows. Add 5.0 mL of 1 N hydrochloric acid, and cool AS = absorbance of the Standard solution
in an ice bath. Add 2.0 mL of 0.1 M sodium nitrite CS = concentration of USP Aminobenzoate Potassium
dropwise, with stirring, allow to stand for 5 min for the RS in the Standard solution (g/mL)
diazotization reaction to be complete, add quickly to CU = nominal concentration of aminobenzoate
10.0 mL of a cold solution of guaiacol (freshly prepared potassium in the Sample solution (mg/mL)
by dissolving 0.20 g of guaiacol in 100 mL of 1 N F = unit conversion factor, 0.001 mg/g
sodium hydroxide), and allow to stand for 30 min.
184 Aminobenzoate / Official Monographs USP 32
Acceptance criteria: 90.0%110.0% Aminobenzoate Potassium Tablets

PERFORMANCE TESTS (Comment on this Monograph)id=m2790=Aminobenzoate
DISSOLUTION 711 Potassium Tablets=A-Monos.pdf)
Medium: Water; 900 mL DEFINITION
Apparatus 1: 100 rpm Aminobenzoate Potassium Tablets contain NLT 90.0% and NMT
Time: 45 min 110.0% of the labeled amount of aminobenzoate potassium
Detector: UV 270 nm (C7H6KNO2).
Standard solution: USP Aminobenzoate Potassium RS in
Medium IDENTIFICATION
Sample solution: Sample per Dissolution 711. Dilute with PROCEDURE
Medium to a concentration that is similar to the Standard Analysis: Dissolve 1 g of finely powdered Tablets in 25 mL
solution. of water, add 5 mL of 3 N hydrochloric acid, and wash the
Tolerances: NLT 75% (Q) of C7H6KNO2 is dissolved. precipitate with two 5-mL portions of cold water.
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Recrystallize from alcohol the precipitate so obtained, and
dry at 110 for 1 h.
ADDITIONAL REQUIREMENTS Acceptance criteria: The p-aminobenzoic acid so obtained
PACKAGING AND STORAGE: Preserve in well-closed containers. melts between 186 and 189.
USP Aminobenzoate Potassium RS ASSAY
PROCEDURE
Standard solution: 5 g/mL of USP Aminobenzoate
Potassium RS
Aminobenzoate Potassium for Oral Sample solution: Transfer an equivalent to 100 mg of
Solution aminobenzoate potassium, from finley powdered tablets
(Comment on this Monograph)id=m2787=Aminobenzoate (NLT 20), to a 200-mL volumetric flask, add 150 mL of
Potassium for Oral Solution=A-Monos.pdf) water, shake by mechanical means for 30 min, dilute with
water to volume and filter. Pipet 2 mL of the filtrate into a
DEFINITION 200-mL volumetric flask, and dilute with water to volume.
Aminobenzoate Potassium for Oral Solution contains NLT Spectrometric conditions
90.0% and NMT 110.0% of the labeled amount of Analytical wavelength: 270 nm
aminobenzoate potassium (C7H6KNO2). Blank: Water
Analysis
IDENTIFICATION Samples: Standard solution and Sample solution
A. ULTRAVIOLET ABSORPTION 197U Calculate the percentage of C7H6KNO2 in the portion of
Sample Solution: 50 g/mL in water Capsule contents taken:
B. Dissolve 400 mg in 10 mL of water, add 1 mL of 3 N
hydrochloric acid, filter, and wash the precipitate with two Result = (AU/AS) (CS/CU) 100
5-mL portions of cold water. Recrystallize from alcohol the
precipitate so obtained, and dry at 110 for 1 h: the p- AU = absorbance of the Sample solution
aminobenzoic acid so obtained melts between 186 and AS = absorbance of the Standard solution
189. CS = concentration of USP Aminobenzoate Potassium
RS in the Standard solution (g/mL)
ASSAY CU = nominal concentration of aminobenzoate
PROCEDURE potassium for the Sample solution (g/mL)
Sample: 100 mg Acceptance criteria: 90.0%110.0%
Analysis: Add 5 mL of hydrochloric acid and 50 mL of
water, cool to 15, and add 25 g of crushed ice. PERFORMANCE TESTS
Titrate with 0.1 M sodium nitrite VS, using a DISSOLUTION 711
calomelplatinum electrode system. Each mL of 0.1 M Medium: Water; 900 mL
sodium nitrite is equivalent to 17.52 mg of C7H6KNO2. Apparatus 1: 100 rpm
Acceptance criteria: 90.0%110.0% Time: 45 min
Detector: UV 270 nm
PERFORMANCE TESTS Standard solution: USP Aminobenzoate Potassium RS in
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements Medium
for a solid packaged in single-unit containers Sample solution: Sample per Dissolution 711. Dilute with
MINIMUM FILL 755: Meets the requirements for a solid Medium to a concentration that is similar to the Standard
packaged in multiple-unit containers solution.
SPECIFIC TESTS Tolerances: NLT 75% (Q) of the labeled amount of
PH 791: 7.09.0, in a solution (1 in 10) C7H6KNO2 is dissolved.
USP Aminobenzoate Potassium RS
USP 32 Official Monographs / Aminobenzoic 185
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements 20.0 mL of water to a third 100-mL beaker to provide
the blank. Treat each as follows. Add 5.0 mL of 1 N
ADDITIONAL REQUIREMENTS hydrochloric acid, and cool in an ice bath. Add 2.0 mL
PACKAGING AND STORAGE: Preserve in well-closed containers. of 0.1 M sodium nitrite dropwise, with stirring, allow to
USP REFERENCE STANDARDS 11 stand for 5 min for the diazotization reaction to be
USP Aminobenzoate Potassium RS complete, add quickly to 10.0 mL of a cold solution of
guaiacol (freshly prepared by dissolving 0.20 g of
guaiacol in 100 mL of 1 N sodium hydroxide), and allow
Aminobenzoate Sodium to stand for 30 min.
Acceptance criteria: The absorbance of the solution
(Comment on this Monograph)id=m2795=Aminobenzoate obtained from the Sample solution does not exceed that of
Sodium=A-Monos.pdf) the solution obtained from the Standard solution,
corresponding to NMT 0.002% of volatile diazotizable
DEFINITION substances, as p-toluidine.
Aminobenzoate Sodium contains NLT 98.5% and NMT 101.0%
of C7H6NNaO2, calculated on the dried basis. SPECIFIC TESTS
IDENTIFICATION LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT
A. ULTRAVIOLET ABSORPTION 197U 1.0% of its weight.
Sample solution: 10 g/mL in 0.001 N sodium hydroxide
B. Dissolve 400 mg in 10 mL of water, add 1 mL of 3 N ADDITIONAL REQUIREMENTS
hydrochloric acid, filter, and wash the precipitate with two PACKAGING AND STORAGE: Preserve in well-closed containers.
5-mL portions of cold water. Recrystallize from alcohol the USP REFERENCE STANDARDS 11
precipitate so obtained, and dry at 110 for 1 h: the p- USP Aminobenzoate Sodium RS
aminobenzoic acid so obtained melts between 186 and
189.
C. IDENTIFICATION TESTSGENERAL, Sodium 191: A solution
(1 in 100) meets the requirements of the flame test for Aminobenzoic Acid
sodium. (Comment on this Monograph)id=m2810=Aminobenzoic
Acid=A-Monos.pdf)
ASSAY
PROCEDURE
Sample: 500 mg of Aminobenzoate Sodium
Analysis: Add 25 mL of water and 25 mL of 3 N
hydrochloric acid and cool in an ice bath. Titrate with 0.1 M
sodium nitrite VS, using a calomel-platinum electrode
system. Each mL of 0.1 M sodium nitrite is equivalent to
15.91 mg of C7H6NNaO2.
Acceptance criteria: 98.5%101.0% C7H7NO2 137.14
Benzoic acid, 4-amino;
IMPURITIES p-Aminobenzoic acid [150-13-0].
CHLORIDE AND SULFATE, Chloride 221: A 1.4-g portion DEFINITION
shows no more chloride than corresponds to 0.4 mL of Aminobenzoic Acid contains NLT 98.5% and NMT 101.5% of
0.020 N hydrochloric acid (0.02%). C7H7NO2, calculated on the dried basis.
CHLORIDE AND SULFATE, Sulfate 221: A 1.4-g portion shows
no more sulfate than corresponds to 0.3 mL of 0.020 N IDENTIFICATION
sulfuric acid (0.02%). A. INFRARED ABSORPTION 197K
HEAVY METALS, Method II 231: NMT 20 ppm B. ULTRAVIOLET ABSORPTION 197U
Organic Impurities Sample solution: 5 g/mL in 0.001 N sodium hydroxide
PROCEDURE: VOLATILE DIAZOTIZABLE SUBSTANCES
Blank: Water ASSAY
Standard stock solution: 10 mg of p-toluidine in 5 mL of PROCEDURE
methanol in a 100-mL volumetric flask, add water to Sample: 250 mg of Aminobenzoic Acid
volume Analysis: Proceed as directed under Nitrite Titration 451.
Standard solution: Transfer 1 mL of Standard stock Each mL of 0.1 M sodium nitrite is equivalent to 13.71 mg
solution to a 100-mL volumetric flask, and dilute with water of C7H7NO2.
to volume. Acceptance criteria: 98.5%101.5%
Sample solution: Transfer 5.0 g of Aminobenzoate Sodium
to a suitable flask, and add a volume of 1.25 N sodium IMPURITIES
hydroxide that is just sufficient to dissolve the sample and Inorganic Impurities
to render the solution just alkaline to phenolphthalein TS. RESIDUE ON IGNITION 281: NMT 0.1%
Dilute with water to 50 mL, and steam-distill the solution, HEAVY METALS, Method II 231: NMT 20 ppm
collecting 95 mL of the distillate in a 100-mL volumetric Organic Impurities
flask. Add water to volume. PROCEDURE 1: VOLATILE DIAZOTIZABLE SUBSTANCES
Spectrometric conditions Blank: water
Mode: UV-Vis Standard stock solution: 10 mg of p-toluidine in 5 mL of
Analytical wavelength: At 405 nm methanol in a 100-mL volumetric flask, add water to
Analysis volume
Samples: Blank, Standard solution, and Sample solution Standard solution: Transfer 1 mL of Standard stock
Transfer 20.0-mL portions of the Standard solution and the solution to a 100-mL volumetric flask, and dilute with water
Sample solution to separate 100-mL beakers, and transfer to volume.
186 Aminobenzoic / Official Monographs USP 32
Sample solution: Transfer 5.0 g of Aminobenzoic Acid to a Standard solution: Standard stock solution, Internal standard
suitable flask, and add a volume of 1.25 N sodium solution, and methanol (1:1:8). Pass through 0.6-m filter
hydroxide that is just sufficient to dissolve the sample and paper. Throughout the preparation, protect against actinic
to render the solution just alkaline to phenolphthalein TS. light.
Dilute with water to 50 mL, and steam-distill the solution, Sample solution: Gel, equivalent to 4.2 mg of
collecting 95 mL of the distillate in a 100-mL volumetric aminobenzoic acid, to a 100-mL volumetric flask, and add
flask. Add water to volume. 10.0 mL of Internal standard solution and 50 mL of
Spectrometric conditions methanol. Shake or sonicate, as necessary, and dilute with
Mode: UV-Vis methanol to volume. Pass, if necessary, through filter paper
Cell: 1 cm (Whatman No. 41 or equivalent). Pass through 0.6-m filter
Analytical wavelength: 450 nm paper. Throughout this preparation, protect against actinic
Analysis light.
Samples: Blank, Standard solution, and Sample solution Chromatographic system
Transfer 20.0-mL portions of the Standard solution and the (See Chromatography 621, System Suitability.)
Sample solution to separate 100-mL beakers, and transfer Mode: LC
20.0 mL of water to a third 100-mL beaker to provide Detector: UV 280 nm
the blank. Treat each as follows. Add 5.0 mL of 1 N Column: 3.9-mm 30-cm; packing L11
hydrochloric acid, and cool in an ice bath. Add 2.0 mL Flow rate: 1 mL/min
of 0.1 M sodium nitrite dropwise, with stirring, allow to Injection size: 15 L
stand for 5 min for the diazotization reaction to be System suitability
complete, add quickly to 10.0 mL of a cold solution of Sample: Standard solution
guaiacol (freshly prepared by dissolving 0.20 g of [NOTEChromatograph replicate 15-L injections of the
guaiacol in 100 mL of 1 N sodium hydroxide), and allow Standard solution until the response ratio variability is
to stand for 30 min. within 1.0% of average.]
Acceptance criteria: The absorbance of the solution [NOTEThe relative retention times of aminobenzoic acid
obtained from the Sample solution does not exceed that of and salicylic acid are about 1.0 and 3.0, respectively.]
the solution obtained from the Standard solution, Suitability requirements
corresponding to NMT 0.002% of volatile diazotizable Resolution: NLT 3.0 between aminobenzoic acid and
substances, as p-toluidine. salicylic acid
PROCEDURE 2: ORDINARY IMPURITIES 466 Analysis
Sample solution: Alcohol Samples: Standard solution and Sample solution
Standard solution: Alcohol Calculate the percentage of C7H7NO2 in the portion of Gel
Eluant: A mixture of toluene, ethyl acetate, and alcohol taken:
(60:20:20), in a nonequilibrated chamber
Visualization: 1 Result = (RU/RS) (CS/CU) 100
SPECIFIC TESTS RU = ratios of the peak responses from the Sample
MELTING RANGE OR TEMPERATURE741: 186189 solution
LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT RS = ratios of the peak responses from the Standard
0.2% of its weight. solution
CS = concentration of USP Aminobenzoic Acid RS in
ADDITIONAL REQUIREMENTS the Standard solution (mg/mL)
PACKAGING AND STORAGE: Preserve in tight, light-resistant CU = nominal concentration for the Sample solution
containers. (mg/mL)
USP Aminobenzoic Acid RS
OTHER COMPONENTS
ALCOHOL DETERMINATION, Method II 611: 42.3%54.0%
(w/w) of C2H5OH
Aminobenzoic Acid Gel
(Comment on this Monograph)id=m2820=Aminobenzoic Acid PERFORMANCE TESTS
Gel=A-Monos.pdf) MINIMUM FILL 755: Meets the requirements
DEFINITION SPECIFIC TESTS

Aminobenzoic Acid Gel contains NLT 90.0% and NMT 110.0% PH 791: 4.06.0
of the labeled amount of aminobenzoic acid (C7H7NO2).
IDENTIFICATION PACKAGING AND STORAGE: Preserve in tight, light-resistant
A. INFRARED ABSORPTION 197K containers
B. ULTRAVIOLET ABSORPTION 197U USP REFERENCE STANDARDS 11
Sample solution: 5 g/mL in alcohol USP Aminobenzoic Acid RS
ASSAY
PROCEDURE
Mobile phase: Mix 300 mL of methanol and 10 mL of Aminobenzoic Acid Topical Solution
glacial acetic acid with 690 mL of water. Allow the mixture (Comment on this Monograph)id=m2830=Aminobenzoic Acid
to cool, and pass, if necessary, through a suitable Topical Solution=A-Monos.pdf)
microporous membrane filter. Degas the solution.
Internal standard solution: 7 mg/mL of salicylic acid in DEFINITION
methanol. Dissolve by sonicating. Aminobenzoic Acid Topical Solution contains, in each mL, NLT
Standard stock solution: 0.42 mg/mL of USP 45 mg and NMT 55 mg of aminobenzoic acid (C7H7NO2).
Aminobenzoic Acid RS in methanol
USP 32 Official Monographs / Aminocaproic 187
IDENTIFICATION Mode: LC
A. To 1 mL of Topical Solution, add 1 mL of 1 N sodium Detector: UV 210 nm
hydroxide, and add, in order, 0.5 mL of potassium iodide Column: 4.6-mm 15-cm; packing L1
TS, 0.5 mL of 3 N hydrochloric acid, and 0.5 mL of sodium Column temperature: 30
hypochlorite TS: a brown precipitate is formed. Flow rate: 0.7 mL/min
B. To 1 mL of Topical Solution, add 2 mL of 3 N Injection size: 20 L
hydrochloric acid, and cool to about 10. Add 1 mL of 10 [NOTERecord chromatograms for NLT two times the
mg/mL sodium nitrite, then add a solution prepared by retention time of aminocaproic acid.]
mixing 50 mg of 2-naphthol with 3 mL of 1.94 M sodium System suitability
hydroxide: a red color is produced. Sample: Standard solution
[NOTEThe relative retention times for aminocaproic acid
ASSAY and methionine are about 0.76 and 1.0, respectively.]
PROCEDURE Suitability requirements
Sample solution: 5 mL of Topical Solution Resolution: NLT 2.0 between aminocaproic acid and
Analysis: Transfer Sample solution to a suitable open vessel, methionine
evaporate on a steam bath to dryness, and proceed as Relative standard deviation: NMT 2.0%
directed under Nitrite Titration 451, beginning with Add Analysis
20 mL of hydrochloric acid. Each mL of 0.1 M sodium Samples: Standard solution and Sample solution
nitrite is equivalent to 13.71 mg of C7H7NO2. Calculate the percentage of C6H13NO2 in the portion of
Acceptance criteria: 4555 mg/mL Aminocaproic Acid taken:
OTHER COMPONENTS Result = (RU/RS) (CS/CU) 100
ALCOHOL DETERMINATION 611: 65%75%
RU = peak response ratio of aminocaproic acid to the
SPECIFIC TESTS internal standard from the Sample solution
SPECIFIC GRAVITY 841: 0.8950.905 RS = peak response ratio of aminocaproic acid to the
ADDITIONAL REQUIREMENTS internal standard from the Standard solution
PACKAGING AND STORAGE: Preserve in tight, light-resistant CS = concentration of USP Aminocaproic Acid in the
containers. Standard solution (mg/mL)
CU = concentration of aminocaproic acid in the
Aminocaproic Acid
IMPURITIES
(Comment on this Monograph)id=m2880=Aminocaproic Inorganic Impurities
Acid=A-Monos.pdf) RESIDUE ON IGNITION 281: NMT 0.1%
SPECIFIC TESTS
PACKAGING AND STORAGE: Preserve in tight containers. Store
at room temperature.
C6H13NO2 131.17 USP REFERENCE STANDARDS 11
Hexanoic acid, 6-amino-; USP Aminocaproic Acid RS
6-Aminohexanoic acid [60-32-2].
DEFINITION
Aminocaproic Acid contains NLT 98.5% and NMT 101.5% of Aminocaproic Acid Injection
C6H13NO2, calculated on the anhydrous basis. (Comment on this Monograph)id=m2910=Aminocaproic Acid
IDENTIFICATION Injection=A-Monos.pdf)
INFRARED ABSORPTION 197K DEFINITION
Aminocaproic Acid Injection is a sterile solution of Aminocaproic
ASSAY Acid in Water for Injection. It contains NLT 95.0% and NMT
PROCEDURE 107.5% of the labeled amount of aminocaproic acid
Solution A: 0.55 mg/mL of sodium 1-heptanesulfonate (C6H13NO2).
Mobile phase: Dissolve 10 g of monobasic potassium
phosphate in 300 mL of Solution A, add 250 mL of IDENTIFICATION
methanol, followed by another 300 mL of Solution A. Adjust INFRARED ABSORPTION 197K
the mixture with phosphoric acid to a pH of 2.2, and dilute Sample: Mix 2 mL of Injection, added dropwise, with 100
with Solution A to 1 L. mL of acetone, rapidly stirring the mixture with a glass rod
Internal standard solution: 1.25 mg/mL of methionine to induce crystallization. Allow the mixture to stand for 15
Standard stock solution: 12.5 mg/mL of USP min, and pass through a medium-porosity, sintered-glass
Aminocaproic Acid RS filter. Wash the crystals with 25 mL of acetone, apply a
Standard solution: Standard stock solution, Internal standard vacuum to remove the solvent, dry at 105 for 30 min, and
solution, and water (5:2:93) cool: the residue is used as the Sample.
Sample stock solution: 12.5 mg/mL of Aminocaproic Acid
Sample solution: Sample stock solution, Internal standard ASSAY
solution, and water (5:2:93) PROCEDURE
Chromatographic system Mobile phase: Transfer 11 g of sodium 1-pentanesulfonate
(See Chromatography 621, System Suitability.) and 40 g of anhydrous sodium sulfate to a 2-L volumetric
188 Aminocaproic / Official Monographs USP 32
flask, and dissolve in 500 mL of water. Add 20 mL of 1 N 1 N hydrochloric acid in a 100-mL beaker. Decant and
sulfuric acid and 30 mL of acetonitrile, and dilute with water discard the hydrochloric acid, and wash the resin with five
to volume. 10-mL portions of water, decanting and discarding the
Standard solution: 2.5 mg/mL of USP Aminocaproic Acid liquid following each washing.
RS in Mobile phase Analysis: Place the washed resin in a glass-stoppered,
System suitability solution: Mix 20 L of benzyl alcohol conical flask, and add a volume of Oral Solution, equivalent
with 100 mL of water. Dilute 1.0 mL of this solution with to 250 mg of aminocaproic acid, and 10 mL of water. Insert
the Standard solution to 10 mL. the stopper in the flask, and shake by mechanical means for
Sample stock solution: Equivalent to 25 mg/mL of 30 min. Transfer the resin slurry to a medium-porosity,
aminocaproic acid, from Injection in water sintered-glass funnel, wash with 100 mL of water, apply
Sample solution: 2.5 mg/mL of Sample stock solution in suction to filter, and discard the washing. Place a beaker
Mobile phase under the stem of the funnel, add 10 mL of 1 N
Chromatographic system hydrochloric acid to the resin, stir for 45 min, and filter by
(See Chromatography 621, System Suitability.) applying suction. Evaporate the filtrate on a steam bath to
Mode: LC dryness, dry at 105 for 1 h, and cool.
Detector: UV 210 nm Acceptance criteria: The residue so obtained meets the
Column: 4-mm 30-cm; packing L1 requirements for the test.
Flow rate: 2 mL/min
Injection size: 50 L ASSAY
System suitability PROCEDURE
Sample: Standard solution and System suitability solution Sample: Equivalent to 250 mg of aminocaproic acid
Suitability requirements Analysis: Add 80 mL of glacial acetic acid and 10 drops of
Resolution: NLT 7.0 between benzyl alcohol and a solution (1 in 500) of crystal violet in chlorobenzene.
aminocaproic acid in the System suitability solution Titrate with 0.1 N perchloric acid in dioxane VS to a blue
[NOTEThe aminocaproic acid peak elutes prior to the endpoint. Perform a blank determination (see Titrimetry
benzyl alcohol peak.] 541). Each mL of 0.1 N perchloric acid is equivalent to
Relative standard deviation: NMT 1.0%, Standard 13.12 mg of C6H13NO2.
solution Acceptance criteria: 95.0%115.0%
Analysis
Calculate the percentage of C6H13NO2 in each mL of the PH 791: 6.06.5
Injection taken: ADDITIONAL REQUIREMENTS
Result = (rU/rS) (CS/CU) 100 USP REFERENCE STANDARDS 11
rU = peak area from the Sample solution USP Aminocaproic Acid RS
rS = peak area from the Standard solution
CS = concentration of USP Aminocaproic Acid RS in
the Standard solution (mg/mL) Aminocaproic Acid Tablets
CU = nominal concentration of aminocaproic acid in
the Sample solution (mg/mL) (Comment on this Monograph)id=m2950=Aminocaproic Acid
Acceptance criteria: 95.0%107.5% Tablets=A-Monos.pdf)
SPECIFIC TESTS DEFINITION

PH 791: 6.07.6 Aminocaproic Acid Tablets contain NLT 95.0% and NMT
OTHER REQUIREMENTS: It meets the requirements under 105.0% of the labeled amount of aminocaproic acid
Injections 1. (C6H13NO2).
BACTERIAL ENDOTOXINS TEST 85: NMT 0.05 USP Endotoxin IDENTIFICATION
Unit/mg of aminocaproic acid INFRARED ABSORPTION 197K
ADDITIONAL REQUIREMENTS Sample solution: Triturate 2 Tablets with 10 mL of water,
PACKAGING AND STORAGE: Preserve in single-dose or and filter into 100 mL of acetone. Swirl the mixture, and
multiple-dose containers, preferably of Type I glass. allow to stand for 15 min to complete crystallization. Pass
USP REFERENCE STANDARDS 11 through a medium-porosity, sintered-glass filter, and wash
USP Aminocaproic Acid RS the crystals with 25 mL of acetone. Apply vacuum to
USP Endotoxin RS remove the solvent, then dry at 105 for 30 min, and cool:
the residue is used as the sample.
ASSAY
Aminocaproic Acid Oral Solution PROCEDURE
Sample solution: Equivalent to 5 mg/mL of aminocaproic
(Comment on this Monograph)id=m2940=Aminocaproic Acid acid from powdered Tablets (NLT 20) in glacial acetic acid.
Oral Solution=A-Monos.pdf) Heat gently to effect solution, and cool.
DEFINITION Analysis: To 100 mL of Sample solution, add 10 drops of a
Aminocaproic Acid Oral Solution contains NLT 95.0% and NMT solution (1 in 500) of crystal violet in chlorobenzene and
115.0% of the labeled amount of aminocaproic acid titrate with 0.1 N perchloric acid in dioxane VS to a blue
(C6H13NO2). endpoint. Perform a blank determination (see Titrimetry
541). Each mL of 0.1 N perchloric acid is equivalent to
IDENTIFICATION 13.12 mg of (C6H13NO2).
Sample: Mix 1 g of ion-exchange resin (strongly acidic
styrene-divinylbenzene cation-exchange resin) with 10 mL of
USP 32 Official Monographs / Aminoglutethimide 189
Acceptance criteria: 95.0%105.0% and add 500 mL of water. Adjust by the addition of either 1
N acetic acid or 1 N potassium hydroxide to a pH of 5.0
PERFORMANCE TESTS 0.1. Dilute with water to volume.
DISSOLUTION 711 Mobile phase: Methanol and Solution A (27:73)
Medium: Water; 900 mL Diluent: Methanol and Solution A (1:1)
Apparatus 1: 100 rpm Standard solution: 0.5 mg/mL of USP Aminoglutethimide
Time: 45 min RS in Diluent
pH 9.5 borate buffer: Dissolve 6.185 g of boric acid and Sample solution: 0.5 mg/mL of Aminoglutethimide in
7.930 g of potassium chloride in 1000 mL of water, and add Diluent [NOTEPass through a 0.45-m or finer porosity
60 mL of 1.0 N sodium hydroxide. Dilute with water to filter, discarding the first 5 mL of the filtrate.]
2000 mL, and add 1.0 N sodium hydroxide, if necessary, to Chromatographic system
adjust to a pH of 9.5 0.1. (See Chromatography 621, System Suitability.)
Standard solution: 0.5 mg/mL of USP Aminocaproic Acid Mode: LC
RS in water Detector: UV 240 nm
Analysis: Into 3 separate 50-mL volumetric flasks pipet (a) 1 Column: 3.9-mm 15-cm; 4-m packing L1
mL of a filtered portion of the solution under test, (b) 1 mL Column temperature: 40
of the Standard solution, and (c) 1 mL of water to provide a Flow rate: 1.3 mL/min
blank. Add 20.0 mL of pH 9.5 borate buffer and 3.0 mL of Injection size: 10 L
freshly prepared -naphthoquinone-4-sodium sulfonate System suitability
solution (1 in 500) to each, swirl to mix, and place the 3 Sample: Standard solution
flasks in a water bath maintained at a temperature of 65 Suitability requirements
5 for 45 min. Cool, and dilute each with water to volume. Tailing factor: NMT 1.7
Determine the amount of C6H13NO2 dissolved from Relative standard deviation: NMT 2.0%
absorbances, at the wavelength of maximum absorbance at Analysis
about 460 nm, obtained from the Sample solution in Samples: Standard solution and Sample solution
comparison with those obtained from the Standard solution, Calculate the percentage of C13H16N2O2 in the portion of
using the blank to set the instrument. Aminoglutethimide taken:
C6H13NO2 is dissolved. Result = (rU/rS) (CS/CU) 100
ADDITIONAL REQUIREMENTS rS = peak area from the Standard solution
PACKAGING AND STORAGE: Preserve in tight containers. CS = concentration of USP Aminoglutethimide RS in
USP REFERENCE STANDARDS 11 the Standard solution (mg/mL)
USP Aminocaproic Acid RS CU = concentration of Aminoglutethimide in the
Aminoglutethimide IMPURITIES
(Comment on this Inorganic Impurities
Monograph)id=m2975=Aminoglutethimide=A-Monos.pdf) RESIDUE ON IGNITION 281: NMT 0.1%
Organic Impurities
PROCEDURE 1: LIMIT OF AZO-AMINOGLUTETHIMIDE
[NOTEUse low-actinic glassware. Conduct this test
promptly under subdued light. Wear protective gloves
resistant to dimethyl sulfoxide to prevent contact with skin.
Use shaking, not sonication or heat, to dissolve the USP
Azo-aminoglutethimide RS and the Sample.]
C13H16N2O2 232.28 Solution A: 150 mL of 0.1 N acetic acid and 50 mL of 0.1
2,6-Piperidinedione, 3-(4-aminophenyl)-3-ethyl-; N potassium hydroxide, diluted in water to 1000 mL
2-(p-Aminophenyl)-2-ethylglutarimide [125-84-8]. Mobile phase: 100 mg of edetate disodium in 350 mL of
Solution A, add 650 mL of methanol and cool to room
DEFINITION temperature. Adjust with glacial acetic acid to a pH of 5.0
Aminoglutethimide contains NLT 98.0% and NMT 102.0% of 0.1.
C13H16N2O2, calculated on the dried basis. Standard solution: 0.5 g/mL of USP Azo-
IDENTIFICATION aminoglutethimide RS in dimethyl sulfoxide
A. INFRARED ABSORPTION 197M Sample solution: 1 mg/mL of Aminoglutethimide in
B. ULTRAVIOLET ABSORPTION 197U dimethyl sulfoxide
Analytical wavelength: 242 nm Chromatographic system
Medium: Methanol (See Chromatography 621, System Suitability.)
Solution: 10 g/mL
Acceptance criteria: Absorptivities differ by NMT 2.0%
ASSAY
PROCEDURE
Solution A: Add 240 mL of 0.1 N acetic acid to 200 mL of
0.1 N potassium hydroxide in a 2000-mL volumetric flask,
190 Aminoglutethimide / Official Monographs USP 32
Mode: LC Acceptance criteria: No turbidity is produced.

Detector: UV 328 nm PH 791
Column: 3.9-mm 15-cm; packing L1 Sample solution: 1 mg/mL in dilute methanol (1 in 20)
Flow rate: 1 mL/min Acceptance criteria: 6.27.3
Injection size: 10 L LOSS ON DRYING 731: Dry at 105 to constant weight: it
System suitability loses NMT 0.5% of its weight.
Suitability requirements ADDITIONAL REQUIREMENTS
Tailing factor: NMT 1.2 PACKAGING AND STORAGE: Preserve in well-closed containers.
Capacity factor: 2.05.0 USP REFERENCE STANDARDS 11
Column efficiency: NLT 800 theoretical plates USP Aminoglutethimide RS
Analysis USP m-Aminoglutethimide RS
Samples: Standard solution and Sample solution USP Azo-aminoglutethimide RS
[NOTEThe aminoglutethimide elutes with the dimethyl
sulfoxide.]
Calculate the percentage of azo-aminoglutethimide in the Aminoglutethimide Tablets
specimen of Aminoglutethimide taken:
(Comment on this Monograph)id=m2985=Aminoglutethimide
Result = (rU/rS) (CS/CU) 100 Tablets=A-Monos.pdf)
rU = peak response from the Sample solution DEFINITION

rS = peak response from the Standard solution Aminoglutethimide Tablets contain NLT 90.0% and NMT
CS = concentration of USP Azo-aminoglutethimide RS 110.0% of the labeled amount of aminoglutethimide
in the Standard solution (g/mL) (C13H16N2O2).
CU = concentration of Aminoglutethimide in the
Acceptance criteria: NMT 0.03% of 3,3-(azodi-4,1- INFRARED ABSORPTION 197M
phenylene)-3,3-dimethylbis-[2,6-piperidinedione] Sample: Transfer 500 mg of finely powdered Tablets to a
(corresponding to azo-aminoglutethimide) suitable container, add 25 mL of acetone, mix, and filter.
PROCEDURE 2: LIMIT OF m-AMINOGLUTETHIMIDE Evaporate the filtrate at room temperature to dryness, and
Solution A, Mobile phase, Diluent, and Chromatographic dry the residue in vacuum over silica gel for 2 h.
system: Prepare as directed in the Assay. ASSAY
Standard solution: 10 g/mL of USP m- PROCEDURE
Aminoglutethimide RS in Diluent Acetate buffer: Prepare a solution in water, containing 120
Sample solution: 1 mg/mL of Aminoglutethimide in mL of 0.1 N acetic acid and 100 mL of 0.1 N potassium
Diluent [NOTEPass through a 0.45-m or finer porosity hydroxide/L of buffer. Before final dilution, adjust by the
filter, discarding the first 5 mL of the filtrate] addition of either 1 N acetic acid or 1 N potassium
Analysis hydroxide to a pH of 5.0 0.1.
Samples: Standard solution and Sample solution Mobile phase: Methanol and Acetate buffer (27:73)
[NOTEThe relative retention times for Diluent: Methanol and Acetate buffer (1:1)
aminoglutethimide and m-aminoglutethimide are about Standard solution: 0.5 mg/mL of USP Aminoglutethimide
0.8 and 1.0, respectively.] RS in Diluent
Calculate the percentage of m-aminoglutethimide in the Sample solution: Finely powder NLT 20 Tablets. Transfer a
specimen of Aminoglutethimide taken: portion of the powder, equivalent to 200 mg of
aminoglutethimide, to a 200-mL volumetric flask. Add 130
Result = (rU/rS) (CS/CU) 100 mL of Diluent, and sonicate for 5 min. Shake by mechanical
rU = peak response from the Sample solution means for 30 min, and dilute with Diluent to volume.
rS = peak response from the Standard solution Centrifuge this solution, transfer 25.0 mL of the clear
CS = concentration of USP m-Aminoglutethimide RS supernatant to a 50-mL volumetric flask, dilute with Diluent
in the Standard solution (mg/mL) to volume and pass through a 0.45-m or finer porosity
CU = concentration of Aminoglutethimide in the filter, discarding the first 5 mL of the filtrate.
Sample solution (mg/mL) Chromatographic system
Acceptance criteria: NMT 2.0% of m-aminoglutethimide (See Chromatography 621, System Suitability.)
is found. Mode: LC
Calculate the percentage of each peak, other than the main Detector: UV 240 nm
peak and the m-aminoglutethimide peak, if present, by Column: 3.9-mm 15-cm; 4-m packing L1
the same formula: Column temperature: 40
Result = (rU/rT) 100 Injection size: 10 L
System suitability
rU = response of each peak Sample: Standard solution
rT = sum of the responses of all the peaks in the Suitability requirements
chromatogram of the Sample solution Tailing factor: NMT 1.7
Acceptance criteria: NMT 1.0% total impurities, other Relative standard deviation: NMT 2.0%
than m-aminoglutethimide Analysis
SPECIFIC TESTS Calculate the percentage of C13H16N2O2 in the portion of
SULFATE Tablets taken:
Sample solution: 1 mg/mL in dilute methanol (1 in 20)
Analysis: To 100 mL of Sample solution, add 1.0 mL of 3 N Result = (rU/rS) (CS/CU) 100
hydrochloric acid and 2.0 mL of barium chloride TS
USP 32 Official Monographs / Aminohippurate 191
rU = peak area from the Sample solution Acceptance criteria: NMT 2.0% total impurities, other
rS = peak area from the Standard solution than m-aminoglutethimide, is found.
CS = concentration of USP Aminoglutethimide RS in
the Standard solution (mg/mL) ADDITIONAL REQUIREMENTS
CU = nominal concentration of aminoglutethimide in PACKAGING AND STORAGE: Preserve in tight, light-resistant
the Sample solution (mg/mL) containers.
Acceptance criteria: 90.0%110.0% USP REFERENCE STANDARDS 11
USP Aminoglutethimide RS
PERFORMANCE TESTS USP m-Aminoglutethimide RS
DISSOLUTION 711
Medium: Dilute hydrochloric acid (7 in 1000); 1000 mL
Time: 30 min Aminohippurate Sodium Injection
Detector: UV 237 nm (Comment on this Monograph)id=m3000=Aminohippurate
Sample solution: Sample per Dissolution 711. Dilute with Sodium Injection=A-Monos.pdf)
pH 7.5 phosphate buffer to a concentration that is similar to
the Standard solution.
Standard solution: USP Aminoglutethimide RS in a mixture
of dilute hydrochloric acid and pH 7.5 phosphate buffer,
having a ratio similar to the Sample solution
C13H16N2O2 is dissolved.
C9H9N2NaO3 216.17
IMPURITIES Glycine, N-(4-aminobenzoyl)-, monosodium salt;
Organic Impurities Monosodium p-aminohippurate [94-16-6].
PROCEDURE
Buffer: Prepare a solution in water, containing 120 mL of DEFINITION
0.1 N acetic acid and 100 mL of 0.1 N potassium Aminohippurate Sodium Injection is a sterile solution of
hydroxide/L of buffer. Before final dilution, adjust by the Aminohippuric Acid in Water for Injection prepared with the
addition of either 1 N acetic acid or 1 N potassium aid of Sodium Hydroxide. It contains NLT 95.0% and NMT
hydroxide to a pH of 5.0 0.1. 105.0% of the labeled amount of C9H9N2NaO3.
Mobile phase: Methanol and Buffer (27:73)
Diluent: Methanol and Buffer (1:1) IDENTIFICATION
Standard solution: 10 g/mL of USP m- A. Procedure
Aminoglutethimide RS in Diluent Analysis: Dilute a volume of Injection, equivalent to 100
Sample solution: Finely powder NLT 20 Tablets. Transfer a mg of aminohippuric acid, to 50 mL and acidify with
portion of the powder, equivalent to 200 mg of hydrochloric acid. Add 0.5 mL of 3 N hydrochloric acid, 0.5
aminoglutethimide, to a 200-mL volumetric flask. Add 130 mL of sodium nitrite solution (1 in 10), and a solution of
mL of Diluent, and sonicate for 5 min. Shake by mechanical 0.20 g of 2-naphthol in 10 mL of 6 N ammonium
means for 30 min, dilute with Diluent to volume, and pass hydroxide.
through a 0.45-m or finer porosity filter, discarding the Acceptance criteria: A red color is produced.
first 5 mL of the filtrate. B. Procedure
Chromatographic system Analysis: Transfer a volume of Injection, equivalent to 200
(See Chromatography 621, System Suitability.) mg of aminohippurate sodium, to a test tube, and add, in
Mode: LC the order named, 2 mL of potassium iodide TS, 10 mL of
Detector: UV 240 nm water, and 5 mL of sodium hypochlorite TS.
Column: 3.9-mm 15-cm; 4-m packing L1 Acceptance criteria: A red color is produced.
Column temperature: 40 C. Identification TestsGeneral, Sodium 191: Meets the
Flow rate: 1.3 mL/min requirements
Sample: Standard solution Sample: Equivalent to 1 g of aminohippurate sodium
Suitability requirements Analysis: Transfer to a 200-mL volumetric flask, and dilute
Tailing factor: NMT 1.7 with water to volume. Transfer 50.0 mL of the solution to a
Relative standard deviation: NMT 2.0% suitable container, add 5 mL of hydrochloric acid, stir, cool
Analysis to 15, slowly titrate with 0.1M sodium nitrate VS, and
Samples: Sample solution and Standard solution proceed as directed under Nitrite Titration 451, beginning
[NOTEThe relative retention times for with Determine the endpoint. Each mL of 0.1 M sodium
aminoglutethimide and m-aminoglutethimide are 0.8 nitrite is equivalent to 19.42 mg of C9H10N2O3.
and 1.0, respectively.] Acceptance criteria: 95.0%105.0%
Calculate the percentage of each peak, other than the
main peak and the m-aminoglutethimide peak, if present: SPECIFIC TESTS
PH 791: 6.77.6
Result = (rU/rT) 100 OTHER REQUIREMENTS: Meets the requirements under
Injections 1
rU = response of each impurity peak in the Sample BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.04 USP
solution Endotoxin Unit/mg of aminohippurate sodium.
rT = sum of the responses of all of the peaks
excluding that of the m-aminoglutethimide
peak in the Sample solution
192 Aminohippurate / Official Monographs USP 32
ADDITIONAL REQUIREMENTS Aminopentamide Sulfate

PACKAGING AND STORAGE: Preserve in tight, light-resistant (Comment on this Monograph)id=m3040=Aminopentamide
containers. Sulfate=A-Monos.pdf)
USP Aminohippuric Acid RS
Aminohippuric Acid
(Comment on this Monograph)id=m3030=Aminohippuric
Acid=A-Monos.pdf)
C19H24N2O H2SO4 394.49
-[2-(Dimethylamino)propyl]--phenylbenzeneacetamide
sulfate [60-46-8].
DEFINITION
Aminopentamide Sulfate contains NLT 95.0% and NMT
103.0% of C19H24 N2O H2SO4.
C9H10N2O3 194.19 IDENTIFICATION
Glycine, N-(4-aminobenzoyl)-; A. INFRARED ABSORPTION 197K
p-Aminohippuric acid [61-78-9]. B. IDENTIFICATION TESTSGENERAL, Sulfate 191
DEFINITION ASSAY
Aminohippuric Acid contains NLT 98.0% and NMT 100.5% of PROCEDURE
C9H10N2O3, calculated on the dried basis. Sample: 500 mg of Aminopentamide Sulfate
Analysis: Add Sample to 100 mL of dimethylformamide in a
IDENTIFICATION suitable container, add 5 drops of thymol blue TS, and
A. INFRARED ABSORPTION 197K titrate with 0.1 N lithium methoxide VS in toluene to a deep
B. PROCEDURE blue endpoint. Perform a blank determination (see Titrimetry
Analysis Dissolve 10 mg in 5 mL of water, and add 0.5 mL 541). Each mL of 0.1 N lithium methoxide is equivalent to
of 3 N hydrochloric acid, 0.5 mL of sodium nitrite solution 19.72 mg of C19H24N2O H2SO4.
(1 in 10), and a solution of 0.20 g of 2-naphthol in 10 mL Acceptance criteria: 95.0%103.0%
of 6 N ammonium hydroxide.
Acceptance criteria: A red color is produced. IMPURITIES
ASSAY RESIDUE ON IGNITION 281: NMT 0.5%
PROCEDURE
Sample: 150 mg of Aminohippuric Acid SPECIFIC TESTS
Analysis: To the Sample, add 5 mL of hydrochloric acid and MELTING RANGE OR TEMPERATURE 741: 179186
50 mL of water, stir until dissolved, cool to 15, and slowly PH 791
titrate with 0.1 M sodium nitrate VS, and proceed as Sample solution: 25 mg/mL
directed under Nitrite Titration 451, beginning with Acceptance criteria: 1.23.0
Determine the endpoint. Each mL of 0.1 M sodium LOSS ON DRYING 731: Dry at 105 for 4 h: it loses NMT
nitrite is equivalent to 19.42 mg of C9H10N2O3. 4.4% of its weight.
Acceptance criteria: 98.0%100.5% CLARITY AND COLOR OF SOLUTION: Dissolve 500 mg in 10 mL
of water: the solution is clear and colorless.
IMPURITIES
Inorganic Impurities ADDITIONAL REQUIREMENTS
RESIDUE ON IGNITION 281: NMT 0.25% PACKAGING AND STORAGE: Preserve in tight containers, and
HEAVY METALS, Method II 231: NMT 10 ppm store at controlled room temperature.
LABELING: Label it to indicate that it is for veterinary use
SPECIFIC TESTS only.
LOSS ON DRYING 731: Dry at 105 for 2 h: it loses NMT USP REFERENCE STANDARDS 11
0.25% of its weight. USP Aminopentamide Sulfate RS
containers.
USP Aminohippuric Acid RS
USP 32 Official Monographs / Aminopentamide 193
Aminopentamide Sulfate Injection PH 791: 2.54.5

(Comment on this Monograph)id=m3045=Aminopentamide OTHER REQUIREMENTS: Meets the requirements under
Sulfate Injection=A-Monos.pdf) Injections 1
BACTERIAL ENDOTOXINS TEST 85: It contains NMT 25 USP
DEFINITION Endotoxin Units/mg of aminopentamide sulfate.
Aminopentamide Sulfate Injection is a sterile solution of
Aminopentamide Sulfate in Water for Injection. It contains NLT ADDITIONAL REQUIREMENTS
90.0% and NMT 110.0% of the labeled amount of PACKAGING AND STORAGE: Preserve in tight, single-dose or
aminopentamide sulfate (C19H24N2O H2SO4). multiple-dose , as described under Injections 1, Containers
for Injections. Store at controlled room temperature.
IDENTIFICATION LABELING: Label Injection to indicate that it is for veterinary
PROCEDURE use only.
Sample: Transfer 10 mL of the Injection to a separator, add USP REFERENCE STANDARDS 11
sodium hydroxide TS until alkaline to litmus, and extract USP Aminopentamide Sulfate RS
with 25 mL of chloroform. Transfer a few drops of the USP Endotoxin RS
chloroform extract to a KRS-5 plate, and allow to dry.
Record the infrared absorption spectrum by the attenuated
total reflectance technique (See Spectrophotometry and Light-
Scattering 851). Aminopentamide Sulfate Tablets
Acceptance criteria: The spectrum thus obtained exhibits (Comment on this Monograph)id=m3050=Aminopentamide
maxima only at the same wavelengths as that of a similar Sulfate Tablets=A-Monos.pdf)
preparation of USP Aminopentamide Sulfate RS,
concomitantly measured. DEFINITION
Aminopentamide Sulfate Tablets contain NLT 95.0% and NMT
ASSAY 105.0% of the labeled amount of aminopentamide sulfate
PROCEDURE (C19H24N2O H2SO4).
Solution A: 28.8 mg/mL of sodium lauryl sulfate in glacial
acetic acid and water (1:4). Pass through a filter having a IDENTIFICATION
0.5-m or finer porosity. A. PROCEDURE
Mobile phase: Acetonitrile, methanol, Solution A, and water Sample: Transfer powdered Tablets, equivalent to 2 mg of
(7:7:1:5) aminopentamide, to a separator, add 20 mL of water and 3
Standard solution: USP Aminopentamide Sulfate RS in mL of 10 N sodium hydroxide. Extract with two 20-mL
water, at a concentration equivalent to the labeled portions of methylene chloride, and evaporate the combined
concentration of aminopentamide sulfate methylene chloride extracts to a volume of about 0.5 mL.
Sample solution: Use the undiluted Injection. Transfer a few drops of the chloroform concentrate to a
Chromatographic system KRS-5 plate, and allow to dry. Record the IR absorption
(See Chromatography 621, System Suitability.) spectrum by the attenuated total reflectance technique (see
Mode: LC Spectrophotometry and Light-Scattering 851).
Detector: UV 254 nm Acceptance criteria: The spectrum thus obtained exhibits
Column: 3.9-mm 30-cm; packing L1 maxima only at the same wavelengths as that of a similar
Temperature: 40 preparation of USP Aminopentamide Sulfate RS,
Flow rate: 1 mL/min concomitantly measured.
Sample: Standard solution Solution A: 28.8 mg/mL of sodium lauryl sulfate in glacial
Suitability requirements acetic acid and water (1:4). Pass through a filter having a
Tailing factor: NMT 2.5 0.5-m or finer porosity.
Relative standard deviation: NMT 2% Mobile phase: Acetonitrile, methanol, Solution A, and water
Analysis (7:7:1:5)
Samples: Sample solution and Standard solution Standard solution: 0.02 mg/mL of USP Aminopentamide
Calculate the percentage of C19H24N2O H2SO4 in each mL Sulfate RS in Mobile phase
of the Injection taken: Sample solution: Equivalent to 0.02 mg/mL of
Result = (rU/rS) (CS/CU) 100 aminopentamide, from Powdered Tablets in Mobile phase
[NOTEFinely powder NLT 10 Tablets. After adding Mobile
rU = peak response in the Sample solution phase sonicate for 5 min, and stir by mechanical means for
rS = peak response in the Standard solution 10 min. Pass this mixture through a filter having a 0.5-m
CS = concentration of USP Aminopentamide Sulfate or finer porosity, discarding the first 5 mL of the filtrate. Use
RS in the Standard solution (mg/mL) the clear filtrate.]
CU = nominal concentration of aminopentamide Chromatographic system
sulfate in the Sample solution (mg/mL) (See Chromatography 621, System Suitability.)
SPECIFIC TESTS
STERILITY TESTS 71: Meets the requirements under Test for
Sterility of the Product to be Examined, Membrane Filtration
194 Aminopentamide / Official Monographs USP 32
Mode: LC anhydrous theophylline (C7H8N4O2), calculated on the

Detector: UV 254 nm anhydrous basis.
Temperature: 40 IDENTIFICATION
Flow rate: 1 mL/min Sample 1: Dissolve 500 mg in 20 mL of water, add, with
Injection size: 50 L constant stirring, 1 mL of 3 N hydrochloric acid, and filter
System suitability (retain the filtrate).
Sample: Standard solution Sample 2: Wash the precipitate from Sample 1 with small
Suitability requirements portions of cold water, and dry at 105 for 1 h.
Column efficiency: NLT 900 theoretical plates A. Sample 2 melts between 270 and 274.
Relative standard deviation: NMT 2% B. PROCEDURE
Analysis Analysis: To 10 mg of Sample 2, in a porcelain dish, add 1
Samples: Sample solution and Standard solution mL of hydrochloric acid and 100 mg of potassium chlorate,
Calculate the percentage of C19H24N2O H2SO4 in the evaporate on a steam bath to dryness, and invert the dish
portion of Tablets taken: over a vessel containing a few drops of 6 N ammonium
hydroxide.
Result = (rU/rS) (CS/CU) 100 Acceptance criteria: The residue acquires a purple color,
which is destroyed by solutions of fixed alkalies.
rU = peak response of the Sample solution C. PROCEDURE
rS = peak responses of the Standard solution Analysis: To Sample 1, add 0.5 mL of benzenesulfonyl
CS = concentration of USP Aminopentamide Sulfate chloride and 5 mL of 1 N sodium hydroxide to render
RS in Standard solution (mg/mL) alkaline, shake by mechanical means for 10 min, add 5 mL
CU = nominal concentration of aminopentamide of 3 N hydrochloric acid to acidify, chill, collect the
sulfate in the Sample solution (mg/mL) precipitated disulfonamide of ethylenediamine, wash with
water, recrystallize from water, and dry at 105 for 1 h.
SPECIFIC TESTS Acceptance criteria: The dried precipitate melts between
LOSS ON DRYING 731: Dry 1 g of powdered Tablets in a 164 and 171.
vacuum at a pressure of 5 mm of mercury or less at 60 for
3 h: it loses NMT 4.0% of its weight. ASSAY
PROCEDURE
PERFORMANCE TESTS Mobile phase: 200 mL of methanol, 960 mg of sodium 1-
DISINTEGRATION 701 pentanesulfonate, and sufficient water to make 1 L. Adjust
Medium: Simulated gastric fluid TS with glacial acetic acid to a pH of 2.9 0.1.
Time: NMT 10 min Diluent: Methanol and water (1:4)
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Standard solution: 80 g/mL of USP Theophylline RS in
Diluent
ADDITIONAL REQUIREMENTS Solution A: 80 g/mL of theobromine in Standard solution
PACKAGING AND STORAGE: Preserve in tight containers, and System suitability solution: 64 g/mL of theophylline and
store at controlled room temperature. 64 g/mL of theobromine, from Solution A in Diluent
LABELING: Label Tablets to indicate that they are for Sample solution: 96 g/mL of Aminophylline in Diluent
veterinary use only. Chromatographic system
USP Aminopentamide Sulfate RS Mode: LC
Detector: UV 254 nm
Aminophylline Flow rate: 1 mL/min
(Comment on this Monograph)id=m3080=Aminophylline=A- System suitability
Monos.pdf) Samples: System suitability solution and Standard solution
[NOTEThe relative retention times for theobromine and
theophylline are 0.65 and 1.0, respectively.]
Change to read: Suitability requirements
Resolution: NLT 3.0 between theobromine and
theophylline, System suitability solution
Tailing factor: NMT 2.0 for theophylline peak, System
solution
Analysis
C16H24N10O4 (anhydrous) 420.43 Aminophylline taken:
1H-Purine-2,6-dione, 3,7-dihydro-1,3-dimethyl-, compd. with
1,2-ethanediamine (2:1); Result = (rU/rS) (CS/CU) 100
Theophylline compound with ethylenediamine (2:1)
[317-34-0]. rU = peak response from the Sample solution
Dihydrate 456.46 rS = peak response from the Standard solution
[5897-66-5USP32]. CS = concentration of USP Theophylline RS in the
DEFINITION CU = concentration of aminophylline in the Sample
Aminophylline is anhydrous or contains NMT two molecules of solution (mg/mL)
water of hydration. It contains NLT 84.0% and NMT 87.4% of
USP 32 Official Monographs / Aminophylline 195
Acceptance criteria: 84.0%87.4% Acceptance criteria: The residue acquires a purple color,
which is destroyed by solutions of fixed alkalies.
OTHER COMPONENTS
ETHYLENEDIAMINE CONTENT ASSAY
Sample solution: 16.67 mg/mL PROCEDURE
Analysis: To 30 mL of Sample solution, add methyl orange Mobile phase: 200 mL of methanol, 960 mg of sodium 1-
TS and titrate with 0.1 N hydrochloric acid VS. Each mL of pentanesulfonate, and sufficient water to make 1 L.
0.1 N hydrochloric acid is equivalent to 3.005 mg of Adjust with glacial acetic acid to a pH of 2.9 0.1.
C2H8N2. Diluent: Methanol and water (1:4)
Acceptance criteria: The content of ethylenediamine Standard solution: 0.08 mg/mL of USP Theophylline RS in
(C2H8N2) is between 157 mg and 175 mg/g of C7H8N4O2 Diluent
found in the Assay. Solution A: 0.08 mg/mL of theobromine in the Standard
solution
IMPURITIES System suitability solution: 64 g/mL of theophylline and
Inorganic Impurities 64 g/mL of theobromine, from Solution A in Diluent
RESIDUE ON IGNITION 281: NMT 0.15% Sample solution: Equivalent to 0.08 mg/mL of
theophylline, from Injection in Diluent
SPECIFIC TESTS Chromatographic system
WATER DETERMINATION, Method I 921: NMT 0.75% (See Chromatography 621, System Suitability.)
(anhydrous form) and NMT 7.9% (hydrous form), Mode: LC
determined on 1.5 g, a mixture of 25 mL of chloroform and Detector: UV 254 nm
25 mL of methanol being used in place of the methanol Column: 3.9-mm 15-cm; packing L1
solvent. Flow rate: 1 mL/min
ADDITIONAL REQUIREMENTS Injection size: 10 L
PACKAGING AND STORAGE: Preserve in tight containers. System suitability
LABELING: Label it to indicate whether it is anhydrous or Samples: System suitability solution and Standard solution
hydrous and also to state the content of anhydrous [NOTEThe relative retention times for theobromine and
theophylline. theophylline are about 0.65 and 1.0, respectively.]
USP REFERENCE STANDARDS 11 Suitability requirements
USP Theophylline RS Resolution: NLT 3.0 between theobromine and
Tailing factor: NMT 2.0 for the theophylline peak, System
Aminophylline Injection Relative standard deviation: NMT 2.0%, Standard
(Comment on this Monograph)id=m3130=Aminophylline solution
Injection=A-Monos.pdf) Analysis
DEFINITION Calculate the percentage of C7H8N4O2 in the portion of
Aminophylline Injection is a sterile solution of Aminophylline in Injection taken:
Water for Injection, or is a sterile solution of Theophylline in
Water for Injection prepared with the aid of Ethylenediamine. Result = (rU/rS) (CS/CU) 100
It contains, in each mL, an amount of aminophylline
equivalent to NLT 93.0% and NMT 107.0% of the labeled rU = peak response of the Sample solution
amount of anhydrous theophylline (C7H8N4O2). rS = peak response of the Standard solution
Aminophylline Injection may contain an excess of CS = concentration of the USP Theophyline RS in the
Ethylenediamine, but no other substance may be added for Standard solution (mg/mL)
the purpose of pH adjustment. CU = nominal concentration of aminophylline in the
[NOTEDo not use the Injection if crystals have separated.] Sample solution (mg/mL)
IDENTIFICATION
Sample 1: Dilute an equivalent of 500 mg of aminophylline, OTHER COMPONENTS
with water to 20 mL, and add, with constant stirring, 1 mL of CONTENT OF ETHYLENEDIAMINE
3 N hydrochloric acid or enough to precipitate the Sample solution: Dilute an equivalent to 500 mg of
theophylline completely, and filter. Aminophylline to 30 mL, if necessary.
Sample 2: Wash the precipitate from Sample 1 with a small Analysis: Add methyl orange TS and titrate with 0.1 N
portion of cold water, and dry at 105 for 1 h. hydrochloric acid VS. Each mL of 0.1 N hydrochloric acid is
A. PROCEDURE equivalent to 3.005 mg of C2H8N2.
Analysis: To Sample 1, add 0.5 mL of benzenesulfonyl Acceptance criteria: 166192 mg of ethylenediamine
chloride and 5 mL of 1 N sodium hydroxide to render (C2H8N2) per g of C7H8N4O2 found in the Assay.
alkaline. Shake by mechanical means for 10 min, add 5 mL
of 3 N hydrochloric acid to acidify, and chill. Collect the SPECIFIC TESTS
precipitated disulfonamide of ethylenediamine, wash with PH 791: 8.69.0
water, recrystallize from water, and dry at 105 for 1 h. PARTICULATE MATTER IN INJECTIONS 788: Meets the
Acceptance criteria: Melts between 164 and 171 requirements for small-volume injections
B. PROCEDURE: Sample 2 melts between 270 and 274. OTHER REQUIREMENTS: Meets the requirements under
C. PROCEDURE Injections 1
Analysis: To 10 mg of Sample 2 contained in a porcelain BACTERIAL ENDOTOXINS TEST 85: It contains NMT 1.0 USP
dish, add 1 mL of hydrochloric acid and 100 mg of Endotoxin Unit/mg of aminophylline.
potassium chlorate, evaporate on a steam bath to dryness,
and invert the dish over a vessel containing a few drops of 6
N ammonium hydroxide.
196 Aminophylline / Official Monographs USP 32
ADDITIONAL REQUIREMENTS Tailing factor: NMT 2.0 for theophylline peak, System
PACKAGING AND STORAGE: Preserve in single-dose containers suitability solution
from which carbon dioxide has been excluded, preferably of Relative standard deviation: NMT 2.0% in the Standard
Type I glass, protected from light. solution
LABELING: Label the Injection to state the content of Analysis
anhydrous theophylline. Samples: Sample solution and Standard solution
USP REFERENCE STANDARDS 11 Calculate the percentage of C7H8N4O2 in each mL of the
USP Endotoxin RS Oral Solution taken:
USP Theophylline RS
rU = peak response of the Sample solution
Aminophylline Oral Solution rS = peak response of the Standard solution
(Comment on this Monograph)id=m3140=Aminophylline Oral CS = concentration of USP Theophylline RS in the
Solution=A-Monos.pdf) Standard solution (mg/mL)
CU = nominal concentration of theophylline in the
DEFINITION Sample solution (mg/mL)
Aminophylline Oral Solution is an aqueous solution of Acceptance criteria: 90.0%110.0%
Aminophylline, prepared with the aid of Ethylenediamine. It
contains an amount of aminophylline equivalent to NLT 90.0% OTHER COMPONENTS
and NMT 110.0% of the labeled amount of anhydrous CONTENT OF ETHYLENEDIAMINE
theophylline (C7H8N4O2). Sample solution: Equivalent to 500 mg of aminophylline to
Aminophylline Oral Solution may contain an excess of 30 mL, dilute with water if necessary.
ethylenediamine, but no other substance may be added for Analysis: Add methyl orange TS, and titrate with 0.1 N
the purpose of pH adjustment. hydrochloric acid VS. Each mL of 0.1 N hydrochloric acid is
equivalent to 3.005 mg of C2H8N2.
IDENTIFICATION Acceptance criteria: 176283 mg of ethylenediamine
Sample 1: To an equivalent to 500 mg of aminophylline, (C2H8N2) per g of C7H8N4O2 found in the Assay
add, with constant stirring, 1 mL of 3 N hydrochloric acid or
an amount sufficient to precipitate the theophylline SPECIFIC TESTS
completely, and filter (retain the filtrate). PH 791: 8.59.7
Sample 2: Wash the precipitate from Sample 1 with small
portions of cold water until free from chloride, and dry at 105 ADDITIONAL REQUIREMENTS
for 1 h. PACKAGING AND STORAGE: Preserve in tight containers.
A. PROCEDURE: Sample 2 melts between 270 and 274. LABELING: Label the Oral Solution to state the content of
B. PROCEDURE anhydrous theophylline.
Analysis: To Sample 1, add 0.5 mL of benzenesulfonyl USP REFERENCE STANDARDS 11
chloride and 5 mL of 1 N sodium hydroxide to render USP Theophylline RS
alkaline, shake by mechanical means for 10 min, add 5 mL
of 3 N hydrochloric acid to acidify, chill, collect the
precipitated disulfonamide of ethylenediamine, wash with Aminophylline Rectal Solution
water, recrystallize from water, and dry at 105 for 1 h.
Acceptance criteria: Melts between 164 and 171 (Comment on this Monograph)id=m3120=Aminophylline Rectal
Solution=A-Monos.pdf)
ASSAY
Mobile phase: 200 mL of methanol, 960 mg of sodium 1- Aminophylline Rectal Solution is an aqueous solution of
pentanesulfonate, and sufficient water to make 1 L. Aminophylline, prepared with the aid of Ethylenediamine. It
Adjust with glacial acetic acid to a pH of 2.9 0.1. contains an amount of aminophylline equivalent to NLT 90.0%
Diluent: Methanol and water (1:4) and NMT 110.0% of the labeled amount of anhydrous
Standard solution: 0.08 mg/mL of USP Theophylline RS in theophylline (C7H8N4O2).
Diluent Aminophylline Rectal Solution may contain an excess of
Solution A: 0.08 mg/mL of theobromine in the Standard ethylenediamine, but no other substance may be added for
solution the purpose of pH adjustment.
System suitability solution: 64 g/mL of theophylline and IDENTIFICATION
64 g/mL of theobromine, from Solution A, in Diluent Sample 1: Dilute an equivalent of 500 mg of aminophylline,
Sample solution: Equivalent to 72 g/mL of anhydrous with water to 20 mL, and add, with constant stirring, 1 mL of
theophylline, from Oral Solution in Diluent 3 N hydrochloric acid or enough to precipitate the
Chromatographic system theophylline completely, and filter.
(See Chromatography 621, System Suitability.) Sample 2: Wash the precipitate from Sample 1 with a small
Mode: LC portion of cold water until free from chloride, and dry at 105
Detector: UV 254 nm for 4 h.
Column: 3.9-mm 15-cm; packing L1 A. INFRARED ABSORPTION 197K
Flow rate: 1 mL/min Sample: Sample 2
Injection size: 10 L B. PROCEDURE
System suitability Analysis: To Sample 1, add 0.5 mL of benzenesulfonyl
Samples: System suitability solution and Standard solution chloride and 5 mL of 1 N sodium hydroxide to render
[NOTEThe relative retention times for theobromine and alkaline, shake by mechanical means for 10 min, add 5 mL
theophylline are about 0.65 and 1.0, respectively.] of 3 N hydrochloric acid to acidify, chill, collect the
Suitability requirements precipitated disulfonamide of ethylenediamine, wash with
Resolution: NLT 3.0 between theobromine and water, recrystallize from water, and dry at 105 for 1 h.
Acceptance criteria: Melts between 164 and 171 Acceptance criteria: Melts between 270 and 274.
Analysis 2: To about 10 mg of the dried precipitate from
ASSAY Sample 1, contained in a porcelain dish, add 1 mL of
PROCEDURE hydrochloric acid and 100 mg of potassium chlorate,
Standard solution: 8 g/mL of USP Theophylline RS in evaporate on a steam bath to dryness, and invert the dish
0.12 M hydrochloric acid over a vessel containing a few drops of 6 N ammonium
Sample stock solution: Equivalent to 1 mg/mL of hydroxide.
aminophylline, from Rectal Solution in water Acceptance criteria: The residue acquires a purple color,
Sample solution: 0.01 mg/mL aminophylline from Sample which is destroyed by solutions of fixed alkalies.
stock solution, in 1.2 M hydrochloric acid, and water (10:89) B. PROCEDURE
[NOTEDilute in hydrochloric acid before diluting with Sample 2: Filtrate from preparation of Sample 1 (above).
water to volume.] Analysis: To Sample 2 add 0.5 mL of benzenesulfonyl
Spectrometric conditions chloride and 5 mL of 1 N sodium hydroxide to render
Cell: 1 cm alkaline, shake by mechanical means for 10 min, add 5 mL
Analytical wavelength: Maxima at 270 nm of 3 N hydrochloric acid to acidify, chill, collect the
Blank: 0.12 M hydrochloric acid precipitated disulfonamide of ethlyenediamine, wash with
Analysis water, recrystallize from water, and dry at 10 for 1 h.
Samples: Standard solution and Sample solution Acceptance criteria: The dried precipitate melts between
Calculate the percentage of C7H8N4O2 in each mL of the 164 and 171.
Rectal Solution taken:
ASSAY
Result = (AU/AS) (CS/CU) 100 PROCEDURE
Sample composite: Tare a small dish and a glass rod, place
AU = absorbance of the Sample solution in the dish NLT 5 Suppositories, and heat on a steam bath
AS = absorbance of the Standard solution until melted. Mix the melt by stirring it with the rod, cool
CS = concentration of USP Theophylline RS in the while stirring, and weigh.
Standard solution (g/mL) Sample stock solution: Weigh a portion of the Sample
CU = nominal concentration of anhydrous composite, equivalent to 1 g of aminophylline, place it in a
theophylline in the Sample solution (mg/mL) beaker, add 60 mL of hot water and 3 mL of nitric acid, and
Acceptance criteria: 90.0%110.0% heat on a steam bath for 15 min with frequent stirring.
Cool, transfer to a separator with the aid of 40 mL of ether,
OTHER COMPONENTS shake well, and allow to separate, using a few mL of alcohol,
ETHYLENEDIAMINE CONTENT if necessary, to bring about separation of any emulsion that
Sample solution: A volume of Rectal Solution equivalent to has formed. Draw the water layer into a 100-mL volumetric
500 mg of Aminophylline. Dilute with water, if necessary, to flask, wash the ether with two 15-mL portions of water,
make 30 mL. adding the washings to the volumetric flask, dilute with
Analysis: Add methyl orange TS, and titrate with 0.1 N water to volume.
hydrochloric acid VS. Each mL of 0.1 N hydrochloric acid is Sample solution: Transfer a portion of the Sample stock
equivalent to 3.005 mg of C2H8N2. solution, equivalent to 250 mg of aminophylline, to a 250-
Acceptance criteria: 218267 mg of ethylenediamine mL conical flask, add 10 mL of 6 N ammonium hydroxide
(C2H8N2) per g of C7H8N4O2 found in the Assay and 20 mL of 0.1 N silver nitrate VS, and heat on a steam
SPECIFIC TESTS bath for 15 min. Cool to between 5 and 10 for 20 min,
PH 791: 9.09.5 then filter, preferably through a filtering crucible of fine
porosity under reduced pressure, and wash the precipitate
ADDITIONAL REQUIREMENTS with small portions of water until the last washing gives not
PACKAGING AND STORAGE: Preserve in tight, single-dose or more than a faint opalescence with hydrochloric acid.
multiple-dose containers, at a controlled room temperature. Dissolve the precipitate by pouring over it small volumes of
LABELING: Label the Rectal Solution to state the content of warm 2 N nitric acid, receiving the solution in a conical
anhydrous theophylline. flask. Wash the filtering crucible a few times with warm
USP REFERENCE STANDARDS 11 water acidified with nitric acid, receiving the washings in the
USP Theophylline RS same flask. Cool, and add 2 mL of ferric ammonium sulfate
TS.
Analysis: Titrate with 0.1 N ammonium thiocyanate VS.
Each mL of 0.1 N ammonium thiocyanate is equivalent to
Aminophylline Suppositories 18.02 mg of C7H8N4O2.
(Comment on this Monograph)id=m3150=Aminophylline Acceptance criteria: 90.0%110.0%
Suppositories=A-Monos.pdf)
OTHER COMPONENTS
DEFINITION CONTENT OF ETHYLENEDIAMINE
Aminophylline Suppositories contain an amount of Sample: Weigh a portion of the Sample composite from the
aminophylline equivalent to NLT 90.0% and NMT 110.0% of Assay, equivalent to 500 mg of aminophylline, and place in
the labeled amount of anhydrous theophylline (C7H8N4O2). a 500-mL conical flask.
Analysis: Add 150 mL of a mixture of equal volumes of
IDENTIFICATION alcohol and ether, and warm gently under reflux for 30 min,
A. PROCEDURE with occasional swirling. Cool to room temperature and
Sample 1: Evaporate a portion of Sample stock solution from titrate with 0.1 N hydrochloric acid VS, using a glass-
the Assay, equivalent to 500 mg of aminophylline, to about modified calomel electrode system (replace the saturated
one-half its volume on a steam bath, adjust with 1 N sodium potassium chloride solution of the calomel electrode with
hydroxide to a pH of 7.0, chill, and filter the crystals of methanol saturated with lithium chloride). Each mL of 0.1 N
theophylline. [NOTESave the filtrate for use in Procedure B.] hydrochloric acid is equivalent to 3.005 mg of
Analysis 1: Wash crystals from Sample 1 with small portions ethylenediamine (C2H8N2).
of ice-cold water, and dry at 105 for 1 h.
198 Aminophylline / Official Monographs USP 32
Acceptance criteria: 152 mg190 mg of C2H8N2 per g of washings with nitric acid, and add an additional 3 mL of the
C7H8N4O2 found in the Assay acid. Cool, and add 2 mL of ferric ammonium sulfate TS.
Analysis: Titrate the excess silver nitrate with 0.1 N
ADDITIONAL REQUIREMENTS ammonium thiocyanate VS. Each mL of 0.1 N silver nitrate is
PACKAGING AND STORAGE: Preserve in well-closed containers, equivalent to 18.02 mg of C7H8N4O2.
in a cold place. Acceptance criteria: 93.0%107.0%
LABELING: Label the Suppositories to state the content of
anhydrous theophylline. OTHER COMPONENTS
ETHYLENEDIAMINE CONTENT
aminophylline from powdered Tablets (NLT 20) to a 100-mL
Aminophylline Tablets conical flask, add 20 mL of water, and digest at 50, with
(Comment on this Monograph)id=m3160=Aminophylline frequent shaking, for 30 min. Cool, filter into a 250-mL
Tablets=A-Monos.pdf) conical flask, and wash with water until the last washing is
neutral to litmus. To the combined filtrate and washings,
DEFINITION add methyl orange TS.
Aminophylline Tablets contain an amount of aminophylline Analysis: Titrate with 0.1 N hydrochloric acid VS. Each mL
equivalent to NLT 93.0% and NMT 107.0% of the labeled of 0.1 N hydrochloric acid is equivalent to 3.005 mg of
amount of theophylline (C7H8N4O2). C2H8N2.
[NOTEThe ammoniacal odor present in the vapor space above Acceptance criteria: 140190 mg of C2H8N2 per g of
Aminophylline Tablets is often quite strong, especially when C7H8N4O2 found in the Assay
bottles having suitably tight closures are newly opened. This is
due to ethylenediamine vapor pressure build-up, a natural PERFORMANCE TESTS
condition in the case of aminophylline.] DISSOLUTION 711
For uncoated or plain coated tablets
IDENTIFICATION Medium: Water; 900 mL
Sample 1: Macerate a quantity of Tablets, equivalent to 500 Apparatus 2: 50 rpm
mg of aminophylline, with 25 mL of water, and filter: the Time: 45 min
filtrate is alkaline to litmus. To the filtrate, add 1 mL of 3 N Standard solution: USP Theophylline RS in Medium.
hydrochloric acid, stir, and chill, if necessary, to precipitate the Sample solutions: Sample per Dissolution 711.
theophylline. Filter, and retain the filtrate, free from washings. Dilute with Medium to a concentration that is similar to
Sample 2: Wash the crystals obtained from Sample 1 on the that of the Standard solution.
filter with small quantities of ice-cold water, and dry at 105 Detector: UV 269 nm
for 1 h. Tolerances: NLT 75% (Q) of the labeled amount of
A. To 10 mg of Sample 2, in a porcelain dish, add 1 mL of C7H8N4O2
hydrochloric acid and 100 mg of potassium chlorate, UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
evaporate on a steam bath to dryness, and invert the dish Procedure for content uniformity
over a vessel containing a few drops of 6 N ammonium Sample solution: Place 1 Tablet in a 250-mL volumetric
hydroxide: the residue acquires a purple color, which is flask, add 200 mL of water, and shake by mechanical
destroyed by solutions of fixed alkalies. means until disintegration is complete. Add water to
B. Recrystallize Sample 2 from water and dry at 105 for 1 volume. Filter a portion of the mixture, discarding the first
h: it melts between 270 and 274. 20 mL of the filtrate.
C. To about 25 mL of Sample 1, add 0.5 mL of Standard solution: 10 g/mL USP Theophylline RS
benzenesulfonyl chloride and 5 mL of 1 N sodium hydroxide Spectrometric conditions
to render alkaline. Shake by mechanical means for 10 min. Cell: 1cm
Add 5 mL of 3 N hydrochloric acid to acidify. Chill, collect Analytical wavelength: 269 nm
the precipitated disulfonamide of ethylenediamine, wash Blank: Water
with water, recrystallize from water, and dry at 105 for 1 h: Analysis
the dried precipitate melts between 164 and 171. Samples: Standard solution and Sample solution
Calculate the percentage of the label claim of C7H8N4O2 in
ASSAY each Tablet:
PROCEDURE
Sample solution: Transfer the equivalent to 2 g of Result = (AU/AS) (CS/CU) 100
aminophylline from powdered Tablets (NLT 20) to a 200-mL
volumetric flask, and add 50 mL of water and 15 mL of 6 N AU = absorbance of the Sample solution
ammonium hydroxide. Allow to stand for 30 min with AS = absorbance of the Standard solution
frequent shaking, warming to 50, if necessary, to dissolve CS = concentration of USP Theophylline RS in the
the aminophylline. Cool the mixture to room temperature if Standard solution (g /mL)
it has been warmed, and add water to volume. Centrifuge CU = concentration of theophylline in the Sample
50 mL of the mixture, and pipet a portion of the clear solution (mg/mL)
supernatant, equivalent to 250 mg of aminophylline, into a
250-mL conical flask, and dilute with water, if necessary, to ADDITIONAL REQUIREMENTS
make 40 mL. Add 8 mL of 6 N ammonium hydroxide and PACKAGING AND STORAGE: Preserve in tight containers.
20.0 mL of 0.1 N silver nitrate VS, mix, heat to boiling, and LABELING: Label the Tablets to state the content of
continue boiling for 15 min. Cool to between 5 and 10 for anhydrous theophylline.
20 min, then filter, preferably through a filtering crucible USP REFERENCE STANDARDS 11
under reduced pressure, and wash the precipitate with three USP Theophylline RS
10-mL portions of water. Acidify the combined filtrate and
Aminophylline Delayed-Release Tablets means until disintegration is complete. Add water to

(Comment on this Monograph)id=m3165=Aminophylline volume. Filter a portion of the mixture, discarding the first
Delayed-Release Tablets=A-Monos.pdf) 20 mL of the filtrate.
DEFINITION Cell: 1 cm
Aminophylline Delayed-Release Tablets contain an amount of Analytical wavelength: 269 nm
aminophylline equivalent to NLT 93.0% and NMT 107.0% of Blank: Water
the labeled amount of anhydrous theophylline (C7H8N4O2). Analysis
[NOTEThe ammoniacal odor present in the vapor space above Samples: Standard solution and Sample solution
Aminophylline Delayed-Release Tablets is often quite strong, Calculate the label claim percentage of C7H8N4O2 in each
especially when bottles having suitably tight closures are newly Tablet:
opened. This is due to ethylenediamine vapor pressure build-
up, a natural condition in the case of aminophylline.] Result = (AU/AS) (CS /CU) 100
ASSAY AU = absorbance of the Sample solution

PROCEDURE AS = absorbance of the Standard solution
Sample solution: Transfer an equivalent to 2 g of CS = concentration of USP Theophylline RS in the
aminophylline, from powdered Tablets (NLT 20), to a 200- Standard solution (g/mL)
mL volumetric flask. Add 50 mL of water and 15 mL of 6 N CU = concentration of theophylline in the Sample
ammonium hydroxide, and allow to stand for 30 min with solution (g/mL)
frequent shaking, warming to 50, if necessary, to dissolve
the aminophylline. Cool the mixture to room temperature if SPECIFIC TESTS
it has been warmed, and add water to volume. Centrifuge OTHER REQUIREMENTS
50 mL of the mixture, and pipet a portion of the clear Sample 1: Macerate a quantity of Tablets, equivalent to 500
supernatant, equivalent to 250 mg of aminophylline, into a mg of aminophylline, with 25 mL of water, and filter: the
250-mL conical flask, and dilute with water, if necessary, to filtrate is alkaline to litmus. To the filtrate add 1 mL of 3 N
make 40 mL. Add 8 mL of 6 N ammonium hydroxide and hydrochloric acid, stir, and chill, if necessary, to precipitate
20.0 mL of 0.1 N silver nitrate VS, mix, heat to boiling, and the theophylline. Filter, and retain the filtrate, free from
continue boiling for 15 min. Cool to between 5 and 10 for washings.
20 min, then filter, preferably through a filtering crucible Sample 2: Wash the crystals obtained from Sample 1 on the
under reduced pressure, and wash the precipitate with three filter with small quantities of ice-cold water, and dry at 105
10-mL portions of water. Acidify the combined filtrate and for 1 h.
washings with nitric acid, and add an additional 3 mL of the Procedure 1
acid. Cool, and add 2 mL of ferric ammonium sulfate TS. Analysis: To 10 mg of Sample 2, in a porcelain dish, add 1
Analysis: Titrate the excess silver nitrate with 0.1 N mL of hydrochloric acid and 100 mg of potassium chlorate,
ammonium thiocyanate VS. Each mL of 0.1 N silver nitrate is evaporate on a steam bath to dryness, and invert the dish
equivalent to 18.02 mg of C7H8N4O2. over a vessel containing a few drops of 6 N ammonium
Acceptance criteria: 93.0%107.0% hydroxide.
Acceptance criteria: The residue acquires a purple color,
OTHER COMPONENTS which is destroyed by solutions of fixed alkalies.
ETHYLENEDIAMINE CONTENT Procedure 2
Sample solution: Weigh a portion of the powdered Tablets Analysis: Recrystallize Sample 2 from water and dry at
prepared in the Assay, equivalent to 350 mg of 105 for 1 h.
aminophylline, transfer to a 100-mL conical flask, add 20 mL Acceptance criteria: It melts between 270 and 274.
of water, and digest at 50, with frequent shaking, for 30 Procedure 3
min. Cool, filter into a 250-mL conical flask, and wash with Analysis: To about 25 mL of Sample 1, add 0.5 mL of
water until the last washing is neutral to litmus. To the benzenesulfonyl chloride and 5 mL of 1 N sodium
combined filtrate and washings add methyl orange TS. hydroxide to render alkaline, and shake by mechanical
Analysis: Titrate with 0.1 N hydrochloric acid VS. Each mL means for 10 min. Add 5 mL of 3 N hydrochloric acid to
of 0.1 N hydrochloric acid is equivalent to 3.005 mg of acidify, chill, collect the precipitated disulfonamide of
C2H8N2. ethylenediamine, wash with water, recrystallize from water,
Acceptance criteria: 140190 mg of C2H8N2 per g of and dry at 105 for 1 h.
C7H8N4O2 found in the Assay Acceptance criteria: The precipitate melts between 164
and 171.
PERFORMANCE TESTS
DISINTEGRATION, Procedure 701: 30 min, determined as ADDITIONAL REQUIREMENTS
directed under Delayed-Release (Enteric-Coated) Tablets PACKAGING AND STORAGE: Preserve in tight containers.
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements LABELING: Label the Tablets to state the content of
Analysis for content uniformity anhydrous theophylline.
Standard solution: 10 g/mL USP Theophylline RS USP REFERENCE STANDARDS 11
Sample solution: Place 1 Tablet in a 250-mL volumetric USP Theophylline RS
flask, add 200 mL of water, and shake by mechanical
200 Aminosalicylate / Official Monographs USP 32
Aminosalicylate Sodium Suitability requirements

(Comment on this Monograph)id=m3340=Aminosalicylate Resolution: NLT 1.7 between aminosalicylic acid and
Sodium=A-Monos.pdf) acetaminophen
Relative standard deviation: NMT 1.0% for the peak
C7H6NNaO3 2H2O 211.15 response ratio of the aminosalicylic acid peak to the
Benzoic acid, 4-amino-2-hydroxy-, monosodium salt, dihydrate; acetaminophen peak
Monosodium 4-aminosalicylate dihydrate [6018-19-5]. Analysis
Anhydrous [133-10-8]. 175.12 Samples: Standard solution and Sample solution
[NOTEAfter use, wash the column for 30 min with
DEFINITION methanol, water, and phosphoric acid (77:23:0.6), and
Aminosalicylate Sodium contains NLT 98.0% and NMT 101.0% then wash for 30 min with methanol and water (50:50).]
of C7H6NNaO3, calculated on the anhydrous basis. Calculate the percentage of C7H6NNaO3 taken:
[CAUTIONPrepare solutions of Aminosalicylate Sodium within
24 h of administration. Under no circumstances use a solution Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100
if its color is darker than that of a freshly prepared solution.]
RU = peak response ratio of aminosalicylate peak to
IDENTIFICATION the acetaminophen peak from the Sample
A. Dissolve 250 mg in 3 mL of 1 N sodium hydroxide, solution
transfer to a 500-mL volumetric flask, and dilute with water RS = peak response ratio of aminosalicylic acid peak
to volume. Transfer a 5-mL aliquot to a 250-mL volumetric to the acetaminophen peak from the Standard
flask containing 12.5 mL of pH 7 phosphate buffer (see solution
Reagents, Indicators, and SolutionsBuffer Solutions), and CS = concentration of USP Aminosalicylic Acid RS in
dilute with water to volume. This solution, when compared the Standard solution (mg/mL)
in a suitable spectrophotometer against a blank of the same CU = concentration of aminosalicylate in the Sample
buffer in the same concentration, exhibits absorbance solution (mg/mL)
maxima at 265 2 nm and 299 2 nm, and the ratio Mr1 = molecular weight of anhydrous aminosalicylate
A265/A299 is between 1.50 and 1.56. sodium, 175.12
B. PROCEDURE Mr2 = molecular weight of aminosalicylic acid, 153.14
Analysis: Place 1-g sample in a small, round-bottom flask, Acceptance criteria: 98.0%101.0%
and add 10 mL of acetic anhydride. Heat the flask on a
steam bath for 30 min, add 40 mL of water, filter, cool, and IMPURITIES
allow to stand until the diacetyl derivative has crystallized. Inorganic Impurities
Collect the precipitate on a filter, wash well with water, and CHLORIDE AND SULFATE, Chloride 221
dry at 105 for 1 h. Sample solution: 25 mg/mL in 4 M nitric acid
Acceptance criteria: The diacetyl derivative melts between Acceptance criteria: The solution shows no more chloride
191 and 197. than corresponds to 0.30 mL of 0.020 N hydrochloric acid
C. PROCEDURE: Dissolve 50 mg in 5 mL of water, add 1 mL (0.042%).
of 3 N hydrochloric acid, and filter if necessary. To the HEAVY METALS, Method II 231: NMT 30 ppm
filtrate add 1 drop of ferric chloride TS: a violet color is Organic Impurities
produced. PROCEDURE: LIMIT OF M-AMINOPHENOL
D. IDENTIFICATIONS TESTSGENERAL, Sodium 191: Meets the Solution A: 12.7 mg/mL tetrabutylammonium hydroxide
requirements in methanol
Mobile phase: Solution A, 0.05 M dibasic sodium
ASSAY phosphate, and 0.05 M monobasic sodium phosphate
PROCEDURE (30:85:85)
Solution A: 12.7 mg/mL tetrabutylammonium hydroxide in Internal standard solution: 5 g/mL of sulfanilamide in
methanol Mobile phase
Mobile phase: Solution A, 0.05 M dibasic sodium Standard stock solution: 12 g/mL of USP m-
phosphate, and 0.05 M monobasic sodium phosphate Aminophenol RS in Mobile phase
(30:85:85) Standard solution: Standard stock solution, Internal
Internal standard solution: 5 mg/mL of acetaminophen in standard solution, and Mobile phase (1:1:8) in a low-actinic
Mobile phase volumetric flask
Standard solution: 0.5 mg/mL of USP Aminosalicylic Acid Sample solution: Use the Sample solution, prepared as
RS and 0.5 mg/mL of acetaminophen from Internal standard directed in the Assay.
solution in Mobile phase [NOTEDissolve in a portion of Chromatographic system
Mobile phase before diluting to final volume. Use a low- (See Chromatography 621, System Suitability.)
actinic volumetric flask.] Mode: LC
Sample solution: 0.69 mg/mL of Aminosalicylate Sodium in Detector: UV 280 nm
Mobile phase and Internal standard solution (9:1) [NOTE Column: 4.6-mm 25-cm; 10-m packing L1
Dissolve in a portion of Mobile phase before diluting to final Flow rate: 1.5 mL/min
volume. Use low-actinic glass ware.] Injection size: 20 L
Chromatographic system System suitability
(See Chromatography 621, System Suitability.) Sample: Standard solution
Mode: LC [NOTEThe relative retention times for sulfanilamide and
Detector: UV 254 nm m-aminophenol are 0.66 and 1.0, respectively.]
Column: 4.6-mm 25-cm; packing L1 Suitability requirements
Flow rate: 1.5 mL/min Resolution: NLT 2.5 between m-aminophenol and
Injection size: 20 L sulfanilamide
System suitability
Sample: Standard solution [NOTEThe relative retention
times for acetaminophen and aminosalicylic acid are 0.83
and 1.0, respectively.]
USP 32 Official Monographs / Aminosalicylate 201
Relative standard deviation: NMT 7% allow to stand until the diacetyl derivative has crystallized.
Analysis Collect the precipitate on a filter, wash well with water, and
Samples: Standard solution and Sample solution dry at 105 for 1 h.
[NOTEAfter use, wash the column for 30 min with Acceptance criteria: The diacetyl derivative melts between
methanol, water, and phosphoric acid (77:23:0.6), and 191 and 197.
then wash for 30 min with methanol and water B. PROCEDURE
(50:50).] Analysis: Shake 0.1 g of Sample with 10 mL of water, and
Calculate the percentage of m-aminophenol, in the filter. To 5 mL of the filtrate, add 1 drop of ferric chloride
portion of Aminosalicylate Sodium taken: TS.
Acceptance criteria: A violet color is produced.
ASSAY
RU = peak response ratio of the m-aminophenol peak PROCEDURE
to the sulfanilamide peak from the Sample Solution A: 12.7 mg/mL of tetrabutylammonium hydroxide
solution in methanol
RS = peak response ratio of the m-aminophenol peak Mobile phase: Solution A, 0.05 M dibasic sodium
to the sulfanilamide peak from the Standard phosphate, and 0.05 M monobasic sodium phosphate
solution (30:85:85)
CS = concentration of USP Aminophenol RS in the Internal standard solution: 5 mg/mL of acetaminophen in
Standard solution (g/mL) Mobile phase
CU = concentration for the Sample solution (mg/mL) Standard solution: 0.5 mg/mL of USP Aminosalicylic Acid
Acceptance criteria: NMT 0.25% RS and 0.5 mg/mL of acetaminophen from the Internal
standard solution in Mobile phase and the Internal standard
SPECIFIC TESTS solution (9:1) [NOTEDissolve in a portion of Mobile phase
PH 791 before diluting to final volume. Use a low-actinic volumetric
Sample solution: 20 mg/mL flask.]
Acceptance criteria: 6.58.5 Sample stock solution: Add the equivalent of 690 mg of
WATER DETERMINATION, Method I 921: aminosalicylate sodium from the powdered Tablets to a 100-
Sample solution: 20 mg/ml mL low-actinic volumetric flask. Add 50 mL of Mobile phase,
Acceptance criteria: 16.0%18.0% and shake for 5 min. Dilute with Mobile phase to volume,
HYDROGEN SULFIDE, SULFUR DIOXIDE, AND AMYL ALCOHOL: and filter.
Dissolve 500 mg in 5 mL of 1 N sodium hydroxide, add 6 Sample solution: 0.69 mg/mL of aminosalicylic acid from
mL of 3 N hydrochloric acid, and stir vigorously: no odor of the Sample stock solution and 0.5 mg/mL of acetaminophen
hydrogen sulfide or sulfur dioxide is perceptible, and not from the Internal standard solution in Mobile phase [NOTE
more than a faint odor of amyl alcohols is perceptible. A Use a low-actinic volumetric flask.]
piece of moistened lead acetate test paper held over the Chromatographic system
mixture does not become discolored. (See Chromatography 621, System Suitability.)
CLARITY AND COLOR OF SOLUTION: One g dissolves in 10 mL Mode: LC
of water to give a clear solution that has not more than a Detector: UV 254 nm
faint yellow color. One g dissolves in a freshly prepared Column: 4.6-mm 25-cm; packing L1
mixture of 5 mL of nitric acid and 45 mL of water to give a Flow rate: 1.5 mL/min
clear solution that has not more than a slight color. Injection size: 20 L
System suitability
ADDITIONAL REQUIREMENTS Sample: Standard solution
PACKAGING AND STORAGE: Preserve in tight, light-resistant [NOTEThe relative retention times for acetaminophen and
containers, protected from excessive heat. aminosalicylic acid are 0.83 and 1.0, respectively.]
USP Aminosalicylic Acid RS Resolution: NLT 1.7 between aminosalicylic acid and
USP m-Aminophenol RS acetaminophen
Relative standard deviation: NMT 1.0%, ratios of the
peak response of aminosalicylic acid to the peak response
Aminosalicylate Sodium Tablets of acetaminophen
Analysis
(Comment on this Monograph)id=m3370=Aminosalicylate Sample: Sample solution and Standard solution
Sodium Tablets=A-Monos.pdf) [NOTEAfter use, wash the column for 30 min with a
DEFINITION mixture of methanol, water, and phosphoric acid
Aminosalicylate Sodium Tablets contain NLT 95.0% and NMT (77:23:0.6), and then wash for 30 min with a mixture of
105.0% of the labeled amount of C7H6NNaO3 2H2O. methanol and water (50:50).]
Calculate the percentage of C7H6NNaO3 2H2O in the
IDENTIFICATION portion of Tablets taken:
Sample: Mix powdered Tablets, equivalent to 3 g of
aminosalicylate sodium, with 40 mL of water, and filter. Add Result = (RU/RS) (CS/CU) (Mr1/Mr2) (100/L)
to the filtrate 15 mL of 1 N acetic acid, and allow to stand
until precipitation has occurred. Collect the precipitate on a RU = peak response ratio of aminosalicylate to
filter, wash well with water, and dry at 105 for 30 min. acetaminophen from the Sample solution
A. PROCEDURE RS = peak response ratio of aminosalicylic acid to
Analysis: Place 1 g of Sample in a small, round-bottom flask, acetaminophen from the Standard solution
and add 10 mL of acetic anhydride. Heat the flask on a CS = concentration of USP Aminosalicylic Acid RS in
steam bath for 30 min, add 40 mL of water, filter, cool, and the Standard solution (mg/mL)
202 Aminosalicylate / Official Monographs USP 32
CU = concentration of aminosalicylate sodium in the RS = peak response ratio of m-aminophenol to

Sample solution sulfanilamide from the Standard solution
(unit/mL) CS = concentration of USP m-Aminophenol RS in the
Mr1 = molecular weight of aminosalicylate sodium Standard solution (g/mL)
dihydrate, 211.15 CU = concentration of aminosalicylate sodium in the
Mr2 = molecular weight of aminosalicylic acid, 153.14 Sample solution, as determined in the Assay
L = label claim (mg/unit) (mg/mL)
Acceptance criteria: 95.0%105.0% F = conversion factor (g/mg)
PERFORMANCE TESTS
DISSOLUTION, Procedure for a Pooled Sample 711 ADDITIONAL REQUIREMENTS
Medium: Water; 900 mL PACKAGING AND STORAGE: Preserve in tight, light-resistant
Apparatus 1: 100 rpm containers, protected from excessive heat.
Time: 45 min USP REFERENCE STANDARDS 11
Analysis: Determine the amount of C7H6NNaO3 2H2O USP Aminosalicylic Acid RS
dissolved, employing the procedure set forth in the Assay, USP m-Aminophenol RS
making any necessary modifications.
Tolerances: NLT 75% (Q) of C7H6NNaO3 2H2O
Aminosalicylic Acid
IMPURITIES (Comment on this Monograph)id=m3400=Aminosalicylic
Organic Impurities Acid=A-Monos.pdf)
LIMIT OF m-AMINOPHENOL
Solution A: 12.7 mg/mL of tetrabutylammonium
hydroxide in methanol
Mobile phase: Solution A, 0.05 M dibasic sodium
phosphate, and 0.05 M monobasic sodium phosphate
(30:85:85)
Internal standard solution: 5 g/mL of sulfanilamide in
Mobile phase
Standard stock solution: 12 g/mL of USP m- C7H7NO3 153.14
Aminophenol RS in Mobile phase Benzoic acid, 4-amino-2-hydroxy-;
Standard solution: Standard stock solution, Internal 4-Aminosalicylic acid [65-49-6].
standard solution, and Mobile phase (1:1:8) in a low-actinic
volumetric flask DEFINITION
Sample stock solution: Add the equivalent of 690 mg of Aminosalicylic Acid contains NLT 98.5% and NMT 100.5% of
aminosalicylate sodium from the powdered Tablets, to a C7H7NO3, calculated on the anhydrous basis.
100-mL low-actinic volumetric flask. Add 50 mL of Mobile
phase, and shake for 5 min. Dilute with Mobile phase to [CAUTIONUnder no circumstances use a solution prepared
volume, and filter. from Aminosalicylic Acid if its color is darker than that of a
Sample solution: 0.69 mg/mL of aminosalicylic acid from freshly prepared solution.]
the Sample stock solution and 0.5 mg/mL of acetaminophen
from the Internal standard solution in Mobile phase [NOTE IDENTIFICATION
Use a low-actinic volumetric flask.] A. Procedure
Chromatographic system Sample solution: Dissolve 250 mg in 3 mL of 1 N sodium
(See Chromatography 621, System Suitability.) hydroxide, transfer to a 500-mL volumetric flask, and dilute
Mode: LC with water to volume.
Detector: UV 280 nm Analysis: Transfer a 5-mL aliquot of the Sample solution to a
Column: 4.6-mm 25-cm; 10-m packing L1 250-mL volumetric flask containing 12.5 mL of pH 7
Flow rate: 1.5 mL/min phosphate buffer (see Reagents, Indicators, and Solutions
Injection size: 20 L Buffer Solutions), and dilute with water to volume. This
System suitability solution, when compared in a suitable spectrophotometer
Sample: Standard solution against a blank of the same buffer in the same
[NOTEThe relative retention times for sulfanilamide and concentration, exhibits absorbance maxima at 265 2 and
m-aminophenol are 0.66 and 1.0, respectively.] 299 2 nm, and the ratio A265/A299 is between 1.50 and
Suitability requirements 1.56.
Resolution: NLT 2.5 between m-aminophenol and B. Place 1 g in a small, round-bottom flask, and add 10 mL
sulfanilamide of acetic anhydride. Heat the flask on a steam bath for 30
Relative standard deviation: NMT 7% min, add 40 mL of water, filter, cool, and allow to stand
Analysis until the diacetyl derivative has crystallized. Collect the
Sample: Sample solution and Standard solution precipitate on a filter, wash well with water, and dry at 105
[NOTEAfter use, wash the column for 30 min with a for 1 h.
mixture of methanol, water, and phosphoric acid Acceptance criteria: The diacetyl derivative so obtained
(77:23:0.6), and then wash for 30 min with a mixture melts between 191 and 197.
of methanol and water (50:50).] C. Shake 0.1 g with 10 mL of water, and filter. To 5 mL of
Calculate the percentage of m-aminophenol in the portion the filtrate, add 1 drop of ferric chloride TS.
of Tablets taken: Acceptance criteria: A violet color is produced.
ASSAY
Result = (RU/RS) (CS/CU) F 100 PROCEDURE
RU = peak response ratio of m-aminophenol to Solution A: 12.7 mg/mL of tetrabutylammonium hydroxide
sulfanilamide from the Sample solution in methanol
USP 32 Official Monographs / Aminosalicylic 203
Mobile phase: Solution A, 0.05 M dibasic sodium Standard stock solution: 12 g/mL of USP m-
phosphate, and 0.05 M monobasic sodium phosphate Aminophenol RS in Mobile phase
(30:85:85) Standard solution: Standard stock solution, Internal
Internal standard solution: 5 mg/mL of acetaminophen in standard solution, and Mobile phase (1:1:8) in a low-actinic
Mobile phase volumetric flask
Standard solution: 0.5 mg/mL of USP Aminosalicylic Acid Sample solution: 0.5 mg/mL of Aminosalicylic Acid in a
RS and 0.5 mg/mL of acetaminophen from Internal standard mixture of Mobile phase and Internal standard solution (9:1)
solution in a mixture of Mobile phase and Internal standard [NOTEDissolve in a portion of Mobile phase before diluting
solution (9:1) to final volume. Use a low-actinic volumetric flask.]
[NOTEDissolve in a portion of Mobile phase before diluting Chromatographic system:
to final volume. Use a low-actinic volumetric flask.] (See Chromatography 621, System Suitability.)
Sample solution: 0.5 mg/mL of Aminosalicylic Acid in a Mode: LC
mixture of Mobile phase and Internal standard solution (9:1) Detector: UV 280 nm
Chromatographic system Column: 4.6-mm 25-cm; 10-m packing L1
(See Chromatography 621, System Suitability.) Flow rate: 1.5 mL/min
Detector: UV 254 nm System suitability
Column: 4.6-mm 25-cm; packing L1 Sample: Standard solution
Flow rate: 1.5 mL/min [NOTERelative retention times for sulfanilamide and for
Injection size: 20 L m-aminophenol are about 0.66 and 1.0, respectively.]
System suitability Suitability requirements
Sample: Standard solution [NOTEThe relative retention Resolution: NLT 2.5 between m-aminophenol and
times for acetaminophen and for aminosalicylic acid are sulfanilamide
0.83 and 1.0, respectively.] Relative standard deviation: NMT 7%
Resolution: NLT 1.7 between aminosalicylic acid and Sample: Sample solution and Standard solution
acetaminophen [NOTEAfter use, wash the column for 30 min with a
Relative standard deviation: NMT 1.0% for the peak mixture of methanol, water, and phosphoric acid
response ratios of the aminosalicylic acid to the (77:23:0.6), and then wash for 30 min with a mixture
acetaminophen peaks of methanol and water (50:50).]
Analysis Calculate the percentage of m-aminophenol, in the
Sample: Sample solution and Standard solution portion of Aminosalicylic Acid taken:
[NOTEAfter use, wash the column for 30 min with a
mixture of methanol, water, and phosphoric acid Result = (RU/RS) (CS/CU) F 100
(77:23:0.6), and then wash for 30 min with a mixture of
methanol and water (50:50).] RU = peak response ratio of m-aminophenol to
Calculate the percentage of C7H7NO3 in the portion of sulfanilamide from the Sample solution
Aminosalicylic Acid taken: RS = peak response ratio of the m-aminophenol to
sulfanilamide from the Standard solution
Result = (RU/RS) (CS/CU) 100 CS = concentration of USP m-Aminophenol RS in the
RU = peak response ratio of aminosalicylic acid to CU = concentration of the Sample solution (mg/mL)
acetaminophen from the Sample solution F = conversion factor (g/mg)
RS = peak response ratio of aminosalicylic acid to Acceptance criteria: NMT 0.25%
acetaminophen from the Standard solution
CS = concentration of USP Aminosalicylic acid RS in SPECIFIC TESTS
the Standard solution (mg/mL) PH 791: 3.03.7, in a saturated solution
CU = concentration of aminosalicylic acid in the WATER DETERMINATION, Method I 921: NMT 0.5%
Sample solution (mg/mL) HYDROGEN SULFIDE, SULFUR DIOXIDE, AND AMYL ALCOHOL
Acceptance criteria: 98.5%100.5% Sample solution: Dissolve 500 mg in 5 mL of 1 N sodium
hydroxide, add 6 mL of 3 N hydrochloric acid, and stir
IMPURITIES vigorously.
Inorganic Impurities Acceptance criteria: No odor of hydrogen sulfide or sulfur
RESIDUE ON IGNITION 281: NMT 0.2% dioxide is perceptible, and NMT a faint odor of amyl alcohol
CHLORIDE AND SULFATE, Chloride 221 is perceptible. A piece of moistened lead acetate test paper
Sample solution: 25 mg/mL in 4 M nitric acid held over the mixture does not become discolored.
Acceptance criteria: The solution shows no more chloride CLARITY AND COLOR OF SOLUTION: One g dissolves in 10 mL
than corresponds to 0.30 mL of 0.020 N hydrochloric acid of sodium bicarbonate solution (1 in 15) to form a clear
(0.042%). solution that has NMT a faint yellow color. One g dissolves
HEAVY METALS, Method II 231: NMT 30 ppm in 50 mL of freshly prepared 1.6 M nitric acid to form a clear
Organic Impurities solution that has NMT a slight color.
LIMIT OF m-AMINOPHENOL
Solution A: 12.7 mg/mL of tetrabutylammonium ADDITIONAL REQUIREMENTS
hydroxide in methanol PACKAGING AND STORAGE: Preserve in tight, light-resistant
Mobile phase: Solution A, 0.05 M dibasic sodium containers, at a temperature not exceeding 30.
phosphate, and 0.05 M monobasic sodium phosphate USP REFERENCE STANDARDS 11
(150:425:425) USP Aminosalicylic Acid RS
Internal standard solution: 5 g/mL of sulfanilamide in USP m-Aminophenol RS
Mobile phase
204 Aminosalicylic / Official Monographs USP 32
Aminosalicylic Acid Tablets (77:23:0.6), and then wash for 30 min with a mixture of
(Comment on this Monograph)id=m3430=Aminosalicylic Acid methanol and water (50:50).]
Tablets=A-Monos.pdf) Calculate the percentage of C7H7NO3 in the portion of
Tablets taken:
DEFINITION
Aminosalicylic Acid Tablets contain NLT 95.0% and NMT Result = (RU/RS) (CS/CU) 100
105.0% of the labeled amount of C7H7NO3.
RU = peak response ratio of aminosalicylic acid to
IDENTIFICATION acetaminophen from the Sample solution
Sample: Mix powdered Tablets, equivalent to 2 g of RS = peak response ratio of aminosalicylic acid to
aminosalicylic acid, with 50 mL of a mixture of acetone and acetaminophen from the Standard solution
chloroform (1:2), and filter. Evaporate the filtrate with a CS = concentration of USP Aminosalicylic Acid RS in
current of warm air to dryness. the Standard solution (mg/mL)
A. PROCEDURE CU = concentration of aminosalicylic acid in the
Analysis: Place 1 g of Sample in a small, round-bottom flask, Sample solution (mg/mL)
and add 10 mL of acetic anhydride. Heat the flask on a Acceptance criteria: 95.0%105.0%
steam bath for 30 min, add 40 mL of water, filter, cool, and
allow to stand until the diacetyl derivative has crystallized. PERFORMANCE TESTS
Collect the precipitate on a filter, wash well with water, and DISSOLUTION 711
dry at 105 for 1 h. Medium: pH 7.5 phosphate buffer; 900 mL (see Reagents,
Acceptance criteria: The diacetyl derivative melts between Indicators, and SolutionsBuffer Solutions)
191 and 197. Apparatus 1: 100 rpm
B. PROCEDURE Time: 45 min
Analysis: Shake 0.1 g of Sample with 10 mL of water, and Analysis: Determine the amount of C7H7NO3 dissolved,
filter. To 5 mL of the filtrate, add 1 drop of ferric chloride employing the procedure in the Assay, making any
TS. necessary modifications.
Acceptance criteria: A violet color is produced. Tolerances: NLT 75% (Q) of C7H7NO3
ASSAY
Solution A: 12.7 mg/mL of tetrabutylammonium hydroxide Organic Impurities
in methanol PROCEDURE: LIMIT OF m-AMINOPHENOL
Mobile phase: Solution A, 0.05 M dibasic sodium Mobile phase and Internal standard solution: Prepare as
phosphate, and 0.05 M monobasic sodium phosphate directed in the Assay.
(30:85:85) Standard stock solution: 12 g/mL of USP m-
Internal standard solution: 5 mg/mL of acetaminophen in Aminophenol RS in Mobile phase
Mobile phase Standard solution: Standard stock solution, Internal
Standard solution: 0.5 mg/mL of USP Aminosalicylic Acid standard solution, and Mobile phase (1:1:8) in a low-actinic
RS and 0.5 mg/mL of acetaminophen from the Internal volumetric flask
standard solution in Mobile phase [NOTEDissolve in a Sample solution: Use the Sample solution in the Assay.
portion of Mobile phase before diluting to final volume. Use Chromatographic system
a low-actinic volumetric flask.] (See Chromatography 621, System Suitability.)
Sample stock solution: Add the equivalent of 500 mg of Mode: LC
aminosalicylic acid from the powdered Tablets, to a 100-mL Detector: UV 280 nm
low-actinic volumetric flask. Add 50 mL of Mobile phase, and Column: 4.6-mm 25-cm; 10-m packing L1
shake for 5 min. Dilute with Mobile phase to volume, and Flow rate: 1.5 mL/min
filter. Injection size: 20 L
Sample solution: 0.5 mg/mL of aminosalicylic acid from System suitability
the Sample stock solution and 0.5 mg/mL of acetaminophen Sample: Standard solution
from the Internal standard solution in Mobile phase [NOTE [NOTEThe relative retention times for sulfanilamide and
Use a low-actinic volumetric flask.] for m-aminophenol are 0.66 and 1.0, respectively.]
Chromatographic system Suitability requirements
(See Chromatography 621, System Suitability.) Resolution: NLT 2.5 between m-aminophenol and
Mode: LC sulfanilamide
Detector: UV 254 nm Relative standard deviation: NMT 7%
Flow rate: 1.5 mL/min Sample: Sample solution and Standard solution
Injection size: 20 L [NOTEAfter use, wash the column for 30 min with a
System suitability mixture of methanol, water, and phosphoric acid
Sample: Standard solution (77:23:0.6), and then wash for 30 min with a mixture
[NOTEThe relative retention times for acetaminophen and of methanol and water (50:50).]
for aminosalicylic acid are 0.83 and 1.0, respectively.] Calculate the percentage of m-aminophenol, in the
Suitability requirements portion of Tablets taken:
Resolution: NLT 1.7 between aminosalicylic acid and
acetaminophen Result = (RU/RS) (CS/CU) F 100
Relative standard deviation: NMT 1.0%, ratios of the RU = peak response ratio of m-aminophenol to
peak response of aminosalicylic acid to the peak response sulfanilamide from the Sample solution
of acetaminophen RS = peak response ratio of m-aminophenol to
Analysis sulfanilamide from the Standard solution
Sample: Sample solution and Standard solution CS = concentration of USP m-Aminophenol RS in the
[NOTEAfter use, wash the column for 30 min with a Standard solution (g/mL)
mixture of methanol, water, and phosphoric acid
USP 32 Official Monographs / Amitraz 205
CU = concentration of aminosalicylic acid in the Temperature

portion of Tablets taken for the Sample solution Column: 250
as determined in the Assay (mg/mL) Detector: 250
F = conversion factor (g/mg) Carrier gas: Dry nitrogen
Acceptance criteria: NMT 1.0% Flow rate: 60 mL/min
Injection size: 5 L
ADDITIONAL REQUIREMENTS System suitability
PACKAGING AND STORAGE: Preserve in tight, light-resistant Sample: Standard solution
containers, at a temperature not exceeding 30. Suitability requirements
USP REFERENCE STANDARDS 11 Resolution: NLT 3.0 between squalane and amitraz
USP Aminosalicylic Acid RS Relative standard deviation: NMT 2.0%
USP m-Aminophenol RS Analysis
Samples: Sample solution, Standard solution, and System
Amitraz Calculate the percentage of C19H23N3 in the portion of
Amitraz taken:
(Comment on this Monograph)id=m3465=Amitraz=A-
Monos.pdf) Result = (RU/RS) (CS/CU) 100
RU = ratio of the responses of the amitraz and
squalane peaks from the Sample solution
RS = ratio of the responses of the amitraz and
squalane peaks from the Standard solution
CS = concentration of USP Amitraz RS in the Standard
solution (mg/mL)
CU = concentration of amitraz in the Sample solution
C19H23N3 293.41 (mg/mL)
Methanimidamide, N-(2,4-dimethylphenyl)-[[N-(2,4- Acceptance criteria: 95.0%101.5%
dimethylphenyl)imino]methyl-N]-methyl-;
N-Methyl-N-2,4-xylyl-N-(N-2,4-xylylformimidoyl)formamidine; IMPURITIES
N-Methylbis(2,4-xylyliminomethyl)amine [33089-61-1]. Inorganic Impurities
DEFINITION Organic Impurities
Amitraz contains NLT 95.0% and NMT 101.5% of C19H23N, PROCEDURE
calculated on the anhydrous basis. Standard solution A: 2.0 mg/mL of USP Amitraz RS in
toluene
IDENTIFICATION Standard solution B: 0.30 mg/mL of 2,4-dimethylaniline
A. INFRARED ABSORPTION 197M in toluene
B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Sample solution: 100 mg/mL of Amitraz in toluene
Sample solution: 2 mg/mL of Amitraz in toluene Chromatographic system
Standard solution: 2 mg/mL of USP Amitraz RS in toluene (See Chromatography 621, Thin-Layer Chromatography.)
Analysis: Proceed as directed for Organic Impurities, Mode: TLC
Procedure. Adsorbent: 0.25-mm layer of chromatographic silica gel
Acceptance criteria: The RF value of the principal spot in mixture
the Sample solution corresponds to that of the Standard Application volume: 2 L
solution. Developing solvent system: Cyclohexane, ethyl acetate,
C. The retention time of the major peak from the System and triethylamine (5:3:2)
suitability solution corresponds to that of the Standard Spray reagent: 0.5% solution of N-(1-naphthyl)
solution, as obtained in the Assay. ethylenediamine dihydrochloride in methanol
ASSAY Analysis
PROCEDURE Samples: Sample solution, Standard solution A, and
Internal standard solution: 10 mg/mL of squalane in Standard solution B
methyl acetate Stand the plate to a depth of 3.5 cm in a solution prepared
Standard solution: 8 mg/mL of USP Amitraz RS in Internal by dissolving 35 g of acetamide in 100 mL of methanol,
standard solution adding 100 mL of triethylamine, and diluting to 250 mL
Sample solution: 8 mg/mL of Amitraz in Internal standard with methanol. Allow the wet plate to stand in a current
solution of cold air for 30 s. Immediately apply Samples separately
System suitability solution: 8 mg/mL of Amitraz in methyl to the plate, at a level about 1 cm below the top of the
acetate impregnated zone. Promptly develop the plate. Examine
Chromatographic system the plate under short-wavelength UV light.
(See Chromatography 621, System Suitability.) Acceptance criteria 1: Any secondary spot from the
Mode: GC Sample solution is not more intense than the corresponding
Detector: Flame ionization spot from the Standard solution A (2.0%).
Column: 4-mm 1.5-m column packed with 3% liquid Expose the plate to the vapor of hydrochloric acid for 10
phase G1 on support S1A min, then expose it to the vapor of nitrogen dioxide
206 Amitraz / Official Monographs USP 32
(prepared by the reaction of nitric acid and zinc) for 10 ASSAY

min, remove any excess nitric oxide by air exhaust, spray PROCEDURE
the plate with Spray reagent, and examine the plate. Internal standard solution: 10 mg/mL of squalane in
Acceptance criteria 2: Any secondary spot from the methyl acetate
Sample solution corresponding to 2,4-dimethylaniline is not Standard solution: 8 mg/mL of USP Amitraz RS in Internal
more intense than the spot from the Standard solution B standard solution
(0.30%). Sample solution: Equivalent to 8 mg/mL of Amitraz, from
the volume of Concentrate in the Internal standard solution
SPECIFIC TESTS System suitability solution: Equivalent to 8 mg/mL of
WATER DETERMINATION, Method I 921: NMT 0.1%, Amitraz, from the volume of the Concentrate in methyl
anhydrous pyridine being used in place of methanol in the acetate
titration vessel Chromatographic system
ADDITIONAL REQUIREMENTS Mode: GC
PACKAGING AND STORAGE: Preserve in well-closed containers. Detector: Flame ionization
LABELING: Label it to indicate that it is for veterinary use Column: 4-mm 1.5-m column packed with 3% liquid
only. phase G1 on support S1A
USP REFERENCE STANDARDS 11 Temperature
USP Amitraz RS Column: 250
Detector: 250
Carrier gas: Dry nitrogen
Amitraz Concentrate for Dip Flow rate: 60 mL/min
Injection size: 5 L
(Comment on this Monograph)id=m3470=Amitraz Concentrate System suitability
for Dip=A-Monos.pdf) Sample: Standard solution
DEFINITION Suitability requirements
Amitraz Concentrate for Dip contains Amitraz in a suitable Resolution: NLT 3.0 between squalane and amitraz
vehicle. It may contain a suitable stabilizing agent. It contains Relative standard deviation: NMT 2.0%
NLT 90.0% and NMT 120.0% of the labeled amount of Analysis
C19H23N3. Samples: Sample solution, Standard solution, and System
IDENTIFICATION Calculate the percentage of C19H23N3 in the Concentrate
A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 taken:
mixture Result = (RU/RS) (CS/CU) 100
Standard solution: 5 mg/mL of USP Amitraz RS in toluene
Sample solution: 5 mg/mL of Amitraz from Concentrate for RU = ratio of the response of the amitraz peak to the
Dip diluted with toluene response of the squalane peak from the Sample
Application volume: 2 L solution
Developing solvent system: Cyclohexane, ethyl acetate, RS = ratio of the response of the amitraz peak to the
and triethylamine (5:3:2) response of the squalane peak from the
Spray reagent: 0.5% solution of N-(1-naphthyl) Standard solution
ethylenediamine dihydrochloride in methanol CS = concentration of USP Amitraz RS in the Standard
Analysis solution (mg/mL)
Samples: Sample solution and Standard solution CU = nominal concentration of amitraz in the Sample
Pre-treatment: Stand the plate to a depth of 3.5 cm in a solution (mg/mL)
solution prepared by dissolving 35 g of acetamide in 100 Acceptance criteria: 90.0%120.0%
mL of methanol, adding 100 mL of triethylamine, and SPECIFIC TESTS
diluting to 250 mL with methanol. Allow the wet plate to WATER DETERMINATION, Method I 921: NMT 0.15%, with
stand in a current of cold air for 30 s. Immediately apply anhydrous pyridine being used in place of methanol in the
Samples separately to the plate, at a level about 1 cm titration vessel
below the top of the impregnated zone. Promptly develop
the plate. ADDITIONAL REQUIREMENTS
Proceed as directed in Chromatography 621, Thin-Layer PACKAGING AND STORAGE: Preserve in well-closed containers.
Chromatography. Examine the plate under short- LABELING: Label it to indicate that it is for veterinary use
wavelength UV light. only, and that it is to be diluted before use. The label states
Acceptance criteria: The RF value of the principal spot of the name and quantity of diluent to be used, the directions
the Sample solution corresponds to that of the Standard for dilution, and the conditions for storage of the constituted
solution. Dip.
B. The retention time of the major peak of the System USP REFERENCE STANDARDS 11
suitability solution corresponds to that of the Standard USP Amitraz RS
solution, as obtained in the Assay.
USP 32 Official Monographs / Amitriptyline 207
Amitriptyline Hydrochloride Mode: LC

(Comment on this Monograph)id=m3480=Amitriptyline Detector: UV 215 nm
Hydrochloride=A-Monos.pdf) Column: 4.6-mm 25-cm; 5-m packing L7
Temperature: 45
System suitability
[NOTEFor relative retention times, see Impurity Table 1.]
Resolution: NLT 1.5 between amitriptyline related
C20H23N HCl 313.86 compound B and nortriptyline, System suitability solution
1-Propanamine, 3-(10,11-dihydro-5H-dibenzo Relative standard deviation: NMT 2.0% for
[a,d]cyclohepten-5-ylidene)-N,N-dimethyl-, hydrochloride; amitriptyline, Standard solution
10,11-Dihydro-N,N-dimethyl-5H-dibenzo[a,d]cycloheptene-5,- Analysis
propylamine hydrochloride [549-18-8]. Samples: Standard solution and Sample solution
[NOTERecord the chromatograms for 40 min.]
DEFINITION Calculate the percentage of C20H23N HCl in the portion of
Amitriptyline Hydrochloride contains NLT 98.0% and NMT Amitriptyline Hydrochloride taken:
102.0% of C20H23N HCl, calculated on the dried basis.
IDENTIFICATION
A. INFRARED ABSORPTION 197K rU = peak response from the Sample solution
B. The retention time of the major peak of the Sample rS = peak response from the Standard solution
solution corresponds to that of the Standard solution, as CS = concentration of USP Amitriptyline Hydrochloride
obtained in the Assay. RS in the Standard solution (mg/mL)
C. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the CU = concentration of amitriptyline hydrochloride in
requirements the Sample solution (mg/mL)
ASSAY
Diluted phosphoric acid: Phosphoric acid and water (1:10) Inorganic Impurities
Buffer: Dissolve 1.42 mg/mL of dibasic sodium phosphate RESIDUE ON IGNITION 281: NMT 0.1%
(Na2HPO4) in water, and adjust with diluted phosphoric acid HEAVY METALS, Method II 231: NMT 10 ppm
to a pH of 7.7. Organic Impurities
Mobile phase: Methanol and Buffer (7:3) PROCEDURE
Standard solution: 0.2 mg/mL of USP Amitriptyline Diluted phosphoric acid, Buffer, Mobile phase,
Hydrochloride RS in Mobile phase Chromatographic system, and System suitability:
Sample stock solution: 1 mg/mL of Amitriptyline Proceed as directed in the Assay.
Hydrochloride in Mobile phase Standard solution: Use the System suitability solution,
Sample solution: Sample stock solution and Mobile phase prepared as directed in the Assay.
(1:5) Sample solution: Use the Sample stock solution, prepared
Stock solution A: 1 mg/mL of USP Amitriptyline Related as directed in the Assay.
Compound A RS in methanol Analysis
Stock solution B: Dissolve USP Amitriptyline Hydrochloride Samples: Sample solution and Standard solution
RS, USP Amitriptyline Related Compound B RS, USP Calculate the percentages of individual amitriptyline
Cyclobenzaprine Hydrochloride RS, and USP Nortriptyline related compounds in the portion of Amitriptyline
Hydrochloride RS in Mobile phase to obtain a solution having Hydrochloride taken:
a known concentration of 0.4 mg/mL of amitriptyline
hydrochloride and 0.6 mg/mL each of amitriptyline related Result = (rU/rS) (CS/CU) 100
compound B, cyclobenzaprine hydrochloride, and
nortriptyline hydrochloride. rU = individual peak response for each amitriptyline
System suitability solution: Dilute suitable volumes of related compound of the Sample solution
Stock solution A, Stock solution B, and the Standard solution rS = response of the corresponding peak in the
with Mobile phase to obtain a solution having 1 g/mL of Standard solution
amitriptyline hydrochloride, 0.5 g/mL of amitriptyline CS = concentration of each of the amitriptyline
related compound A, and 1.5 g/mL each of amitriptyline related compounds in the Standard solution
related compound B, cyclobenzaprine hydrochloride, and (mg/mL)
nortriptyline hydrochloride.
208 Amitriptyline / Official Monographs USP 32
CU = concentration of amitriptyline hydrochloride in Acceptance criteria: The UV absorption spectrum of this

the Sample solution (mg/mL) solution exhibits a maximum at the same wavelength as
[NOTEDiscard any peak with a relative retention time of that of a similar solution of USP Amitriptyline Hydrochloride
less than 0.22. Use the response of the amitriptyline RS, concomitantly measured.
hydrochloride peak of the Standard solution and the B. The retention time of the major peak of the Sample
concentration of amitriptyline hydrochloride in the solution corresponds to that of the Standard solution, as
Standard solution to calculate the percentage of obtained in the Assay.
unknown individual impurities.] C. IDENTIFICATIONORGANIC NITROGENOUS BASES 181
Acceptance criteria: See Impurity Table 1. Analysis: Pipet a volume of Injection, equivalent to 50 mg
of amitriptyline hydrochloride, into a separator containing
Impurity Table 1 25 mL of water. Proceed as directed in the chapter,
beginning with In a second separator, and using water in
Relative Relative Acceptance place of 0.01 N hydrochloric acid in the Reference Standard
Retention Response Criteria, solution.
Name Time Factor NMT (%) Acceptance criteria: The Sample solution so obtained meets
Amitriptyline 0.35 0.05 the requirements of the test.
related
compound A ASSAY
PROCEDURE
Amitriptyline 0.52 0.15 Buffer: 11.04 g of monobasic sodium phosphate in 900 mL
related of water, adjust with phosphoric acid to a pH of 2.5 0.5,
compound B and dilute with water to make 1000 mL.
Nortriptyline 0.60 0.15 Mobile phase: Acetonitrile and Buffer (21:29)
hydrochloride Standard solution: 0.2 mg/mL of USP Amitriptyline
Cyclobenzaprine 0.76 0.15 Hydrochloride RS
hydrochloride Sample solution: Nominally equivalent to 0.2 mg/mL of
amitriptyline hydrochloride, from Injection diluted with
Amitriptyline 1.0 water
hydrochloride Chromatographic system
Any unspecified 0.10 each (See Chromatography 621, System Suitability.)
impurity Mode: LC
Total impurities 1.0 Detector: UV 254 nm
Column: 4-mm 30-cm; packing L1
Flow rate: 2 mL/min
MELTING RANGE OR TEMPERATURE 741: 195199 System suitability
PH 791: 5.06.0, in a solution (1 in 100) Sample: Standard solution
LOSS ON DRYING 731: Dry a sample at a pressure not Suitability requirements
exceeding 5 mm of mercury at 60 to constant weight: it Column efficiency: NLT 800 theoretical plates
loses NMT 0.5% of its weight. Tailing factor: NMT 2.0
ADDITIONAL REQUIREMENTS Analysis
PACKAGING AND STORAGE: Preserve in well-closed containers. Sample: Sample solution and Standard solution
USP REFERENCE STANDARDS 11 Calculate the percentage of C20H23N HCl in each mL of
USP Amitriptyline Hydrochloride RS Injection taken:
USP Amitriptyline Related Compound A RS
USP Amitriptyline Related Compound B RS Result = (rU/rS) (CS/CU) 100
USP Cyclobenzaprine Hydrochloride RS
USP Nortriptyline Hydrochloride RS rU = peak response from the Sample solution
CS = concentration, in mg/mL, of USP Amitriptyline
Hydrochloride RS in the Standard solution
Amitriptyline Hydrochloride Injection CU = nominal concentration of amitriptyline
(Comment on this Monograph)id=m3510=Amitriptyline hydrochloride in the Sample solution (mg/mL)
Hydrochloride Injection=A-Monos.pdf) Acceptance criteria: 90.0%110.0%

Amitriptyline Hydrochloride Injection is a sterile solution of PYROGEN 151: Amitriptyline Hydrochloride Injection,
Amitriptyline Hydrochloride in Water for Injection. It contains diluted with Sodium Chloride Injection containing 0.9% of
NLT 90.0% and NMT 110.0% of the labeled amount of NaCl to a concentration of 2.5 mg of amitriptyline
amitriptyline hydrochloride (C20H23N HCl). hydrochloride/mL, meets the requirements, the test dose
being 1 mL/kg.
IDENTIFICATION PH 791: 4.06.0
A. UV ABSORPTION OTHER REQUIREMENTS: Meets the requirements under
Sample solution: Pipet 1 mL of Injection into a 125-mL Injections 1
separator containing 10 mL of water and 1 mL of 1 N
sodium hydroxide, mix, extract with two 10-mL portions of ADDITIONAL REQUIREMENTS
methylene chloride, and evaporate the extracts on a steam PACKAGING AND STORAGE: Preserve in single-dose or
bath just to dryness. Dissolve the residue in methanol, add 1 multiple-dose containers, preferably of Type I glass.
mL of 1.2 N hydrochloric acid, then add methanol to make USP REFERENCE STANDARDS 11
100 mL. Dilute 10 mL of this solution with methanol to 100 USP Amitriptyline Hydrochloride RS
mL.
USP 32 Official Monographs / Amlodipine 209
Amitriptyline Hydrochloride Tablets Acceptance criteria: 90.0%110.0%

(Comment on this Monograph)id=m3550=Amitriptyline PERFORMANCE TESTS
Hydrochloride Tablets=A-Monos.pdf) DISSOLUTION 711
DEFINITION Medium: 0.1 N hydrochloric acid; 900 mL
Amitriptyline Hydrochloride Tablets contain NLT 90.0% and Apparatus 1: 100 rpm
NMT 110.0% of the labeled amount of amitriptyline Time: 45 min
hydrochloride (C20H23N HCl). Spectrometric conditions
Analytical wavelength: UV 239 nm
IDENTIFICATION Standard solution: USP Amitriptyline Hydrochloride RS in
A. PROCEDURE Medium
Analysis: Transfer a quantity of finely powdered Tablets, Sample solution: Sample per Dissolution 711
equivalent to 10 mg of amitriptyline hydrochloride, to a Analysis: Determine the amount of C20H23N HCl dissolved
100-mL volumetric flask, add 50 mL of methanol, shake from UV absorbances of the Sample solution, suitably diluted
well, then add methanol to volume. Filter a portion of this with Medium, if necessary, in comparison with a Standard
solution, and dilute 10 mL of the filtrate with methanol to solution having a known concentration.
100 mL. Tolerances: NLT 75% (Q) of the labeled amount of C20H23N
Acceptance criteria: The UV absorption spectrum of this HCl
solution exhibits a maximum at the same wavelength as UNIFORMITY OF DOSAGE UNITS: Meet the requirements
that of a similar solution of USP Amitriptyline Hydrochloride
RS, concomitantly measured. ADDITIONAL REQUIREMENTS
B. The retention time of the major peak of the Sample PACKAGING AND STORAGE: Preserve in well-closed containers.
solution corresponds to that of the Standard solution, as USP REFERENCE STANDARDS 11
obtained in the Assay. USP Amitriptyline Hydrochloride RS
ASSAY
PROCEDURE
Buffer: 11.04 g of monobasic sodium phosphate in 900 mL Amlodipine Besylate
of water, adjust with phosphoric acid to a pH of 2.5 0.5, (Comment on this Monograph)id=m3570=Amlodipine
dilute to make 1000 mL Besylate=A-Monos.pdf)
Mobile phase: Acetonitrile and Buffer (21:29)
Standard solution: 0.2 mg/mL of USP Amitriptyline
Hydrochloride RS in Mobile phase
Sample solution: Transfer 20 Tablets to a 500-mL
volumetric flask, add 250 mL of Mobile phase, and shake the
mixture for 1 h or until the tablets have disintegrated. Add
Mobile phase to volume and filter. Dilute a measured volume
of the clear filtrate with Mobile phase to obtain a solution
containing 0.2 mg/mL of amitriptyline hydrochloride.
Chromatographic system C20H25ClN2O5 C6H6O3S 567.05
(See Chromatography 621, System Suitability.) 3,5-Pyridinedicarboxylic acid, 2-[(2-aminoethoxy)methyl]-4-(2-
Mode: LC chlorophenyl)-1,4-dihydro-6-methyl-, 3-ethyl 5-methyl ester,
Detector: UV 254 nm ()-, monobenzenesulfonate;
Column: 4-mm 30-cm; packing L1 3-Ethyl 5-methyl ()-2-[(2-aminoethoxy)methyl]-4-(o-
Flow rate: 2 mL/min chlorophenyl)-1,4-dihydro-6-methyl-3,5-pyridinedicarboxylate,
Injection size: 20 L monobenzenesulfonate [111470-99-6].
System suitability
Sample: Standard solution DEFINITION
Suitability requirements Amlodipine Besylate contains NLT 97.0% and NMT 102.0% of
Column efficiency: NLT 800 theoretical plates C20H25ClN2O5 C6H6O3S, calculated on the anhydrous basis.
Relative standard deviation: NMT 2.0% IDENTIFICATION
Analysis A. INFRARED ABSORPTION 197M
Samples: Sample solution and Standard solution B. The retention time of the major peak of the Sample
Calculate the percentage of C20H23N HCl in each Tablet solution corresponds to that of the Standard solution, as
taken: obtained in the Assay.
Result = (rU/rS) (CS/CU) 100 ASSAY

PROCEDURE
rU = peak response from the Sample solution Buffer: Dissolve 7.0 mL of triethylamine in 800 mL of
rS = peak response from the Standard solution water. Adjust with phosphoric acid to a pH of 3.0 0.1, and
CS = concentration of USP Amitriptyline Hydrochloride dilute with water to 1 L.
RS in the Standard solution (mg/mL) Mobile phase: Methanol, acetonitrile, and Buffer (7:3:10)
CU = nominal concentration of amitriptyline Standard solution: 0.05 mg/mL of USP Amlodipine
hydrochloride in the Sample solution (mg/mL) Besylate RS in Mobile phase
210 Amlodipine / Official Monographs USP 32
Sample solution: 0.05 mg/mL of Amlodipine Besylate in System suitability solution: Dissolve 5 mg of Amlodipine
Mobile phase Besylate in 5 mL of hydrogen peroxide, and heat at 70 for
Chromatographic system 45 min.
(See Chromatography 621, System Suitability.) Standard solution: 0.003 mg/mL of USP Amlodipine
Mode: LC Besylate RS in Mobile phase
Detector: UV 237 nm Sample solution: 1 mg/mL of Amlodipine Besylate in
Column: 3.9-mm 15-cm; packing L1 Mobile phase
Flow rate: 1.0 mL/min Chromatographic system: Proceed as directed in the
Injection size: 10 L Assay.
System suitability (See Chromatography 621, System Suitability.)
Sample: Standard solution System suitability
Suitability requirements Sample: System suitability solution and Standard solution
Relative standard deviation: NMT 2.0% [NOTEThe relative retention times for benzene
Analysis sulfonate, amlodipine impurity A, and amlodipine are
Samples: Standard solution and Sample solution 0.2, 0.5, and 1.0, repectively. Amlodipine impurity A is
Calculate the percentage of C20H25ClN2O5 C6H6O3S in the 3-ethyl 5-methyl 2-[(2-aminoethoxy)methyl]-4-(2-
portion of Amlodipine Besylate taken: chlorophenyl)-6-methylpyridine-3,5-dicarboxylate.]
Result = (rU/rS) (CS/CU) 100 Resolution: NLT 4.5 between amlodipine impurity A and
amlodipine, System suitability solution
rU = peak response from the Sample solution Relative standard deviation: NMT 10%, Standard
rS = peak response from the Standard solution solution
CS = concentration of USP Amiodipine Besylate RS in Analysis
the Standard solution (mg/mL) Samples: Standard solution and Sample solution
CU = concentration of amiodipine besylate in the Record the chromatograms for a period of time that is 3
Sample solution (mg/mL) times the retention time of amlodipine.
Acceptance criteria: 97.0%102.0% Calculate the percentage of each impurity in the portion
of Amlodipine Besylate taken:
IMPURITIES
Inorganic Impurities Result = (rU/rS) (CS/CU) (1/F) 100
HEAVY METALS, Method II 231: NMT 20 ppm rU = peak response for each impurity from the
Organic Impurities Sample solution
PROCEDURE 1 rS = peak response for amlodipine besylate from the
Standard stock solution: 7 mg/mL of USP Amlodipine Standard solution
Besylate RS in methanol CS = concentration of USP Amiodipine Besylate RS in
Standard solution 1: 0.21 mg/mL from Standard stock the Standard solution (mg/mL)
solution and methanol CU = concentration of amiodipine besylate in the
Standard solution 2: 0.07 mg/mL from Standard stock Sample solution (mg/mL)
solution and methanol F = relative response factor, 0.5 for amlodipine
Sample solution: 70 mg/mL of Amlodipine Besylate in impurity A and to 1.0 for other impurities
methanol Acceptance criteria
System suitability solution: 70 mg of USP Amlodipine Amlodipine impurity A: NMT 0.3%
Besylate RS in methanol Total other impurities: NMT 0.3%
Mode: TLC [NOTEDisregard any peak less than 0.03%, and
Adsorbent: 0.25-mm layer of chromatographic silica gel disregard any peak due to benzene sulfonate.]
mixture
Application volume: 10 L SPECIFIC TESTS
Developing solvent system: Use the upper layer of a OPTICAL ROTATION 781: 0.10 to +0.10, measured at 20
mixture of methyl isobutyl ketone, water, and glacial acetic Sample solution: 10 mg/mL, in methanol
acid (2:1:1). WATER DETERMINATION, Method I 921: NMT 0.5%
Analysis
Samples: Standard solution 1 and 2, Sample solution, and ADDITIONAL REQUIREMENTS
System suitability solution PACKAGING AND STORAGE: Preserve in tight containers,
Proceed as directed for Chromatography 621, Thin-Layer protected from light. Store at room temperature.
Chromatography. Dry the plate for 15 min at 80. USP REFERENCE STANDARDS 11
Examine the plate under UV light at 254 nm and at 365 USP Amlodipine Besylate RS
nm. The chromatogram from the System suitability
solution shows two clearly separated minor spots with RF
values of 0.18 and 0.22. Compare the intensities of any Aromatic Ammonia Spirit
secondary spots observed in the chromatogram of the
Sample solution with those of the principal spots in the (Comment on this Monograph)id=m3630=Aromatic Ammonia
chromatograms of the Standard solutions. Spirit=A-Monos.pdf)
Acceptance criteria: Any spot obtained from the Sample
solution, except for the principal spot, is not greater in size DEFINITION
than the spot obtained from Standard solution 1 (0.3%), Aromatic Ammonia Spirit is a hydroalcoholic solution that
and at most two spots are more intense than the spot contains, in each 100 mL, NLT 1.7 g and NMT 2.1 g of total
obtained from Standard solution 2 (0.1%). NH3 and ammonium carbonate corresponding to NLT 3.5 g
PROCEDURE 2 and NMT 4.5 g of (NH4)2CO3.
Buffer and Mobile phase: Prepare as directed in the
Assay.
USP 32 Official Monographs / Ammonium 211
ASSAY HEAVY METALS, Method I 231: NMT 10 ppm

TOTAL NH3 LIMIT OF THIOCYANATE
Sample: 10.0 mL Sample solution: 100 mg/mL
Analysis: Transfer to a 250-mL conical flask containing 50 Analysis: Acidify 10 mL of the Sample solution with
mL of water. Add 30.0 mL of 0.5 N sulfuric acid VS, and boil hydrochloric acid, and add a few drops of ferric chloride TS.
until the solution becomes clear. Cool, and add methyl red Acceptance criteria: No orange-red color is produced.
TS. Titrate the excess acid with 0.5 N sodium hydroxide VS.
Perform a blank determination (see Titrimetry 541, Residual SPECIFIC TESTS
Titrations). Each mL of 0.5 N sulfuric acid is equivalent to PH 791
8.515 mg of NH3. Sample solution: 50 mg/mL
AMMONIUM CARBONATE Acceptance criteria: 4.66.0
Sample: 10.0 mL LOSS ON DRYING 731: Dry over silica gel for 4 h: it loses
Analysis: Transfer to a flask of 300-mL capacity. Add 30 mL NMT 0.5% of its weight.
of 0.5 N sodium hydroxide, and boil the mixture, replacing
the water lost by evaporation, until the vapors no longer ADDITIONAL REQUIREMENTS
turn moistened red litmus paper blue. Cool, dilute with 100 PACKAGING AND STORAGE: Preserve in tight containers.
mL of cold, carbon dioxide-free water, add 6 drops of
phenolphthalein TS, then add just enough 0.5 N sulfuric
acid VS to discharge the color of the phenolphthalein. Add Ammonium Chloride Injection
methyl orange TS. Titrate with 0.5 N sulfuric acid VS.
Perform a blank determination (see Titrimetry 541, Residual (Comment on this Monograph)id=m3720=Ammonium Chloride
Titrations). Each mL of 0.5 N sulfuric acid consumed in the Injection=A-Monos.pdf)
titration with methyl orange TS is equivalent to 48.04 mg of DEFINITION
(NH4)2CO3. Ammonium Chloride Injection is a sterile solution of
Acceptance criteria: In each 100 mL, NLT 1.7 g and NMT Ammonium Chloride in Water for Injection. It contains NLT
2.1 g of total NH3 and ammonium carbonate corresponding 95.0% and NMT 105.0% of the labeled amount of NH4Cl.
to NLT 3.5 g and NMT 4.5 g of (NH4)2CO3 Hydrochloric acid may be added to adjust the pH.
OTHER COMPONENTS IDENTIFICATION
ALCOHOL DETERMINATION, Method I 611: 62.0%68.0% A. IDENTIFICATION TESTSGENERAL, Ammonium 191: Meets
ADDITIONAL REQUIREMENTS the requirements
PACKAGING AND STORAGE: Preserve in tight, light-resistant B. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the
containers, at a temperature not exceeding 30. requirements
ASSAY
PROCEDURE
Ammonium Chloride Sample: A volume of Injection equivalent to 200 mg of
ammonium chloride
(Comment on this Monograph)id=m3690=Ammonium Analysis: Transfer Sample to a 500-mL Kjeldahl flask. Dilute
Chloride=A-Monos.pdf) with water to 200 mL, and add 50 mL of sodium hydroxide
NH4Cl 53.49 solution (2 in 5). Immediately connect the flask by means of
Ammonium chloride [12125-02-9]. a distillation trap to a well-cooled condenser, the delivery
tube of which dips into 40 mL of boric acid solution (1 in
DEFINITION 25) contained in a suitable receiver. Heat to boiling, and
Ammonium Chloride contains NLT 99.5% and NMT 100.5% of distill 200 mL. Cool the liquid in the receiver, if necessary,
NH4Cl, calculated on the dried basis. then add methyl red TS. Titrate with 0.1 N sulfuric acid VS.
Perform a blank determination (see Titrimetry 541). Each
IDENTIFICATION mL of 0.1 N sulfuric acid is equivalent to 5.349 mg of
IDENTIFICATION TESTSGENERAL, Ammonium and Chloride NH4Cl.
191: Meets the requirements Acceptance criteria: 95.0%105.0%
OTHER COMPONENTS
ASSAY CHLORIDE CONTENT
PROCEDURE Sample solution: A measured volume of Injection,
Sample: 100 mg of Ammonium Chloride evaporated, if necessary, equivalent to 2 g of ammonium
Analysis: Add 10 mL of water to the Sample, and swirl to chloride
dissolve. Add 10 mL of glacial acetic acid, 75 mL of Analysis: Transfer Sample solution to a 200-mL volumetric
methanol, and 0.5 mL of eosin Y TS. Titrate, with shaking, flask. Dilute with water to volume. Transfer 10.0 mL of this
with 0.1 N silver nitrate VS to a pink endpoint. Each mL of solution to a conical flask, add 10 mL of glacial acetic acid,
0.1 N silver nitrate is equivalent to 5.349 mg of NH4Cl. 75 mL of methanol, and 0.5 mL of eosin Y TS. Titrate, with
Acceptance criteria: 99.5%100.5% shaking, with 0.1 N silver nitrate VS to a pink endpoint.
IMPURITIES Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of
Inorganic Impurities Cl.
RESIDUE ON IGNITION 281 Acceptance criteria: 63.0%70.3%
Sample: 2 g SPECIFIC TESTS
Analysis: Add 1 mL of sulfuric acid to the Sample, and PH 791
heat the mixture gently until volatilization is complete. Sample solution: A concentration of NMT 100 mg/mL of
Ignite the residue. ammonium chloride
Acceptance criteria: The residue is white, and when
ignited, NMT 0.1% of nonvolatile substance remains.
212 Ammonium / Official Monographs USP 32
Acceptance criteria: 4.06.0 Acceptance criteria: No reddish-orange color is produced.

OTHER REQUIREMENTS: It meets the requirements under
Injections 1. ADDITIONAL REQUIREMENTS
PARTICULATE MATTER IN INJECTIONS 788: Meets the PACKAGING AND STORAGE: Preserve in tight containers.
requirements for small-volume injections
BACTERIAL ENDOTOXINS TEST 85: It contains NMT 1.72 USP
Endotoxin Units/mEq of chloride. Ferric Ammonium Citrate
ADDITIONAL REQUIREMENTS (Comment on this Monograph)id=m3761=Ferric Ammonium
PACKAGING AND STORAGE: Preserve in single-dose or Citrate=A-Monos.pdf)
multiple-dose containers, preferably of Type I or Type II
glass. DEFINITION
LABELING: The label states the content of ammonium Ferric Ammonium Citrate contains NLT 16.5% and NMT 18.5%
chloride in terms of weight and milliequivalents in a given of iron (Fe).
volume. The label states also the total osmolar concentration
in mOsmol/L or mOsmol/mL. The label states that the IDENTIFICATION
Injection is not for direct injection but is to be diluted with A. Ignite 0.5 g: it chars, and leaves a residue of iron oxide.
Sodium Chloride Injection to the appropriate strength before B. To 10 mL of a solution of Ferric Ammonium Citrate (1 in
use. 100) add 6 N ammonium hydroxide dropwise: the solution
USP REFERENCE STANDARDS 11 darkens, but no precipitate forms.
USP Endotoxin RS C. To 5 mL of a solution of Ferric Ammonium Citrate (1 in
100) add 0.3 mL of potassium permanganate TS and 4 mL
of mercuric sulfate TS, and heat the mixture to boiling: a
white precipitate forms.
Ammonium Chloride Delayed-Release ASSAY
Tablets PROCEDURE
(Comment on this Monograph)id=m3760=Ammonium Chloride Sample solution: Transfer 1 g of Ferric Ammonium Citrate
Delayed-Release Tablets=A-Monos.pdf) to a 250-mL conical flask, and dissolve in 25 mL of water
and 5 mL of hydrochloric acid.
DEFINITION Analysis: Add 4 g of potassium iodide, insert the stopper,
Ammonium Chloride Delayed-Release Tablets contain NLT and allow to stand protected from light for 15 min. Add
94.0% and NMT 106.0% of the labeled amount of NH4Cl. 100 mL of water. Titrate the liberated iodine with 0.1 N
Ammonium Chloride Delayed-Release Tablets are enteric- sodium thiosulfate VS, using starch TS as the indicator.
coated. Perform a blank determination, and make any necessary
IDENTIFICATION correction (see Titrimetry 541). Each mL of 0.1 N sodium
A. IDENTIFICATION TESTSGENERAL, Ammonium and Chloride thiosulfate is equivalent to 5.585 mg of iron (Fe).
191: Meets the requirements Acceptance criteria: 16.5%18.5%
Sample solution: A filtered solution of finely powdered IMPURITIES
Tablets, equivalent to 100 mg/mL ammonium chloride Inorganic Impurities
ASSAY CHLORIDE AND SULFATE, Sulfate 221
PROCEDURE Sample solution: Dissolve 100 mg in 1 mL of 2.7 N
Sample: Finely powder NLT 20 Tablets. Transfer a portion of hydrochloric acid, and dilute with water to 30 mL. Add 3
the powder, equivalent to 200 mg of ammonium chloride, mL of barium chloride TS, and dilute with water to 50 mL.
to a 500-mL Kjeldahl flask. Acceptance criteria: Any turbidity formed after 10 min is
Analysis: Add 200 mL of water and 50 mL of sodium NMT that produced in a similarly treated control solution
hydroxide solution (2 in 5) to the Sample. Immediately containing 0.31 mL of 0.020 N sulfuric acid (0.3%).
connect the flask by means of a distillation trap to a well- LIMIT OF LEAD
cooled condenser, the delivery tube of which dips into 40 Standard stock solution: Dissolve 159.8 mg of lead
mL of boric acid solution (1 in 25) contained in a suitable nitrate in 100 mL of water containing 1 mL of nitric acid.
receiver. Heat to boiling, and distill 200 mL. Cool the liquid Dilute with water to 1000 mL.
in the receiver, if necessary, then add methyl red TS. Titrate Standard solution: [NOTEPrepare this solution on the
with 0.1 N sulfuric acid VS. Perform a blank determination, day of use.]
and make any necessary correction. Each mL of 0.1 N Transfer 10 mL of Standard stock solution to a 500-mL
sulfuric acid is equivalent to 5.349 mg of NH4Cl. volumetric flask, and dilute with water to volume. Each
Acceptance criteria: 94.0%106.0% mL contains the equivalent of 2 g of lead (Pb).
Sample solution: Transfer 15 g of Ferric Ammonium
PERFORMANCE TESTS Citrate to a 100-mL volumetric flask (previously rinsed with
DISINTEGRATION 701: 2 h, determined as directed for nitric acid and water), dissolve in a mixture of 50 mL of
Enteric-Coated Tablets water and 1 mL of nitric acid, and dilute with water to
volume.
IMPURITIES Analysis
Inorganic Impurities Samples: Standard solution and Sample solution
LIMIT OF THIOCYANATE Blank: Water
Sample solution: Powder and dissolve in water a sufficient Spectrometric conditions
number of Tablets to make 25 mL of a solution containing (See Spectrophotometry and Light-Scattering 851.)
100 mg/mL of ammonium chloride solution, and filter. Mode: Atomic absorption spectrophotometer equipped
Analysis: Acidify 10 mL of the Sample solution with with a deuterium arc background corrector, a digital
hydrochloric acid, and add a few drops of ferric chloride readout device, and a burner head capable of handling
TS. 15% solids content
USP 32 Official Monographs / Ammonium 213
Analysis Ferric Ammonium Citrate for Oral

Samples: Standard solution and Sample solution Solution
Perform a blank determination with water, following the
manufacturers operating instructions. Separately aspirate (Comment on this Monograph)id=m3762=Ferric Ammonium
portions of the Standard solution and the Sample solution, Citrate for Oral Solution=A-Monos.pdf)
and record the absorbances. DEFINITION
Calculate the lead content, in g per g, in the portion of Ferric Ammonium Citrate for Oral Solution contains Ferric
Ferric Ammonium Citrate taken: Ammonium Citrate and an effervescent mixture of a suitable
organic acid and an alkali metal bicarbonate. It contains NLT
(AU/AS) CS/CU 90.0% and NMT 110.0% of the labeled amount of Fe. It may
AU = absorbance of the Sample solution contain one or more suitable flavors, colors, or stabilizing
AS = absorbance of the Standard solution agents.
CS = concentration of lead in the Standard solution IDENTIFICATION
(g/mL) A 6-g portion dissolves in 600 mL of water with
CU = concentration of Ferric Ammonium Citrate effervescence. The collected gas meets the requirements of
(g/ml) the test for Identification TestsGeneral, Bicarbonate 191,
Acceptance criteria: NMT 10 g/g and the resulting solution meets the requirements of the
MERCURY tests for Identification TestsGeneral, Iron 191, and for
Mercury Stock Solution and Standard Mercury solution: Identification TestsGeneral, Citrate 191.
Proceed as directed for Mercury 261, Method I.
Mercury Detection Instrument, Aeration Apparatus, and ASSAY
Stannous Chloride Solution: Proceed as directed for PROCEDURE
Mercury 261, Method IIa and Method IIb. Sample solution: Transfer 6 g of Ferric Ammonium Citrate
Standard solutions: Transfer 0.25, 0.50, 1.0, and 3.5 mL for Oral Solution to a 250-mL conical flask, and dissolve in
of Mercury Stock Solution to four separate glass-stoppered 100 mL of water. Allow the gas to escape, add 5 mL of
bottles, such as biological oxygen-demand bottles, of about hydrochloric acid and 4 g of potassium iodide, insert the
300-mL capacity. Dilute the contents of each bottle with stopper, and allow to stand protected from light for 15 min.
water to 100 mL. These solutions contain the equivalent of Add 25 mL of water.
2.5, 5.0, 10.0, and 35.0 g/mL of mercury, respectively. Analysis: Titrate the liberated iodine with 0.1 N sodium
Sample solution: Transfer 1.000 g of Ferric Ammonium thiosulfate VS, using starch TS as the indicator. Perform a
Citrate to a 200-mL centrifuge bottle with a polytef-lined blank determination (see Titrimetry 541). Each mL of 0.1 N
screw cap, and add 5 mL of nitric acid and 5 mL of sodium thiosulfate is equivalent to 5.585 mg of Fe.
hydrochloric acid. Close the bottle tightly, digest on a Acceptance criteria: 90.0%110.0%
steam bath for 1 h, and cool. Quantitatively transfer the
solution to a suitable glass-stoppered bottle, dilute with PERFORMANCE TESTS
water to 100 mL, and bubble air through the solution for 2 UNIFORMITY OF DOSAGE UNITS 905: It meets the
min. Prepare a reagent blank in the same manner. requirements for powder packaged in single-unit containers.
Analysis
Add 5 mL of Stannous Chloride Solution (1 in 10) to each DELIVERABLE VOLUME 698: It meets the requirements for
solution, and immediately insert the bubbler of the powder packaged in multiple-unit containers.
Aeration Apparatus. Obtain the absorbances as directed ADDITIONAL REQUIREMENTS
by the instrument manufacturers operating instructions. PACKAGING AND STORAGE: Preserve in tight, light-resistant
Perform a blank determination, and make any necessary containers, and store in a cool place.
correction. Plot the absorbances of the Standard solutions
versus concentrations, in g/mL, of mercury, and draw
the straight line best fitting the plotted points. From the
graph so obtained, determine the concentration, in g/g, Ammonium Molybdate
of mercury in the Sample solution.
(Comment on this Monograph)id=m3795=Ammonium
Acceptance criteria: NMT 10 g/g
Molybdate=A-Monos.pdf)
SPECIFIC TESTS (NH4)6Mo7O24 4H2O 1235.86
FERRIC CITRATE: To a solution of Ferric Ammonium Citrate (1 Molybdate (Mo7O246), hexaammonium, tetrahydrate;
in 100) add potassium ferrocyanide TS: no blue precipitate is Hexaammonium molybdate tetrahydrate [12054-85-2]
formed.
OXALATE: Transfer 1 g to a 125-mL separator, dissolve in 10 DEFINITION
mL of water, add 2 mL of hydrochloric acid, and extract Ammonium Molybdate contains NLT 99.3% and NMT 101.8%
successively with one 50-mL portion and one 20-mL portion of (NH4)6Mo7O24 4H2O.
of ether. Transfer the combined ether extracts to a 150-mL
beaker, add 10 mL of water, and remove the ether by IDENTIFICATION
evaporation on a steam bath. Add 1 drop of glacial acetic PROCEDURE
acid and 1 mL of calcium acetate solution (1 in 20): no Sample: 0.6 g
turbidity is produced within 5 min. Analysis: Dissolve the Sample in 1.4 mL of water and 1.45
mL of ammonium hydroxide. Cool this mixture, and add
ADDITIONAL REQUIREMENTS slowly, with mixing, 7.2 mL of a well-cooled mixture of 3.2
PACKAGING AND STORAGE: Preserve in tight, light-resistant mL of nitric acid and 4 mL of water. Allow to stand for 24
containers, in a cool place.
214 Ammonium / Official Monographs USP 32
to 48 h, and filter through a sintered-glass filter. To 5 mL of INSOLUBLE SUBSTANCES

the filtrate, add 2 mL of dibasic sodium phosphate TS. Sample: 20 g
Acceptance criteria: A yellow precipitate is formed, and it Analysis: Dissolve the Sample in 200 mL of water in a
is soluble in an excess of 6 N ammonium hydroxide. beaker, heat to boiling, cover, and heat on a steam bath
for 1 h. Filter the hot solution through a tared filtering
ASSAY crucible, wash the insoluble residue with hot water, and
PROCEDURE dry at 105 for 2 h.
Sample solution: 1 g Acceptance criteria: The weight of the residue so
Analysis: Dissolve the Sample solution in a mixture of 10 mL obtained is NMT 1 mg (0.005%).
of water and 1 mL of ammonium hydroxide in a 250-mL LIMIT OF NITRATE
volumetric flask, and dilute with water to volume. Filter the Sample: 1 g
solution, and transfer 50.0 mL of the filtrate to a 600-mL Analysis: Dissolve the Sample in 10 mL of water containing
beaker. Add 250 mL of water, 20 g of ammonium chloride, 5 mg of sodium chloride, and add 0.10 mL of a 1 in 1000
15 mL of hydrochloric acid, and 0.15 mL of methyl orange solution of indigo carmine in 3.6 N sulfuric acid.
TS, heat nearly to boiling, and add 18 mL of lead acetate Acceptance criteria: The blue color is not completely
TS. To the hot solution add, slowly and with constant discharged in 5 min.
stirring, a saturated solution of ammonium acetate until the ARSENATE, PHOSPHATE, AND SILICATE
color turns yellow, and then add 15 mL of lead acetate TS. Sample: 2.5 g
Cover the beaker, and heat just below the boiling Analysis: Dissolve the Sample in 70 mL of water in a
temperature until the precipitate has settled. Filter through a container other than one of glass. Adjust with 1.2 N
tared, porous porcelain crucible, wash with seven or eight hydrochloric acid to a pH of between 3 and 4, transfer to a
successive portions of a mixture of water, saturated glass container, add 2 mL of bromine TS, and adjust with
ammonium acetate solution, and nitric acid (890:100:10), 1.2 N hydrochloric acid to a pH of 1.8 0.1. Heat almost
and finally wash with three successive portions of hot water. to boiling, and cool to room temperature. Dilute with
Ignite to constant weight at 560 to 625. Each mg of the water to 90 mL, add 10 mL of hydrochloric acid, and
lead molybdate so obtained is equivalent to 0.4809 mg of transfer to a separator. Add 1 mL of butyl alcohol and 30
(NH4)6Mo7O24 4H2O. mL of 4-methyl-2-pentanone, shake vigorously, and allow
Acceptance criteria: 99.3%101.8% the phases to separate. Discard the aqueous phase, and
wash the ketone phase with three successive 10-mL
IMPURITIES portions of 1.2 N hydrochloric acid, discarding the
Inorganic Impurities washings. To the washed ketone phase, add 10 mL of 1.2
CHLORIDE AND SULFATE, Chloride 221: A 0.5-g portion N hydrochloric acid to which has just been added 0.2 mL
shows no more chloride than 0.30 mL of 0.001 N of a freshly prepared 1 in 50 solution of stannous chloride
hydrochloric acid (NMT 20 ppm). in hydrochloric acid. Similarly treat a control solution
CHLORIDE AND SULFATE, Sulfate 221: A 0.25-g portion prepared by dissolving 0.5 g of specimen in 70 mL of
shows no more sulfate than corresponds to 1.0 mL of 0.001 water and adding 2 mL of sodium silicate solution (1 in
N sulfuric acid (NMT 200 ppm). 20,000).
HEAVY METALS 231 Acceptance criteria: Any blue color in the test solution
Stock solution: Dissolve 2.0 g Ammonium Molybdate in does not exceed that in the control solution.
20 mL of water, add 10 mL of 2.5 N sodium hydroxide MAGNESIUM AND ALKALI SALTS
and 2 mL of ammonium hydroxide, and dilute with water Sample: 5.0 g
to 40 mL. Analysis: Dissolve the Sample in 50 mL of water, and filter.
Control solution: To 10 mL of Stock solution add 1.0 mL To the filtrate, add 0.5 g of sodium carbonate and 25 mL
of Standard Lead Solution, prepared as directed under Heavy of 2.5 N sodium hydroxide. Boil the solution gently for 5
Metals 231, and dilute with water to 40 mL. min, cool, and filter through an ignited and tared filter.
Sample solution: Dilute the remaining 30-mL portion of Wash the filter with 1 N ammonium hydroxide. Ignite the
the Stock solution with water to 40 mL. filter at 800 25 for 30 min.
Analysis: To both solutions, add 1.2 mL of thioacetamide- Acceptance criteria: The weight of the residue so
glycerin base TS and 2 mL of pH 3.5 Acetate Buffer, and obtained does not exceed 1 mg (NMT 0.200 ppm).
allow to stand for 5 min: any color in the Sample solution
does not exceed that in the Control solution. ADDITIONAL REQUIREMENTS
Acceptance criteria: NMT 10 ppm PACKAGING AND STORAGE: Preserve in tight containers.
PHOSPHATE
Sample: 20 g
Analysis: Dissolve the Sample in 100 mL of 3 N
ammonium hydroxide, add 3.5 mL of ferric nitrate solution Ammonium Molybdate Injection
(1 in 10), and allow to stand for 15 min. Warm gently to (Comment on this Monograph)id=m3798=Ammonium
coagulate the precipitate, and filter. Wash the filter several Molybdate Injection=A-Monos.pdf)
times with 1.5 N ammonium hydroxide, then wash the
filter with 60 mL of warm 4 N nitric acid to dissolve the DEFINITION
residue on the filter, collecting the filtrate in a glass- Ammonium Molybdate Injection is a sterile solution of
stoppered, 250-mL conical flask. Add 13 mL of ammonium Ammonium Molybdate in Water for Injection. It contains NLT
hydroxide, warm to 40, add 50 mL of ammonium 85.0% and NMT 115.0% of the labeled amount of
molybdate TS, shake for 5 min, and allow to stand at 40 molybdenum (Mo).
for 2 h. Similarly treat 100 mL of a Standard solution
prepared by dissolving 143.3 mg of dried monobasic IDENTIFICATION
potassium phosphate in water to make 1000 mL, and then A. The Sample solution, prepared as directed in the Assay,
diluting 1.0 mL of this solution with 3 N ammonium exhibits an absorption maximum at about 313 nm when
hydroxide to 100 mL. tested as directed for Analysis in the Assay.
Acceptance criteria: Any yellow precipitate formed from
the Sample solution does not exceed that obtained from the
Standard solution (5 ppm).
USP 32 Official Monographs / Amobarbital 215
B. PROCEDURE ADDITIONAL REQUIREMENTS

Sample: 5 mL of Injection PACKAGING AND STORAGE: Preserve in single-dose or
Analysis: Add 0.3 mL of alkaline mercuric-potassium iodide multiple-dose containers, preferably of Type I or Type II
TS to the Sample. glass.
Acceptance criteria: A reddish-brown color develops. LABELING: Label the Injection to indicate that it is to be
C. PROCEDURE diluted to the appropriate strength with Sterile Water for
Sample: 50 mL of Injection Injection or other suitable fluid before administration.
Analysis: Evaporate the Sample on a steam bath to a
volume of 0.3 mL, and add 0.3 mL of ammonium
hydroxide. Cool, and add slowly, with mixing, a well-cooled
mixture of 1 mL of nitric acid and 1.2 mL of water. Allow to Amobarbital Sodium
stand for 2448 h, and pass through a sintered-glass filter. (Comment on this Monograph)id=m3950=Amobarbital
To the filtrate add 0.5 mL of dibasic sodium phosphate TS. Sodium=A-Monos.pdf)
Acceptance criteria: A yellow precipitate is formed, and it C11H17N2NaO3 248.25
dissolves in an excess of 6 N ammonium hydroxide. 2,4,6(1H,3H,5H)-Pyrimidinetrione, 5-ethyl-5-(3-methyl-
ASSAY butyl)-, monosodium salt;
PROCEDURE Sodium 5-ethyl-5-isopentylbarbiturate [64-43-7].
Diluent: Ammonium hydroxide and water (1:24) DEFINITION
[NOTEStore in a plastic bottle.] Amobarbital Sodium contains NLT 98.5% and NMT 100.5% of
Solution A: 10 mg/mL of sodium sulfate. C11H17N2NaO3, calculated on the dried basis.
Molybdenum stock solution: Transfer 1.84 g of previously
assayed Ammonium Molybdate to a 1000-mL volumetric IDENTIFICATION
flask, and dilute with Diluent to volume. This solution A. INFRARED ABSORPTION 197K
contains the equivalent of 1000 g/mL of molybdenum. Sample: Residue obtained in the Assay
Standard solutions: Transfer 0, 1, 2, 3, and 4 mL, B. IDENTIFICATION TESTSGENERAL, Sodium 191
respectively, of Molybdenum stock solution to separate 100- Sample: 200 mg
mL volumetric flasks, and to the respective flasks add 5, 4, Analysis: Ignite Sample
3, 2, and 1 mL of Diluent. To each flask add 10 mL of Acceptance criteria: The residue effervesces with acid and
Solution A, and dilute with water to volume. These Standard responds to the test for sodium.
solutions contain, respectively, 0, 10, 20, 30, and 40 g of
molybdenum/mL. ASSAY
Sample solution: A volume of Injection, equivalent to 500 PROCEDURE
g of molybdenum, to a 25-mL volumetric flask. Add 1.25 Sample solution: 500 mg of Amobarbital Sodium, in 15
mL of Diluent and 2.5 mL of Solution A, and dilute with mL of water in a separator
water to volume. Analysis: To the Sample solution add 2 mL of hydrochloric
Spectrometric conditions acid, shake, and completely extract the liberated
Mode: Atomic absorption spectrophotometer amobarbital with 25-mL portions of chloroform. Test for
(See Spectrophotometry and Light-Scattering 851.) completeness of extraction by extracting with an additional
Analytical wavelength: Molybdenum emission line of 10-mL portion of chloroform and evaporating the solvent:
313.3 nm NMT 0.5 mg of residue remains. Filter the combined extract
Lamp: Molybdenum hollow-cathode lamp through a glass filter funnel into a tared beaker, and wash
Flame: Nitrous oxideacetylene reducing flame the separator and the filter with several small portions of
Blank: Water chloroform. Evaporate the combined filtrate and washings
Analysis on a steam bath with the aid of a current of air, dry the
Samples: Standard solutions and the Sample solution residue at 105 for 30 min, cool, and weigh. The weight of
Plot the absorbances of the Standard solutions versus the residue, multiplied by 1.097, represents the weight of
concentration, in g/mL, of molybdenum, and draw the C11H17N2NaO3.
straight line best fitting the five plotted points. From the Acceptance criteria: 98.5%100.5%
graph so obtained, determine the concentration, in
g/mL, of molybdenum in the Sample solution. IMPURITIES
Calculate the percentage of Mo in each mL of the Injection Inorganic Impurities
taken: HEAVY METALS, Method II 231: NMT 30 ppm
Result = 25 (C/V) 100 SPECIFIC TESTS

COMPLETENESS OF SOLUTION 641: 1.0 g in 10 mL of carbon
C = concentration of molybdenum in the Sample dioxide-free water: after 1 min, the solution is clear and free
solution (g/mL) from undissolved solid.
V = volume of Injection taken (mL) PH 791: NMT 11.0, in the solution prepared for the test
Acceptance criteria: 85.0%115.0% for Completeness of solution
LOSS ON DRYING 731: Dry 1 g, at 105 for 4 h: it loses
SPECIFIC TESTS NMT 2.0% of its weight.
PYROGEN TEST 151: Meets the requirements, the test dose OTHER REQUIREMENTS: Where the label states that
being 10 mL of Injection/kg Amobarbital Sodium is sterile, it meets the requirements for
PH 791: 3.06.0 Sterility Tests 71 and it contains NMT 0.4 USP Endotoxin
PARTICULATE MATTER IN INJECTIONS 788: Meets the Unit/mg of amobarbital sodium (Bacterial Endotoxins Test
requirements for small-volume injections 85.) Where the label states that Amobarbital Sodium must
OTHER REQUIREMENTS: Meets the requirements under be subjected to further processing during the preparation of
Injections 1
216 Amobarbital / Official Monographs USP 32
injectable dosage forms, it meets the above requirements for ADDITIONAL REQUIREMENTS
Bacterial Endotoxins. PACKAGING AND STORAGE: Preserve as described under
Injections 1, Containers for Sterile Solids.
ADDITIONAL REQUIREMENTS USP REFERENCE STANDARDS 11
PACKAGING AND STORAGE: Preserve in tight containers. USP Amobarbital RS
LABELING: Where it is intended for use in preparing USP Endotoxin RS
injectable dosage forms, the label states that it is sterile or
must be subjected to further processing during the
preparation of injectable dosage forms.
USP REFERENCE STANDARDS 11 Amodiaquine
USP Amobarbital RS (Comment on this Monograph)id=m4020=Amodiaquine=A-
USP Endotoxin RS Monos.pdf)
Amobarbital Sodium for Injection

(Comment on this Monograph)id=m3960=Amobarbital Sodium
for Injection=A-Monos.pdf)
DEFINITION
Amobarbital Sodium for Injection is Amobarbital Sodium C20H22ClN3O 355.86
suitable for parenteral use. It contains NLT 98.5% and NMT Phenol, 4-[(7-chloro-4-quinolinyl)amino]-2-[(diethylamino)-
100.5% of the labeled amount of C11H17N2NaO3, calculated on methyl]-;
the dried basis. 4-[(7-Chloro-4-quinolyl)amino]--(diethylamino)-o-cresol
[86-42-0].
ASSAY
Sample solution: A portion of Injection, equivalent to Amodiaquine contains NLT 97.0% and NMT 103.0% of
about 500 mg of amobarbital sodium, in 15 mL of water in C20H22ClN3O, calculated on the anhydrous basis.
a separator
Analysis: To the solution, add 2 mL of hydrochloric acid, IDENTIFICATION
shake, and completely extract the liberated amobarbital with A. INFRARED ABSORPTION 197K
25-mL portions of chloroform. Test for completeness of Standard solution: 20 mg of USP Amodiaquine
extraction by extracting with an additional 10-mL portion of Hydrochloride RS in 10 mL of water in a separator, add 1
chloroform and evaporating the solvent: NMT 0.5 mg of mL of ammonium hydroxide, and extract by shaking with
residue remains. Filter the combined extract through a glass 25 mL of chloroform. Draw off and evaporate the
filter funnel into a tared beaker, and wash the separator and chloroform extract, and dry the residue at 105 for 2 h.
the filter with several small portions of chloroform. B. ULTRAVIOLET ABSORPTION 197U
Evaporate the combined filtrate and washings on a steam Sample solution: 10 g/mL in 0.1 N hydrochloric acid
bath with the aid of a current of air, dry the residue at 105
for 30 min, cool, and weigh. The weight of the residue, ASSAY
multiplied by 1.097, represents the weight of C11H17N2NaO3. PROCEDURE
Acceptance criteria: 98.5%100.5% Standard solution: 15 g/mL of USP Amodiaquine
Hydrochloride RS in 0.1 N hydrochloric acid
PERFORMANCE TESTS Sample solution: 15 g/mL of Amodiaquine in 0.1 N
UNIFORMITY OF DOSAGE UNITS 905: Meets the hydrochloric acid
requirements Spectrometric conditions
Cell: 1 cm
IMPURITIES Analytical wavelength: 342 nm
Inorganic Impurities Blank: 0.1 N hydrochloric acid
HEAVY METALS, Method II 231: NMT 30 ppm Analysis
SPECIFIC TESTS Calculate the percentage of C20H22ClN3O in the portion of
INJECTIONS, Constituted Solutions 1: At the time of use, it Amodiaquine taken:
meets the requirements.
LOSS ON DRYING 731: Dry 1 g, at 105 for 4 h: it loses Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100
BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.4 USP AU = absorbance of the Sample solution
Endotoxin Unit/mg of amobarbital sodium. AS = absorbance of the Standard solution
STERILITY TESTS 71: Meets the requirements CS = concentration of USP Amodiaquine
COMPLETENESS OF SOLUTION 641: 1.0 g in 10 mL of carbon Hydrochloride RS in the Standard solution
dioxide-free water: after 1 min, the solution is clear and free (g/mL)
from undissolved solid. CU = concentration of amodiaquine hydrochloride in
PH 791: NMT 11.0, in the solution prepared for the the Sample solution (g/mL)
Completeness of Solution 641 test. Mr1 = molecular weight of amodiaquine, 355.87
OTHER REQUIREMENTS: It meets the requirements of the Mr2 = molecular weight of anhydrous aminodiaquine
Identification tests under Amobarbital Sodium. It meets the hydrochloride, 428.79
requirements under Injections 1, Labeling.
USP 32 Official Monographs / Amodiaquine 217
Acceptance criteria: 97.0%103.0% B. ULTRAVIOLET ABSORPTION 197U

Sample solution: 10 g/mL in dilute hydrochloric acid (1 in
IMPURITIES 100)
Inorganic Impurities C. IDENTIFICATION TESTSGENERAL, Chloride 191: meets the
RESIDUE ON IGNITION 281: NMT 0.2% requirements.
Organic Impurities
PROCEDURE ASSAY
Standard solution A: To 20 mg of USP Amodiaquine PROCEDURE
Hydrochloride RS in a glass-stoppered test tube add 1.0 mL Sample solution: 15 g/mL of Amodiaquine Hydrochloride
of chloroform (saturated with ammonium hydroxide), and in dilute hydrochloric acid (1 in 100)
shake vigorously for 2 min. Allow the solids to settle, and Standard solution: 15 g/mL of USP Amodiaquine
decant the liquid into a second test tube. Hydrochloride RS in dilute hydrochloric acid (1 in 100)
Standard solution B: Standard solution A and chloroform Spectrometric conditions
(saturated with ammonium hydroxide) (1:199) Cell: 1 cm
Sample solution: 15 mg/mL of Amodiaquine in Analytical wavelength: 342 nm
chloroform (saturated with ammonium hydroxide) Blank: Dilute hydrochloric acid (1 in 100)
Adsorbent: 0.25-mm layer of chromatographic silica gel Analysis
mixture Samples: Standard solution and Sample solution
Application volume: 10 L Calculate the percentage of C20H22ClN3O 2HCl in the
Developing solvent system: Chloroform (saturated with portion of Amodiaquine Hydrochloride taken:
ammonium hydroxide) and dehydrated alcohol (9:1)
Analysis Result = (AU/AS) (CS/CU) 100
Samples: Standard solution A, Standard solution B, and
and Sample solution AU = absorbance of the Sample solution
Proceed as directed for Chromatography 621, Thin-Layer AS = absorbance of the Standard solution
Chromatography. Develop the chromatogram until the CS = concentration of USP Amodiaquine
solvent front has moved about three-fourths of the Hydrochloride RS in the Standard solution
length of the plate. Remove the plate from the (g/mL)
developing chamber, mark the solvent front, allow the CU = nominal concentration of the Sample solution
solvent to evaporate, and examine the plate under short- (g/mL)
wavelength UV light. Acceptance criteria: 97.0%103.0%
Acceptance criteria: The chromatograms show principal
spots at about the same RF value, and no secondary spot, if PERFORMANCE TESTS
present in the chromatogram from the Sample solution, is COMPLETENESS OF SOLUTION 641: A solution of 200 mg in
more intense than the principal spot obtained from 10 mL of water is clear.
Standard solution B. IMPURITIES
WATER DETERMINATION, Method I 921: NMT 0.5% RESIDUE ON IGNITION 281: NMT 0.2%
Organic Impurities
ADDITIONAL REQUIREMENTS PROCEDURE
PACKAGING AND STORAGE: Preserve in tight containers. Standard solution A: To 20 mg of USP Amodiaquine
USP REFERENCE STANDARDS 11 Hydrochloride RS in a glass-stoppered test tube, add 1.0
USP Amodiaquine Hydrochloride RS mL of chloroform (saturated with ammonium hydroxide),
and shake vigorously for 2 min. Allow the solids to settle,
and decant the liquid into a second test tube.
Standard solution B: Dilute 1.0 mL of Standard solution A
Amodiaquine Hydrochloride to 200 mL with chloroform (saturated with ammonium
(Comment on this Monograph)id=m4040=Amodiaquine hydroxide).
Hydrochloride=A-Monos.pdf) Sample solution: To 200 mg of Amodiaquine
Hydrochloride in a glass-stoppered test tube, add 10 mL of
C20H22ClN3O 2HCl 2H2O 464.81 chloroform (saturated with ammonium hydroxide). Shake
Phenol, 4-[(7-chloro-4-quinolinyl)amino]-2-[(diethylamino)- vigorously for 2 min. Allow the solids to settle, and decant
methyl]-, dihydrochloride, dihydrate; the liquid into a second test tube.
4-[(7-Chloro-4-quinolyl)amino]--(diethylamino)-o-cresol Developing solvent system: Chloroform (saturated with
dihydrochloride dihydrate [6398-98-7]. ammonium hydroxide) and dehydrated alcohol (9:1)
Anhydrous 428.79 Chromatographic system
[69-44-3]. (See Chromatography 621, Thin-Layer Chromatography.)
DEFINITION Mode: TLC
Amodiaquine Hydrochloride contains NLT 97.0% and NMT Adsorbent: 0.25-mm layer of chromatographic silica gel
103.0% of C20H22ClN3O 2HCl, calculated on the anhydrous mixture
basis. Application volume: 10 L
Analysis
IDENTIFICATION Samples: Standard solution A, Standard solution B, and
A. INFRARED ABSORPTION 197K Sample solution
Sample: Dissolve 20 mg of Amodiaquine Hydrochloride in Develop the chromatogram in the Developing solvent
10 mL of water in a separator, add 1 mL of ammonium system until the solvent front has moved three-fourths of
hydroxide, and extract by shaking with 25 mL of the length of the plate. Remove the plate from the
chloroform. Draw off and evaporate the chloroform extract, developing chamber, mark the solvent front, allow the
and dry the residue at 105 for 2 h.
218 Amodiaquine / Official Monographs USP 32
solvent to evaporate, and examine the plate under short- acid (1 in 100). Combine the acid extracts in a 200-mL
wavelength UV light. volumetric flask, add dilute hydrochloric acid (1 in 100) to
Acceptance criteria: The chromatograms show principal volume. Pipet 20 mL of this solution into a 100-mL
spots at about the same RF value, and no secondary spot, if volumetric flask, add dilute hydrochloric acid (1 in 100) to
present in the chromatogram from the Sample solution, is volume.
more intense than the principal spot of Standard solution B. Standard solution: 15 g/mL of undried USP Amodiaquine
Hydrochloride RS in dilute hydrochloric acid (1 in 100)
SPECIFIC TESTS Spectrometric conditions
WATER DETERMINATION, Method I 921: 7.0%9.0% Cell: 1 cm
Analytical wavelength: 342 nm
ADDITIONAL REQUIREMENTS Blank: Dilute hydrochloric acid (1 in 100).
PACKAGING AND STORAGE: Preserve in tight containers. Analysis
USP REFERENCE STANDARDS 11 Samples: Sample solution and Standard solution
USP Amodiaquine Hydrochloride RS Calculate the percentage of C20H22ClN3O in the portion of
Tablets taken:
Amodiaquine Hydrochloride Tablets Result = (AU/AS) (CS/CU) F 100

(Comment on this Monograph)id=m4050=Amodiaquine AU = absorbance of the solution from the Tablets
Hydrochloride Tablets=A-Monos.pdf) AS = absorbance of the solution from Standard
solution
DEFINITION CS = concentration calculated, on the anhydrous
Amodiaquine Hydrochloride Tablets contain an amount of basis, of the USP Amodiaquine Hydrochloride
amodiaquine hydrochloride (C20H22ClN3O 2HCl 2H2O) RS in the Standard solution (g/mL)
equivalent to NLT 93.0% and NMT 107.0% of the labeled CU = nominal concentration of amodiaquine in the
amount of amodiaquine (C20H22ClN3O). Sample solution (g/mL)
IDENTIFICATION F = molecular weight correction factor, 0.7656
A. INFRARED ABSORPTION 197K Acceptance criteria: 93.0%107.0%
Sample: Powder one or more Tablets, and transfer a PERFORMANCE TESTS
portion of the powder, equivalent to 50 mg of DISSOLUTION 711
amodiaquine, to a 125-mL separator. Add 20 mL of water, Medium: Water; 900 mL
and shake for 1 min. Add 25 mL of chloroform and 1 mL of Apparatus 2: 50 rpm
ammonium hydroxide, shake for 2 min, and when settled, Time: 30 min
filter the chloroform extract through cotton that previously Detector: UV 342 nm
has been rinsed with chloroform, collecting the extract in a Sample solution: Sample per Dissolution 711.
vessel suitable for evaporation. Evaporate the chloroform, Standard solution: USP Amodiaquine Hydrochloride RS in
and dry the residue at 105 for 1 h. Medium
B. ULTRAVIOLET ABSORPTION 197U Analysis
Sample solution: Prepare as directed in the Assay. Samples: Sample solution and Standard solution
ASSAY Determine the amount of C20H22ClN3O 2HCl 2H2O
PROCEDURE dissolved from UV absorbances on filtered portions of the
Sample solution: Finely powder NLT 20 Tablets. Transfer a solution under test, suitably diluted with water, if
portion of the powder, nominally equivalent to 300 mg of necessary, in comparison with a Standard solution having a
amodiaquine, to a 250-mL beaker, add 100 mL of dilute known concentration of USP Amodiaquine Hydrochloride
hydrochloric acid (1 in 100), and heat on a steam bath for RS.
about 15 min with occasional stirring. Cool the mixture to Tolerances: An amount of C20H22ClN3O 2HCl 2H2O
room temperature, transfer to a 200-mL volumetric flask, equivalent to NLT 75% (Q) of the labeled amount of
add dilute hydrochloric acid (1 in 100) to volume. Pipet 10 C20H22ClN3O
mL of the clear supernatant into a 125-mL separator. Add UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
10 mL of dilute hydrochloric acid (1 in 100), and wash with ADDITIONAL REQUIREMENTS
20 mL of chloroform, discarding the washing. Add 4.5 mL PACKAGING AND STORAGE: Preserve in tight containers.
of 1 N sodium hydroxide, and extract with four 25-mL USP REFERENCE STANDARDS 11
portions of chloroform. Extract the combined chloroform USP Amodiaquine Hydrochloride RS
solutions with three 50-mL portions of dilute hydrochloric
USP 32 Official Monographs / Amoxapine 219
Amoxapine SPECIFIC TESTS

(Comment on this Monograph)id=m4070=Amoxapine=A- MELTING RANGE OR TEMPERATURE, Class I 741: 177181
Monos.pdf) LOSS ON DRYING 731: Dry it at 105 for 4 h: it loses NMT
0.5% of its weight.
USP Amoxapine RS
C17H16ClN3O
Dibenz[b,f][1,4]oxazepine, 2-chloro-11-(1-piperazinyl)-;
313.78 Amoxapine Tablets
2-Chloro-11-(1-piperazinyl)dibenz[b,f][1,4]oxazepine (Comment on this Monograph)id=m4080=Amoxapine
[14028-44-5]. Tablets=A-Monos.pdf)
DEFINITION DEFINITION
Amoxapine contains NLT 98.5% and NMT 101.0% of Amoxapine Tablets contain NLT 90.0% and NMT 110.0% of
C17H16ClN3O, calculated on the dried basis. the labeled amount of amoxapine (C17H16ClN3O).
IDENTIFICATION IDENTIFICATION
INFRARED ABSORPTION 197K Sample: Triturate a quantity of finely ground Tablets,
ASSAY equivalent to 50 mg of amoxapine, with 10 mL of
PROCEDURE chloroform, and filter. Evaporate the filtrate on a steam bath
Sample: 300 mg to dryness (about 30 min).
Analysis: Transfer the Sample to a 250-mL flask, dissolve in ASSAY
50 mL of glacial acetic acid, and add 3 drops of crystal PROCEDURE
violet TS. Titrate with 0.1 N perchloric acid VS to an emerald Solution A: 1.38 mg/mL of monobasic sodium phosphate
green endpoint. Perform a blank determination, and make in water
any necessary correction (see Titrimetry 541). Each mL of Solution B: 113 mg/mL of tetramethylammonium chloride
0.1 N perchloric acid is equivalent to 15.69 mg of in water
C17H16ClN3O. Mobile phase: Acetonitrile, 0.01 M monobasic sodium
Acceptance criteria: 98.5%101.0% phosphate, 1 M tetramethylammonium chloride, and dilute
IMPURITIES phosphoric acid (1 in 5). (180:309:10:1).
Inorganic Impurities Standard stock solution: 1 mg/mL of USP Amoxapine RS
RESIDUE ON IGNITION 281: NMT 0.1% in acetonitrile. Shake by mechanical means to dissolve, and
Organic Impurities then dilute with acetonitrile to volume.
PROCEDURE Standard solution: 0.1 mg/mL from Standard stock solution
Standard solution A: 0.50 mg/mL of USP Amoxapine RS diluted with Mobile phase
in chloroform Sample solution: Finely powder NLT 20 Tablets. Transfer a
Standard solution B: 0.25 mg/mL USP Amoxapine RS in portion of the powder, equivalent to 50 mg of amoxapine,
chloroform; from Standard solution A to a 50-mL volumetric flask. Add 40 mL of Mobile phase,
Sample solution: 50 mg/mL of Amoxapine in chloroform and shake vigorously by mechanical means for 20 min.
Developing solvent system: Chloroform, methanol, and Dilute with Mobile phase to volume and filter. Pipet 5.0 mL
ammonium hydroxide (18:2:0.1) of the filtrate into a 50-mL volumetric flask, and dilute with
Chromatographic system Mobile phase to volume.
(See Chromatography 621, Thin-Layer Chromatography.) Chromatographic system
Mode: TLC (See Chromatography 621, System Suitability.)
Adsorbent: 0.2-mm layer of chromatographic silica gel Mode: LC
mixture Detector: UV 254 nm
Application volume: 5 L Column: 4.6-mm 25-cm; packing L1
Analysis Flow rate: 1.5 mL/min
Samples: Standard solution A, Standard solution B, and Injection size: 10 L
Sample solution System suitability
Develop the chromatogram in the Developing solvent Sample: Standard solution
system until the solvent front has moved three-fourths of Suitability requirements
the length of the plate. Remove the plate from the Column efficiency: NLT 1200 theoretical plates from the
developing chamber, examine it under short-wavelength analyte peak
UV light, and compare the intensities of any secondary Tailing factor: NMT 1.8 for the analyte peak
spots observed in the chromatogram of the Sample Relative standard deviation: NMT 2.0%
solution with those of the principal spots in the Analysis
chromatogram of the Standard solutions. Samples: Sample solution and Standard solution
Acceptance criteria: No secondary spot from the Calculate the percentage of C17H16ClN3O in the portion of
chromatogram of the Sample solution is larger or more Tablets taken:
intense than the principal spot of Standard solution B Result = (rU/rS) (CS/CU) 100
(0.5%), and the sum of the intensities of the secondary
spots of the Sample solution corresponds to NMT 1.0%.
220 Amoxapine / Official Monographs USP 32
rU = amoxapine peak area from the Sample solution Standard solution: 1.2 mg/mL of USP Amoxicillin RS in
rS = amoxapine peak area from the Standard solution Diluent. [NOTEUse this solution within 6 h.]
CS = concentration of USP Amoxapine RS in the Sample solution: 1.2 mg/mL of Amoxicillin in Diluent.
Standard solution (mg/mL) [NOTEUse this solution within 6 h.]
CU = nominal concentration of amoxapine in the Chromatographic system
Sample solution (mg/mL) (See Chromatography 621, System Suitability.)
Acceptance criteria: 90.0%110.0% Mode: LC
Detector: UV 230 nm
PERFORMANCE TESTS Column: 4-mm 25-cm; packing L1
DISSOLUTION 711 Flow rate: 1.5 mL/min
Medium: Simulated gastric fluid (without enzyme); 900 mL. Injection size: 10 L
Apparatus 2: 50 rpm System suitability
Time: 30 min Sample: Standard solution
Sample solution: Sample per Dissolution 711. Suitability requirements
Standard solution: USP Amoxapine RS in Medium Capacity factor: 1.12.8
Spectrometric conditions Column efficiency: NLT 1700 theoretical plates
Analytical wavelength: 294 nm Tailing factor: NMT 2.5
Analysis: Determine the amount of C17H16ClN3O dissolved Relative standard deviation: NMT 2.0%
from UV absorbances of filtered portions of the Sample Analysis
solution, suitably diluted with Medium, if necessary, in Samples: Standard solution and Sample solution
comparison with a Standard solution having a known Calculate the quantity, in g, of C16H19N3O5S/mg taken:
concentration of USP Amoxapine RS.
Acceptance criteria: NLT 80% (Q) of the labeled amount Result = (rU/rS) (CS/CU) P
of C17H16ClN3O
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements rU = peak response from the Sample solution
ADDITIONAL REQUIREMENTS CS = concentration of USP Amoxicillin RS in the
PACKAGING AND STORAGE: Preserve in well-closed containers. Standard solution (mg/mL)
USP REFERENCE STANDARDS 11 CU = concentration of Sample solution (mg/mL)
USP Amoxapine RS P = stated amoxicillin content of USP Amoxicillin RS
(g/mg)
Acceptance criteria: 9001050 g
Amoxicillin SPECIFIC TESTS
(Comment on this Monograph)id=m4100=Amoxicillin=A- CRYSTALLINITY 695: Meets the requirements
Monos.pdf) DIMETHYLANILINE 223: Meets the requirement
PH 791: 3.56.0
Sample solution: 2 mg/ml
WATER DETERMINATION, Method I 921: 11.5%14.5%
STERILITY TESTS 71: Where the label states that Amoxicillin
is sterile, it meets the requirements when tested as directed
in Test for Sterility of the Product to Be Examined, Direct
Inoculation of the Culture Medium, except to use Fluid
Thioglycollate Medium containing polysorbate 80 solution (1
C16H19N3O5S 3H2O 419.45 in 200) and an amount of sterile penicillinase sufficient to
4-Thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, 6-[ inactivate the amoxicillin in each tube, to use
[amino(4-hydroxyphenyl)acetyl]amino-3,3-dimethyl-7-oxo-, SoybeanCasein Digest Medium containing polysorbate 80
trihydrate [2S-[2,5,6(S*)]]-; solution (1 in 200) and an amount of sterile penicillinase
(2S,5R,6R)-6-[(R)-(-)-2-Amino-2-(p- sufficient to inactivate the amoxicillin in each tube, and to
hydroxyphenyl)acetamido]-3,3-dimethyl-7-oxo-4-thia-1- shake the tubes once daily.
azabicyclo[3.2.0]heptane-2-carboxylic acid trihydrate BACTERIAL ENDOTOXINS TEST 85: Where the label states that
[61336-70-7]. Amoxicillin is sterile or Amoxicillin must be subjected to
Anhydrous 365.41 further processing during the preparation of injectable
[26787-78-0]. dosage forms, it contains NMT 0.25 Endotoxin Unit/mg of
amoxicillin.
DEFINITION
Amoxicillin contains NLT 900 g and NMT 1050 g of ADDITIONAL REQUIREMENTS
C16H19N3O5S per mg, calculated on the anhydrous basis. PACKAGING AND STORAGE: Preserve in tight containers, and
IDENTIFICATION LABELING: Where it is intended for use in preparing
INFRARED ABSORPTION 197K injectable dosage forms, the label states that it is intended
for veterinary use only and that it is sterile or must be
ASSAY subjected to further processing during the preparation of
PROCEDURE injectable dosage forms. Label all other Amoxicillin to
Diluent: 6.8 g/L of monobasic potassium phosphate in indicate that it is to be used in the manufacture of
water, and adjust with a 45% (w/w) solution of potassium nonparenteral drugs only.
hydroxide to a pH of 5.0 0.1 USP REFERENCE STANDARDS 11
Mobile phase: Acetonitrile and Diluent (1:24). Decrease the USP Amoxicillin RS
acetonitrile concentration to increase the retention time of USP Endotoxin RS
amoxicillin.
USP 32 Official Monographs / Amoxicillin 221
Amoxicillin Boluses rS = amoxicillin peak response from the Standard

(Comment on this Monograph)id=m4105=Amoxicillin solution
Boluses=A-Monos.pdf) CS = concentration of USP Amoxicillin RS in the
DEFINITION CU = nominal concentration of the Sample solution
Amoxicillin Boluses contain NLT 90.0% and NMT 110.0% of (mg/mL)
the labeled amount of amoxicillin (C16H19N3O5S). P = stated content of USP Amoxicillin RS (mg/mg)
IDENTIFICATION
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 PERFORMANCE TESTS
Adsorbent: 0.25-mm layer of chromatographic silica gel DISINTEGRATION 701
mixture Medium: Simulated gastric fluid being used instead of
Standard solution: 4 mg/mL of USP Amoxicillin RS in 0.1 N water
hydrochloric acid. [NOTEUse within 10 min of Time: 30 min
preparation.]
Sample solution: 4 mg/mL of amoxicillin, from powdered SPECIFIC TESTS
Boluses in 0.1 N hydrochloric acid WATER DETERMINATION, Method I 921: NMT 7.5%
Application volume: 5 L ADDITIONAL REQUIREMENTS
Developing solvent system: Methanol, chloroform, PACKAGING AND STORAGE: Preserve in tight containers, and
pyridine, and water (9:8:1:3) store at controlled room temperature.
Spray reagent: 3 mg/mL of ninhydrin in alcohol LABELING: Label Boluses to indicate that they are for
Analysis veterinary use only.
Samples: Standard solution and Sample solution USP REFERENCE STANDARDS 11
When the solvent front has moved about three-fourths of USP Amoxicillin RS
the length of the plate, remove the plate from the
chamber, and dry with warm air for 10 min. Locate the
spots on the plate by spraying lightly with Spray reagent,
and dry at 110 for 15 min. Amoxicillin Capsules
Acceptance criteria: The RF value of the principal spot of (Comment on this Monograph)id=m4110=Amoxicillin
the Sample solution corresponds to that of the Standard Capsules=A-Monos.pdf)
solution.
DEFINITION
ASSAY Amoxicillin Capsules contain the equivalent of NLT 90.0% and
PROCEDURE NMT 120.0% of the labeled amount of amoxicillin
Diluent: 6.8 mg/mL of monobasic potassium phosphate in (C16H19N3O5S).
water. Adjust with a 45% (w/w) solution of potassium
hydroxide to a pH of 5.0 0.1. IDENTIFICATION
Mobile phase: Acetonitrile and Diluent (1:24). Decrease the THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
acetonitrile concentration to increase the retention time of Standard solution: 4 mg/mL of USP Amoxicillin RS in 0.1 N
amoxicillin. hydrochloric acid
Standard solution: 1.2 mg/mL of USP Amoxicillin RS in [NOTEUse within 10 min after preparation.]
Diluent. [NOTEUse this solution within 6 h.] Sample solution: Equivalent to 4 mg/mL of amoxicillin,
Sample solution: Weigh and finely powder NLT 5 Boluses. from Capsule powder in 0.1 N hydrochloric acid
Transfer a portion of the powder, equivalent to 250 mg of Chromatographic system
amoxicillin, to a 250-mL volumetric flask, add Diluent to (see Chromatography 621, Thin-layer chromatography.)
volume, and mix. Sonicate if necessary to ensure complete Mode: TLC
dissolution of the amoxicillin. Pass a portion of this solution Adsorbent: 0.25-mm layer of chromatographic silica gel
through a filter of 1-m or finer porosity. mixture
[NOTEUse this solution within 6 h.] Application volume: 5 L
Chromatographic system Developing solvent system: Methanol, chloroform,
(See Chromatography 621, System Suitability.) pyridine, and water (9:8:1:3)
Mode: LC Spray reagent: 3 mg/mL of ninhydrin in alcohol
Column: 4-mm 25-cm; packing L1 Samples: Standard solution and Sample solution
Flow rate: 1.5 mL/min When the solvent front has moved three-fourths of the
Injection size: 10 L length of the plate, remove the plate from the chamber,
System suitability and dry with warm air for 10 min. Locate the spots on
Sample: Standard solution the plate by spraying lightly with Spray reagent and dry at
Suitability requirements 110 for 15 min.
Capacity factor: 1.12.8 Acceptance criteria: The RF value of the principal spot of
Column efficiency: NLT 1700 theoretical plates the Sample solution corresponds to that of the Standard
Tailing factor: NMT 2.5 solution.
Analysis ASSAY
Samples: Standard solution and Sample solution PROCEDURE
Calculate the percentage of C16H19N3O5S in the portion of Diluent: Dissolve 6.8 g/L of monobasic potassium
Boluses taken: phosphate in water, and adjust with a 45% (w/w) solution
of potassium hydroxide to a pH of 5.0 0.1.
Result = (rU/rS) (CS/CU) 100 P Mobile phase: Acetonitrile and Diluent (1:24). Decrease the
acetonitrile concentration to increase the retention time of
rU = amoxicillin peak response from the Sample amoxicillin.
solution
222 Amoxicillin / Official Monographs USP 32
Standard solution: 1.2 mg/mL of USP Amoxicillin RS in UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Diluent. [NOTE Use this solution within 6 h.]
Sample solution: Remove, as completely as possible, the SPECIFIC TESTS
contents of NLT 20 Capsules, mix the combined contents, WATER DETERMINATION, Method I 921: NMT 14.5%
and transfer a quantity, equivalent to 200 mg of anhydrous
amoxicillin, to a 200-mL volumetric flask, and add Diluent to ADDITIONAL REQUIREMENTS
volume. Sonicate if necessary to ensure complete PACKAGING AND STORAGE: Preserve in tight containers, and
dissolution. Pass a portion of this solution through a suitable store at controlled room temperature.
filter having a 1-m or finer porosity, and use the filtrate. LABELING: When more than one Dissolution test is given, the
[NOTEUse this solution within 6 h.] labeling states the Dissolution test used only if Test 1 is not
Chromatographic system used.
(See Chromatography 621, System Suitability.) USP REFERENCE STANDARDS 11
Mode: LC USP Amoxicillin RS
Detector: UV 230 nm
Flow rate: 1.5 mL/min Amoxicillin Intramammary Infusion
System suitability (Comment on this Monograph)id=m4120=Amoxicillin
Sample: Standard solution Intramammary Infusion=A-Monos.pdf)
Capacity factor: 1.12.8 Amoxicillin Intramammary Infusion is a suspension of
Column efficiency: NLT 1700 theoretical plates Amoxicillin in a suitable vegetable oil vehicle. It contains NLT
Tailing factor: NMT 2.5 90.0% and NMT 120.0% of the labeled amount of amoxicillin
Relative standard deviation: NMT 2.0% (C16H19N3O5S). It contains a suitable dispersing agent and
Analysis preservative.
Calculate the percentage of C16H19N3O5S in the portion of IDENTIFICATION
Capsules taken: THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
Standard solution: 4 mg/mL of USP Amoxicillin RS in 0.1 N
Result = (rU/rS) (CS/CU) P 100 hydrochloric acid. [NOTEUse within 10 min after
preparation.]
rU = peak response of the Sample solution Sample solution: Transfer a quantity of Intramammary
rS = peak response of the Standard solution Infusion, equivalent to 60 mg of amoxicillin, to a 50-mL
CS = concentration of USP Amoxicillin RS in the centrifuge tube. Add 25 mL of toluene, and centrifuge.
Standard solution (mg/mL) Decant and discard the toluene. Wash the residue with four
CU = nominal concentration of amoxicillin in the 25-mL portions of toluene, sonicating for 30 s after each
Sample solution (mg/mL) addition of toluene. Dry the residue in a vacuum over silica
P = stated amoxicillin content of USP Amoxicillin RS gel. Add 15 mL of 0.1 N hydrochloric acid to the residue.
(mg/mL) Chromatographic system
Acceptance criteria: 90.0%120.0% (see Chromatography 621, Thin-layer chromatography.)
PERFORMANCE TESTS mixture (see Chromatography 621)
DISSOLUTION 711 Application volume: 5 L
Test 1 Developing solvent system: Methanol, chloroform,
Medium: Water; 900 mL pyridine, water, and (9:8:1:3)
Apparatus 1: 100 rpm, for Capsules containing 250 mg Spray reagent: 3 mg/mL of ninhydrin in alcohol
Apparatus 2: 75 rpm, for Capsules containing 500 mg Analysis
Time: 60 min Samples: Standard solution and Sample solution
Analytical wavelength: UV 272 nm When the solvent front has moved about three-fourths of
Standard solution: USP Amoxicillin RS in Medium the length of the plate, remove the plate from the
Sample solution: Sample per Dissolution 711; dilute with chamber, and dry with warm air for 10 min. Locate the
Medium to a concentration that is similar to Standard spots on the plate by spraying lightly with Spray reagent
solution. and dry at 110 for 15 min.
Tolerances: NLT 80% (Q) of the labeled amount of Acceptance criteria: The RF value of the principal spot from
C16H19N3O5S the Sample solution corresponds to that from the Standard
Test 2 (If the product complies with this test, the labeling solution.
indicates that it meets USP Dissolution Test 2.)
Medium: Water; 900 mL ASSAY
Apparatus 1: 100 rpm PROCEDURE
Time: 90 min Analysis: Proceed as directed for amoxicillin under
Analytical wavelength: UV 272 nm AntibioticsMicrobial Assays 81. Expel the contents of 1
Standard solution: USP Amoxicillin RS in Medium syringe of Intramammary Infusion into a high-speed glass
Sample solution: Sample per Dissolution 711; dilute with blender jar containing 499.0 mL of Buffer No. 3 and 1.0 mL
Medium to concentration that is similar to Standard of polysorbate 80, and blend for 35 min. Allow to stand for
solution. 10 min, and dilute a measured volume of the aqueous
Tolerances: NLT 80% (Q) of the labeled amount of phase quantitatively and stepwise with Buffer No. 3 to obtain
C16H19N3O5S
a Sample Dilution having a concentration assumed to be finer porosity, and use the filtrate as Sample solution A.
equal to the median dose level of the Standard. [NOTEUse this solution within 6 h.]
Acceptance criteria: 90.0%120.0% Sample solution B (where the label states the quantity of
amoxicillin in a given volume of constituted suspension):
SPECIFIC TESTS Constitute Amoxicillin for Injectable Suspension as directed
WATER DETERMINATION, Method I 921: NMT 1.0%, 20 mL in the labeling. Quantitatively dilute a measured volume of
of a mixture of toluene and methanol (7:3) being used in the constituted suspension with Diluent to obtain a solution
place of methanol in the titration vessel containing 1 mg/mL of anhydrous amoxicillin. Pass a
portion of this solution through a suitable filter of 1-m or
ADDITIONAL REQUIREMENTS finer porosity, and use the filtrate as Sample solution B.
PACKAGING AND STORAGE: Preserve in well-closed disposable [NOTEUse this solution within 6 h.]
syringes. Chromatographic system
LABELING: Label it to indicate that it is intended for (See Chromatography 621, System Suitability.)
veterinary use only. Mode: LC
USP Amoxicillin RS Column: 4-mm 25-cm; packing L1
Amoxicillin for Injectable Suspension System suitability
(Comment on this Monograph)id=m4129=Amoxicillin for Suitability requirements
Injectable Suspension=A-Monos.pdf) Capacity factor: 1.12.8
DEFINITION Column efficiency: NLT 1700 theoretical plates
Amoxicillin for Injectable Suspension is a sterile mixture of Tailing factor: NMT 2.5
Amoxicillin and one or more suitable buffers, preservatives, Relative standard deviation: NMT 2.0%
stabilizers, and suspending agents. It contains NLT 90.0% and Analysis
NMT 120.0% of the labeled amount of amoxicillin Samples: Standard solution and Sample solutions
(C16H19N3O5S). Calculate the percentage of C16H19N3O5S in the container,
or in the portion of constituted Amoxicillin for Injectable
IDENTIFICATION Suspension taken:
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
Standard solution: 4 mg/mL of USP Amoxicillin RS in 0.1 N Result = (rU/rS) (Cs/CU) P 100
hydrochloric acid. [NOTEUse within 10 min after
preparation.] rU = amoxicillin peak response of the Sample solution
Sample solution: Equivalent of 4 mg/mL of amoxicillin, rS = amoxicillin peak response of the Standard
from Amoxicillin for Injectable Suspension, in 0.1 N solution
hydrochloric acid. Allow to stand for 5 min before use. CS = concentration of USP Amoxicillin RS in the
Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution
mixture CU = nominal concentration, in mg of anhydrous
Application volume: 5 L amoxicillin/mL, of Sample solution A or of
Developing solvent system: Methanol, chloroform, Sample solution B on the basis of the labeled
pyridine, and water (9:8:1:3) quantity in the container or in the portion of
Spray reagent: 3 mg/mL of ninhydrin in alcohol constituted suspension taken, respectively, and
Analysis the extent of dilution
Samples: Standard solution and Sample solution P = stated content of USP Amoxicillin RS (mg/mg)
When the solvent front has moved about three-fourths of Acceptance criteria: 90.0%120.0%
the length of the plate, remove the plate from the SPECIFIC TESTS
chamber, and dry with warm air for 10 min. Locate the STERILITY TESTS 71: It meets the requirements when tested
spots on the plate by spraying lightly with Spray reagent, as directed in Test for Sterility of the Product to Be Examined,
and dry at 110 for 15 min. Direct Inoculation of the Culture Medium, except to use Fluid
Acceptance criteria: The RF value of the principal spot Thioglycollate Medium containing polysorbate 80 solution (1
obtained from the Sample solution corresponds to that in 200) and an amount of sterile penicillinase sufficient to
obtained from the Standard solution. inactivate the amoxicillin in each tube, to use
ASSAY SoybeanCasein Digest Medium containing polysorbate 80
PROCEDURE solution (1 in 200) and an amount of sterile penicillinase
Diluent: 6.8 mg/mL of monobasic potassium phosphate in sufficient to inactivate the amoxicillin in each tube, and to
water. Adjust with a 45% (w/w) solution of potassium shake the tubes once daily.
hydroxide to a pH of 5.0 0.1. PH 791: 5.07.0, in the suspension constituted as directed
Mobile phase: Acetonitrile and Diluent (1:24). Decrease the in the labeling
acetonitrile concentration to increase the retention time of WATER DETERMINATION, Method I 921: 11.0%14.0%
amoxicillin. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.25
Standard solution: 1.2 mg/mL of USP Amoxicillin RS in Endotoxin Unit/mg of amoxicillin.
Diluent. [NOTEUse this solution within 6 h.] ADDITIONAL REQUIREMENTS
Sample solution A (where it is represented as being in a PACKAGING AND STORAGE: Preserve as described in Injections
single-dose container): Constitute Amoxicillin for Injectable 1, Containers for Sterile Solids.
Suspension as directed in the labeling. Withdraw all of the LABELING: Label it to indicate that it is for veterinary use
withdrawable contents, using a hypodermic needle and only.
syringe, and quantitatively dilute with Diluent to obtain a USP REFERENCE STANDARDS 11
solution containing 1 mg/mL of anhydrous amoxicillin. Pass USP Amoxicillin RS
a portion of this solution through a suitable filter of 1-m or
USP Endotoxin RS ADDITIONAL REQUIREMENTS

PACKAGING AND STORAGE: Preserve in multiple-dose
containers equipped with a suitable dosing pump.
Amoxicillin Oral Suspension only.
(Comment on this Monograph)id=m4130=Amoxicillin Oral USP REFERENCE STANDARDS 11
Suspension=A-Monos.pdf) USP Amoxicillin RS
DEFINITION
Amoxicillin Oral Suspension is a suspension of Amoxicillin in
Soybean Oil. It contains NLT 90.0% and NMT 120.0% of the Amoxicillin for Oral Suspension
labeled amount of amoxicillin (C16H19N3O5S). (Comment on this Monograph)id=m4140=Amoxicillin for Oral
Suspension=A-Monos.pdf)
IDENTIFICATION
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST DEFINITION
Standard solution: 4 mg/mL of USP Amoxicillin RS in 0.1 N Amoxicillin for Oral Suspension contains the equivalent of NLT
hydrochloric acid. [NOTEUse within 10 min after 90.0% and NMT 120.0% of the labeled amount of amoxicillin
preparation.] (C16H19N3O5S). It contains one or more suitable buffers, colors,
Sample solution: Shake a portion of Oral Suspension with a flavors, preservatives, stabilizers, sweeteners, and suspending
mixture of acetone and 0.1 N hydrochloric acid (4:1) to agents.
obtain a solution containing 1 mg/mL of amoxicillin.
(See Chromatography 621, Thin-layer chromatography.) THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution: 4 mg/mL of USP Amoxicillin RS in 0.1 N
mixture (See Chromatography 621.) hydrochloric acid. [NOTEUse within 10 min after
Application volume: 5 L preparation.]
Developing solvent system: Methanol, chloroform, Sample solution: Equivalent of 4 mg/mL of amoxicillin,
pyridine and water, (9:8:1:3) from Oral Suspension in 0.1 N hydrochloric acid. [NOTE
Spray reagent: 3 mg/mL of ninhydrin in alcohol Allow the solution to stand for 5 min before use.]
Analysis Chromatographic system
Samples: Sample solution and Standard solution (see Chromatography 621, System Suitability.)
When the solvent front has moved about three-fourths of Mode: TLC
the length of the plate, remove the plate from the Adsorbent: 0.25-mm layer of chromatographic silica gel
chamber, and dry with warm air for 10 min. Locate the mixture
spots on the plate by spraying lightly with Spray reagent Application volume: 5 L
and dry at 110 for 15 min. Developing solvent system: Methanol, chloroform,
Acceptance criteria: The RF value of the principal spot pyridine, and water (9:1:8:3)
from the Sample solution corresponds to that from the Spray reagent: 3 mg/mL of ninhydrin in alcohol
Standard solution. Analysis
ASSAY When the solvent front has moved three-fourths of the
PROCEDURE length of the plate, remove the plate from the chamber,
Standard solution: Prepare as directed for Standard and dry with warm air for 10 min. Locate the spots on
Preparation under Iodometric AssayAntibiotics 425, using the plate by spraying lightly with Spray reagent and dry at
USP Amoxicillin RS. 110 for 15 min.
Sample solution: Using the dosing pump, deliver a number Acceptance criteria: The RF value of the principal spot of
of doses of Oral Suspension, equivalent to 250 mg of the Sample solution corresponds to that of the Standard
amoxicillin, to a separator containing 100 mL of hexanes, solution.
and shake vigorously. Add 140 mL of water, and shake for 5
min. Allow the layers to separate, and drain the lower, ASSAY
aqueous layer into a 250-mL volumetric flask. Repeat the PROCEDURE
extraction with two 50-mL portions of water. Combine the Diluent: Dissolve 6.8 g/L of monobasic potassium
aqueous extracts in the volumetric flask, dilute with water to phosphate in water, and adjust with a 45% (w/w) solution
volume. of potassium hydroxide to a pH of 5.0 0.1.
Analysis: Proceed with Oral Suspension as directed for Mobile phase: Acetonitrile and Diluent (1:24). Decrease the
Procedure under Iodometric AssayAntibiotics 425, using acetonitrile concentration to increase the retention time of
USP Amoxicillin RS. amoxicillin.
Calculate the percentage of C16H19N3O5S in each dose of Standard solution: 1.2 mg/mL of USP Amoxicillin RS in
Oral Suspension taken: Diluent. [NOTEUse this solution within 6 h.]
Sample solution: Dilute a measured volume of Amoxicillin
(250/N)(F/2000)(B I) for Oral Suspension, constituted as directed in the labeling,
freshly mixed and free from air bubbles, quantitatively and
N = the number of doses taken, and the other terms stepwise in Diluent to obtain a solution containing nominally
are as defined therein. 1 mg/mL of anhydrous amoxicillin. Filter a portion of this
Acceptance criteria: 90.0%120.0% solution through a suitable filter of 1-m or finer porosity,
and use the filtrate as the Sample solution. [NOTEUse this
SPECIFIC TESTS solution within 6 h.]
WATER DETERMINATION, Method I 921: NMT 2.0%, 20 mL Chromatographic system
of a mixture of toluene and methanol (7:3) being used in (See Chromatography 621, System Suitability.)
place of methanol in the titration vessel.
Mode: LC Spray reagent: 3 mg/mL of ninhydrin in alcohol

Column: 4-mm 25-cm; packing L1 Samples: Standard solution and Sample solution
Flow rate: 1.5 mL/min Dry the plate with the aid of a current of warm air for 10
Injection size: 10 L min. Locate the spots on the plate by spraying lightly with
System suitability Spray reagent, and dry at 110 for 15 min.
Sample: Standard solution Acceptance criteria: The RF value of the principal spot of
Suitability requirements the Sample solution corresponds to that of the Standard
Capacity factor: 1.12.8 solution.
Tailing factor: NMT 2.5 ASSAY
Relative standard deviation: NMT 2.0% PROCEDURE
Analysis Diluent: 6.8 g/L of monobasic potassium phosphate in
Samples: Standard solution and Sample solution water, and adjust with a 45% (w/w) solution of potassium
Calculate the percentage of C16H19N3O5S in each mL of the hydroxide to a pH of 5.0 0.1
constituted Amoxicillin for Oral Suspension taken: Mobile phase: Acetonitrile and Diluent (1:24). Decrease the
acetonitrile concentration to increase the retention time of
Result = (rU/rS) (CS/CU) P 100 amoxicillin.
Standard solution: 1.2 mg/mL of USP Amoxicillin RS in
rU = peak response of the Sample solution Diluent. [NOTEUse this solution within 6 h.]
rS = peak response of the Standard solution Sample solution: Place NLT 5 Tablets in a high-speed glass
CS = concentration of USP Amoxicillin RS in the blender jar containing Diluent sufficient to yield a
Standard solution (mg/mL) concentration of 1 mg of anhydrous amoxicillin/mL, blend
CU = nominal concentration of anhydrous amoxicillin for 4 1 min, allow to stand for 5 min, and centrifuge a
in the Sample solution (mg/mL) portion of the mixture.
P = stated amoxicillin content of USP Amoxicillin RS [NOTEWhere the volume of Diluent required would
(g/mg) exceed 500 mL, place 5 Tablets in a volumetric flask of
Acceptance criteria: 90.0%120.0% such capacity that when finally diluted to volume, a
concentration of 1 mg of anhydrous amoxicillin per mL
PERFORMANCE TESTS would be obtained. Add a volume of Diluent equivalent to
UNIFORMITY OF DOSAGE UNITS 905 three-fourths of the capacity of the volumetric flask, and
For solids packaged in single-unit containers: Meets the sonicate for 5 min. Dilute with Diluent to volume, add a
requirements magnetic stirring bar, and stir for 30 min. Centrifuge a
DELIVERABLE VOLUME 698: Meets the requirements portion of this solution.]
Pass a portion of the clear supernatant through a suitable
SPECIFIC TESTS filter having a 1-m or finer porosity, and use the filtrate.
PH 791: 5.07.5, in the suspension constituted as directed [NOTEUse this solution within 6 h.]
in the labeling Chromatographic system
WATER DETERMINATION, Method I 921: NMT 3.0%, except (See Chromatography 621, System Suitability.)
where it is labeled as containing 80 mg of amoxicillin per Mode: LC
mL after constitution, the limit is NMT 4.0%. Detector: UV 230 nm
ADDITIONAL REQUIREMENTS Column: 4-mm 25-cm; packing L1
PACKAGING AND STORAGE: Preserve in tight containers, and Flow rate: 1.5 mL/min
store at a controlled room temperature. Injection size: 10 L
USP REFERENCE STANDARDS 11 System suitability
USP Amoxicillin RS Sample: Standard solution
Capacity factor: 1.12.8
Amoxicillin Tablets Tailing factor: NMT 2.5
(Comment on this Monograph)id=m4170=Amoxicillin Relative standard deviation: NMT 2.0%
Tablets=A-Monos.pdf) Analysis
DEFINITION Calculate the percentage of C16H19 N3O5S in each Tablet
Amoxicillin Tablets contain NLT 90.0% and NMT 120.0% of the taken:
labeled amount of amoxicillin (C16H19N3O5S).
Result = (rU/rS) (CS/CU) P 100
IDENTIFICATION
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 rU = peak response of amoxicillin from the Sample
Sample solution: 4 mg/mL, from powdered Tablets in 0.1 solution
N hydrochloric acid. [NOTEUse within 10 min after rS = peak response of amoxicillin from the Standard
preparation.] solution
Standard solution: 4 mg/mL of USP Amoxicillin RS in 0.1 N CS = concentration of USP Amoxicillin RS in the
hydrochloric acid Standard solution (mg/mL)
Adsorbent: 0.25-mm layer of chromatographic silica gel CU = nominal concentration of amoxicillin in the
mixture Sample solution (mg/mL)
Application volume: 5 L P = stated content of USP Amoxicillin RS (mg/mg)
Developing solvent system: Methanol, chloroform,
pyridine, and water (9:8:1:3)
Acceptance criteria: 90.0%120.0% For veterinary products: Proceed as directed above, except
to use Apparatus 2 at 100 rpm.
PERFORMANCE TESTS
DISSOLUTION 711 ADDITIONAL REQUIREMENTS
Medium: Water; 900 mL PACKAGING AND STORAGE: Preserve in tight containers, and
Apparatus 2: 75 rpm store at controlled room temperature.
Time: 30 min LABELING: Label chewable Tablets to indicate that they are
Determine the amount of C16H19N3O5S dissolved by to be chewed before swallowing. Tablets intended solely for
employing the following method. veterinary use are so labeled.
pH 5.0 Buffer: 27.2 g of monobasic potassium phosphate USP REFERENCE STANDARDS 11
in 3 L of water, adjust with a 45% (w/w) solution of USP Amoxicillin RS
potassium hydroxide to a pH of 5.0 0.1, and dilute with
water to obtain 4 L of solution
Mobile phase: Acetonitrile and pH 5.0 Buffer (1:39), and
pass through a filter having a 0.5-m or finer porosity Amoxicillin Tablets for Oral Suspension
Standard solution: 0.05 mg/mL of USP Amoxicillin RS in (Comment on this Monograph)id=m4173=Amoxicillin Tablets
pH 5.0 Buffer. [NOTEUse this solution within 6 h.] for Oral Suspension=A-Monos.pdf)
Sample solution: Pass a portion of the sample through a
filter having a 0.5-m or finer porosity. Quantitatively dilute DEFINITION
a volume of the filtrate with water to obtain a concentration Amoxicillin Tablets for Oral Suspension contain NLT 90.0% and
of 0.045 mg/mL of amoxicillin. NMT 110.0% of the labeled amount of amoxicillin
Chromatographic system (C16H19N3O5S).
Detector: UV 230 nm THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
Column Sample solution: An aqueous dispersion of Amoxicillin
Analytical: 3.9-mm 30-cm; packing L1 Tablets for Oral Suspension in 0.1 N hydrochloric acid
Guard: 2-mm 2-cm; packing L2 containing 4 mg/mL of amoxicillin. Use within 10 min of
Temperature: Analytical column is maintained at a preparation.
constant temperature of 40 1 Standard solution: 4 mg/mL of USP Amoxicillin RS in 0.1 N
Flow rate: 0.7 mL/min hydrochloric acid
Injection size: 10 L Chromatographic system
System suitability (see Chromatography 621, Thin-Layer Chromatography.)
Sample: Standard solution Mode: TLC
Suitability requirements Adsorbent: 0.25-mm layer of chromatographic silica gel
Capacity factor: 1.12.8 mixture
Column efficiency: NLT 1700 theoretical plates Application volume: 5 L
Tailing factor: NMT 2.5 Developing solvent system: Methanol, chloroform,
Relative standard deviation: NMT 1.5% pyridine, and water (9:8:1:3)
Analysis Spray reagent: 3 mg/mL of ninhydrin in alcohol
Samples: Standard solution and Sample solution Analysis
Calculate the percentage of C16H19N3O5S dissolved by the Samples: Standard solution and Sample solution
formula: Dry the plate with the aid of a current of warm air for 10
min. Locate the spots on the plate by spraying lightly with
Result = (rU/rS) (CS D V P (100/L) Spray reagent, and dry at 110 for 15 min.
Acceptance criteria: The RF value of the principal spot of
rU = peak response of amoxicillin from the Sample the Sample solution corresponds to that of the Standard
solution solution.
rS = peak response of amoxicillin from the Standard
solution ASSAY
CS = concentration of USP Amoxicillin RS in the PROCEDURE
Standard solution (mg/mL) Diluent: 6.8 g/L of monobasic potassium phosphate in
V = volume of the dissolution medium, 900 mL water, and adjust with a 45% (w/w) solution of potassium
D = dilution factor for the Sample solution hydroxide to a pH of 5.0 0.1
P = stated content of USP Amoxicillin RS (mg/mg) Mobile phase: Acetonitrile and Diluent (1:24). Decrease the
L = label claim (mg/tablet) acetonitrile concentration to increase the retention time of
Time: 30 min amoxicillin.
Tolerances: NLT 75% (Q) of the labeled amount of Standard solution: 1.2 mg/mL of USP Amoxicillin RS in
C16H19N3O5S Diluent. [NOTEUse this solution within 6 h.]
For products labeled as chewable tablets: Proceed as Sample solution: Prepare a dispersion of 20 Tablets for Oral
directed above. Suspension using an accurately measured volume of water.
For chewable tablets labeled to contain 200 mg or 400 Quantitatively dilute a portion of the dispersion with Diluent
mg to obtain a solution containing 1.2 mg/mL of amoxicillin.
Time: 20 min Pass a portion of the solution through a filter having a 1-m
Tolerances: NLT 70% (Q) of the labeled amount of or finer porosity, and use the filtrate. [NOTEUse this
C16H19N3O5S solution within 6 h.]
For chewable tablets labeled to contain 125 mg or 250 Chromatographic system
mg (See Chromatography 621, System Suitability.)
Time: 90 min
C16H19N3O5S
Mode: LC Flow rate: 0.7 mL/min

Detector: UV 230 nm Injection size: 10 L
Column: 4-mm 25-cm; packing L1 System suitability
Flow rate: 1.5 mL/min Sample: Standard solution
Injection size: 10 L Suitability requirements
System suitability Capacity factor: 1.12.8
Sample: Standard solution Column efficiency: NLT 1700 theoretical plates
Suitability requirements Tailing factor: NMT 2.5
Capacity factor: 1.12.8 Relative standard deviation: NMT 1.5%
Column efficiency: NLT 1700 theoretical plates Analysis
Tailing factor: NMT 2.5 Samples: Sample solution and Standard solution
Relative standard deviation: NMT 2.0% Calculate the percentage of C16H19N3O5S dissolved:
Analysis
Samples: Standard solution and Sample solution Result = (rU/rS) (CS D V P (100/L)
Calculate the percentage of C16H19N3O5S in each
Amoxicillin Tablet for Oral Suspension taken: rU = amoxicillin peak responses from the Sample
solution
Result = (rU/rS) (CS/CU) P 100 rS = amoxicillin peak responses from the Standard
solution
rU = amoxicillin peak response from the Sample CS = concentration of USP Amoxicillin RS in the
solution Standard solution (mg/mL)
rS = amoxicillin peak response from the Standard V = volume of medium, 900
solution D = dilution factor required to prepare the Sample
CS = concentration of USP Amoxicillin RS in the solution
Standard solution (mg/mL) P = stated content of USP Amoxicillin RS (g/mg)
CU = nominal concentration of the Sample solution L = label claim (mg/tablet)
(mg/mL) Tolerances: NLT 80% (Q) of the labeled amount of
P = stated content of USP Amoxicillin RS (mg/mg) C16H19N3O5S
Acceptance criteria: 90.0%110.0% UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
PERFORMANCE TESTS SPECIFIC TESTS
DISINTEGRATION 701 DISPERSION FINENESS: Place 2 Tablets for Oral Suspension in
Medium: Water at 20 5 100 mL of water, and stir until completely dispersed. A
Time: 3 min smooth dispersion is obtained that passes through a No. 25
DISSOLUTION 711 sieve.
Apparatus 2: 75 rpm ADDITIONAL REQUIREMENTS
Time: 30 min PACKAGING AND STORAGE: Preserve in tight containers.
Determine the amount of C16H19N3O5S dissolved by USP REFERENCE STANDARDS 11
employing the following method. USP Amoxicillin RS
pH 5.0 buffer: 27.2 g of monobasic potassium phosphate
in 3 L of water, adjust with a 45% (w/w) solution of
potassium hydroxide to a pH of 5.0 0.1, dilute with water Amoxicillin and Clavulanate Potassium
to obtain 4 L of solution
Mobile phase: Acetonitrile and pH 5.0 buffer (1:39), and for Oral Suspension
pass through a filter having a 0.5-m or finer porosity (Comment on this Monograph)id=m4190=Amoxicillin and
Standard solution: 0.05 mg/mL of USP Amoxicillin RS in Clavulanate Potassium for Oral Suspension=A-Monos.pdf)
pH 5.0 buffer. [NOTEUse this solution within 6 h.]
Sample solution: Pass a portion of the sample through a DEFINITION
filter having a 0.5-m or finer porosity. Quantitatively dilute Amoxicillin and Clavulanate Potassium for Oral Suspension
an accurately measured volume of the filtrate with water to contains the equivalent of NLT 90.0% and NMT 120.0% of
obtain a concentration of 0.045 mg/mL of amoxicillin. the labeled amount of amoxicillin (C16H19N3O5S) and the
Chromatographic system equivalent of NLT 90.0% and NMT 125.0% of the labeled
(See Chromatography 621, System Suitability.) amount of clavulanic acid (C8H9NO5). It contains one or more
Mode: LC suitable buffers, colors, flavors, preservatives, stabilizers,
Detector: UV 230 nm sweeteners, and suspending agents.
Column
Analytical column: 3.9-mm 30-cm; packing L1 IDENTIFICATION
Guard column: 2-mm 2-cm; packing L2 The retention times of the major peaks of the Sample solution
Temperature: Analytical column is maintained at a correspond to those of the Standard solution, as obtained in
constant temperature of 40 1 the Assay.

PROCEDURE DELIVERABLE VOLUME 698
Buffer: 7.8 g of monobasic sodium phosphate in 900 mL of For powder packaged in multiple-unit containers: Meets
water, adjust with phosphoric acid or 10 N sodium the requirements
hydroxide to a pH of 4.4 0.1, and dilute with water to UNIFORMITY OF DOSAGE UNITS 905
1000 mL. For powder packaged in single-unit containers: Meets
Mobile phase: Methanol and Buffer (1:19), and pass the requirements
through a filter having a 0.5-m or finer porosity.
Standard solution: 0.5 mg/mL of USP Amoxicillin RS and SPECIFIC TESTS
0.2 mg/mL of USP Clavulanate Lithium RS in water PH 791: 3.86.6, in the suspension constituted as directed
Sample solution: Dilute an accurately measured volume of in the labeling, the test being performed immediately after
Amoxicillin and Clavulanic Acid for Oral Suspension, constitution.
constituted as directed in the labeling, quantitatively with
water to obtain a solution containing 0.5 mg/mL of ADDITIONAL REQUIREMENTS
amoxicillin. Stir by mechanical means for 10 min, and filter. PACKAGING AND STORAGE: Preserve in tight containers, at
Use the filtrate as the Sample solution within 1 h of the controlled room temperature.
dilution of the suspension. USP REFERENCE STANDARDS 11
Chromatographic system USP Amoxicillin RS
(See Chromatography 621, System Suitability.) USP Clavulanate Lithium RS
Mode: LC
Detector: UV 220 nm
Column: 4-mm 30-cm; 3- to 10-m packing L1 Amoxicillin and Clavulanate Potassium
Flow rate: 2 mL/min Tablets
System suitability (Comment on this Monograph)id=m4195=Amoxicillin and
Sample: Standard solution Clavulanate Potassium Tablets=A-Monos.pdf)
[NOTEThe relative retention times for clavulanic acid and
amoxicillin are about 0.5 and 1.0, respectively.] DEFINITION
Suitability requirements Amoxicillin and Clavulanate Potassium Tablets contain the
Resolution: NLT 3.5 between the amoxicillin and equivalent of NLT 90.0% and NMT 120.0% of the labeled
clavulanic acid peaks amounts of amoxicillin (C16H19N3O5S) and clavulanic acid
Column efficiency: NLT 550 theoretical plates from each (C8H9NO5).
analyte peak IDENTIFICATION
Tailing factor: NMT 1.5 for each analyte peak The retention times of the major peaks of the Sample solution
Relative standard deviation: NMT 2.0% correspond to those of the Standard solution, as obtained in
Analysis the Assay.
Calculate the percentage of C16H19N3O5S in each mL of the ASSAY
constituted Amoxicillin and Clavulanate Potassium for Oral PROCEDURE
Suspension taken: Buffer: 7.8 g of monobasic sodium phosphate in 900 mL of
water, adjust with phosphoric acid or 10 N sodium
Result = (rU/rS) (CS/CU) P 100 hydroxide to a pH of 4.4 0.1, and dilute with water to
1000 mL
rU = amoxicillin peak response from the Sample Mobile phase: Methanol and Buffer (1:19), and pass
solution through a filter having a 0.5-m or finer porosity
rS = amoxicillin peak response from the Standard Standard solution: 0.5 mg/mL of USP Amoxicillin RS and
solution 0.2 mg/mL of USP Clavulanate Lithium RS in water
CS = concentration of USP Amoxicillin RS in the Sample stock solution: Dissolve NLT 10 Tablets in water
Standard solution (mg/mL) with the aid of mechanical stirring, transfer to a suitable
CU = nominal concentration of amoxicillin in the volumetric flask, and dilute with water to volume. Filter a
Sample solution (mg/mL) portion of this solution, discarding the first 10 mL of the
P = stated content of USP Amoxicillin RS (mg/mg) filtrate.
Calculate the percentage of C8H9NO5 in each mL of the Sample solution: Dilute an accurately measured volume of
constituted Amoxicillin and Clavulanate Potassium for Oral the Sample stock solution filtrate with water to obtain a
Suspension taken: solution containing 0.5 mg/mL of amoxicillin. Use this
Sample solution within 1 h.
Result = (rU/rS) (CS/CU) P 100 Chromatographic system
rU = clavulanic acid peak response from the Sample (See Chromatography 621, System Suitability.)
solution Mode: LC
rS = clavulanic acid peak response from the Standard Detector: UV 220 nm
solution Column: 4-mm 30-cm; 3- to 10-m packing L1
CS = concentration of USP Clavulanate Lithium RS in Flow rate: 2 mL/min
the Standard solution (mg/mL) Injection size: 20 L
CU = nominal concentration of clavulanate in the System suitability
Sample solution (mg/mL) Sample: Standard solution
P = stated content of USP Clavulanate Potassium RS [NOTEThe relative retention times for clavulanic acid and
(mg/mg) amoxicillin are about 0.5 and 1.0, respectively.]
Acceptance criteria: 90.0%120.0% of the labeled amount Suitability requirements
of C16H19N3O5S and 90.0%125.0% of the labeled amount Resolution: NLT 3.5 between the amoxicillin and
of C8H9NO5 clavulanic acid peaks
USP 32 Official Monographs / Amphetamine 229
Column efficiency: NLT 550 theoretical plates from each NMT 8.0% where the labeled amount of amoxicillin in each
analyte peak Tablet is more than 125 mg;
Tailing factor: NMT 1.5 for each analyte peak Where the Tablets are labeled for veterinary use only
Relative standard deviation: NMT 2.0% NMT 10.0%
Analysis
Calculate the percentage of C16H19N3O5S in each Tablet PACKAGING AND STORAGE: Preserve in tight containers.
taken: LABELING: Label chewable Tablets to include the word
chewable in juxtaposition to the official name. The
Result = (rU/rS) (CS/CU) P 100 labeling indicates that chewable Tablets may be chewed
before being swallowed or may be swallowed whole. Tablets
rU = amoxicillin peak response from the Sample intended for veterinary use only are so labeled.
solution USP REFERENCE STANDARDS 11
rS = amoxicillin peak response from the Standard USP Amoxicillin RS
solution USP Clavulanate Lithium RS
CS = concentration of USP Amoxicillin RS in the
CU = nominal concentration of amoxicillin in the
Sample solution (mg/mL) Amphetamine Sulfate
P = stated content of USP Amoxicillin RS (mg/mg) (Comment on this Monograph)id=m4260=Amphetamine
Calculate the percentage of C8H9NO5 in each Tablet taken: Sulfate=A-Monos.pdf)
rU = clavulanic acid peak response from the Sample

solution
rS = clavulanic acid peak response from the Standard
solution
CS = concentration of USP Clavulanate Lithium RS in
the Standard solution (mg/mL) (C9H13N)2 H2SO4 368.49
CU = nominal concentration of clavulanate in the Benzeneethanamine, -methyl-, sulfate (2:1), ()-;
Sample solution (mg/mL) ()--Methylphenethylamine sulfate (2:1) [60-13-9].
P = stated content of USP Clavulanate Lithium RS
(mg/mg) DEFINITION
Amphetamine Sulfate, dried at 105 for 2 h, contains NLT
Acceptance criteria: 90.0%120.0% 98.0% and NMT 102.0% of (C9H13N)2 H2SO4.
PERFORMANCE TESTS IDENTIFICATION
DISINTEGRATION 701: Tablets labeled for veterinary use A. MELTING RANGE OR TEMPERATURE, Class I 741
only; 30 min, simulated gastric fluid TS being substituted for Sample solution: 20 mg/mL Amphetamine Sulfate
water in the test. Analysis: To 5 mL of Sample solution add 5 mL of 1 N
DISSOLUTION 711 sodium hydroxide, cool to 10, add 1 mL of a mixture of
[NOTETablets labeled for veterinary use only are exempt absolute ether and benzoyl chloride (2:1), insert the stopper,
from this requirement.] and shake for 3 min. Filter the precipitate, wash with 10 mL
Medium: Water; 900 mL of cold water, and recrystallize from diluted alcohol.
Apparatus 2: 75 rpm Acceptance criteria: The crystals of the benzoyl derivative
Time: 30 min; or 45 min where the Tablets are labeled as of amphetamine so obtained, after drying at 80 for 3 h,
chewable. melt between 131 and 135.
Analysis: Determine the amount of C16H19N3O5S and B. IDENTIFICATION TESTSGENERAL, Sulfate 191: Meets the
C8H9NO5 dissolved, employing the Analysis set forth in the requirements of the tests
Assay, making any necessary volumetric adjustments. Sample solution: 100 mg/mL
C16H19N3O5S and NLT 80% (Q) of the labeled amount of ASSAY
C8H9NO5 PROCEDURE
For tablets labeled as chewable: NLT 80% (Q) of the Sample: 500 mg of amphetamine
labeled amounts of C16H19N3O5S and C8H9NO5 is dissolved Analysis: Dissolve Sample in 50 mL of glacial acetic acid,
in 45 min. and titrate with 0.1 N perchloric acid VS. Perform a blank
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements determination (see Titrimetry 541). Each mL of 0.1 N
for Weight Variation with respect to amoxicillin and for perchloric acid is equivalent to 36.85 mg of (C9H13N)2
Content Uniformity with respect to clavulanic acid. H2SO4.
SPECIFIC TESTS
WATER DETERMINATION, Method I 921 IMPURITIES
NMT 7.5% where the labeled amount of amoxicillin in each Inorganic Impurities
Tablet is 250 mg or less; RESIDUE ON IGNITION 281: NMT 0.2%
NMT 10.0% where the labeled amount of amoxicillin in Organic Impurities
each Tablet is more than 250 mg but less than or equal to PROCEDURE 1
500 mg; Buffer solution: Dissolve 2.16 g of sodium 1-
NMT 11.0% where the labeled amount of amoxicillin in octanesulfonate in 1000 mL of water. Add 1.0 mL of
each Tablet is more than 500 mg. triethylamine, and adjust with phosphoric acid to pH of
Where Tablets are labeled as chewable 2.5.
NMT 6.0% where the labeled amount of amoxicillin in each
Tablet is 125 mg or less;
230 Amphetamine / Official Monographs USP 32
Mobile phase: Acetonitrile, methanol, and Buffer solution water for 30 min, and filter into a small flask. To the filtrate
(37:19:144) add 3 mL of 1 N sodium hydroxide. Cool to 10 to 15, add
Diluent: 3.12 mL of phosphoric acid to 1000 mL with 1 mL of a mixture of 1 volume of benzoyl chloride and 2
water volumes of absolute ether, insert the stopper, and shake well
Standard stock solution: 0.3 mg/mL of USP for 3 min. Filter the precipitate, wash with 15 mL of cold
Dextroamphetamine Sulfate RS in Diluent water, and recrystallize twice from diluted alcohol.
Standard solution: 0.003 mg/mL of dextroamphetamine Acceptance criteria: The crystals of the benzoyl derivative
from Standard stock solution diluted with Diluent of amphetamine so obtained, after drying at 80 for 2 h,
Sample solution: 0.3 mg/mL of Amphetamine Sulfate in melt between 131 and 135.
Diluent. [NOTESonicate for 5 min and then dilute with
Diluent to volume.] ASSAY
(See Chromatography 621, System Suitability.) Standard solution: Prepare as directed under Amphetamine
Mode: LC Assay 331.
Detector: UV 215 nm Sample solution: Finely powder NLT 20 Tablets. Transfer a
Column: 4.6-mm 15-cm; 5-m packing L1 portion of the powder, equivalent to 5 mg of amphetamine
Flow rate: 1 mL/min sulfate, to a 100-mL beaker, add 2 mL of hydrochloric acid
Injection size: 50 L solution (1 in 100), swirl gently to wet the powder
System suitability thoroughly, warm on a steam bath for 1 min, with
Samples: Standard stock solution and Sample solution occasional gentle swirling, and cool. Add 3 g of purified
Suitability requirements siliceous earth, and mix until a fluffy mixture is obtained.
Resolution: NLT 1.5, between amphetamine and any Analysis: Proceed as directed under Amphetamine Assay
adjacent peak, if any, Sample solution 331.
Tailing factor: NMT 2.0, Standard stock solution Calculate the quantity, in mg, of (C9H13N)2 H2SO4 in the
Relative standard deviation: NMT 2.0%, Standard stock portion of Tablets taken:
solution
Analysis Result = 0.01 C [(AU257 AU280)/(AS257 AS280)]
Calculate the percentage of each impurity in the portion C = concentrationof USP Dextroamphetamine Sulfate
of Amphetamine Sulfate taken: RS in the Standard solution (g/mL)
Result = (rI/rS) (CS/CU) 100 PERFORMANCE TESTS
rI = peak response for each impurity from the DISSOLUTION, Procedure for a Pooled Sample 711
Sample solution Medium: Water; 500 mL
rS = peak response for amphetamine from the Apparatus 1: 100 rpm
Standard solution Time: 45 min
CS = concentration of USP Dextroamphetamine Standard solution: USP Dextroamphetamine Sulfate RS in
Sulfate RS in the Standard solution (mg/mL) Medium
CU = concentration of Amphetamine Sulfate in the Mobile phase: 1.1 g of sodium 1-heptanesulfonate in 575
Sample solution (mg/mL) mL of water. Add 25 mL of dilute glacial acetic acid (14 in
Acceptance criteria 100) and 400 mL of methanol. Adjust by the dropwise
Individual impurities: NMT 0.1% addition of glacial acetic acid to a pH of 3.3 0.1, if
Total impurities: NMT 0.5% necessary, filter, and degas the solution.
PROCEDURE 2: DEXTROAMPHETAMINE: A solution (20 mg/mL) Chromatographic system
is optically inactive (See Chromatography 621, System Suitability.)
Mode: LC
SPECIFIC TESTS Detector: UV 254 nm
LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT Column: 3.9-mm 30-cm; packing L1
1.0% of its weight. Flow rate: 1 mL/min
PACKAGING AND STORAGE: Preserve in well-closed containers. Sample: Standard solution
USP Dextroamphetamine Sulfate RS Relative standard deviation: NMT 2.0%
Analysis
Sample: Filtered portion of the solution under test
Calculate the percentage of (C9H13N)2 H2SO4 dissolved in
Amphetamine Sulfate Tablets comparison with a Standard solution having a known
(Comment on this Monograph)id=m4290=Amphetamine concentration of USP Dextroamphetamine Sulfate RS and
Sulfate Tablets=A-Monos.pdf) similarly chromatographed.
Acceptance criteria: NLT 75% (Q) is dissolved
DEFINITION UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Amphetamine Sulfate Tablets contain NLT 93.0% and NMT
107.0% of the labeled amount of (C9H13N)2 H2SO4. ADDITIONAL REQUIREMENTS
IDENTIFICATION USP REFERENCE STANDARDS 11
MELTING RANGE OR TEMPERATURE, Class I 741 USP Dextroamphetamine Sulfate RS
Sample solution: Macerate a quantity of powdered Tablets,
equivalent to 50 mg of amphetamine sulfate, with 10 mL of
USP 32 Official Monographs / Amphotericin 231
Amphotericin B a 50-mL volumetric flask, and dilute with methanol to

(Comment on this Monograph)id=m4340=Amphotericin B=A- volume. Transfer 4.0 mL of this solution to a 50-mL
Monos.pdf) volumetric flask, and dilute with methanol to volume.
[NOTEPrepare this solution fresh daily.]
Cell: 1 cm
Measurement wavelength: 304 nm and 282 nm
Blank: 16 mg/mL of dimethyl sulfoxide in methanol
Analysis: Concomitantly determine the absorbances of the
Nystatin standard solution and Amphotericin B standard
solution and the Sample solution against the Blank. Calculate
C47H73NO17 924.08 the percentage of amphotericin A taken:
Amphotericin B;
14,39-Dioxabicyclo[33.3.1]nonatriaconta-19,21,23,25,27,29,31- Result = 25 WN[(AB282 AU304) (AB304 AU282)]/[(AB282 AN304)
heptaene-36-carboxylic acid, 33-[(3-amino-3,6-dideoxy--D- (AB304 AN282)] WU
mannopyranosyl)oxy]-1,3,5,6,9,11,17,37-
octahydroxy-15,16,18-trimethyl-13-oxo-, WN = weight of USP Nystatin RS taken (mg)
(1R,3S,5R,6R,9R,11R,15S,16R,17R,18S, AB282 = absorbance of the Amphotericin B standard
19E,21E,23E,25E,27E,29E,31E,33R,35S,36R,37S)-; solution at 282 nm
[1R- AU304 = absorbance of the Sample solution at 304 nm
AB304 = absorbance of the Amphotericin B standard
(1R*,3S*,5R*,6R*,9R*,11R*,15S*,16R*,17R*,18S*,19E,21E,23E,25E,27E,29E,31E,33R*,35S*,36R*,37S*)]-33-
[(3-Amino-3,6-dideoxy--D- solution at 304 nm
mannopyranosyl)oxy]-1,3,5,6,9,11,17,37- AU282 = absorbance of the Sample solution at 282 nm
octahydroxy-15,16,18-trimethyl-13-oxo-14,39-dioxabicyclo AN304 = absorbance of the Nystatin standard solution at
[33.3.1]nonatriaconta-19,21,23,25,27,29,31-heptaene-36- 304 nm
carboxylic acid [1397-89-3]. AN282 = absorbance of the Nystatin standard solution at
282 nm
DEFINITION WU = weight of the Amphotericin B taken (mg)
Amphotericin B has a potency of NLT 750 g of C47H73NO17/ Acceptance criteria: NMT 5%
mg, calculated on the dried basis. [NOTEAmphotericin B intended for use in preparing
dermatological creams, lotions, ointments, oral
IDENTIFICATION suspensions, and capsules contains NMT 15% of
ULTRAVIOLET ABSORPTION 197U amphotericin A, calculated on the dried basis.]
Analytical wavelength I: 240320 nm
Solution I: Prepare as directed for Sample solution in the SPECIFIC TESTS
Limit of amphotericin A, and compare its absorbance to that LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered
of the Amphotericin B standard solution. An extra peak may bottle in vacuum at a pressure not exceeding 5 mm of
occur at 304 nm in the spectrum of this solution. mercury at 60 for 3 h: it loses NMT 5.0% of its weight.
Analytical wavelength II: 320400 nm
Solution II: Prepare as directed for Sample solution in the ADDITIONAL REQUIREMENTS
Limit of amphotericin A, and then dilute with 9 volumes of PACKAGING AND STORAGE: Preserve in tight, light-resistant
methanol. Compare its absorbance to that of a similar containers, and store in a cold place.
dilution of the Amphotericin B standard solution. LABELING: Label it to state whether it is intended for use in
preparing dermatological and oral dosage forms or
ASSAY parenteral dosage forms.
PROCEDURE USP REFERENCE STANDARDS 11
Proceed with Amphotericin B as directed under Antibiotics USP Amphotericin B RS
Microbial Assays 81. USP Nystatin RS
Acceptance criteria: NLT 750 g of C47H73NO17 per mg
IMPURITIES
Inorganic Impurities Amphotericin B Cream
RESIDUE ON IGNITION 281: NMT 0.5%, the charred residue (Comment on this Monograph)id=m4350=Amphotericin B
being moistened with 2 mL of nitric acid and 5 drops of Cream=A-Monos.pdf)
sulfuric acid
[NOTEAmphotericin B intended for use in preparing DEFINITION
dermatological creams, lotions, ointments, oral suspensions, Amphotericin B Cream contains NLT 90.0% and NMT 125.0%
and capsules, yields NMT 3.0%.] of the labeled amount of Amphotericin B.
Organic Impurities
PROCEDURE: LIMIT OF AMPHOTERICIN A ASSAY
Sample solution: Dissolve 50 mg of Amphotericin B in PROCEDURE
10.0 mL of dimethyl sulfoxide in a 50-mL volumetric flask, Sample stock solution: Blend a suitable portion of Cream
and dilute with methanol to volume. Transfer 4.0 mL of in a high-speed blender with a sufficient volume of dimethyl
this solution to a 50-mL volumetric flask, and dilute with sulfoxide to give a convenient concentration. Quantitatively
methanol to volume. dilute a volume of this solution with dimethyl sulfoxide to
Nystatin standard solution: Dissolve 20 mg of USP obtain a concentration of 20 g/mL of amphotericin B.
Nystatin RS in 40.0 mL of dimethyl sulfoxide in a 200-mL Sample solution: Quantitatively dilute a volume of Sample
volumetric flask, and dilute with methanol to volume. stock solution with Buffer No. 10 to obtain a Test Dilution
Transfer 4.0 mL of this solution to a 50-mL volumetric flask, having a concentration assumed to be equal to the median
and dilute with methanol to volume. dose level of the Standard.
Amphotericin B standard solution: Dissolve 50 mg of Analysis: Proceed as directed for amphotericin B under
USP Amphotericin B RS in 10.0 mL of dimethyl sulfoxide in AntibioticsMicrobial Assays 81.
232 Amphotericin / Official Monographs USP 32
Acceptance criteria: 90.0%125.0% the solution should be protected from light during
administration.
PERFORMANCE TESTS USP REFERENCE STANDARDS 11
MINIMUM FILL 755: It meets the requirements. USP Amphotericin B RS
USP Endotoxin RS
PACKAGING AND STORAGE: Preserve in collapsible tubes or
other well-closed containers.
USP REFERENCE STANDARDS 11 Amphotericin B Lotion
USP Amphotericin B RS (Comment on this Monograph)id=m4370=Amphotericin B
Lotion=A-Monos.pdf)
Amphotericin B for Injection DEFINITION

Amphotericin B Lotion contains NLT 90.0% and NMT 125.0%
(Comment on this Monograph)id=m4360=Amphotericin B for of the labeled amount of amphotericin B.
Injection=A-Monos.pdf)
ASSAY
DEFINITION PROCEDURE
Amphotericin B for Injection is a sterile complex of Sample stock solution: Quantitatively dissolve a suitable
Amphotericin B, deoxycholate sodium, and one or more volume of Lotion in sufficient dimethyl sulfoxide to give a
suitable buffers. It contains NLT 90.0% and NMT 120.0% of convenient concentration. Quantitatively dilute an accurately
the labeled amount of C47H73NO17. measured volume of this solution with dimethyl sulfoxide to
obtain a concentration of 20 g/mL of amphotericin B.
ASSAY Sample solution: Quantitatively dilute an accurately
PROCEDURE measured volume of the Sample stock solution with Buffer
Sample solution A (where it is packaged as a single-dose No. 10 to obtain a Test Dilution having a concentration
container): Constitute Amphotericin B for Injection as assumed to be equal to the median dose level of the
directed in the labeling. Withdraw all of the withdrawable Standard.
contents, using a suitable hypodermic needle and syringe, Procedure: Proceed as directed for amphotericin B under
and dilute quantitatively and stepwise with dimethyl AntibioticsMicrobial Assays 81.
sulfoxide to obtain a solution containing 20 g/mL of Acceptance criteria: 90.0%125.0%
amphotericin B.
Sample solution B (where the labeling states the quantity of PERFORMANCE TESTS
amphotericin B in a given volume of constituted solution): MINIMUM FILL 755: Meets the requirements
Constitute Amphotericin B for Injection as directed in the
labeling. Withdraw a volume of the resultant solution, using SPECIFIC TESTS
a suitable hypodermic needle and syringe, and dilute PH 791: 5.07.0
quantitatively and stepwise with dimethyl sulfoxide to obtain
a solution containing 20 g/mL of amphotericin B. ADDITIONAL REQUIREMENTS
Analysis: Proceed as directed for amphotericin B under PACKAGING AND STORAGE: Preserve in well-closed containers.
AntibioticsMicrobial Assays 81, using a volume of Sample USP REFERENCE STANDARDS 11
solution diluted quantitatively and stepwise with Buffer No. USP Amphotericin B RS
10 to obtain a Test Dilution having a concentration assumed
to be equal to the median dose level of the Standard.
Acceptance criteria: 90.0%120.0% Amphotericin B Ointment
SPECIFIC TESTS (Comment on this Monograph)id=m4380=Amphotericin B
STERILITY TESTS 71: It meets the requirements when tested Ointment=A-Monos.pdf)
as directed in Test for Sterility of the Product to be Examined,
Membrane Filtration, 50 mg from each container being DEFINITION
tested. Amphotericin B Ointment is Amphotericin B in a suitable
PH 791: 7.28.0 ointment base. It contains NLT 90.0% and NMT 125.0% of
Sample solution: 10 mg/mL of amphotericin B the labeled amount of amphotericin B.
LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered
bottle in a vacuum at a pressure not exceeding 5 mm of ASSAY
mercury at 60 for 3 h: it loses NMT 8.0% of its weight. PROCEDURE
OTHER REQUIREMENTS: It meets the requirements for Sample stock solution: Mix a portion of Ointment,
Uniformity of Dosage Units 905 and for InjectionsLabels equivalent to 30 mg of amphotericin B, with 10.0 mL of
and Labeling 1. ether in a suitable glass-stoppered conical flask and allow to
BACTERIAL ENDOTOXINS TEST 85: It contains NMT 5.0 USP stand, with intermittent shaking, for 1 h. Add 20.0 mL of
Endotoxin Units/mg of amphotericin B. For products used or dimethyl sulfoxide and shake by mechanical means for 10
labeled for intrathecal injection, it contains NMT 0.9 USP min. Dilute quantitatively and stepwise with dimethyl
Endotoxin Unit/mg of amphotericin B. sulfoxide to a concentration of 20 g/mL.
Sample solution: Quantitatively dilute an accurately
ADDITIONAL REQUIREMENTS measured volume of Sample stock solution with Buffer No. 10
PACKAGING AND STORAGE: Preserve in Containers for Sterile to obtain a Test Dilution having a concentration assumed to
Solids as described under Injections 1, in a refrigerator and be equal to the median dose level of the Standard.
protected from light. Analysis: Proceed as directed for amphotericin B under
LABELING: Label it to indicate that it is intended for use by AntibioticsMicrobial Assays 81.
intravenous infusion to hospitalized patients only, and that
USP 32 Official Monographs / Ampicillin 233
Acceptance criteria: 90.0%125.0% Pre-column: 4-mm 5-cm; 5- to 10-m packing L1

Flow rate: 2 mL/min
MINIMUM FILL 755: Meets the requirements System suitability
WATER DETERMINATION, Method I 921: NMT 1.0%, 20 mL Samples: System suitability solution and Standard solution
of a mixture of toluene and methanol (7:3) being used in [NOTEThe relative retention times for ampicillin and
place of methanol in the titration vessel caffeine are 0.5 and 1.0, respectively, System suitability
solution.]
PACKAGING AND STORAGE: Preserve in collapsible tubes, or Resolution: NLT 2.0 between the caffeine and ampicillin
other well-closed containers. peaks, System suitability solution.
USP REFERENCE STANDARDS 11 Tailing factor: NMT 1.4 , Standard solution
USP Amphotericin B RS Capacity factor, k: NMT 2.5, Standard solution
Relative standard deviation: NMT 2.0% from replicate
injections, Standard solution
Ampicillin Analysis
(Comment on this Monograph)id=m4430=Ampicillin=A- Calculate the quantity, in g, of C13H19NO4S in each mg of
Monos.pdf) Ampicillin taken:
Result = (rU/rS (CS P/W) 100
rS = peak response of the Standard solution
CS = concentration of USP Ampicillin RS in the
W = weight, in mg, of Ampicillin taken for the Sample
C16H19N3O4S (anhydrous) 349.41 solution (mg/mL)
4-Thia-1-azabicyclo[3.2.0]heptane-2 carboxylic acid, [6- P = stated content of USP Ampicillin RS (g/ml)
(aminophenylacetyl)amino]-3,3-dimethyl-7-oxo-, [2S- Acceptance criteria: NLT 900 g and NMT 1050 g
[2,5,6(S*)]]-;
(2S,5R,6R)-6-[(R)-2-Amino-2-phenylacetamido]-3,3-dimethyl-7- SPECIFIC TESTS
oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid STERILITY TESTS 71: Where the label states that Ampicillin is
[69-53-4]. sterile, it meets the requirements when tested as directed for
Trihydrate 403.46 Test for Sterility of the Product to be Examined, Membrane
[7177-48-2]. Filtration, except to dissolve 6 g in 800 mL of Fluid D
containing sufficient sterile penicillinase to inactivate the
DEFINITION ampicillin and to swirl the vessel until solution is complete
Ampicillin is anhydrous or contains three molecules of water of before filtering.
hydration. It contains NLT 900 g and NMT 1050 g of CRYSTALLINITY 695: Meets the requirements
C16H19N3O4S per mg, calculated on the dried basis. PH 791: 3.56.0
IDENTIFICATION Sample solution: 10 mg/mL
INFRARED ABSORPTION 197K: except that where the DIMETHYLANILINE 223: Meets the requirements
specimen under test is the trihydrate, both it and the USP WATER DETERMINATION, Method I 921: NMT 2.0% where
Ampicillin Trihydrate RS are undried. it is labeled as Ampicillin (anhydrous); between 12.0% and
15.0% where it is labeled as Ampicillin (trihydrate)
ASSAY BACTERIAL ENDOTOXINS TEST 85: Where the label states that
PROCEDURE Ampicillin is sterile or must be subjected to further
Mobile phase: Acetonitrile, water, 1 M monobasic processing during the preparation of injectable dosage
potassium phosphate, and 1 N acetic acid (80:909:10:1) forms, it contains NMT 0.15 USP Endotoxin Unit/mg of
Diluent: Water, 1 M monobasic potassium phosphate, and ampicillin.
1 N acetic acid (989:10:1)
Standard solution: 1 mg/mL of USP Ampicillin RS in ADDITIONAL REQUIREMENTS
Diluent. [NOTEDissolve the sample by shaking and PACKAGING AND STORAGE: Preserve in tight containers.
sonication, if necessary, to dissolve. Use this solution LABELING: Label to indicate whether it is anhydrous or is the
promptly after preparation.] trihydrate. Where the quantity of ampicillin is indicated in
System suitability solution: 0.12 mg/mL of caffeine in the labeling of any preparation containing Ampicillin, this
Standard solution shall be understood to be in terms of anhydrous ampicillin
Sample solution: Equivalent to 1 mg/mL of anhydrous (C16H19N3O4S). Where it is intended for use in preparing
ampicillin from Diluent. [NOTEAdd a sufficient quantity of injectable dosage forms, the label states that it is the
Diluent, shake and sonicate, if necessary, to dissolve and trihydrate and that it is sterile or must be subjected to
then dilute to volume. Use this solution promptly after further processing during the preparation of injectable
preparation.] dosage forms.
Chromatographic system USP REFERENCE STANDARDS 11
(See Chromatography 621, System Suitability.) USP Ampicillin RS
Mode: LC USP Ampicillin Trihydrate RS
Detector: UV 254 nm USP Endotoxin RS
Column: 4-mm 30-cm analytical column containing 5-
to 10-m packing L1
234 Ampicillin / Official Monographs USP 32
Ampicillin Boluses Acceptance criteria: 90.0%120.0%

(Comment on this Monograph)id=m4433=Ampicillin Boluses=A- PERFORMANCE TESTS
Monos.pdf) UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Ampicillin Boluses contain an amount of ampicillin (as the LOSS ON DRYING 731: NMT 5.0%
trihydrate) equivalent to NLT 90.0% and NMT 120.0% of the
labeled amount of ampicillin (C16H19N3O4S). ADDITIONAL REQUIREMENTS
IDENTIFICATION LABELING: Label the Boluses to indicate that they are for
PROCEDURE veterinary use only.
Standard solution: 5 mg/mL of USP Ampicillin RS in USP REFERENCE STANDARDS 11
acetone and 0.1 N hydrochloric acid (4:1) USP Ampicillin RS
Sample solution: Equivalent of 10 mg/mL of ampicillin,
from 1 or more powdered Boluses, in acetone and 0.1 N
hydrochloric acid (4:1)
Chromatographic system Ampicillin Capsules
(See Chromatography 621, Thin-Layer Chromatography.) (Comment on this Monograph)id=m4440=Ampicillin
Mode: TLC Capsules=A-Monos.pdf)
mixture DEFINITION
Application volume: 2 L Ampicillin Capsules contain an amount of ampicillin (anhydrous
Developing solvent system: Acetone, toluene, glacial or as the trihydrate) equivalent to NLT 90.0% and NMT
acetic acid, and water (26:4:1:4) 120.0% of the labeled amount of ampicillin (C16H19N3O4S).
Spray reagent: 3 mg/mL of ninhydrin in alcohol
Analysis IDENTIFICATION
Samples: Standard solution and Sample solution THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
When the solvent front has moved about three-fourths of Standard solution: 5 mg/mL of USP Ampicillin RS in a
the length of the plate, remove the plate from the mixture of acetone and 0.1 N hydrochloric acid (4:1)
chamber, mark the solvent front, and allow to air-dry. Sample solution: 5 mg/mL of ampicillin, from Powdered
Locate the spots on the plate by spraying lightly with Capsules, in a mixture of acetone and 0.1 N hydrochloric
Spray reagent and dry at 90 for 15 min. acid (4:1)
Acceptance criteria: The RF value of the principal spot of Chromatographic system
the Sample solution corresponds to that of the Standard (See Chromatography 621, Thin-Layer Chromatography.)
solution. Mode: TLC
ASSAY mixture
PROCEDURE Application volume: 2 L
Standard solution: Prepare as directed for Standard Developing solvent system: Acetone, toluene, glacial
Preparation under Iodometric AssayAntibiotics 425, using acetic acid, and water (26:4:1:4)
USP Ampicillin RS. Spray reagent: 3 mg/mL of ninhydrin in alcohol
Sample solution: Place NLT 5 Boluses in a high-speed glass Analysis
blender jar containing a volume of water, and blend for 4 Samples: Standard solution and Sample solution
1 min. Dilute an accurately measured volume of this stock Apply the Sample solution and the Standard solution and
solution quantitatively and stepwise with water to obtain a develop the chromatogram. When the solvent front has
Sample solution containing 1.25 mg/mL of ampicillin. moved about three-fourths of the length of the plate,
Analysis remove the plate from the chamber, mark the solvent
Samples: Standard solution and Sample solution front, and allow to air-dry. Locate the spots on the plate
Proceed as directed for Procedure under Iodometric Assay by spraying lightly with Spray reagent and dry at 90 for
Antibiotics 425. 15 min.
Calculate as A the quantity, in mg, of C16H19N3O4S in each Acceptance criteria: The RF value of the principal spot of
Bolus taken: the Sample solution corresponds to that of the Standard
solution.
A = (T/D)(F/2000)(B I)
ASSAY
T = labeled quantity of ampicillin in each Bolus (mg) PROCEDURE
D = concentration of ampicillin in the Sample solution Standard solution: Prepare as directed for Standard
on the basis of the labeled quantity in each Preparation under Iodometric AssayAntibiotics 425, using
Bolus and the extent of dilution (mg/mL) USP Ampicillin RS.
Calculate the quantity, in percentage, of C16H19N3O4S in Sample solution: Place NLT 5 Capsules in a high-speed
each Bolus taken: glass blender jar containing an accurately measured volume
of water, and blend for 4 1 min. Dilute an accurately
Result = (A/L) 100 measured volume of this stock solution quantitatively and
stepwise with water to obtain a Sample solution containing
A = as defined above 1.25 mg/mL of ampicillin.
L = label claim (mg)
Analysis: Proceed as directed for Procedure under Iodometric Constitute Ampicillin for Injection in a volume of Diluent,
AssayAntibiotics 425. corresponding to the volume of solvent specified in the
Calculate the quantity, in mg, of C16H19N3O4S in each labeling. Withdraw all of the withdrawable contents, using
Capsule taken: a suitable hypodermic needle and syringe, and
quantitatively dilute with Diluent. [NOTEUse this solution
Result = (T/D) (F/2000) (B I) promptly after preparation.]
Sample solution B (where the label states the quantity of
T = labeled quantity, in mg, of ampicillin in each ampicillin in a given volume of constituted solution): 1
Capsule mg/mL of ampicillin in Diluent
D = concentration of ampicillin in the Sample solution Constitute 1 container of Ampicillin for Injection in a volume
on the basis of the labeled quantity in each of Diluent, corresponding to the volume of solvent specified
Capsule and the extent of dilution (mg/mL) in the labeling. Quantitatively dilute an accurately
Acceptance criteria: 90.0%120.0% measured portion of the constituted solution with Diluent.
[NOTEUse this solution promptly after preparation.]
PERFORMANCE TESTS Chromatographic system
DISSOLUTION, Procedure for a Pooled Sample 711 (See Chromatography 621, System Suitability.)
Medium: Water; 900 mL Mode: LC
Apparatus 1: 100 rpm Detector: UV 254 nm
Time: 45 min Column: 4-mm 30 cm analytical column containing 5-
Standard solution: L/900 mg/mL of USP Ampicillin RS in to 10-m packing L1
water, L being the labeled amount, in mg, of Precolumn: 4-mm 5-cm; 5- to 10-m packing L1
ampicillin/Capsule Flow rate: 2 mL/min
Analysis: Proceed as directed for Procedure in Automated Injection size: 20 L
Methods of Analysis 16, AntibioticsHydroxylamine Assay, System suitability
using a filtered portion of the solution under test as the Samples: System suitability solution and Standard solution
Sample solution. [NOTEThe relative retention times for ampicillin and
Calculate the quantity, in mg, of C16H19N3O4S dissolved: caffeine are 0.5 and 1.0, respectively, System suitabilty
0.9 C P (AU/AS) solution. ]
Tolerances: NLT 75% (Q) of the labeled amount of Resolution: NLT 2.0 between the caffeine and the
C16H19N3O4S is dissolved. ampicillin, System suitabilty solution.
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements System suitability solution: 0.12 mg/mL of caffeine in
Standard solution
SPECIFIC TESTS Tailing factor: NMT 1.4, Standard solution
WATER DETERMINATION, Method I 921: NMT 4.0% where Capacity factor: NMT 2.5, Standard solution
the Capsules contain anhydrous ampicillin, or between Relative standard deviation: NMT 2.0%, Standard
10.0% and 15.0% where the Capsules contain ampicillin solution
trihydrate. Analysis
ADDITIONAL REQUIREMENTS Calculate the quantity, in mg, of ampicillin (C16H19N3O4S) in
PACKAGING AND STORAGE: Preserve in tight containers. the container and in the volume of constituted solution
LABELING: Label to indicate whether the ampicillin therein is taken:
in the anhydrous form or is the trihydrate.
USP REFERENCE STANDARDS 11 Result = (rU/rS) (CS P/1000) (L/D)
USP Ampicillin RS
CS = concentration of USP Ampicillin RS in the
Ampicillin for Injection Standard solution (mg/mL)
(Comment on this Monograph)id=m4443=Ampicillin for D = concentration, in mg of ampicillin
Injection=A-Monos.pdf) (C16H19N3O4S)/mL, of Sample solution A or
Sample solution B, based on the labeled
DEFINITION quantity in the container or in the portion of
Ampicillin for Injection contains an amount of Ampicillin constituted solution taken, respectively, and the
Sodium equivalent to NLT 90.0% and NMT 115.0% of the extent of dilution
labeled amount of ampicillin (C16H19N3O4S). L = labeled quantity of ampicillin (C16H19 N3O4S) in
the container, or in the volume of constituted
ASSAY solution taken (mg)
PROCEDURE P = stated content of USP Ampicillin RS (g/mg)
Mobile phase: Acetonitrile, water, 1 M monobasic [NOTEWhere the test for Uniformity of dosage units has
potassium phosphate, and 1 N acetic acid (80:909:10:1) been performed using the Procedure for content uniformity,
Diluent: Water, 1 M monobasic potassium phosphate, and use the average of these determinations as the Assay
1 N acetic acid (989:10:1) value.]
Standard solution: 1 mg/mL of USP Ampicillin RS in
Diluent Acceptance criteria: 90.0%115.0%
[NOTEDissolve the Standard by shaking and sonication, if PERFORMANCE TESTS
necessary, to dissolve. Use this solution promptly after UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements.
preparation.] Procedure for content uniformity: Perform the assay on
System suitability solution: 0.12 mg/mL of caffeine in individual containers using Sample solution A or Sample
Standard solution solution B, or both, as appropriate.
Sample solution A (where it is represented as being in a
single-dose container): 1 mg/mL of ampicillin in Diluent.
SPECIFIC TESTS Analysis: Proceed as directed under Iodometric Assay

OTHER REQUIREMENTS: It meets the requirements of the test Antibiotics 425.
for Identification under Ampicillin Sodium. It also meets the Calculate the quantity, in mg, of C16H19N3O4S in the portion
requirements under Injections 1, Labeling. of Soluble Powder taken:
CRYSTALLINITY 695: Meets the requirements.
[NOTEAmpicillin Sodium in the freeze-dried form is exempt (F/20)(B I)
from this requirement.]
PH 791: 8.010.0, in a solution containing 10.0 mg/mL of Terms as defined underIodometric AssayAntibiotics 425
ampicillin Acceptance criteria: 90.0%120.0%
Sample solution: 10.0 mg/mL
WATER DETERMINATION, Method I 921: NMT 2.0% SPECIFIC TESTS
PARTICULATE MATTER IN INJECTIONS 788: Meets the PH 791: 3.56.0
requirements for small-volume injections. Sample solution: equivalent of 20 mg/mL of ampicillin
STERILITY TESTS 71: Meets the requirements. WATER DETERMINATION, Method I 921: NMT 5.0%
BACTERIAL ENDOTOXINS TEST 85: NMT 0.15 USP Endotoxin ADDITIONAL REQUIREMENTS
Unit/mg of ampicillin PACKAGING AND STORAGE: Preserve in tight containers.
CONSTITUTED SOLUTION: At the time of use, it meets the LABELING: Label it to indicate that it is for veterinary use
requirements for InjectionsConstituted Solutions 1. only.
PACKAGING AND STORAGE: Preserve in Containers for Sterile USP Ampicillin RS
Solids as described under Injections 1. Protect the
constituted solution from freezing.
USP REFERENCE STANDARDS 11 Ampicillin for Injectable Suspension
USP Ampicillin RS
USP Ampicillin Sodium RS (Comment on this Monograph)id=m4474=Ampicillin for
USP Endotoxin RS Injectable Suspension=A-Monos.pdf)
DEFINITION
Ampicillin for Injectable Suspension is a dry mixture of
Ampicillin Soluble Powder ampicillin trihydrate and one or more suitable buffers,
preservatives, stabilizers, and suspending agents. It contains
(Comment on this Monograph)id=m4448=Ampicillin Soluble the equivalent of NLT 90.0% and NMT 120.0% of the labeled
Powder=A-Monos.pdf) amount of ampicillin (C16H19N3O4S).
DEFINITION IDENTIFICATION
Ampicillin Soluble Powder is a dry mixture of Ampicillin (as the THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
trihydrate) and one or more suitable diluents and stabilizing Standard solution: 5 mg/mL of USP Ampicillin RS in
agents. It contains NLT 90.0% and NMT 120.0% of the acetone and 0.1 N hydrochloric acid (4:1)
labeled amount of ampicillin (C16H19N3O4S). Sample solution: 5 mg/mL of ampicillin in acetone and 0.1
IDENTIFICATION N hydrochloric acid (4:1)
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Chromatographic system
Standard solution: 5 mg/mL of USP Ampicillin RS in (See Chromatography 621, Thin-Layer Chromatography.)
acetone and 0.1 N hydrochloric acid (4:1) Mode: TLC
Sample solution: 10 mg/mL of ampicillin from Soluble Adsorbent: 0.25-mm layer of chromatographic silica gel
Powder in acetone and 0.1 N hydrochloric acid (4:1) mixture
Chromatographic system Application volume: 2 L
(See Chromatography 621, Thin-Layer Chromatography.) Developing solvent system: Acetone, toluene, glacial
Adsorbent: 0.25-mm layer of chromatographic silica gel acetic acid, and water (26:4:1:4)
mixture Spray reagent: 3 mg/mL of ninhydrin in alcohol
Application volume: 2 L Analysis
Developing solvent system: Acetone, toluene, glacial Samples: Sample solution and Standard solution
acetic acid, and water (26:4:1:4) When the solvent front has moved three-fourths of the
Spray reagent: 3 mg/mL of ninhydrin in alcohol length of the plate, remove the plate from the chamber,
Analysis mark the solvent front, and allow to air-dry. Locate the
Samples: Sample solution and Standard solution spots on the plate by spraying lightly with Spray reagent,
When the solvent front has moved three-fourths of the and dry at 90 for 15 min.
length of the plate, remove the plate from the chamber, Acceptance criteria: The RF value of the principal spot from
mark the solvent front, and allow to air-dry. Locate the the Sample solution corresponds to that from the Standard
spots on the plate by spraying lightly with Spray reagent solution.
and dry at 90 for 15 min. ASSAY
Acceptance criteria: The RF value of the principal spot PROCEDURE
obtained from the solution under test corresponds to that Phosphate buffer solution: 136 mg/mL of monobasic
obtained from the Standard solution. potassium phosphate
ASSAY Mobile phase: Acetonitrile, glacial acetic acid, water, and
PROCEDURE Phosphate buffer solution (90:1:900:10). Pass through a 0.45-
Standard solution: Prepare as directed under Iodometric m nylon filter, and degas.
AssayAntibiotics 425, using USP Ampicillin RS. Standard solution: 0.5 mg/mL of USP Ampicillin RS.
Sample solution: Transfer a quantity of Soluble Powder, Dissolve with sonication. Pass through a 0.45-m PTFE filter,
equivalent to 125 mg of ampicillin, to a 100-mL volumetric discarding the first 3 mL of the filtrate.
flask, dissolve in and dilute with water to volume.
Caffeine solution: 30 mg of caffeine in a 50-mL volumetric ADDITIONAL REQUIREMENTS

flask. Add 25 mL of water, sonicate to dissolve, and dilute PACKAGING AND STORAGE: Preserve in Containers for Sterile
with water to volume. Pass through a 0.45-m PTFE filter, Solids as described under Injections 1.
discarding the first 3 mL of the filtrate. USP REFERENCE STANDARDS 11
System suitability solution: Caffeine solution and Standard USP Ampicillin RS
solution (1:9) USP Endotoxin RS
Sample solution: Dilute a volume of Ampicillin for
Injectable Suspension, constituted as directed in the
labeling, with water to obtain a solution containing 0.5
mg/mL of ampicillin. Pass through a 0.45-m PTFE filter, Ampicillin for Oral Suspension
discarding the first 3 mL of the filtrate. (Comment on this Monograph)id=m4480=Ampicillin for Oral
Chromatographic system Suspension=A-Monos.pdf)
Mode: LC DEFINITION
Detector: UV 254 nm Ampicillin for Oral Suspension contains an amount of Ampicillin
Column: 3.9-mm 30-cm; 10-m packing L1 (anhydrous or as the trihydrate) equivalent to NLT 90.0% and
Temperature: 40 NMT 120.0% of the labeled amount of C16H19N3O4S, when
Flow rate: 2 mL/min constituted as directed. It contains one or more suitable
Injection size: 20 L buffers, colors, flavors, preservatives, and sweetening
System suitability ingredients.
Order of elution: Ampicillin followed by caffeine THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
Resolution: Greater than 2 between ampicillin and Standard solution: 5 mg/mL of USP Ampicillin RS in a
caffeine, System suitability solution mixture of acetone and 0.1 N hydrochloric acid (4:1)
Column efficiency: NLT 2000 theoretical plates for the Sample solution: 5 mg/mL of ampicillin in a mixture of
ampicillin peak, Standard solution acetone and 0.1 N hydrochloric acid (4:1)
Tailing factor: NMT 1.4, Standard solution Chromatographic system
Relative standard deviation: NMT 2.0%, Standard (See Chromatography 621, Thin-Layer Chromatography.)
solution Mode: TLC
Samples: Sample solution and Standard solution mixture
Calculate the percentage of C16H19N3O4S in each mL of the Application volume: 2 L
constituted solution of Ampicillin for Injectable Suspension Developing solvent system: Acetone, toluene, glacial
taken: acetic acid, and water (26:4:1:4)
Spray reagent: 3 mg/mL of ninhydrin in alcohol
Result = (rU/rS) (CS/CU) 100 Analysis
rU = average peak response of ampicillin from the Apply the Standard solution and the Sample solution and
Sample solution develop the chromatogram. When the solvent front has
rS = average peak response of ampicillin from the moved about three-fourths of the length of the plate,
Standard solution remove the plate from the chamber, mark the solvent
CS = concentration of USP Ampicillin RS in the front, and allow to air-dry. Locate the spots on the plate
Standard solution (mg/mL) by spraying lightly with Spray reagent and dry at 90 for
CU = nominal concentration of ampicillin in the 15 min: the RF value of the principal spot of the Sample
Sample solution (mg/mL) solution corresponds to that of the Standard solution.
ASSAY
PERFORMANCE TESTS PROCEDURE
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements Standard solution: Prepare as directed for Standard solution
under Iodometric AssayAntibiotics 425, using USP
SPECIFIC TESTS Ampicillin RS.
STERILITY TESTS 71: It meets the requirements when tested Sample solution: Dilute an accurately measured volume of
as directed for Antibiotic Solids, Bulks, and Blends in the Ampicillin for Oral Suspension, constituted as directed in the
section Test for Sterility of the Product to Be Examined, labeling, freshly mixed and free from air bubbles,
Membrane Filtration, except to use Fluid D, to which has quantitatively and stepwise with water to obtain a solution
been added sufficient sterile penicillinase to inactivate the containing 1.25 mg/mL of ampicillin.
ampicillin and to swirl the vessel until solution is complete Analysis: Proceed as directed for Iodometric Assay
before filtering. If it does not dissolve completely, proceed as Antibiotics 425, Procedure.
directed for Solids in the section Test for Sterility of the Calculate the quantity, in mg, of C16H19N3O4S in each mL of
Product to be Examined, Direct Inoculation of the Culture the constituted suspension prepared from Ampicillin for
Medium, except to use Fluid Thioglycollate Medium and Oral Suspension taken:
SoybeanCasein Digest Medium containing sufficient
penicillinase to inactivate the ampicillin in each vessel. Result = (T/D) (F/2000) (BI)
BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.15
Endotoxin Unit/mg of ampicillin. T = the labeled quantity of ampicillin in the
PH 791: 5.07.0, in the suspension constituted as directed constiuted suspension (mg/ml)
in the labeling D = concentration of ampicillin in the Sample solution
WATER DETERMINATION Method I 921: 11.4%14.0% on the basis of the labeled quantity in the
OTHER REQUIREMENTS: Meets the requirements for Labeling constiuted suspension and the extent of
under Injections 1. dilution (mg/mL)

PERFORMANCE TESTS Proceed as directed under Procedure.
DELIVERABLE VOLUME 698: Meets the requirements Calculate the quantity, in mg, of C16H19N3O4S in each Tablet
UNIFORMITY OF DOSAGE UNITS 905 taken:
For solid packaged in single-unit containers: Meets the
requirements Result = (T/D) (F/2000) (BI)
SPECIFIC TESTS T = labeled quantity of ampicillin in each Tablet (mg)
PH 791: 5.07.5, in the suspension constituted as directed D = concentration of ampicillin in the Assay
in the labeling Preparation on the basis of the labeled quantity
WATER DETERMINATION, Method I 921: NMT 2.5%, or NMT in each Tablet and the extent of dilution
5.0% if it contains ampicillin trihydrate and contains the (mg/mL)
equivalent of 100 mg/mL of ampicillin when constituted as F = conversion factor (g)
directed in the labeling B = volume of 0.01 N sodium thiosulfate consumed
in the Blank Determination (g)
ADDITIONAL REQUIREMENTS I = volume of 0.01 N sodium thiosulfate consumed
PACKAGING AND STORAGE: Preserve in tight containers. in the Inactivation and Titration (mL)
LABELING: Label to indicate whether the ampicillin therein is Acceptance criteria: 90.0%120.0%
in the anhydrous form or is the trihydrate.
USP REFERENCE STANDARDS 11 PERFORMANCE TESTS
USP Ampicillin RS DISSOLUTION, Procedure for a Pooled Sample 711
Ampicillin Tablets Time: 45 min
Standard solution: L/900 mg/mL of USP Ampicillin RS in
(Comment on this Monograph)id=m4490=Ampicillin Tablets=A- water, L being the labeled amount, in mg, of ampicillin per
Monos.pdf) Tablet
Analysis
DEFINITION Samples: Standard solution and Sample solution
Ampicillin Tablets contain an amount of Ampicillin (anhydrous Proceed as directed under Procedure in the section
form or trihydrate form) equivalent to NLT 90.0% and NMT Automated Methods of Analysis 16, Antibiotics
120.0% of the labeled amount of ampicillin (C16H19N3O4S). Hydroxylamine Assay, using a filtered portion of the
IDENTIFICATION solution under test as the Sample solution.
A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Calculate the quantity, in mg, of C16H19N3O4S dissolved:
Standard solution: 5 mg/mL of USP Ampicillin RS in a
mixture of acetone and 0.1 N hydrochloric acid (4:1) 0.9 C P (AU/AS)
Sample solution: 5 mg/mL of ampicillin, from Powdered C = concentration of USP Ampicillin RS in the
Tablets, in a mixture of acetone and 0.1 N hydrochloric acid Standard solution (mg/mL)
(4:1) P = stated content of USP Ampicillin RS (g/mg)
Chromatographic system AU = absorbance of the Sample solution
(See Chromatography 621, Thin-Layer Chromatography.) AS = absorbance of the Standard solution
Adsorbent: 0.25-mm layer of chromatographic silica gel Tolerances: NLT 75% (Q) of the labeled amount of
mixture C16H19N3O4S is dissolved
Application volume: 2 L UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Developing solvent system: Acetone, toluene, glacial
acetic acid, and water (26:4:1:4) SPECIFIC TESTS
Spray reagent: 3 mg/mL of ninhydrin in alcohol WATER DETERMINATION, Method I 921: NMT 4.0% where
Analysis nonchewable Tablets contain anhydrous Ampicillin; between
Samples: Standard solution and Sample solution 9.5% and 12.0% where nonchewable Tablets contain
Apply the Standard solution and the Sample solution and Ampicillin trihydrate; NMT 3.0% where chewable Tablets
develop the chromatogram using the Developing solvent contain anhydrous Ampicillin; NMT 5.0% where chewable
system, until the solvent front has moved about three- Tablets contain Ampicillin trihydrate; and NMT 13.0% where
fourths of the length of the plate. Locate the spots on the Tablets are labeled for veterinary use only and contain
plate by spraying lightly with the Spray reagent and dry at Ampicillin trihydrate.
90 for 15 min.
Acceptance criteria: The RF value of the principal spot from ADDITIONAL REQUIREMENTS
the Sample solution corresponds to that from the Standard PACKAGING AND STORAGE: Preserve in tight containers, and
solution. store at controlled room temperature.
LABELING: Label the Tablets to indicate whether the
ASSAY ampicillin therein is in the anhydrous form or is the
IODOMETRIC ASSAYANTIBIOTICS 425 trihydrate. Label chewable Tablets to indicate that they are
Standard solution: Prepare as directed under Standard to be chewed before swallowing. Tablets intended for
preparation, using USP Ampicillin RS. veterinary use only are so labeled.
Sample solution: Place NLT 5 Tablets in a high-speed glass USP REFERENCE STANDARDS 11
blender jar containing an accurately measured volume of USP Ampicillin RS
water, and blend for 4 1 min. Dilute an accurately
measured volume of this stock solution with water to obtain
a concentration of 1.25 mg/mL of ampicillin.
Ampicillin and Probenecid for Oral AU = absorbance of the Sample solution

Suspension AS = absorbance of the Standard solution
CS = concentration of USP Probenecid RS in the
(Comment on this Monograph)id=m4499=Ampicillin and Standard solution (mg/mL)
Probenecid for Oral Suspension=A-Monos.pdf) CU = nominal concentration of probenecid in the
DEFINITION Sample solution (mg/mL)
Ampicillin and Probenecid for Oral Suspension contains an Acceptance criteria: 90.0%110.0%
amount of Ampicillin (as the trihydrate) equivalent to NLT PERFORMANCE TESTS
90.0% and NMT 120.0% of the labeled amount of ampicillin UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
(C16H19N3O4S) and NLT 90.0% and NMT 110.0% of the for Solids in Single-Unit Containers under Content Uniformity
labeled amount of probenecid (C13H19NO4S). It contains one or with respect to ampicillin and probenecid.
more suitable colors, flavors, and suspending agents. DELIVERABLE VOLUME 698: Meets the requirements
AMPICILLIN PH 791: 5.07.5, in the suspension constituted as directed
Standard solution: Prepare as directed for Standard in the labeling
Preparation under Iodometric AssayAntibiotics 425, using WATER DETERMINATION, Method I 921: NMT 5.0%
USP Ampicillin RS.
Sample stock solution: Constitute Ampicillin and ADDITIONAL REQUIREMENTS
Probenecid for Oral Suspension as directed in the labeling, PACKAGING AND STORAGE: Preserve in tight, unit-dose
and mix. Transfer the resulting suspension to a high-speed containers.
glass blender jar containing sufficient water to make 500.0 USP REFERENCE STANDARDS 11
mL, and blend for 10 min. USP Ampicillin RS
Sample solution: Quantitatively dilute a measured volume USP Probenecid RS
of the Sample stock solution with water to obtain a Sample
solution containing nominally 1.25 mg/mL of ampicillin.
Analysis Proceed as directed for Procedure under Iodometric
AssayAntibiotics 425. Ampicillin Sodium
Calculate the quantity, in mg, of C16H19N3O4S in the portion (Comment on this Monograph)id=m4510=Ampicillin
of Ampicillin and Probenecid for Oral Suspension taken: Sodium=A-Monos.pdf)
Result = (F/CU) (VB VU) (1/VS) (100/L) C16H18N3NaO4S 371.39
4-Thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, [6-
F = equivalent of ampicillin/mL of titrant (g/mL) (aminophenylacetyl)amino]-3,3-dimethyl-7-oxo-,
CU = nominal concentration of ampicillin in the Sample monosodium salt, [2S-[2,5,6(S*)]]-;
solution (mg/mL) Monosodium D-(-)-6-(2-amino-2-phenylacetamido)-3,3-
VB = volume of Blank titer (mL) dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-
VU = volume of Sample titer (mL) carboxylate [69-52-3].
VS = volume of Sample solution used in the titration
L = label claim of ampicillin in the Ampicillin and DEFINITION
Probenecid for Oral Suspension (g/mg) Ampicillin Sodium has a potency equivalent to NLT 845 g and
Acceptance criteria: 90.0%120.0% NMT 988 g of (C16H19N3O4S) per mg, calculated on the
PROBENECID anhydrous basis.
Standard solution: 1 mg/mL of USP Probenecid RS in IDENTIFICATION
sodium carbonate solution (1 in 100) A. INFRARED ABSORPTION 197M
Sample solution: Constitute Ampicillin and Probenecid for
Oral Suspension as directed in the labeling, and mix. B. IDENTIFICATION TESTSGENERAL, Sodium 191
Quantitatively dilute the resulting suspension with the
sodium carbonate solution (1 in 100) to obtain a solution ASSAY
containing nominally 1 mg/mL of probenecid. PROCEDURE
Transfer 2.0 mL of the clear Sample solution to a 125-mL Diluent: Water, 1 M monobasic potassium phosphate, and
separator, and add 8.0 mL of 1.0 N hydrochloric acid. 1 N acetic acid (989:10:1)
Extract this solution with four 20-mL portions of Mobile phase: Acetonitrile, water, 1 M monobasic
chloroform, filtering each extract through a glass wool potassium phosphate, and 1 N acetic acid (80:909:10:1)
pledget and 6 g of chloroform-washed anhydrous sodium Standard solution: 1 mg/mL of USP Ampicillin RS in
sulfate into a 100-mL volumetric flask. Wash the pledget Diluent using shaking and sonication, if necessary, to
and the sodium sulfate with chloroform, collecting the dissolve. Use this solution promptly after preparation.
washings in the 100-mL volumetric flask, dilute with System suitability solution: 0.12 mg/mL of caffeine in
chloroform to volume, and mix. Treat 2.0 mL of the Standard solution
Standard solution in the same manner. Sample solution: [NOTEAmpicillin Sodium is hygroscopic.
Spectrometric conditions Minimize exposure to the atmosphere, and weigh
Mode: Spectrophotometry promptly.]
Analytical wavelength: 257 nm Equivalent to 1 mg/mL of anhydrous ampicillin in Diluent
Blank: Chloroform washed with sodium carbonate solution [NOTEUse this solution promptly after preparation.]
(1 in 100) Chromatographic system
Analysis (See Chromatography 621, System Suitability.)
Calculate the percentage of C13H19NO4S in the Ampicillin
and Probenecid for Oral Suspension taken:
Mode: LC RU = ratio of the response of the methylene chloride

Detector: UV 254 nm peak to that of the dioxane peak of the Sample
Column solution
Pre-column: 4-mm 5-cm; 5- to 10-m packing L1 RS = ratio of the response of the methylene chloride
Analytical column: 5- to 10-m; packing L1 peak to that of the dioxane peak of the
Flow rate: 2 mL/min Standard solution
Injection size: 20 L CS = concentration (mg/mL), of methylene chloride
System suitability in the Standard solution
Samples: System suitability solution and Standard solution CU = concentration for the Sample solution (mg/mL)
[NOTEThe relative retention times for ampicillin and Acceptance criteria: The limit is 0.2%.
caffeine are 0.5 and 1.0, respectively, System suitability DIMETHYLANILINE 223: Meets the requirement, the Internal
solution.] standard solution, Standard solution, and Sample solution
Suitability requirements being prepared as follows.
Resolution: NLT 2.0, between the caffeine and the Internal standard solution: Dissolve 75 mg of N,N-
ampicillin peaks, System suitability solution diethylaniline in 25 mL of 1 N hydrochloric acid, and dilute
Tailing factor: NMT 1.4, Standard solution quantitatively and stepwise with water to obtain a solution
Capacity factor: NMT 2.5, Standard solution containing 30 g/mL.
Relative standard deviation: NMT 2.0%, Standard Standard solution: Transfer 50.0 mg of N,N-dimethylaniline
solution to a 50-mL volumetric flask, add 25 mL of 1 N hydrochloric
Analysis acid, swirl to dissolve. Dilute with water to volume. Transfer
Samples: Standard solution and Sample solution 2.0 mL of the resulting solution to a 100-mL volumetric
Calculate the quantity, of C16H19N3O4S (g) in each mg of flask. Dilute with water to volume. To a suitable centrifuge
Ampicillin Sodium taken: tube add 1.0 mL of this solution, 1.0 mL of 1.25 N sodium
hydroxide, 1.0 mL of Internal standard solution, and 1.0 mL
Result = (rU/rS) (CS/CU) P of cyclohexane, shake vigorously for 1 min, and centrifuge.
Use the clear supernatant.
rU = peak response of the Sample solution Sample solution: Transfer 1.0 g of Ampicillin Sodium to a
rS = peak response of the Standard solution centrifuge tube, add 2 mL of 1.25 N sodium hydroxide,
CS = concentration, (mg/mL) of USP Ampicillin RS in swirl to dissolve the sample, add 1.0 mL of Internal standard
the Standard solution solution and 1.0 mL of cyclohexane, shake vigorously for 1
CU = concentration for the Sample solution min, and centrifuge. Use the clear supernatant.
(mg/mL)
P = stated content of USP Ampicillin RS (g/mg) SPECIFIC TESTS
Acceptance criteria: 845988 g per mg CRYSTALLINITY 695: Meets the requirements.
[NOTEAmpicillin Sodium in the freeze-dried form is exempt
IMPURITIES from this requirement.]
Organic Impurities PH 791: 8.010.0
PROCEDURE: LIMIT OF METHYLENE CHLORIDE Sample solution: 10.0 mg/mL
Internal standard solution: 2.1 mg/mL of dioxane in WATER DETEMINATION, Method I 921: NMT 2.0%
dimethyl sulfoxide STERILITY TESTS 71: Where the label states that Ampicillin
Standard solution: 0.33 mg/mL of methylene chloride in Sodium is sterile meets the requirements.
Internal standard solution BACTERIAL ENDOTOXINS TEST 85: Where the label states that
Sample solution: 166.7 mg/mL of Ampicillin Sodium in Ampicillin Sodium is sterile or the label states that Ampicillin
Internal standard solution Sodium must be subjected to further processing during the
Chromatographic system processing of injectable dosage forms contains NMT 0.15
(See Chromatography 621, System Suitability.) USP Endotoxin Unit/mg of ampicillin.
Mode: GC
Detector: Flame ionization ADDITIONAL REQUIREMENTS
Column: 1.8-m 4-mm glass column packed with a 10% PACKAGING AND STORAGE: Preserve in tight containers.
phase G39 on unsilanized support S1A LABELING: Where it is intended for use in preparing
Temperature injectable dosage forms, the label states that it is sterile or
Column: 65 must be subjected to further processing during the
Injection port: 100 preparation of injectable dosage forms.
Detector block: 260 USP REFERENCE STANDARDS 11
Carrier gas: Nitrogen USP Ampicillin RS
Flow rate: 60 mL/min USP Ampicillin Sodium RS
Injection size: 1 L USP Endotoxin RS
System suitability
[NOTEThe relative retention times for methylene
chloride and dioxane are 0.5 and 1.0, respectively] Ampicillin and Sulbactam for Injection
Suitability requirements (Comment on this Monograph)id=m4528=Ampicillin and
Resolution: NLT 4 Between the methylene chloride peak Sulbactam for Injection=A-Monos.pdf)
and the dioxane peak
Relative standard deviation: NMT 5% DEFINITION
Analysis Ampicillin and Sulbactam for Injection is a sterile, dry mixture of
Samples: Standard solution and Sample solution Ampicillin Sodium and Sulbactam Sodium. It contains the
Calculate the percentage of methylene chloride in the equivalent of NLT 90.0% and NMT 115.0% of the labeled
portion of Ampicillin Sodium taken: amounts of ampicillin (C16H19N3O4S) and sulbactam
(C8H11NO5S), the labeled amounts representing proportions of
Result = (RU/RS) (CS/CU) 100 ampicillin to sulbactam of 2:1. It contains NLT 563 g of
ampicillin and 280 g of sulbactam per mg, calculated on the Suitability requirements
anhydrous basis. Resolution: NLT 4.0 between ampicillin and sulbactam
alkaline degradation product, System suitability solution
IDENTIFICATION Column efficiency: NLT 3500 theoretical plates, from
The retention time of the major peaks of the Sample solution the sulbactam peak, Standard solution
correspond to those of the Standard solution, as obtained in Tailing factor: NMT 1.5, Standard solution
the Assay. Relative standard deviation: NMT 2.0%, Standard
solution
ASSAY Analysis
PROCEDURE Samples: Standard solution and the appropriate Sample
0.005 M Tetrabutylammonium hydroxide: Dilute 6.6 mL solution
of a 40% solution of tetrabutylammonium hydroxide with Calculate the percentage of C16H19N3O4S and of C8H11NO5S
water to obtain 1800 mL of solution. Adjust with 1 M in the portion of Ampicillin and Sulbactam for Injection
phosphoric acid to a pH of 5.0 0.1, and dilute with water taken:
to 2000 mL.
Mobile phase: Acetonitrile and 0.005 M Result = (rU/rS) (CS/CU) P F 100
Tetrabutylammonium hydroxide (7:33)
Standard solution: 0.6 mg/mL of ampicillin and 0.3 rU = peak area for the appropriate analyte from
mg/mL of sulbactam, obtained from USP Ampicillin RS and Sample solution A
USP Sulbactam RS in Mobile phase. [NOTEInject this rS = peak area from the Standard solution
solution promptly.] CS = concentration of the appropriate USP Reference
System suitability stock solution: Prepare a 0.3 mg/mL Standard in the Standard solution (mg/mL)
solution of USP Sulbactam RS in 0.01 N sodium hydroxide, CU = concentration of Sample solution A (mg/mL)
and allow to stand for 30 min. Adjust with phosphoric acid P = stated content of the appropriate USP Reference
to a pH of 5.0 0.1. Transfer 5 mL of the solution to a 25- Standard (g/mg)
mL volumetric flask, add 4.25 mL of acetonitrile, and dilute F = unit conversion factor; 0.001 mg/g
with 0.005 M Tetrabutylammonium hydroxide to volume. Calculate the quantities of C16H19N3O4S and of C8H11NO5S
System suitability solution: Transfer 1 mL of System withdrawn from the container, or in the volume of
suitability stock solution to a 25-mL volumetric flask, add 15 constituted solution taken:
mg of USP Ampicillin RS, and dilute with Mobile phase to
volume. [NOTEInject this solution promptly.] Result = (rU/rS) (CS/CU) P F 100
Sample solution A: Mix the contents of a container of
Ampicillin and Sulbactam for Injection. Quantitatively rU = peak area for the appropriate analyte from
dissolve a portion of the powder in the Mobile phase to Sample solution B or Sample solution C
obtain a solution having a concentration of 1 mg of the rS = peak area from the Standard solution
powder per mL. [NOTEInject this solution promptly.] CS = concentration of the appropriate USP Reference
Sample solution B (where it is represented as being in a Standard in the Standard solution (mg/mL)
single-dose container): Constitute a container of Ampicillin CU = nominal concentration of ampicillin or sulbactam
and Sulbactam for Injection with a volume of water (mg/mL)
corresponding to the volume of solvent specified in the P = stated content of the appropriate USP Reference
labeling. Withdraw the total withdrawable contents from the Standard (g/mg)
container, using a suitable hypodermic needle and syringe, F = unit conversion factor; 0.001 mg/g
and dilute quantitatively, and stepwise if necessary, with Acceptance criteria: 90.0%115.0%
Mobile phase to obtain a solution containing nominally 0.6
mg/mL of ampicillin and 0.3 mg/mL of sulbactam. [NOTE PERFORMANCE TESTS
Inject this solution promptly.] UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Sample solution C (where the label states the quantities of
ampicillin and sulbactam in a given volume of constituted SPECIFIC TESTS
solution): Constitute a container of Ampicillin and STERILITY TESTS 71: It meets the requirements when tested
Sulbactam for Injection with a volume of water as directed for Test for Sterility of the Product to be Examined,
corresponding to the volume of solvent specified in the Membrane Filtration.
labeling. Dilute a volume of the constituted solution PH 791: 8.010.0, in a solution containing 10 mg of
quantitatively, and stepwise if necessary, with the Mobile ampicillin and 5 mg of sulbactam per mL.
phase to obtain a solution containing 0.6 mg/mL of Sample solution: 10 mg/mL ampicillin and 5 mg/ml
ampicillin and nominally 0.3 mg/mL of sulbactam. [NOTE sulbactam
Inject this solution promptly.] WATER DETERMINATION, Method I 921: NMT 2.0%
Chromatographic system PARTICULATE MATTER IN INJECTIONS 788: Meets the
(See Chromatography 621, System Suitability.) requirements for small-volume injections
Mode: LC BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.17 USP
Detector: UV 230 nm Endotoxin Unit in a portion equivalent to 1 mg of a mixture
Column: 4-mm 30-cm; packing L1 of ampicillin and sulbactam (0.67 and 0.33 mg,
Flow rate: 2 mL/min respectively).
Injection size: 10 L CONSTITUTED SOLUTION: At the time of use, it meets the
System suitability requirements for InjectionsConstituted Solutions 1.
Samples: System suitability solution and Standard solution OTHER REQUIREMENTS: It meets the requirements for
[NOTEThe relative retention times for ampicillin for InjectionsLabeling 1.
sulbactam alkaline degradation product are 0.7 and 1.0,
respectively, System suitability solution; for ampicillin and ADDITIONAL REQUIREMENTS
sulbactam are 0.35 and 1.0, respectively, Standard PACKAGING AND STORAGE: Preserve as described in
solution.] InjectionsContainers for Sterile Solids 1.
USP REFERENCE STANDARDS 11 rU = amprolium peak response from the Sample

USP Ampicillin RS solution
USP Endotoxin RS rS = amprolium peak response from the Standard
USP Sulbactam RS solution
CS = concentration of USP Amprolium RS in the
CU = concentration of Amprolium in the Sample
Amprolium solution (mg/mL)
(Comment on this Monograph)id=m4540=Amprolium=A- Acceptance criteria: 97.0%101.0%
Monos.pdf)
SPECIFIC TESTS
LOSS ON DRYING 731: Dry it at a pressure not exceeding 5
mm of mercury at 100 for 3 h: it loses NMT 1.0% of its
weight.
C14H19ClN4 HCl 315.24 only.
1-[(4-Amino-2-propyl-5-pyrimidinyl)methyl]-2-methylpyridinium USP REFERENCE STANDARDS 11
chloride monohydrochloride; USP Amprolium RS
1-[(4-Amino-2-propyl-5-pyrimidinyl)methyl]-2-picolinium
chloride monohydrochloride [121-25-5].
DEFINITION Amprolium Soluble Powder
Amprolium contains NLT 97.0% and NMT 101.0% of (Comment on this Monograph)id=m4547=Amprolium Soluble
amprolium (C14H19ClN4 HCl), calculated on the dried basis. Powder=A-Monos.pdf)
IDENTIFICATION DEFINITION
A. INFRARED ABSORPTION 197K: Previously dried Amprolium Soluble Powder contains NLT 95.0% and NMT
B. ULTRAVIOLET ABSORPTION 197U 105.0% of the labeled amount of amprolium (C14H19ClN4
Analytical wavelength: 246 nm HCl).
Sample solution: 10 g/mL in 0.1 N hydrochloric acid
Acceptance criteria: Absorptivities calculated on the dried IDENTIFICATION
basis do not differ by more than 3.0%. ULTRAVIOLET ABSORPTION 197U
Sample solution: 10 g/mL (filtered) in 0.1 N hydrochloric
ASSAY acid
PROCEDURE
Diluent: Methanol, acetonitrile, and water (9:1:10) ASSAY
Mobile phase: 6 g of sodium 1-heptanesulfonate in 500 mL PROCEDURE
of water, add 12 mL of glacial acetic acid, 2.0 mL of Diluent: Methanol, acetonitrile, and water (9:1:10)
triethylamine, 450 mL of methanol, and 50 mL of Mobile phase: 6 g of sodium 1-heptanesulfonate in 500 mL
acetonitrile. Pass through a suitable filter of 0.5-m or finer of water, add 12 mL of glacial acetic acid, 2.0 mL of
porosity. triethylamine, 450 mL of methanol, and 50 mL of
System suitability solution: 0.5 mg/mL of USP Amprolium acetonitrile
RS and 0.2 mg/mL of 2-picoline in Diluent Pass through a suitable filter of 0.5-m or finer porosity.
Standard solution: 0.5 mg/mL of USP Amprolium RS in System suitability solution: 0.5 mg/mL of USP Amprolium
Diluent RS and 0.2 mg/mL of 2-picoline in Diluent
Sample solution: 0.5 mg/mL of Amprolium in Diluent Standard solution: 0.5 mg/mL of USP Amprolium RS in
Chromatographic system Diluent
(See Chromatography 621, System Suitability.) Sample solution: Nominally equivalent to 0.5 mg/mL of
Mode: LC amprolium from Soluble Powder in Diluent
Detector: UV 254 nm [NOTEFirst add a sufficient quantity of Diluent to Soluble
Column: 4.6-mm 25-cm; packing L13 Powder and sonicate for 10 min. Allow to cool to room
Flow rate: 0.6 mL/min temperature, dilute with Diluent to volume. Pass through a
Injection size: 10 L suitable filter of 0.5-m or finer porosity, and use the clear
System suitability filtrate.]
Sample: System suitability solution Chromatographic system
Suitability requirements (See Chromatography 621, System Suitability.)
Resolution: NLT 7 between amprolium and 2-picoline Mode: LC
Column efficiency: NLT 6500 theoretical plates from the Detector: UV 254 nm
amprolium peak Column: 4.6-mm 25-cm; packing L13
Tailing factor: NMT 2.3 for the analyte peak Flow rate: 0.6 mL/min
Relative standard deviation: NMT 1.0% for amprolium Injection size: 10 L
Analysis System suitability
Samples: Standard solution and Sample solution Sample: System suitability solution
Calculate the percentage of C14H19ClN4 HCl in the portion Suitability requirements
of Amprolium taken: Resolution: NLT 7 between amprolium and 2-picoline
Column efficiency: NLT 6500 theoretical plates from the
Result = (rU/rS) (CS/CU) 100 amprolium peak
USP 32 Official Monographs / Amyl 243
Tailing factor: NMT 2.3 for the analyte peak rS = amprolium peak response from the Standard
Relative standard deviation: NMT 1.0% amprolium solution
Analysis CS = concentration of USP Amprolium RS in the
Samples: Sample solution and Standard solution Standard solution (mg/mL)
Calculate the percentage of C14H19ClN4 HCl in the portion CU = nominal concentration of amprolium oral
of Soluble Powder taken: solution in the Sample solution (mg/mL)
SPECIFIC TESTS
rU = amprolium peak response obtained from the PH 791: 2.53.0
Sample solution
rS = amprolium peak response obtained from the ADDITIONAL REQUIREMENTS
Standard solution PACKAGING AND STORAGE: Preserve in tight containers,
CS = concentration of USP Amprolium RS in the protected from light. Store at a temperature between 5 and
Standard solution (mg/mL) 30, in a dry place.
CU = nominal concentration of amprolium in the LABELING: Label it to indicate that it is for veterinary use
Sample solution (mg/mL) only.
USP Amprolium RS
only. Amyl Nitrite
USP REFERENCE STANDARDS 11 (Comment on this Monograph)id=m4570=Amyl Nitrite=A-
USP Amprolium RS Monos.pdf)
C5H11NO2 117.15
Mixture of nitrous acid, 2-methylbutyl ester, and nitrous acid,
Amprolium Oral Solution 3-methylbutyl ester [8017-89-8; 110-46-3].
(Comment on this Monograph)id=m4548=Amprolium Oral DEFINITION
Solution=A-Monos.pdf) Amyl Nitrite is a mixture of the nitrite esters of 3-methyl-1-
butanol and 2-methyl-1-butanol. It contains NLT 85.0% and
DEFINITION NMT 103.0% of C5H11NO2.
Amprolium Oral Solution contains NLT 93.0% and NMT [CAUTIONAmyl Nitrite is very flammable. Do not use where it
107.0% of the labeled amount of amprolium (C14H19ClN4 may be ignited.]
HCl).
IDENTIFICATION
IDENTIFICATION A. The NMR spectrum recorded as directed in the Assay
ULTRAVIOLET ABSORPTION 197U exhibits, among other peaks, a doublet with a band
Solution: 10 g/mL (filtered), in 0.1 N hydrochloric acid centered at about 1 ppm and a multiplet with a band
ASSAY centered at about 4.8 ppm representing methyl protons and
PROCEDURE methylene protons alpha to the nitrite group, respectively,
Mobile phase: To 4.5 g of sodium 1-hexanesulfonate add both relative to the tetramethylsilane singlet at 0 ppm.
1500 mL of water, 400 mL of methanol, and 100 mL of B. To a few drops of it, add a mixture of 1 mL of ferrous
acetonitrile, mix, and allow to cool to room temperature. sulfate TS and 5 mL of 3 N hydrochloric acid.
Adjust with phosphoric acid to a pH of 5.1, and pass Acceptance criteria: A greenish brown color is produced.
through a filter having a 0.5-m or finer porosity. ASSAY
Standard solution: 0.5 mg/mL of USP Amprolium RS PROCEDURE
Sample solution: Equivalent to 0.48 mg/mL of amprolium, Diluent: Carbon tetrachloride
from Oral Solution diluted with water Internal standard: USP Benzyl Benzoate RS
Chromatographic system Analysis: Transfer 4 to 5 mEq of Internal standard to a
(See Chromatography 621, System Suitability.) semimicro sampling tube, add 2 to 3 mL of Diluent, apply a
Mode: LC sampling valve and septum,* thereby sealing the tube, and
Detector: UV 268 nm determine the weight of the sealed assembly. Open the
Column: 3.9-mm 30-cm; packing L11 valve, introduce about 500 L of Amyl Nitrite with a syringe,
Temperature: 45 close the valve, and determine the weight of the sealed
Flow rate: 1 mL/min assembly when it has attained constant weight. Shake the
Injection size: 10 L sampling tube and valve assembly, and transfer about 500
System suitability L of the solution to a precision NMR tube as directed for
Sample: Standard solution Nuclear Magnetic Resonance 761, Absolute Method of
Suitability requirements Quantitation. With no spinning, or with the spinning
Tailing factor: NMT 1.5 adjusted so that the spinning side bands of neither the
Relative standard deviation: NMT 2.0% substance under assay nor the Internal standard interfere
Analysis with the regions to be integrated, record as AS the average
Samples: Standard solution and Sample solution area of the Internal standard singlet appearing at about 5.3
Calculate the percentage of C14H19ClN4 HCl in each mL of ppm, representing the methylene protons of benzyl
the Oral Solution taken:
*Suitable sampling tubes, sampling valves, and septums are available,
Result = (rU/rS) (CS/CU) 100 respectively, as catalog Nos. K-749000, K-749100, and K-749102 (50
septums) or K-749101 (100 septums), from Kontes Glass Company, Vineland,
rU = amprolium peak response from the Sample NJ 08360.
solution
244 Amyl / Official Monographs USP 32
benzoate, and record as AU the average area of the multiplet ASSAY

with a band center at about 4.8 ppm, representing the PROCEDURE
alpha methylene protons of amyl nitrite, with reference to Diluent: Carbon tetrachloride
the tetramethylsilane singlet at 0 ppm. Internal standard: USP Benzyl Benzoate RS
Calculate the quantity of C5H11NO2 in the Amyl Nitrite Analysis: Remove the gauze or other covering from one or
taken, using 58.57 as the equivalent weight of amyl nitrite more Inhalant ampuls containing a total of 300 to 400 L of
(EWU) and 106.12 as that of benzyl benzoate (EWS). amyl nitrite. Weigh accurately the clean and dry intact glass
Acceptance criteria: 85.0%103.0% ampul(s), and place the weighed specimen in a freezer for
NLT 15 min. Transfer the chilled specimen to a glass-
OTHER COMPONENTS stoppered, 25-mL conical flask containing a solution of 4 to
CONTENT OF TOTAL NITRITES 5 mEq of Internal standard in 1 to 2 mL of Diluent. Break the
Sample: A portion of Amyl Nitrite ampul(s) with a glass rod, and rinse any sample or glass
Chromatographic system fragments adhering to the glass rod with 1 mL of Diluent
(See Chromatography 621, System Suitability.) into the main Assay solution. Insert the stopper in the flask
Mode: GC immediately and proceed as directed for Nuclear Magnetic
Detector: Thermal conductivity Resonance 761, Absolute Method of Quantitation, beginning
Column: Under typical conditions, the instrument contains with When dissolution has been completed. With no
a 3-mm 2-m column packed with a methyl polysiloxane spinning, or with the spinning adjusted so that the spinning
oil, 25% by weight on suitable calcined diatomite. side bands of neither the substance under Assay nor the
Column temperature: 80 Internal standard interfere with the regions to be integrated,
Injection port and detector block temperature: record as AS the average area of the Internal standard singlet
Maintained at about 10 above the temperature of the appearing at about 5.3 ppm, representing the methylene
column protons of benzyl benzoate, and record as AU the average
Carrier gas: Helium area of the multiplet with a band center at about 4.8 ppm,
Flow rate: 60 mL/min representing the alpha methylene protons of amyl nitrite,
Injection size: NMT 2 L with reference to the tetramethylsilane singlet at 0 ppm.
Analysis: From the area under the curve, calculate the Calculate the quantity of C5H11NO2 in the Inhalant taken,
percentage (a/a) of total nitrites, represented by the area using 58.57 as the equivalent weight of amyl nitrite (EWU)
under the main peak of the chromatogram, in the Amyl and 106.12 as that of benzyl benzoate (EWS). Rinse the
Nitrite taken. flask containing the assay preparation with three 5-mL
Acceptance criteria: NLT 97.0% portions of ether, decanting each rinsing carefully to avoid
loss of glass fragments, and evaporate any remaining ether
IMPURITIES with the aid of a current of dry air. Transfer the dry glass
Inorganic Impurities fragments to a tared watch glass, weigh, and subtract the
LIMIT OF NONVOLATILE RESIDUE weight of the glass fragments from that of the intact
Sample: 10 mL ampul(s) to obtain the weight of the Inhalant taken.
Analysis: Evaporate at room temperature in a tared Acceptance criteria: 80.0%105.0%
evaporating dish, in a well-ventilated hood, and dry the
residue at 105 for 1 h. OTHER COMPONENTS
Acceptance criteria: The weight of the residue does not CONTENT OF TOTAL NITRITES
exceed 2 mg (0.02%). Sample: Remove the gauze or other covering, place the
glass container of Inhalant upright in a dry iceacetone
SPECIFIC TESTS slurry, and cool for 10 min. Dry the container of Inhalant,
SPECIFIC GRAVITY 841: 0.8700.876 place it in a pointed glass tube, and break the container
ACIDITY: To 0.30 mL in a glass-stoppered cylinder, add a with a glass rod.
mixture of 0.60 mL of 0.1 N sodium hydroxide, 10 mL of Chromatographic system
water, and 1 drop of phenolphthalein TS, and invert the (See Chromatography 621, System Suitability.)
cylinder three times. Mode: GC
Acceptance criteria: The red tint of the water layer is still Detector: Thermal conductivity
perceptible. Column: Under typical conditions, the instrument contains
ADDITIONAL REQUIREMENTS a 3-mm 2-m column packed with a methyl polysiloxane
PACKAGING AND STORAGE: Preserve in tight containers, and oil, 25% by weight on suitable calcined diatomite.
store in a cool place, protected from light. Column temperature: 80
USP REFERENCE STANDARDS 11 Injection port and detector block temperature: Maintain
USP Benzyl Benzoate RS at about 10 above the temperature of the column.
Carrier gas: Helium
Flow rate: 60 mL/min
Injection size: NMT 2 L
Amyl Nitrite Inhalant Analysis: From the area under the curve, calculate the
(Comment on this Monograph)id=m4600=Amyl Nitrite percentage (a/a) of total nitrites, represented by the area
Inhalant=A-Monos.pdf) under the main peak of the chromatogram, in the Amyl
Nitrite taken.
DEFINITION Acceptance criteria: NLT 95.0%
Amyl Nitrite Inhalant contains a mixture of the nitrite esters of
3-methyl-1-butanol and 2-methyl-1-butanol. It contains NLT SPECIFIC TESTS
80.0% and NMT 105.0% of C5H11NO2. It contains a suitable SPECIFIC GRAVITY 841: 0.8700.880
stabilizer. ACIDITY
[CAUTIONAmyl Nitrite Inhalant is very flammable. Do not use Analysis: To 0.30 mL in a glass-stoppered cylinder, add a
where it may be ignited.] mixture of 0.60 mL of 0.1 N sodium hydroxide, 10 mL of
USP 32 Official Monographs / Anileridine 245
water, and 1 drop of phenolphthalein TS, and invert the Acceptance criteria: 98.5%101.0%
cylinder three times.
Acceptance criteria: The red tint of the water layer is still IMPURITIES
perceptible. Inorganic Impurities
OTHER REQUIREMENTS: It meets the requirements of the RESIDUE ON IGNITION 281: NMT 0.1%
Identification tests under Amyl Nitrite. CHLORIDE AND SULFATE, Chloride 221
Sample: 180 mg
ADDITIONAL REQUIREMENTS Analysis: Dissolve the Sample in a mixture of 1 mL of nitric
PACKAGING AND STORAGE: Preserve in tight, unit-dose glass acid and 40 mL of water.
containers, wrapped loosely in gauze or other suitable Acceptance criteria: NMT 400 ppm; the solution shows
material, and store in a cool place, protected from light. no more chloride than corresponds to 0.10 mL of 0.020 N
USP REFERENCE STANDARDS 11 hydrochloric acid.
USP Benzyl Benzoate RS
SPECIFIC TESTS
Anileridine ADDITIONAL REQUIREMENTS

(Comment on this Monograph)id=m4730=Anileridine=A- PACKAGING AND STORAGE: Preserve in tight, light-resistant
Monos.pdf) containers. Store at room temperature.
Anileridine Injection
(Comment on this Monograph)id=m4760=Anileridine
DEFINITION
C22H28N2O2 352.47 Anileridine Injection is a sterile solution of Anileridine in Water
4-Piperidinecarboxylic acid, 1-[2-(4-aminophenyl)ethyl]-4- for Injection, prepared with the aid of Phosphoric Acid. It
phenyl-, ethyl ester; contains NLT 90.0% and NMT 115.0% of the labeled amount
Ethyl 1-(p-aminophenethyl)-4-phenylisonipecotate [144-14-9]. of anileridine (C22H28N2O2), as the phosphate.
DEFINITION IDENTIFICATION
Anileridine contains NLT 98.5% and NMT 101.0% of A. PROCEDURE
C22H28N2O2, calculated on the anhydrous basis. Sample solution: 0.25 mg/mL Anileridine from Injection
diluted with water
IDENTIFICATION Analysis: To 5 mL of the Sample solution, add 2 mL of a
A. PROCEDURE solution (1 in 100) of p-dimethylaminobenzaldehyde in
Sample stock solution: Dissolve 40 mg of Anileridine in 2.3 alcohol.
mL of 0.1 N hydrochloric acid in a 100-mL volumetric flask, Acceptance criteria: A yellow color develops immediately.
and dilute with water to volume. B. A volume of Injection, diluted with water to a
Buffer solution: Dissolve 5.68 g of anhydrous dibasic concentration of 25 mg of anileridine in 1000 mL, exhibits
sodium phosphate and 3.63 g of monobasic potassium absorbance maxima at 234 1 and 285 2 nm.
phosphate in water to make 1000 mL: the pH is 7.0 0.05.
Sample solution A: Sample stock solution, Buffer solution, ASSAY
and water (4:25:71) PROCEDURE
Sample solution B: Sample stock solution, Buffer solution, Standard solution: 250 g/mL of USP Anileridine
and water (20:25:55) Hydrochloride RS in 0.1 N hydrochloric acid
Acceptance criteria: The UV absorption spectrum of Sample Each mg of anileridine hydrochloride is equivalent to 0.8286
solution A exhibits a maximum at 234 1 nm; and the UV mg of anileridine.
absorption spectrum of Sample solution B exhibits a [NOTEPrepare on the day of the assay.]
maximum at 285 2 nm. The ratio 5A234/A285 is 8.8. Sample solution: Nominally equivalent to 200 g/mL of
B. PROCEDURE anileridine, from a volume of Injection diluted with 0.1 N
Sample solution: 0.2 mg/mL of Anileridine in 0.1 N hydrochloric acid
hydrochloric acid. Blank: 0.1 N hydrochloric acid
Analysis: To 5 mL of the Sample solution, add 2 mL of a Spectrometric conditions
solution of p-dimethylaminobenzaldehyde in alcohol (1 in Analytical wavelength: 560 nm
100). Cell: 1 cm
Acceptance criteria: A yellow color develops immediately. Analysis
Samples: Sample solution, Standard solution, and Blank
ASSAY Transfer 5.0 mL each of the Standard solution, the Sample
PROCEDURE solution, and Blank to separate 200-mL volumetric flasks.
Sample: 350 mg of Anileridine To each flask add 25 mL of water, 5 mL of 1 N
Analysis: Dissolve the Sample in 50 mL of glacial acetic hydrochloric acid, and 5 mL of sodium nitrite solution (1
acid, add 1 drop of crystal violet TS and titrate with 0.1 N in 1000). Allow to stand for 2 min, then add to each flask
perchloric acid VS to a blue-green endpoint. Perform a blank 5 mL of ammonium sulfamate solution (1 in 200). Allow
determination (See Titrimetry 541). Each mL of 0.1 N to stand for 3 min, then add 5 mL of N-(1-
perchloric acid is equivalent to 17.62 mg of C22H28N2O2. naphthyl)ethylenediamine dihydrochloride solution (1 in
246 Anileridine / Official Monographs USP 32
1000). Allow to stand for 1 h, and dilute with water to ASSAY

volume. PROCEDURE
[NOTEUse the reagent blank to set the instrument.] Sample: 200 mg of Anileridine Hydrochloride
Calculate the percentage of C22H28N2O2 in each mL of the Analysis: Dissolve the Sample in 10 mL of glacial acetic acid
Injection taken: by heating it on a steam bath. Cool immediately in a cold
water bath, add 5 mL of mercuric acetate TS, 20 mL of
Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 acetone, and 0.5 mL of indicator solution (70 mg of -
naphtholbenzein, 10 mg of crystal violet, and 40 mg of
AU = absorbance of the Sample solution quinaldine red in 100 mL of glacial acetic acid). Titrate with
AS = absorbance of the the Standard solution 0.1 N perchloric acid VS to a gray-green endpoint. Perform
CS = concentration of USP Anileridine Hydrochloride a blank determination (see Titrimetry 541). Each mL of 0.1
RS in the Standard solution (g/mL) N perchloric acid is equivalent to 21.27 mg of C22H28N2O2
CU = nominal concentration of anileridine in the 2HCl.
Sample solution (g/mL) Acceptance criteria: 96.0%102.0%
Mr1 = molecular weight of anileridine, 352.48
Mr2 = molecular weight of anileridine hydrochloride, OTHER COMPONENTS
425.40 CONTENT OF CHLORIDE
Acceptance criteria: 90.0%115.0% Sample: 200 mg
Analysis: Dissolve the Sample in 50 mL of water in a glass-
SPECIFIC TESTS stoppered flask. Add 25.0 mL of 0.1 N silver nitrate VS, then
PH 791: 4.55.0 add 5 mL of 2 N nitric acid and 5 mL of nitrobenzene, shake
BACTERIAL ENDOTOXINS TEST 85: It contains NMT 7.2 USP vigorously, and add 2 mL of ferric ammonium sulfate TS.
Endotoxin Units/mg of anileridine. Titrate the excess silver nitrate with 0.1 N ammonium
OTHER REQUIREMENTS: It meets the requirements under thiocyanate VS. Each mL of 0.1 N silver nitrate is equivalent
Injections 1. to 3.545 mg of Cl.
PACKAGING AND STORAGE: Preserve in single-dose or IMPURITIES
multiple-dose containers, preferably of Type I glass, Inorganic Impurities
protected from light. RESIDUE ON IGNITION 281: NMT 0.1%
USP Anileridine Hydrochloride RS SPECIFIC TESTS
USP Endotoxin RS PH 791: 2.53.0
LOSS ON DRYING 731: Dry it at a pressure below 5 mm of
mercury at 100 for 2 h: it loses NMT 1.0% of its weight.
Anileridine Hydrochloride
(Comment on this Monograph)id=m4790=Anileridine ADDITIONAL REQUIREMENTS
Hydrochloride=A-Monos.pdf) PACKAGING AND STORAGE: Preserve in tight, light-resistant
containers.
C22H28N2O2 2HCl 425.39 USP REFERENCE STANDARDS 11
4-Piperidinecarboxylic acid, 1-[2-(4-aminophenyl)ethyl]-4- USP Anileridine Hydrochloride RS
phenyl-, ethyl ester, dihydrochloride;
Ethyl 1-(p-aminophenethyl)-4-phenylisonipecotate
dihydrochloride [126-12-5].
Anileridine Hydrochloride Tablets
DEFINITION
Anileridine Hydrochloride contains NLT 96.0% and NMT (Comment on this Monograph)id=m4820=Anileridine
102.0% of C22H28N2O2 2HCl, calculated on the dried basis. Hydrochloride Tablets=A-Monos.pdf)
A. INFRARED ABSORPTION 197K Anileridine Hydrochloride Tablets contain an amount of
B. PROCEDURE anileridine hydrochloride (C22H28N2O2 2HCl) equivalent to
Sample stock solution: 0.5 mg/mL in water NLT 95.0% and NMT 105.0% of the labeled amount of
pH 7.0 Buffer solution: 5.68 g of anhydrous dibasic sodium anileridine (C22H28N2O2).
phosphate and 3.63 g of monobasic potassium phosphate in IDENTIFICATION
water to make 1000 mL: the pH, determined A. PROCEDURE
potentiometrically, is 7.0 0.05. Sample solution: Transfer an equivalent to 50 mg of
Sample solution A: Sample stock solution, pH 7.0 Buffer anileridine, from finely powdered Tablets, to a 250-mL
solution, and water (4:25:71) volumetric flask. Add 100 mL of water, and heat on a steam
Sample solution B: Sample stock solution, pH 7.0 Buffer bath. Cool, dilute to volume, and filter.
solution, and water (20:25:55) Analysis: To 5 mL of the Sample solution add 2 mL of a
Acceptance criteria: The UV absorption spectrum of Sample solution (1 in 100) of p-dimethylaminobenzaldehyde in
solution A exhibits a maximum at 234 1 nm, and the UV alcohol.
absorption spectrum of Sample solution B exhibits a Acceptance criteria: A yellow color develops immediately.
maximum at 285 2 nm. B. PROCEDURE
C. PROCEDURE Sample stock solution: Transfer an equivalent to 50 mg of
Analysis: To 5 mL of a solution (1 in 5000) add 2 mL of a 1 anileridine, from finely powdered Tablets, to a 100-mL
in 100 solution of p-dimethylaminobenzaldehyde in alcohol. volumetric flask. Add 30 mL of water, and heat on a steam
Acceptance criteria: A yellow color develops immediately. bath. Cool, dilute to volume, and filter.
D. IDENTIFICATION TESTSGENERAL, Chloride 191: 10
mg/mL
USP 32 Official Monographs / Antazoline 247
Buffer solution: 5.68 g of anhydrous dibasic sodium Tolerances: NLT 65% (Q) of the labeled amount of
phosphate and 3.63 g of monobasic potassium phosphate in C22H28N2O2 is dissolved
water to make 1000 mL: the pH is 7.0 0.05. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Sample solution A: Standard stock solution, Buffer solution,
and water (4:25:71) ADDITIONAL REQUIREMENTS
Sample solution B: Standard stock solution, Buffer solution, PACKAGING AND STORAGE: Preserve in tight, light-resistant
and water (20:25:55) containers.
Acceptance criteria: The UV absorption spectrum of Sample USP REFERENCE STANDARDS 11
solution A exhibits a maximum at 234 1 nm, and the UV USP Anileridine Hydrochloride RS
absorption spectrum of Sample solution B exhibits a
maximum at 285 2 nm.
ASSAY Antazoline Phosphate
PROCEDURE (Comment on this Monograph)id=m4920=Antazoline
Standard solution: 250 g/mL of USP Anileridine Phosphate=A-Monos.pdf)
Hydrochloride RS in 0.1 N hydrochloric acid. (Each mg of
anileridine hydrochloride is equivalent to 0.8286 mg of
anileridine.)
[NOTEPrepare on the day of the assay.]
anileridine, from finely powdered Tablets (NLT 20), to a 250-
mL volumetric flask. Add 25 mL of 1 N hydrochloric acid
and 100 mL of water, and heat on a water bath. Cool, and
dilute to volume. Filter the solution, discarding the first 25 C17H19N3 H3PO4 363.35
mL of the filtrate. 1H-Imidazole-2-methanamine, 4,5-dihydro-N-phenyl-N-
Blank: 0.1 N hydrochloric acid (phenylmethyl)-, phosphate (1:1);
Spectrometric system 2-[(N-Benzylanilino)methyl]-2-imidazoline phosphate (1:1)
Analytical wavelength: 560 nm [154-68-7].
Cell: 1 cm
Analysis DEFINITION
Samples: Standard solution, Sample solution, and Blank Antazoline Phosphate contains NLT 98.0% and NMT 101.0% of
Transfer 5.0 mL each of the Standard solution, Sample C17H19N3 H3PO4, calculated on the dried basis.
solution, and Blank to separate 200-mL volumetric flasks. IDENTIFICATION
To each flask add 25 mL of water, 5 mL of 1 N A. INFRARED ABSORPTION 197M
hydrochloric acid, and 5 mL of sodium nitrite solution (1 B. The RF value of the principal spot of the Identification
in 1000). Allow to stand for 2 min, then add to each flask solution corresponds to that of Standard solution A as
5 mL of ammonium sulfamate solution (1 in 200). Allow obtained in the Procedure under Impurities.
to stand for 3 min, then add 5 mL of N-(1-
naphthyl)ethylenediamine dihydrochloride solution (1 in ASSAY
1000). Allow to stand for 1 h, and dilute to volume. PROCEDURE
[NOTEUse the reagent blank to set the instrument.] Sample solution: 750 mg of Antazoline Phosphate in 50
Calculate the percentage of C22H28N2O2 in the portion of mL of glacial acetic acid
Tablets taken: Analysis: Titrate with 0.1 N perchloric acid VS using a glass
electrode and a calomel electrode containing a saturated
Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 solution of lithium chloride in glacial acetic acid (see
Titrimetry 541). Perform a blank determination. Each mL of
AU = absorbance of the solution from the Sample 0.1 N perchloric acid is equivalent to 36.34 mg of C17H19N3
solution H3PO4.
AS = absorbance of the solution from the Standard Acceptance criteria: 98.0%101.0%
solution
CS = concentration of USP Anileridine Hydrochloride IMPURITIES
RS in the Standard solution (g/mL) Organic Impurities
CU = nominal concentration of anileridine in the PROCEDURE
Sample solution (g/mL) Standard stock solution: 0.10 mg/mL of USP Antazoline
Mr1 = molecular weight of anileridine, 352.48 Phosphate RS in methanol
Mr2 = molecular weight of anileridine hydrochloride, Standard solutions: Dilute the Standard stock solution with
425.40 methanol to obtain five Standard solutions having the
Acceptance criteria: 95.0%105.0% following compositions:
PERFORMANCE TESTS
DISSOLUTION 711 Percentage
Medium: 0.01 N hydrochloric acid; 900 mL Standard Concentration (for Comparison
Apparatus 1: 100 rpm Solution Dilution (g RS/mL) with Sample)
Time: 45 min A (1 in 2) 50 0.5
Standard solution: USP Anileridine Hydrochloride RS in B (2 in 5) 40 0.4
Medium
Sample solution: Filtered portion of the solution under test C (3 in 10) 30 0.3
Analysis: Determine the amount of C22H28N2O2 dissolved, D (1 in 5) 20 0.2
using the Analysis set forth in the Assay and in comparison E (1 in 10) 10 0.1
to a Standard solution having a known concentration of USP
Anileridine Hydrochloride RS.
248 Antazoline / Official Monographs USP 32
Sample solution: 10 mg/mL of Antazoline Phosphate in IDENTIFICATION

methanol A. INFRARED ABSORPTION 197K: Meets the requirements
Identification solution: 50 g/mL from Sample solution in B. ULTRAVIOLET ABSORPTION 197U
methanol Sample solution: 10 g/mL in chloroform
(See Chromatography 621, Thin-Layer Chromatography.) ASSAY
Mode: TLC PROCEDURE
Adsorbent: 0.25-mm layer of chromatographic silica gel [NOTEUse low-actinic glassware.]
mixture Mobile phase: n-Hexane, dichloromethane, and glacial
Application volume: 10 L acetic acid (41:6:3)
Developing solvent system: Methanol, diethylamine, Internal standard solution: 500 g/mL of o-nitroaniline in
and ethyl acetate (2:1:17) n-hexane [NOTEFirst dissolve o-nitroaniline in a small
Analysis quantity of dichloromethane, and then dilute with n-
Samples: Each Standard solution, Sample solution, and hexane.]
Identification solution System suitability stock solution: 0.1 mg/mL of USP
Proceed as directed in the chapter. Position the plate in a Anthralin RS and 0.2 mg/mL of danthron in
chromatographic chamber, and develop the dichloromethane
chromatograms in a solvent system, until the solvent front System suitability solution: System suitability stock solution,
has moved about three-fourths of the length of the plate. n-hexane, and Mobile phase (1:1:3)
Examine the plate under short-wavelength UV light, and Solvent blank solution: Mobile phase, n-hexane, and
compare the intensities of any secondary spots observed dichloromethane (3:1:1)
in the chromatogram of the Sample solution with those of Standard stock solution: 250 g/mL of USP Anthralin RS in
the principal spots from the Standard solutions. [NOTE dichloromethane
Disregard any spots observed at the origins of the Standard solution: Standard stock solution, Internal standard
chromatograms.] solution, and Mobile phase (1:1:3)
Acceptance criteria: No secondary spot from the Sample Sample stock solution: 250 g/mL of Anthralin in
solution is larger or more intense than the principal spot of dichloromethane
Standard solution A (0.5%), and the sum of the intensities of Sample solution: Pipet 10 mL of Sample stock solution into
all secondary spots of the Sample solution corresponds to a 100-mL volumetric flask, dilute with dichloromethane to
NMT 1.0%. volume, and mix. Pipet 5 mL of this solution and 5 mL of
the Internal standard solution into a 25-mL volumetric flask
SPECIFIC TESTS and dilute with mobile phase to volume.
MELTING RANGE OR TEMPERATURE, Class Ia 741: 194198, Chromatographic system
with decomposition (See Chromatography 621, System Suitability.)
PH 791: 4.05.0, in a solution (1 in 50) Mode: LC
LOSS ON DRYING 731: Dry it at 105 for 4 h: it loses NMT Detector: UV 354 nm
0.5% of its weight. Column: 4.6-mm 25-cm; packing L3
Flow rate: 2 mL/min
PACKAGING AND STORAGE: Preserve in tight containers. System suitability
USP REFERENCE STANDARDS 11 Samples: System suitability solution, Solvent blank solution,
USP Antazoline Phosphate RS and Standard solution
[NOTEThe relative retention times for anthralin, danthron
(if present), dianthrone (if present), and o-nitroaniline are
Anthralin 1.0, 1.2, 1.7, and 2.3, respectively.]
(Comment on this Monograph)id=m4970=Anthralin=A- Resolution: NLT 1.3, System suitability solution
Monos.pdf) Tailing factor: NMT 1.5, System suitability solution
Relative standard deviation: NMT 2.0% of the ratio of
the peak responses, Standard solution
[NOTEChromatograph Solvent blank solution: no effect on
the baseline is discernible at the retention time of
anthralin.]
Analysis
Calculate the percentage of C14H10O3 in the portion taken:
C14H10O3 226.23
9(10H)-Anthracenone, 1,8-dihydroxy-; Result = (RU/RS) (CS/CU) 100
1,8-Dihydroxy-9-anthrone [1143-38-0; 480-22-8].
RU = response ratio of the anthralin peak to the o-
DEFINITION nitroaniline peak from the Sample solution
Anthralin contains NLT 97.0% and NMT 102.0% of C14H10O3,
calculated on the dried basis.
USP 32 Official Monographs / Anthralin 249
RS = response ratio of the anthralin peak to the o- Standard stock solution: 0.25 mg/mL of USP Anthralin RS
nitroaniline peak from the Standard solution in dichloromethane
CS = concentration of USP Anthralin RS in the Standard solution: Standard stock solution, Internal standard
Standard solution (g/mL) solution, and Mobile phase (2:2:21)
CU = nominal concentration of Anthralin in the Sample Sample solution: 5 g of Cream in a tared 100-mL beaker.
solution (g/mL) Add 20 mL of dichloromethane and 10 mL of glacial acetic
Acceptance criteria: 97.0%102.0% acid, and stir to disperse the Cream. Transfer the contents
of the beaker to a filter paper (Whatman No. 4, or
IMPURITIES equivalent) with the aid of dichloromethane, and filter into
Inorganic Impurities a 100-mL volumetric flask. Thoroughly wash the precipitate
RESIDUE ON IGNITION 281: NMT 0.1% with dichloromethane, and allow the washings to drain
into the flask. Dilute with dichloromethane to volume.
SPECIFIC TESTS Pipet a volume of this solution, equivalent to 0.5 mg of
MELTING RANGE OR TEMPERATURE, Class I 741: 178181 anthralin, and 2 mL of Internal standard solution into a 25-
LOSS ON DRYING 731: Dry Anthralin over silica gel for 4 h: mL volumetric flask, and dilute with Mobile phase to
it loses NMT 0.5% of its weight. volume.
ACIDITY OR ALKALINITY: Suspend Anthralin in water, and Chromatographic system
filter: the filtrate is neutral to litmus. (See Chromatography 621, System Suitability.)
CHLORIDE AND SULFATE, Chloride 221: Add 1 g of Anthralin Mode: LC
to 15 mL of water, mix, and filter. Acidify 5 mL of the filtrate Detector: UV 354 nm
with nitric acid, and add a few drops of silver nitrate TS: no Column: 4.6-mm 25-cm; packing L3
more opalescence is produced immediately than is present in Flow rate: 2 mL/min
a 5-mL portion of the filtrate to which nothing has been Injection size: 10 L
added. System suitability
CHLORIDE AND SULFATE, Sulfate 221: To 5 mL of the Samples: System suitability solution, Solvent blank solution,
untreated filtrate obtained in the test for Chloride add 3 and Standard solution
drops of 3 N hydrochloric acid and 5 drops of barium [NOTEThe relative retention times for anthralin, danthron
chloride TS: no more turbidity is produced than is present in (if present), dianthrone (if present), and o-nitroaniline are
a 5-mL portion of the filtrate to which nothing has been about 1.0, 1.2, 1.7, and 2.3, respectively.]
added. Suitability requirements
ADDITIONAL REQUIREMENTS Resolution: NLT 1.3, System suitability solution
PACKAGING AND STORAGE: Preserve in tight containers in a Tailing factor: NMT 1.5, System suitability solution
cool place. Protect from light. Relative standard deviation: NMT 2.0% of the ratio of
USP REFERENCE STANDARDS 11 the peak responses, Standard solution
USP Anthralin RS [NOTEChromatograph Solvent blank solution: no effect on
the baseline is discernible at the retention time of
anthralin.]
Analysis
Anthralin Cream Samples: Standard solution and Sample solution
(Comment on this Monograph)id=m4975=Anthralin Cream=A- Calculate the percentage of C14H10O3 in the portion of
Monos.pdf) Cream taken:
DEFINITION Result = (RU/RS) (CS/CU) 100

Anthralin Cream is Anthralin in an aqueous (oil-in-water) or oily
(water-in-oil) cream vehicle. Anthralin Cream labeled to RU = response ratio of the anthralin peak to the o-
contain more than 0.1% of anthralin contains NLT 90.0% and nitroaniline peak from the Sample solution
NMT 115.0% of the labeled amount of C14H10O3; and Cream RS = response ratio of the anthralin peak to the o-
labeled to contain 0.1% or less of anthralin contains NLT nitroaniline peak from the Standard solution
90.0% and NMT 130.0% of the labeled amount of C14H10O3. CS = concentration of USP Anthralin RS in the
ASSAY CU = nominal concentration of anthralin in the Sample
PROCEDURE solution (g/mL)
[NOTEUse low-actinic glassware.] Acceptance criteria: Cream labeled to contain more than
Mobile phase: n-Hexane, dichloromethane, and glacial 0.1% of anthralin contains 90.0%115.0% of the labeled
acetic acid (41:6:3) amount of C14H10O3; and Cream labeled to contain 0.1% or
Internal standard solution: 500 g/mL of o-nitroaniline in less of anthralin contains 90.0%130.0% of the labeled
n-hexane [NOTEFirst dissolve o-nitroaniline in a small amount of C14H10O3.
quantity of dichloromethane, and then dilute with n-
hexane.] ADDITIONAL REQUIREMENTS
System suitability stock solution: 0.1 mg/mL of USP PACKAGING AND STORAGE: Preserve in tight containers, in a
Anthralin RS and 0.2 mg/mL of danthron in cool place. Protect from light.
dichloromethane LABELING: Label it to indicate whether the cream vehicle is
System suitability solution: System suitability stock solution, aqueous or oily.
n-hexane, and Mobile phase (1:1:3) USP REFERENCE STANDARDS 11
Solvent blank solution: Mobile phase, n-hexane, and USP Anthralin RS
dichloromethane (3:1:1)
250 Anthralin / Official Monographs USP 32
Anthralin Ointment RU = response ratios from the anthralin peak to the o-

(Comment on this Monograph)id=m5000=Anthralin nitroaniline peak from the Sample solution
Ointment=A-Monos.pdf) RS = response ratios from the anthralin peak to the o-
nitroaniline peak from the Standard solution
DEFINITION CS = concentration of USP Anthralin RS in the
Anthralin Ointment is Anthralin in a petrolatum or other Standard solution (g/mL)
oleaginous vehicle. Anthralin Ointment labeled to contain CU = nominal concentration of anthralin in the Sample
more than 0.1% of anthralin contains NLT 90.0% and NMT solution (g/mg)
115.0% of the labeled amount of C14H10O3; and Ointment Acceptance criteria: Ointment labeled to contain more
labeled to contain 0.1% or less of anthralin contains NLT than 0.1% of anthralin contains 90.0%115.0% of the
90.0% and NMT 130.0% of the labeled amount of C14H10O3. labeled amount of C14H10O3; and Ointment labeled to
contain 0.1% or less of anthralin contains 90.0%130.0% of
ASSAY the labeled amount of C14H10O3.
PROCEDURE
[NOTEUse low-actinic glassware.] ADDITIONAL REQUIREMENTS
Mobile phase: n-Hexane, dichloromethane, and glacial PACKAGING AND STORAGE: Preserve in tight containers, in a
acetic acid (41:6:3) cool place. Protect from light.
Internal standard solution: 500 g/mL of o-nitroaniline in USP REFERENCE STANDARDS 11
n-hexane [NOTEFirst dissolve o-nitroaniline in a small USP Anthralin RS
quantity of dichloromethane, and then dilute with n-
hexane.]
System suitability stock solution: 0.1 mg/mL of USP
Anthralin RS and 0.2 mg/mL of danthron in Anthrax Vaccine Adsorbed
dichloromethane (Comment on this Monograph)id=m5010=Anthrax Vaccine
System suitability solution: System suitability stock solution, Adsorbed=A-Monos.pdf)
n-hexane, and Mobile phase (1:1:3)
Solvent blank solution: Mobile phase, n-hexane, and DEFINITION
dichloromethane (3:1:1) Anthrax Vaccine Adsorbed is a sterile, milky-white suspension
Standard stock solution: 0.25 mg/mL of USP Anthralin RS made from cell-free filtrates of microaerophilic cultures of an
in dichloromethane avirulent, nonencapsulated strain of Bacillus anthracis. The final
Standard solution: Standard stock solution, Internal standard product contains no dead or live bacteria. The production
solution, and Mobile phase (2:2:21) cultures are grown in a chemically defined protein-free
Sample solution: 5 g of Ointment in a tared 100-mL medium containing amino acids, vitamins, inorganic salts, and
beaker. Add 20 mL of dichloromethane and 10 mL of glacial sugars. The sterile filtrate is adsorbed on sterile aluminum
acetic acid, and stir to disperse the Ointment. Transfer the hydroxide, concentrated 10-fold, and resuspended in sterile
contents of the beaker to a filter paper (Whatman No. 4, or physiological saline containing formaldehyde with
equivalent) with the aid of dichloromethane, and filter into a benzethonium chloride as a preservative. Sublots may be
100-mL volumetric flask. Thoroughly wash the precipitate combined to produce final lots. The product meets potency
with dichloromethane, and allow the washings to drain into requirements when tested against the U.S. Reference Standard
the flask. Dilute with dichloromethane to volume. Pipet a Anthrax Vaccine, in accordance with approved procedures
volume of this solution, equivalent to 0.5 mg of anthralin, (guinea pig intracutaneous challenge models).
and 2 mL of Internal standard solution into a 25-mL IDENTIFICATION
volumetric flask, and dilute with Mobile phase to volume. [NOTEPerform analyses on the filtration.]
(See Chromatography 621, System Suitability.) Trichloroacetic acid solution: Prepare a solution of
Mode: LC trichloroacetic acid (see Reagents, Indicators, and Solutions
Detector: UV 354 nm Reagent Specifications) in water containing 100 g
Column: 4.6-mm 25-cm; packing L3 trichloroacetic acid per 100 mL of the solution.
Flow rate: 2 mL/min Sample buffer: Prepare a solution containing 141 mM
Injection size: 10 L tris(hydroxymethyl)aminomethane, 106 mM
System suitability tris(hydroxymethyl)aminomethane hydrochloride, 0.51 mM
Samples: System suitability solution, Solvent blank solution, edetate disodium, 2% (w/v) dodecyl lithium sulfate, 10%
and Standard solution glycerol, 0.22 mM Coomassie blue G-250, and 0.175 mM
[NOTEThe relative retention times for anthralin, danthron phenolsulfonphthalein. If necessary, adjust with hydrochloric
(if present), dianthrone (if present), and o-nitroaniline are acid or sodium hydroxide to a pH of 8.5.
1.0, about 1.2, about 1.7, and about 2.3, respectively.] Running buffer: Prepare a solution containing 25 mM
Suitability requirements tris(hydroxymethyl)aminomethane, 192 mM glycine, and
Resolution: NLT 1.3, System suitability solution 0.1% (w/v) dodecyl sodium sulfate (see Reagents, Indicators,
Tailing factor: NMT 1.5, System suitability solution and SolutionsReagent Specifications) in water. If necessary,
Relative standard deviation: NMT 2.0% of the ratio of adjust with hydrochloric acid or sodium hydroxide to a pH
the peak responses , Standard solution of 8.5.
[NOTEChromatograph Solvent blank solution: no effect on Transblotting buffer: Prepare a solution containing 12.5
the baseline is discernible at the retention time of mM tris(hydroxymethyl)aminomethane, 96 mM glycine, and
anthralin.] 10% methanol. If necessary, adjust with hydrochloric acid or
Analysis sodium hydroxide to a pH of 8.0.
Samples: Standard solution and Sample solution Blocking buffer: Prepare a solution containing 10 mM
Calculate the percentage of C14H10O3 in the portion of monobasic sodium phosphate, 150 mM sodium chloride,
Ointment taken: 5% (w/v) nonfat dry milk, and 0.05% (w/v) Polysorbate 20.
Result = (RU/RS) (CS/CU) 100 Adjust with sodium hydroxide to a pH of 7.4.
USP 32 Official Monographs / Anthrax 251
Primary antibody solutions: Prepare suitable monoclonal Sample solution, and mark the strips as PA, LF, and EF at
antibodies raised against the Protective Antigen (PA), the the top. Place each strip in a heat-sealable bag, add 5 mL
Lethal Factor (LF), and the Edema Factor (EF), respectively, of Blocking buffer, and seal the bag. Incubate for 30 min
of Bacillus anthracis in murine ascites cells, harvested, and with constant agitation. Open each bag, and pour out the
used without further purification. Immediately before use, Blocking buffer. Add 9 mL of the diluted Primary antibody
dilute each of the murine ascites fluids containing the solution against PA to the bag containing the strip marked
monoclonal antibodies 1:1000 with the Blocking buffer. PA. Similarly, add 9 mL of the diluted Primary antibody
Secondary antibody solution: Immediately before use, solution against LF and EF to the bags containing strips
dissolve according to the manufacturers instructions, if labeled LF and EF, respectively. Seal the bags, and incubate
necessary, and dilute the stock horseradish peroxidase under agitation for 2 h at room temperature or overnight
conjugated to goat anti-mouse IgG solution 1:1000 with at 2 to 8. Remove the strips from the plastic bags, and
Blocking buffer. place in separate plastic boxes. Add sufficient Blocking
Chromogenic visualization solution: 150 mg/mL of 4- buffer so that each strip is completely immersed. Agitate for
chloro-1-naphthol in water. at least 30 min at room temperature with two changes of
Sample solution: Use Anthrax Vaccine Filtrate as is. Blocking buffer. Remove the strips, and place each strip in a
Analysis: In a suitable centrifuge tube transfer 30/c mL of new heat-sealable plastic bag. Add 9 mL of the Secondary
the Sample solution, where c is the total protein antibody solution to each plastic bag. Seal the bags, and
concentration, in g/mL, of the solution as determined in incubate for 1 h at room temperature under agitation.
the test for Total protein. Add 16.5/c mL of Trichloroacetic Remove the strips from the plastic bags, and place in
acid solution, and incubate for at least 10 min. Centrifuge at separate plastic boxes. Add sufficient Blocking buffer so that
9000 g for 10 min, decant off the supernatant, and hold the each strip is completely immersed. Agitate for at least 30
tube inverted to drain on a filter paper. Dissolve the pellet in min at room temperature with two changes of the Blocking
60 L of Sample buffer, and transfer the solution to a buffer. Transfer each strip into a new heat-sealable plastic
polypropylene microfuge tube that has a lid. Close the lid bag, add 9 mL of Chromogenic visualization solution and 10
tightly, secure with a lid-lock, and heat at 100 for 5 min. L of 30% hydrogen peroxide, and seal the bags. Incubate
Allow the solution to cool to room temperature, and for 30 min under agitation. Transfer the strips into separate
centrifuge at 10,000 g for 15 s to collect the liquids. In a plastic boxes, and remove the excess 4-chloro-1-naphthol
suitable device for polyacrylamide-gel electrophoresis (see by incubating with water under agitation for 10 min. Visual
Electrophoresis 726 and Biotechnology-Derived Articles observation indicates a strong positive band on the strip
Polyacrylamide Gel Electrophoresis 1056) add appropriate labeled PA, a faintly detectable band on the strip labeled
volumes of the Running buffer in the upper and the lower LF, and no detectable band on the strip labeled EF.
buffer chambers. Attach a 4%20% gradient tris-glycine 83 kDA PROTEIN
polyacrylamide slab gel sandwiched between two glass Trichloroacetic acid solution, Sample buffer, Running
plates, such that the wells for sample application are buffer, and Sample solution: Prepare as directed under
exposed to the Running buffer in the upper buffer chamber. Identification, Procedure.
Apply about 20-L aliquots of the treated Sample solution in Staining solution: Prepare a solution of Coomassie blue
three alternate lanes. [NOTEDo not apply any solution in G-250 having a concentration of 1.25 g/L in a mixture of
the outside lanes. ] Connect the lower buffer chamber water, methanol, and acetic acid (5:4:1).
electrode to the positive terminal and the upper buffer Protein molecular weight standard solution: Reconstitute
chamber electrode to the negative terminal of a suitable a vial of protein molecular weight standard mixture
power supply unit, and carry out the electrophoresis at a containing proteins of molecular weights at least in the
constant current of about 40 mA. When the dye-front is range of 14200 kDa, according to manufacturers
about 1 cm from the bottom of the gel (about 40 min), instruction. Dilute the solution with Sample buffer such that
stop the current, and remove the gel from the gel assembly. the concentration of each protein in the solution is about
[NOTEDo not touch the gel with bare hands. Use gloves.] 0.5 g/L.
Place three to four filter papers, cut to the size of the gel Analysis: In a suitable centrifuge tube transfer 10/c mL of
and soaked in the Transblotting buffer, on the anode plate the Sample solution, where c is the total protein
of a suitable semidry electroblotter. Cut a nitrocellulose concentration, in g/mL, of the solution as determined by
membrane to the same size as the gel plus 12 mm on the test for Total protein (see below). Add 5.5/c mL of
each side, and wet the membrane by immersing it into Trichloroacetic acid solution, and incubate for at least 10 min.
the Transblotting buffer for about 15 s, such that there is no Centrifuge at 9000 g for about 10 min, decant off the
air-bubble between the buffer and the membrane. Place supernatant, and hold the tube inverted to drain on a filter
the wet membrane immediately on the stack of filter paper. Dissolve the pellet in 20 L of Sample buffer, and
papers, and remove all air bubbles between the membrane transfer the solution to a polypropylene microfuge tube with
and filter paper by rolling a pipet, or equivalent, gently a lid. Transfer 20 L of Protein molecular weight standard
over the surface of the membrane. Place a few drops of the solution to another polypropylene microfuge tube with a lid.
Transblotting buffer on the membrane, and then carefully Close the lids tightly, secure with lid-locks, and heat both
place the gel on it. Gently roll a pipet, or equivalent, over solutions at 100 for 5 min. Allow the solutions to cool to
the surface of the gel to ensure intimate contact between room temperature, and centrifuge at 10,000 g for 15 s to
the gel and the membrane, making sure that there are no collect the liquids. Apply the solutions to two consecutive
air bubbles in between. Place a filter paper cut to the size lanes of a 4%20% gradient tris-glycine polyacrylamide slab
of the gel and soaked in the Transblotting buffer, such that gel [NOTEDo not apply any solution in the outside lanes.],
there is no air-bubble between the filter paper and the gel. and electrophorese as directed under Identification (see
Place two to three additional filter papers, prepared in a Electrophoresis 726 and Biotechnology-Derived Articles
similar manner, on the top, and complete the transfer stack Polyacrylamide Gel Electrophoresis 1056). When the dye-
by placing the cathode plate on the top. Apply a current of front is about 1 cm from the bottom of the gel (about 40
about 250 mA, and continue transfer for 90 min. min), stop the current, and remove the gel from the gel
Remove the membrane, and wash it quickly by immersing assembly. Soak the gel in a suitable volume of the Staining
into water for 15 s. [NOTEDo not touch the membrane solution for at least 1 h, such that the gel is completely
with bare hands. Use gloves.] Cut the membrane into three immersed in the Staining solution during staining. [NOTE
strips such that each strip contains a lane containing the
252 Anthrax / Official Monographs USP 32
Use disposable gloves.] Destain the gel with a large volume Vaccine Adsorbed with respect to the corresponding U.S.
of water under constant agitation with repeated changes of Reference Standard Anthrax Vaccine is the antilog of the
water until the background of the gel is completely color horizontal distance between the two parallel lines.
free. Using the molecular weights of the proteins in Protein Acceptance criteria: The relative potency of Anthrax
molecular weight standard solution, identify the band Vaccine Adsorbed is acceptable if it is between 0.53 and
corresponding to the Protective Antigen (MW about 83 kDa) 1.79, both values inclusive.
in the Sample solution lane. [NOTEThis band is also the
predominant band in the lane of the Sample solution.] OTHER COMPONENTS
Scan the gel, and determine the relative amount (by peak [NOTEPerform analysis on final product.]
area) of the 83 kDa band by densitometry in the lane of ALUMINUM Not a test
the Sample solution. The content of 83 kDa band is NLT STANDARD SOLUTIONS: Prepare as directed under Aluminum
35% of the total peak area. 206, Standard Preparations, except to prepare solutions
TOTAL PROTEIN containing 10, 20, 30, 40, and 50 g/mL of aluminum.
Standard solution A: Prepare a solution of albumin bovine Sample solution: Mix Anthrax Vaccine Adsorbed, Final
serum (see Reagents, Indicators, and SolutionsReagent Product well, and transfer 0.2 mL to a 10-mL volumetric
Specifications) in water to obtain a known concentration of flask. Add 0.5 mL of concentrated sulfuric acid and 0.5 mL
2.0 mg/mL. of concentrated nitric acid, and mix gently. Incubate at
Standard solutions B, C, D, and E: Dilute Standard solution room temperature for 30 min or until the solution becomes
A with water to obtain solutions having protein essentially clear. Dilute with water to volume.
concentrations of 4, 8, 16, and 24 g/mL, respectively. Analysis: Proceed as directed under Aluminum 206,
Sample solution: Use Anthrax Vaccine Filtrate as is. Procedure. Plot the absorbances versus the content of
Analysis (see Biotechnology-Derived ArticlesTotal Protein aluminum, in g/mL, for the Standard solutions, and draw a
Assay 1057, Method 3): To a series of test tubes transfer best-fit straight line through the points using a linear
800 L each of Standard solutions B, C, D, and E and the regression model. Calculate the amount of aluminum in
Sample solution. Also transfer 800 L of water to be used as Anthrax Vaccine Adsorbed, in mg/mL. The aluminum
the blank. Add 200 L of Coomassie blue G-250 dye concentration is between 0.8 and 1.5 mg/mL.
solution (see Reagents, Indicators, and SolutionsReagent
Specifications) to each tube, and mix without foaming. IMPURITIES
Determine absorbances of the solutions at 595 nm using a [NOTEPerform analyses on final product.]
suitable spectrophotometer (see Spectrophotometry and Organic Impurities
Light-Scattering 851), using the blank to set the instrument PROCEDURE 1 FORMALDEHYDE:
to zero. Potassium ferricyanide solution: 25 mg/mL of potassium
[NOTEDo not use quartz (silica) spectrophotometer cells; ferricyanide
the dye binds to silica.] Phenylhydrazine hydrochloride solution: 4 g of
Construct a standard curve by plotting the absorbances phenylhydrazine hydrochloride in 100 mL of absolute
versus protein concentrations, in g/mL, of Standard alcohol, add 2 mL of water
solutions B, C, D, and E and by drawing a best-fit straight Standard stock solution: To prepare a stock solution,
line using the linear regression method. From the standard proceed as directed. Determine the concentration of
curve, determine the total protein concentration of the formaldehyde in percent (w/v) as follows:
Sample solution using the absorbance value. The protein Sample solution: 3 mL of Formaldehyde Solution to a tared
concentration is between 5 and 20 g/mL. flask containing 10 mL of water, insert the stopper in the
flask tightly, and accurately determine the weight of the
ASSAY Formaldehyde Solution taken. Slowly and quantitatively
[NOTEPerform analysis on the final product.] add a mixture of 50.0 mL of 1 N sodium hydroxide VS and
RELATIVE POTENCY 50 mL of hydrogen peroxide TS that has been previously
Standard solutions: Dilute approved U.S. Reference neutralized to bromothymol blue TS with 1 N sodium
Standard Anthrax Vaccine 1:1.6, 1:4, 1:10, and 1:25 hydroxide. Heat the contents of the flask cautiously on a
aseptically with a sterile 0.9% sodium chloride solution. steam bath for 15 min, shaking it occasionally with a rotary
Sample solutions: Dilute Anthrax Vaccine Adsorbed, Final motion. Allow the mixture to cool, rinse the funnel and the
Product 1:1.6, 1:4, 1:10, and 1:25 aseptically with a sterile inner wall of the flask with water, and after allowing it to
0.9% sodium chloride solution. stand for 30 min, add 25 drops of bromothymol blue TS.
Analysis Analysis: Titrate the excess alkali with 1 N sulfuric acid VS.
Samples: Standard solutions and Sample solutions Perform a blank determination (see Titrimetry 541,
Assign each dilution to a set of 12 randomly selected Residual Titrations). Also make a correction based upon the
guinea pigs, strain Mdh:S(RA), 6 males and 6 females, acidity found in the test for Acidity under Formaldehyde
each weighing 315385 g on the day of vaccination. Solution. Each mL of 1 N sodium hydroxide is equivalent to
Inject the animals subcutaneously in the ventral abdomen 30.03 mg of Formaldehyde Solution (CH2O).
with 0.5 mL of the assigned dilutions. On the 14th day Acceptance criteria: 37.0%, by weight, of CH2O for bulk
post-vaccination, challenge the animals with containers; 36.5%, by weight, of CH2O for small containers
approximately 1000 spores of Bacillus anthracis strain Standard solutions: Dilute the Standard stock solution in
Vollum 1B, and record the deaths daily for a 10-day water to obtain solutions having concentrations of 0.005%,
observation period. Record the numbers of surviving 0.01%, and 0.02% (w/v).
animals for each of the Standard solutions and the Sample Sample solution: Use Anthrax Vaccine Adsorbed, Final
solutions at the end of the test. Perform calculations by Product as is.
estimating best-fit lines for the Standard solutions and the Analysis: To suitable glass centrifuge tubes transfer 1.0 mL
Sample solutions using a logistic regression model that each of water, the Standard solutions, and the Sample
utilizes the number of animals that survived at the end of solution. To each tube add 1.0 mL of Potassium ferricyanide
the test and the time to death for the animals that died. solution, 4.0 mL of 18% (w/v) hydrochloric acid and 2.0 mL
Evaluate statistically the lines corresponding to the of Phenylhydrazine hydrochloride solution. Mix after each
Standard solutions and the Sample solutions for parallelism. addition. Incubate for 5060 min at room temperature.
Determine the common slope, and draw the parallel lines Centrifuge the solutions at 10,000 g for at least 10 min, and
using the common slope. The relative potency of Anthrax measure absorbances of the supernatants at 540 nm using a
USP 32 Official Monographs / Anticoagulant 253
suitable spectrophotometer (see Spectrophotometry and coupled with a standard silversilver chloride reference
Light-Scattering 851). Plot the absorbances versus electrode. Plot the voltage readings versus concentration of
concentrations of formaldehyde, in mg/mL, in the Standard chloride, in mg/mL, for Standard solutions A and B, and
solutions, and draw the best-fit straight line through the draw a straight line joining the points.
points. Calculate the concentration of chloride ion in the Sample
Calculate the amount of formaldehyde in the sample in solution from the voltage reading. Assuming that the
percent (w/v). chloride ion comes entirely from sodium chloride, calculate
Acceptance criteria: The concentration of formaldehyde in the concentrations of sodium chloride in the Sample
Anthrax Vaccine Adsorbed is less than 0.02% (w/v). solution.
PROCEDURE 2: BENZETHONIUM CHLORIDE Acceptance criteria: The concentration of sodium chloride
Citrate buffer: 25 g of citric acid monohydrate in 60 mL of in Anthrax Vaccine Adsorbed is between 0.75% and 0.95%
water, and adjust with a solution of sodium hydroxide to a (w/v).
pH of 4.5. Transfer the solution to a 100-mL volumetric
flask. Dilute with water to volume. ADDITIONAL REQUIREMENTS
Dye solution: 50 mg of 2,4,5,7-tetrabromofluorescein in PACKAGING AND STORAGE: Preserve in multiple-dose tight
100 mL of water, and mix. Dilute 1 mL of this solution with Type I glass containers. Store at a temperature between 2
water to 100 mL. and 8. Do not freeze.
Docusate sodium solution: 50 g/mL of docusate sodium LABELING: Label it to state that it is to be well shaken before
Standard solution A: 0.5 g of benzethonium chloride in a use and that it is not to be frozen.
100-mL volumetric flask, dissolve in 60 mL water, and dilute EXPIRATION DATE: The expiration date is 18 months from
with water to volume the date of manufacture.
Standard solutions B, C, D, and E: Dilute Standard solution
A with water to obtain solutions having concentrations of
0.001%, 0.002%, 0.003%, and 0.004% (w/v), respectively. Anticoagulant Citrate Dextrose
Sample solution: Use Anthrax Vaccine Adsorbed, Final
Product as is. Solution
Analysis: Transfer 4.0 mL each of Standard solutions B, C, D, (Comment on this Monograph)id=m5100=Anticoagulant Citrate
and E and the Sample solution to suitable glass centrifuge Dextrose Solution=A-Monos.pdf)
tubes. Add 1.0 mL of Citrate buffer and 0.4 mL of the Dye
solution to each tube. Add 4.0 mL of 1,1,2,2- DEFINITION
tetrachloroethane to each tube, and vigorously mix on a Anticoagulant Citrate Dextrose Solution is a sterile solution of
vortex mixer for 1 min. Centrifuge at about 1000 g for at Citric Acid, Sodium Citrate, and Dextrose in Water for
least 15 min to separate the organic layer from the aqueous Injection. It contains in each 1000 mL:
layer. Transfer 2.0 mL of the organic layer from the tubes to
another set of glass tubes. Add 4.0 mL of water and 0.5 mL Solution A Solution B
of Citrate buffer to each tube, and mix on a vortex mixer for Total Citrate, expressed as citric acid, anhydrous (C6H8O7)
about 1 min. Titrate the benzethonium chloride-dye
complex in each tube with the Docusate sodium solution (see NLT 20.59 g NLT 12.37 g
Titrimetry 541) to the colorimetric endpoint indicated by NMT 22.75 g NMT 13.67 g
the disappearance of the pink color of the organic layer. Dextrose (C6H12O6H2O)
[NOTEVigorously mix the solution on a vortex mixer after
each addition of the Docusate sodium solution.] Plot the NLT 23.28 g NLT 13.96 g
volumes of Docusate sodium solution required versus the NMT 25.73 g NMT 15.44 g
concentrations of benzethonium chloride in Standard Sodium (Na)
solutions B, C, D, and E, and draw a best-fit straight line
through the points. Determine the concentration of NLT 4.90 g NLT 2.94 g
benzethonium chloride in the Sample solution from the NMT 5.42 g NMT 3.25 g
volume of Docusate sodium solution required to titrate the
Sample solution. It contains no antimicrobial agents.
Acceptance criteria: The concentration of benzethonium Prepare Anticoagulant Citrate Dextrose Solution as follows:
chloride in Anthrax Vaccine Adsorbed is between 0.0015%
and 0.0030% (w/v). Solution A Solution B
SPECIFIC TESTS Citric Acid (anhydrous) 7.3 g 4.4 g
[NOTEPerform tests on final product.] Sodium Citrate (dihydrate) 22.0 g 13.2 g
SAFETY: It meets the requirements when tested as directed
in Biological Reactivity Tests, In Vivo 88, Safety Tests Dextrose (monohydrate) 24.5 g 14.7 g
Biologicals. Water for Injection 1000 mL 1000 mL
STERILITY TESTS 71: It meets the requirements when tested
as directed under Test for Sterility of the Product to Be Dissolve the ingredients, and mix. Filter the solution until clear,
Examined, Direct Inoculation of the Culture Medium. place immediately in suitable containers, and sterilize.
PH 791: 7.58.5 If desired, 8 g and 4.8 g of monohydrated citric acid may be
SODIUM CHLORIDE used instead of the indicated, respective amounts of
Standard solutions A and B: Prepare two solutions of anhydrous citric acid; 19.3 g and 11.6 g of anhydrous sodium
sodium chloride in water having concentrations of 0.2 mM citrate may be used instead of the indicated, respective
and 2.0 mM, respectively. amounts of dihydrated sodium citrate; and 22.3 g and 13.4 g
Sample solution: Transfer 0.5 mL of Anthrax Vaccine of anhydrous dextrose may be used instead of the indicated,
Adsorbed, Final Product to a 50-mL volumetric flask. Dilute respective amounts of monohydrated dextrose.
Analysis: Determine the voltage readings of Standard IDENTIFICATION
solutions A and B and the Sample solution using an ion- Add a few drops of solution (1 in 20) to 5 mL of hot alkaline
specific electrode specific for the chloride ion electrically cupric tartrate TS: a copious red precipitate of cuprous oxide
254 Anticoagulant / Official Monographs USP 32
is formed. When concentrated to one-half its volume, it A = 100 mm divided by the length of the polarimeter
meets the requirements of the tests for Identification Tests tube (mm)
Citrate 191 and for Identification TestsSodium 191 R = observed rotation (degrees)
Where the Solution is labeled to contain dextrose
ASSAY monohydrate, calculate the percentage (g/100 mL)
TOTAL CITRATE C6H12O6 H2O in the portion of Solution taken:
Mobile phase, Standard solution A, and Chromatographic
system: Proceed as directed under Assay for Citric Result = (100/F) (Mr1/Mr2) A R
Acid/Citrate and Phosphate 345.
Sample solution: Pipet 5 mL of Solution into a suitable 100 = percentage
volumetric flask, and proceed as directed for Assay F = midpoint of the specific rotation range for
Preparation for Citric Acid/Citrate Assay under General anhydrous dextrose (degrees), 52.9
Chapter 345. Mr1 = molecular weight for dextrose monohydrate,
Analysis: Proceed as directed for Analysis under General 198.17
Chapter 345. Mr2 = molecular weight for anhydrous dextrose, 180.16
Calculate the quantity, in mg, of anhydrous citric acid A = 100 mm divided by the length of the polarimeter
(C6H8O7) in the volume of Solution taken: tube (mm)
R = observed rotation (degrees)
Result = (Mr1/Mr2) CS (rU/rS) 0.001 D
IMPURITIES
Mr1 = molecular weight of anhydrous citric acid, Inorganic Impurities
192.12 CHLORIDE AND SULFATE, Chloride 221: A 10-mL portion
Mr2 = molecular weight of citrate (C6H5O7), 189.10 shows no more chloride than corresponds to 0.50 mL of
CS = concentration of citrate in Standard solution A 0.020 N hydrochloric acid (0.0035%).
(g/mL)
rU = citrate peak area of the Sample solution SPECIFIC TESTS
rS = citrate peak area of Standard solution A PH 791: 4.55.5
D = dilution factor OTHER REQUIREMENTS: Meets the requirements under
SODIUM Injections 1
Solution A: Add 1.04 g of lithium nitrate to a 1000-mL BACTERIAL ENDOTOXINS TEST 85: It contains NMT 5.56 USP
volumetric flask, add a suitable nonionic surfactant, and add Endotoxin Units/mL.
water to volume. This solution contains 15 mEq of
lithium/1000 mL. ADDITIONAL REQUIREMENTS
Standard solution: Add 8.18 g of sodium chloride, PACKAGING AND STORAGE: Preserve in single-dose containers,
previously dried at 105 for 2 h to a 1000-mL volumetric of colorless, transparent Type I or Type II glass, or of a
flask, and dilute with water to volume. This solution contains suitable plastic material (see Transfusion and Infusion
140 mEq of sodium/1000 mL. Transfer 50 L of this solution Assemblies and Similar Medical Devices 161).
to a 10-mL volumetric flask, and dilute with Solution A to LABELING: Label to indicate the number of mL of Solution
volume. required/100 mL of whole blood or the number of mL of
Sample solution: Add 25 mL of Solution into a 50-mL Solution required/volume of whole blood to be collected.
volumetric flask, and dilute with water to volume. Transfer USP REFERENCE STANDARDS 11
50 L of this solution to a 10-mL volumetric flask, and dilute USP Citric Acid RS
with Solution A to volume. USP Endotoxin RS
Analysis: Using a suitable flame photometer, adjusted to
read zero with Solution A, concomitantly determine the
sodium flame emission readings for the Standard solution Anticoagulant Citrate Phosphate
and the Sample solution at the wavelength of maximum Dextrose Solution
emission at 589 nm.
Calculate the quantity, in g, of Na in 1000 mL of Solution (Comment on this Monograph)id=m5130=Anticoagulant Citrate
taken: Phosphate Dextrose Solution=A-Monos.pdf)
Result = W (Ar/Mr) (rU/rS) DEFINITION

Anticoagulant Citrate Phosphate Dextrose Solution is a sterile
W = weight of sodium chloride taken to make the solution of Citric Acid, Sodium Citrate, Monobasic Sodium
Standard solution (g), 8.18 Phosphate, and Dextrose in Water for Injection. It contains, in
Ar = atomic weight of sodium, 22.99 each 1000 mL, NLT 2.11 g and NMT 2.33 g of monobasic
Mr = molecular weight of sodium chloride, 58.44 sodium phosphate (NaH2PO4 H2O); NLT 24.22 g and NMT
rU = sodium emission readings of the Sample solution 26.78 g of dextrose (C6H12O6 H2O); NLT 19.16 g and NMT
rS = sodium emission readings of the Standard 21.18 g of total citrate, expressed as citric acid, anhydrous
solution (C6H8O7); and NLT 6.21 g and NMT 6.86 g of Sodium (Na). It
DEXTROSE contains no antimicrobial agents.
Analysis: Determine the angular rotation of Solution in a Prepare Anticoagulant Citrate Phosphate Dextrose Solution as
suitable polarimeter tube (see Optical Rotation 781). follows.
Where the Solution is labeled to contain anhydrous
dextrose, calculate the percentage (g/100 mL) of C6H12O6 Citric Acid (anhydrous) 2.99 g
in the portion of Solution taken:
Sodium Citrate (dihydrate) 26.3 g
Result = (100/F) A R Monobasic sodium phosphate 2.22 g
(monohydrate; NaH2PO4H2O)
100 = percentage Dextrose (monohydrate) 25.5 g
F = midpoint of the specific rotation range for
anhydrous dextrose (degrees), 52.9
Water for Injection A sufficient quantity the beaker. Heat the beaker and contents over a burner
To make 1000 mL that has been adjusted to cause boiling of the solution to
start in 3.54 min. Boil the solution for 2 min, accurately
Dissolve the ingredients, and mix. Filter the solution until clear, timed, and filter immediately through the tared crucible,
place immediately in suitable containers, and sterilize. taking care to transfer all of the boiling chips or glass beads
If desired, 3.27 g of monohydrated citric acid may be used to the crucible. Wash the precipitate with hot water and 10
instead of the indicated amount of anhydrous citric acid; mL of alcohol. Dry the crucible and contents at 110 to
23.06 g of anhydrous sodium citrate may be used instead of constant weight. Perform a blank determination, and
the indicated amount of dihydrated sodium citrate; 1.93 g of correct the weight of the precipitate from the sample for
anhydrous monobasic sodium phosphate may be used instead any precipitate obtained in the blank.
of the indicated amount of monohydrated monobasic sodium Each mg of cuprous oxide precipitate of the substance
phosphate; and 23.2 g of anhydrous dextrose may be used under assay is equivalent to 0.496 mg of C6H12O6 H2O.
instead of the indicated amount of monohydrated dextrose. SODIUM
Solution A: Transfer 1.04 g of lithium nitrate to a 1000-mL
IDENTIFICATION volumetric flask, add a suitable nonionic surfactant, and add
It meets the requirements for Identification TestsGeneral water to volume. This solution contains 15 mEq of
191, Phosphates and the following test. Add a few drops of lithium/1000 mL.
a solution (1 in 20) to 5 mL of hot alkaline cupric tartrate Standard solution B: Transfer 8.18 g of sodium chloride,
TS: a copious red precipitate of cuprous oxide is formed. previously dried at 105 for 2 h to a 1000-mL volumetric
When concentrated to one-half its volume, meets the flask, and dilute with water to volume. This solution contains
requirements for Identification TestsGeneral 191, Citrate 140 mEq of sodium/1000 mL. Transfer 50 L of this solution
and 191, Sodium. to a 10-mL volumetric flask, and dilute with Solution A to
volume.
ASSAY Sample solution: Transfer 25 mL of Solution into a 50-mL
TOTAL CITRATE AND TOTAL PHOSPHATE volumetric flask, and dilute with water to volume. Transfer
Mobile phase, Standard solution B, and Chromatographic 50 L of this solution to a 10-mL volumetric flask, and dilute
system: Proceed as directed under Assay for Citric with Solution A to volume.
Acid/Citrate and Phosphate 345. Analysis: Using a suitable flame photometer, adjusted to
Sample solution for total citrate: Pipet 10 mL of Solution read zero with Solution A, concomitantly determine the
into a suitable volumetric flask, and proceed as directed for sodium flame emission readings for the Standard solution
Assay for Citric Acid/Citrate and Phosphate 345, Assay and the Sample solution at the wavelength of maximum
Preparation for Citric Acid/Citrate Assay. emission at 589 nm.
Sample solution for total phosphate: Pipet 5 mL of Calculate the quantity, in g, of Na in 1000 mL of Solution
Solution into a suitable volumetric flask, and proceed as taken:
directed for Assay for Citric Acid/Citrate and Phosphate 345,
Assay Preparation for Phosphate Assay. Result = (rU/rS) (Ar/Mr) W
Analysis: Proceed as directed for Assay for Citric Acid/Citrate
and Phosphate 345, Procedure. rU = sodium emission readings from the Sample
Calculate the quantity, in mg, of C6H8O7 in the volume of solution
Solution taken: rS = sodium emission readings from the Standard
solution
Result = (rU/rS) CS (Mr1/Mr2) 0.001 D Ar = atomic weight of sodium, 22.99
Mr = molecular weight of sodium chloride, 58.44
rU = citrate peak area from the Sample solution for W = weight of sodium chloride taken to make the
total citrate Standard solution (g), 8.18
rS = citrate peak area from Standard solution B
CS = concentration of citrate in Standard solution B
(g/mL) IMPURITIES
Mr1 = molecular weight of anhydrous citric acid, Inorganic Impurities
192.12 CHLORIDE AND SULFATE, Chloride 221: A 10-mL portion
Mr2 = molecular weight of citrate (C6H5O7), 189.10 shows no more chloride than corresponds to 0.50 mL of
D = dilution factor 0.020 N hydrochloric acid (0.0035%).
Calculate the quantity of phosphate, in mg, expressed as
NaH2PO4 H2O, in the volume of Solution taken: SPECIFIC TESTS
PH 791: 5.06.0
Result = (rU/rS) CS (Mr1/Mr2) BACTERIAL ENDOTOXINS TEST 85: It contains NMT 5.56 USP
Endotoxin Units/mL.
rU = phosphate peak area from the Sample solution for OTHER REQUIREMENTS: It meets the requirements under
total phosphate Injections 1.
rS = phosphate peak area from Standard solution B
CS = concentration of phosphate in Standard solution B ADDITIONAL REQUIREMENTS
(g/mL) PACKAGING AND STORAGE: Preserve in single-dose containers,
Mr1 = molecular weight of monobasic sodium of colorless, transparent, Type I or Type II glass, or of a
phosphate monohydrate, 137.99 suitable plastic material (see Transfusion and Infusion
Mr2 = molecular weight of phosphate (PO4), 94.97 Assemblies and Similar Medical Devices 161).
DEXTROSE LABELING: Label it to indicate the number of mL of Solution
Tare a clean, medium-porosity filtering crucible containing required/100 mL of whole blood or the number of mL of
several carborundum boiling chips or glass beads. Pipet 50 Solution required/volume of whole blood to be collected.
mL of freshly mixed alkaline cupric tartrate TS into a 400- USP REFERENCE STANDARDS 11
mL beaker. Add the boiling chips or glass beads from the USP Citric Acid RS
tared crucible, 45 mL of water, and 5.0 mL of Solution to USP Endotoxin RS
Anticoagulant Citrate Phosphate Calculate the quantity of phosphate, in mg, expressed as

Dextrose Adenine Solution NaH2PO4 H2O, in the volume of Solution taken:
(Comment on this Monograph)id=m5140=Anticoagulant Citrate Result = (rU/rS) CS (Mr1/Mr2) 0.001 D
Phosphate Dextrose Adenine Solution=A-Monos.pdf)
rU = phosphate peak area from the Sample solution for
DEFINITION total phosphate
Anticoagulant Citrate Phosphate Dextrose Adenine Solution is a rS = phosphate peak area from Standard solution B
sterile solution of Citric Acid, Sodium Citrate, Monobasic CS = concentration of phosphate in Standard solution B
Sodium Phosphate, Dextrose, and Adenine in Water for (g/mL)
Injection. It contains, in each 1000 mL, NLT 2.11 g and NMT Mr1 = molecular weight of monobasic sodium
2.33 g of monobasic sodium phosphate (NaH2PO4 H2O); NLT phosphate monohydrate, 137.99
30.30 g and NMT 33.50 g of dextrose (C6H12O6 H2O); NLT Mr2 = molecular weight of phosphate (PO4), 94.97
19.16 g and NMT 21.18 g of total citrate, expressed as citric D = dilution factor
acid, anhydrous (C6H8O7); NLT 6.21 g and NMT 6.86 g of SODIUM
sodium (Na); and NLT 0.247 g and NMT 0.303 g of adenine Solution A: Transfer 1.04 g of lithium nitrate to a 1000-mL
(C5H5N5). It contains no antimicrobial agents. volumetric flask, add a suitable nonionic surfactant, and add
Prepare Anticoagulant Citrate Phosphate Dextrose Adenine water to volume. This solution contains 15 mEq of
Solution as follows. lithium/1000 mL.
Standard solution: Transfer 8.18 g of sodium chloride,
Citric Acid (anhydrous) 2.99 g previously dried at 105 for 2 h to a 1000-mL volumetric
Sodium Citrate (dihydrate) 26.3 g flask, and dilute with water to volume. This solution contains
140 mEq of sodium/1000 mL. Transfer 50 L of this solution
Monobasic Sodium Phosphate 2.22 g to a 10-mL volumetric flask, and dilute with Solution A to
(monohydrate; NaH2PO4H2O) volume.
Dextrose (monohydrate) 31.9 g Sample solution: Transfer 25 mL of Solution into a 50-mL
C5H5N5 0.275 g volumetric flask, and dilute with water to volume. Transfer
50 L of this solution to a 10-mL volumetric flask, and dilute
Water for Injection A sufficient quantity with Solution A to volume.
To make 1000 mL Analysis: Using a suitable flame photometer, adjusted to
read zero with Solution A, concomitantly determine the
Dissolve the ingredients, and mix. Filter the solution until clear, sodium flame emission readings for the Standard solution
place immediately in suitable containers, and sterilize. and the Sample solution at the wavelength of maximum
If desired, 3.27 g of monohydrated citric acid may be used emission at 589 nm.
instead of the indicated amount of anhydrous citric acid; Calculate the quantity, in g, of Na in 1000 mL of Solution
23.06 g of anhydrous sodium citrate may be used instead of taken:
the indicated amount of dihydrated sodium citrate; 1.93 g of
anhydrous monobasic sodium phosphate may be used instead Result = (rU/rS) (Ar/Mr) W 2
of the indicated amount of monohydrated monobasic sodium
phosphate; and 29.0 g of anhydrous dextrose may be used rU = sodium emission readings from the Sample
instead of the indicated amount of monohydrated dextrose. solution
rS = sodium emission readings from the Standard
ASSAY solution
TOTAL CITRATE AND TOTAL PHOSPHATE Ar = atomic weight of sodium, 22.99
Mobile phase, Standard solution B, and Chromatographic Mr = molecular weight of sodium chloride, 58.44
system: Proceed as directed under Assay for Citric W = weight of sodium chloride taken to make the
Acid/Citrate and Phosphate 345. Standard solution (g), 8.18
Sample solution for total citrate: Pipet 10 mL of Solution DEXTROSE
into a suitable volumetric flask, and proceed as directed for Tare a clean, medium-porosity filtering crucible containing
Assay for Citric Acid/Citrate and Phosphate 345, Assay several carborundum boiling chips or glass beads. Pipet 50
Preparation for Citric Acid/Citrate Assay. mL of freshly mixed alkaline cupric tartrate TS into a 400-
Sample solution for total phosphate: Pipet 5 mL of mL beaker. Add the boiling chips or glass beads from the
Solution into a suitable volumetric flask, and proceed as tared crucible, 45 mL of water, and 5.0 mL of Solution to
directed for under Assay for Citric Acid/Citrate and Phosphate the beaker. Heat the beaker and contents over a burner
345, Assay Preparation for Phosphate Assay. that has been adjusted to cause boiling of the solution to
Analysis: Proceed as directed for Assay for Citric Acid/Citrate start in 3.54 min. Boil the solution for 2 min, accurately
and Phosphate 345, Procedure. timed, and filter immediately through the tared crucible,
Calculate the quantity, in mg, of C6H8O7 in the volume of taking care to transfer all of the boiling chips or glass beads
Solution taken: to the crucible. Wash the precipitate with hot water and 10
mL of alcohol. Dry the crucible and contents at 110 to
Result = (rU/rS) CS (Mr1/Mr2) 0.001 D constant weight. Perform a blank determination, and make
any necessary correction.
rU = citrate peak area from the Sample solution for Each mg of cuprous oxide precipitate obtained is equivalent
total citrate to 0.496 mg of C6H12O6 H2O.
rS = citrate peak area from Standard solution B ADENINE
CS = concentration of citrate in Standard solution B Mobile phase: To 3.45 g of ammonium dihydrogen
(g/mL) phosphate in 950 mL of water, add 10 mL of glacial acetic
Mr1 = molecular weight of anhydrous citric acid, acid, and dilute with water to 1000 mL.
192.12 Standard solutions: Quantities of USP Adenine RS in dilute
Mr2 = molecular weight of citrate (C6H5O7), 189.10 hydrochloric acid (1 in 120) in three separate volumetric
D = dilution factor
flasks, dilute with the dilute hydrochloric acid solution to Heparin Sodium 75,000 Units
volume to obtain Standard solutions having known Sodium Chloride Injection A sufficient quantity
concentrations of 0.25, 0.275, and 0.30 mg of adenine/mL,
respectively. Protect from light. To make 1000 mL
System suitability solution: 0.275 mg/mL of each USP
Adenine RS and purine, in dilute hydrochloric acid (1 in Add the Heparin Sodium, in solid form or in solution, to the
120) Sodium Chloride Injection, mix, filter if necessary, and sterilize.
Chromatographic system ASSAY
(See Chromatography 621, System Suitability.) HEPARIN SODIUM
Mode: LC Standard solution: Determine by preliminary trial, if
Detector: UV 254 nm necessary, approximately the minimum quantity of USP
Column: 4-mm 30-cm stainless steel; packing L9 Heparin Sodium RS which, when added in 0.8 mL of saline
Flow rate: 2 mL/min TS, maintains fluidity in 1 mL of prepared plasma for 1 h
Injection size: 20 L after the addition of 0.2 mL of calcium chloride solution (1
System suitability in 100). This quantity is usually between 1 and 3 USP
Sample: System suitability solution (NLT four injections) Heparin Units. On the day of the assay prepare a Standard
Suitability requirements solution such that it contains, in each 0.8 mL of saline TS,
Resolution: NLT 3.0 between adenine and purine the above-determined quantity of the Reference Standard.
Relative standard deviation: NMT 2.5% for adenine Sample solution: Dilute the Solution in sufficient saline TS
peak and NMT 2.0% for the retention time of adenine to give a concentration estimated to correspond to that of
peak the Standard solution.
Analysis Preparation of plasma: Collect blood from sheep directly
Samples: Standard solution and Sample solution into a vessel containing 8% sodium citrate solution in the
Plot the responses against the concentrations, in mg, of proportion of one volume to each 19 volumes of blood to
USP Adenine RS/mL of the Standard solutions. be collected. Mix immediately by gentle agitation and
Calculate the quantity, in mg, of C5H5N5 in each mL of the inversion of the vessel. Promptly centrifuge the blood, and
Solution taken as the value read directly from the pool the separated plasma. To a 1-mL portion of the pooled
Standard curve corresponding to the response obtained plasma in a clean test tube, add 0.2 mL of calcium chloride
from the portion of the Solution chromatographed. solution (1 in 100). Consider the plasma suitable for use if a
IMPURITIES solid clot forms within 5 min. To store plasma for future use,
Inorganic Impurities subdivide the pooled lot into portions not exceeding 100
CHLORIDE AND SULFATE, Chloride 221: A 10-mL portion mL in volume, and store in the frozen state, preventing even
shows no more chloride than corresponds to 0.50 mL of partial thawing prior to use. For use in the assay, thaw the
0.020 N hydrochloric acid (0.0035%). frozen plasma in a water bath at a temperature not
exceeding 37. Remove particulate matter by straining the
SPECIFIC TESTS thawed plasma through a coarse filter.
PH 791: 5.06.0 Analysis
BACTERIAL ENDOTOXINS TEST 85: It contains NMT 5.56 USP Samples: Standard solution and Sample solution
Endotoxin Units/mL. To meticulously clean 13-mm 100-mm test tubes, add
OTHER REQUIREMENTS: It meets the requirements under graded amounts of the Standard solution, selecting the
Injections 1. amounts so that the largest does not exceed 0.8 mL, and
so that they correspond roughly to a geometric series in
ADDITIONAL REQUIREMENTS which each step is approximately 5% greater than the
PACKAGING AND STORAGE: Preserve in single-dose containers, next lower. To each tube so prepared, add sufficient saline
of colorless, transparent, Type I or Type II glass, or of a TS to make the total volume 0.8 mL. Add 1.0 mL of
suitable plastic material (see Transfusion and Infusion prepared plasma to each tube. Then add 0.2 mL of
Assemblies and Similar Medical Devices 161). calcium chloride solution (1 in 100), note the time,
LABELING: Label it to indicate the number of mL of solution immediately insert a suitable stopper in each tube, and
required/100 mL of whole blood or the number of mL of mix the contents by inverting three times in such a way
solution required/volume of whole blood to be collected. that the entire inner surface of the tube is wet.
USP REFERENCE STANDARDS 11 In the same manner set up a series using the Sample
USP Adenine RS solution, completing the entire process of preparing and
USP Citric Acid RS mixing the tubes of both the Standard solution and the
USP Endotoxin RS Sample solution within 20 min after the addition of the
prepared plasma. In one h, accurately timed, after the
addition of the calcium chloride, determine the extent of
clotting in each tube, recognizing three grades (0.25, 0.50,
Anticoagulant Heparin Solution and 0.75) between zero and full clotting (1.0). If the series
(Comment on this Monograph)id=m5190=Anticoagulant does not contain two tubes graded more than 0.5 and two
Heparin Solution=A-Monos.pdf) tubes graded less than 0.5, repeat the Assay, using
appropriately modified Standard solution and Sample
DEFINITION solution.
Anticoagulant Heparin Solution is a sterile solution of Heparin Calculation: Convert to logarithms the volumes of Standard
Sodium in Sodium Chloride Injection. Its potency is NLT solution used in the successive five or six tubes that bracket
90.0% and NMT 110.0% of the potency stated on the label in a grade of clotting of 0.5, including at least two tubes with
terms of USP Heparin Units. It contains NLT 0.85% and NMT a larger and two tubes with a smaller grade than 0.5.
0.95% of sodium chloride (NaCl). It may be buffered. It Number and list the tubes serially, and tabulate for each the
contains no antimicrobial agents. grade of clotting observed in each tube. From the log-
Prepare Heparin solution as follows:
volumes, x, and separately from their corresponding grades Sodium Citrate (dihydrate) 40 g
of clotting, y, compute the paired averages xi and yi of Water for Injection A sufficient quantity
Tubes 1, 2, and 3; of Tubes 2, 3, and 4; of Tubes 3, 4, and
5; and, where the series consists of 6 tubes, of Tubes 4, 5, To make 1000 mL
and 6, respectively. If for one of these paired averages the
average grade, yi, is exactly 0.50, the corresponding xi is the [NOTEAnhydrous sodium citrate (35.1 g) may be used instead
median log-volume of the Standard solution, xs. Otherwise, of the dihydrate.]
interpolate xs from the paired values of yi, xi and yi +1, xi +1 Dissolve the Sodium Citrate in sufficient Water for Injection to
that fall immediately below and above grade 0.5 as: make 1000 mL, and filter until clear. Place the solution in
suitable containers, and sterilize.
xS = xi + (yi 0.5)(xi +1 xi)/(yi yi+1)
IDENTIFICATION
From the paired data on the tubes of the Sample solution, IDENTIFICATION TESTSGENERAL, Sodium 191 and Citrate
compute similarly its median log-volume u. 191: When evaporated to a concentration of 1 in 20, it
The log potency of the Sample solution is: meets the requirements.
M = xS xU + log R ASSAY
PROCEDURE
where R = vS/vU is the ratio of the USP Heparin Units (vS)/mL Mobile phase, Standard solution A, and Chromatographic
of the Standard solution to the mg (vU) of Anticoagulant system: Proceed as directed under Assay for Citric
Heparin Solution/mL of the Sample solution. Acid/Citrate and Phosphate 345.
Repeat the assay independently, and average the two or Sample solution: Pipet 10 mL of Solution into a suitable
more values of M to obtain M. If the second determination volumetric flask, and proceed as directed for Assay
of M differs by more than 0.05 from the first Preparation for Citric Acid/Citrate Assay under General
determination, continue the Assay until the log confidence Chapter 345.
interval computed as directed under Design and Analysis of Analysis: Proceed as directed for Analysis under General
Biological Assays 111, Confidence Intervals for Individual Chapter 345.
Assays does not exceed 0.20. The potency of Solution in Calculate the quantity, in mg, of C6H5Na3O7 2H2O in the
USP Heparin Units/mL is P* = antilog M. volume of Solution taken:
SODIUM CHLORIDE Result = CS (rU/rS) (Mr1/Mr2) D 0.001
Sample solution: Solution and potassium chromate TS CS = concentration of citrate in Standard solution A
(5:1) (g/mL)
Analysis: Titrate with 0.1 N silver nitrate VS. Each mL of 0.1 rU = citrate peak area of the Sample solution
N silver nitrate is equivalent to 5.844 mg of NaCl. rS = citrate peak area of Standard solution A
Acceptance criteria: 0.85%0.95% Mr1 = molecular weight of anhydrous citric acid,
SPECIFIC TESTS 294.10
PH 791: 5.07.5 Mr2 = molecular weight of citrate (C6H5O7), 189.10
BACTERIAL ENDOTOXINS TEST 85: It contains NMT 2.5 USP D = dilution factor
Endotoxin Units/mL. Acceptance criteria: 3.80 g4.20 g
INJECTIONS 1: Meets the requirements SPECIFIC TESTS
ADDITIONAL REQUIREMENTS PH 791: 6.47.5
PACKAGING AND STORAGE: Preserve in single-dose containers, BACTERIAL ENDOTOXINS TEST 85: It contains NMT 5.56 USP
of colorless, transparent Type I or Type II glass, or of a Endotoxin Units/mL.
suitable plastic material (see Transfusion and Infusion OTHER REQUIREMENTS: It meets the requirements under
Assemblies and Similar Medical Devices 161). Injections 1.
LABELING: Label it in terms of USP Heparin Units, and to ADDITIONAL REQUIREMENTS
indicate the number of mL of Solution required per 100 mL PACKAGING AND STORAGE: Preserve in single-dose containers,
of whole blood. preferably of Type I or Type II glass.
USP REFERENCE STANDARDS 11 USP REFERENCE STANDARDS 11
USP Endotoxin RS USP Citric Acid RS
USP Heparin Sodium RS USP Endotoxin RS
Anticoagulant Sodium Citrate Solution Antihemophilic Factor

(Comment on this Monograph)id=m5220=Anticoagulant (Comment on this Monograph)id=m5250=Antihemophilic
Sodium Citrate Solution=A-Monos.pdf) Factor=A-Monos.pdf)
Anticoagulant Sodium Citrate Solution is a sterile solution of Antihemophilic Factor conforms to the regulations of the FDA
Sodium Citrate in Water for Injection. It contains, in each 100 concerning biologics (see Biologics 1041). It is a sterile,
mL, NLT 3.80 g and NMT 4.20 g of sodium citrate dihydrate freeze-dried powder containing the Factor VIII fraction
(C6H5Na3O7 2H2O). It contains no antimicrobial agents.
USP 32 Official Monographs / Antimony 259
prepared from units of human venous plasma that have been Antimony Potassium Tartrate
tested for the absence of hepatitis B surface antigen, obtained (Comment on this Monograph)id=m5290=Antimony Potassium
from whole-blood donors and pooled. It may contain Heparin Tartrate=A-Monos.pdf)
Sodium or Sodium Citrate. It meets the requirements of the
test for potency, by comparison with the U.S. Standard
Antihemophilic Factor (Factor VIII) or with a working reference
that has been calibrated with it, in containing NLT 80% and
NMT 120% of the potency stated on the label, the stated
potency being NLT 100 Antihemophilic Factor Units/g of
protein. It meets the requirements of the test for pyrogen, the
test dose being 10 Antihemophilic Factor Units/kg.
C8H4K2O12Sb2 3H2O 667.87
SPECIFIC TESTS Antimonate(2-), bis[-[2,3-dihydroxybutanedioato(4-)-
EXPIRATION DATE: The expiration date is not later than 2 O1,O2:O3,O4]]-di-, dipotassium, trihydrate, stereoisomer;
years from date of manufacture, within which time it may be Dipotassium bis[-[L-(+)-tartrato(4-)]]diantimonate(2-)
stored at room temperature and used within 6 months of trihydrate [28300-74-5].
the time of such storage. Anhydrous 613.82
ADDITIONAL REQUIREMENTS [11071-15-1].
PACKAGING AND STORAGE: Preserve in hermetic containers, in DEFINITION
a refrigerator, unless otherwise indicated. Antimony Potassium Tartrate contains NLT 99.0% and NMT
LABELING: Label it to state that it is to be used within 4 h 103.0% of C8H4K2O12Sb2 3H2O.
after constitution, that it is for intravenous administration,
and that a filter is to be used in the administration IDENTIFICATION
equipment. A. When heated to redness, it chars, emits an odor
resembling that of burning sugar, and leaves a blackened
residue. This residue has an alkaline reaction, and when a
small fragment of it is held in a nonluminous flame, the
Cryoprecipitated Antihemophilic Factor flame is tinted violet.
(Comment on this Monograph)id=m5260=Cryoprecipitated B. In a solution (1 in 20), acidified with hydrochloric acid,
Antihemophilic Factor=A-Monos.pdf) hydrogen sulfide TS produces an orange-red precipitate,
which is soluble in ammonium sulfide TS and in 1 N sodium
DEFINITION hydroxide.
Cryoprecipitated Antihemophilic Factor conforms to the
regulations of the FDA concerning biologics (640.50 to C. IDENTIFICATION TESTSGENERAL 191, Tartrate
640.57) (see Biologics 1041). It is a sterile, frozen concentrate ASSAY
of human antihemophilic factor prepared from the Factor VIII- PROCEDURE
rich cryoprotein fraction of human venous plasma obtained Sample: 500 mg of Antimony Potassium Tartrate
from suitable whole-blood donors from a single unit of plasma Analysis: Dissolve in 50 mL of water, add 5 g of potassium
derived from whole blood or by plasmapheresis, collected and sodium tartrate, 2 g of sodium borate, and 3 mL of starch
processed in a closed system. It contains no preservative. It TS, and immediately titrate with 0.1 N iodine VS to the
meets the requirements of the test for potency by comparison production of a persistent blue color. Each mL of 0.1 N
with the U.S. Standard Antihemophilic Factor (Factor VIII) or iodine is equivalent to 16.70 mg of C8H4K2O12Sb2 3H2O.
with a working reference that has been calibrated with it, in Acceptance criteria: 99.0%103.0%
having an average potency of NLT 80 Antihemophilic Factor
Units/container, made at intervals of NMT 1 month during the IMPURITIES
dating period. Inorganic Impurities
ARSENIC 211
SPECIFIC TESTS Sample solution: Dissolve 100 mg in 5 mL of hydrochloric
EXPIRATION DATE: The expiration date is not later than 1 acid. Add 10 mL of a recently prepared solution of 20 g of
year from the date of collection of source material. stannous chloride in 30 mL of hydrochloric acid.
ADDITIONAL REQUIREMENTS Analysis: Transfer the Sample solution to a color-
PACKAGING AND STORAGE: Preserve in hermetic containers at comparison tube, and allow to stand for 30 min. Viewed
a temperature of 18 or lower. downward over a white surface, the color of the solution
LABELING: Label it to indicate the ABO blood group appears no deeper than that of a blank to which has been
designation and the identification number of the donor from added 15 g of arsenic (0.015%).
whom the source material was obtained. Label it also with LEAD 251: NMT 20 ppm
the type and result of a serologic test for syphilis, or to SPECIFIC TESTS
indicate that it was non-reactive in such test; with the type COMPLETENESS OF SOLUTION 641: Meets the requirements,
and result of a test for hepatitis B surface antigen, or to using a 750-mg specimen and water as the solvent
indicate that it was non-reactive in such test; with a warning LOSS ON DRYING 731: Dry at 105 to constant weight: it
not to use it if there is evidence of breakage or thawing; loses NMT 2.7% of its weight.
with instructions to thaw it before use to a temperature ACIDITY OR ALKALINITY
between 20 and 37, after which it is to be stored at room Sample solution: 1.0 g in 50 mL of carbon dioxide-free
temperature and used as soon as possible but within 6 h water
after thawing; to state that it is to be used within 4 h after Analysis: Titrate with 0.010 N hydrochloric acid or 0.010 N
the container is entered; and to state that it is for sodium hydroxide to a pH of 4.5.
intravenous administration, and that a filter is to be used in
the administration equipment.
260 Antimony / Official Monographs USP 32
Acceptance criteria: NMT 2.0 mL is required. Analysis: Titrate with 0.010 N hydrochloric acid or 0.010 N
sodium hydroxide to a pH of 4.5
ADDITIONAL REQUIREMENTS Acceptance criteria: NMT 2.0 mL is required.
Antimony Sodium Tartrate
(Comment on this Monograph)id=m5300=Antimony Sodium
Tartrate=A-Monos.pdf) Antipyrine
(Comment on this Monograph)id=m5350=Antipyrine=A-
Monos.pdf)
C8H4Na2O12Sb2 581.61
Antimonate(2-), bis[-[2,3-dihydroxybutanedioato(4-)-
O1,O2:O3,O4]]di-, disodium, stereoisomer; C11H12N2O 188.23
Disodium bis[-[L-(+)-tartrato(4-)]]diantimonate(2-) 1,2-Dihydro-1,5-dimethyl-2-phenyl-3H-pyrazol-3-one;
[34521-09-0]. 2,3-Dimethyl-1-phenyl-3-pyrazolin-5-one [60-80-0].
Antimony Sodium Tartrate contains NLT 98.0% and NMT Antipyrine contains NLT 99.0% and NMT 100.5% of
101.0% of C8H4Na2O12Sb2, calculated on the dried basis. C11H12N2O, calculated on the dried basis.
IDENTIFICATION TESTSGENERAL, Antimony 191, Sodium 191, A. INFRARED ABSORPTION 197K
and Tartrate 191 B. ULTRAVIOLET ABSORPTION 197U
Wavelength: 266 nm
ASSAY Sample solution: 20 g/mL in methanol
PROCEDURE Acceptance criteria: Absorptivities calculated on the dried
Sample: 500 mg of Antimony Potassium Tartrate basis, do not differ by more than 3.0%
Analysis: Dissolve in 50 mL of water, add 5 g of potassium C. PROCEDURE
sodium tartrate, 2 g of sodium borate, and 3 mL of starch Analysis: Add tannic acid TS to a solution of it.
TS, and immediately titrate with 0.1 N iodine VS to the Acceptance criteria: A white precipitate is formed.
production of a persistent blue color. Each mL of 0.1 N
iodine is equivalent to 14.54 mg of C8H4Na2O12Sb2. ASSAY
Acceptance criteria: 98.0%101.0% PROCEDURE
Sample: 150 mg of Antipyrine
IMPURITIES Analysis: Dissolve Sample in 25 mL of water in a 250-mL
Inorganic Impurities iodine flask. Add 2 g of sodium acetate, 1 mL of diluted
ARSENIC, Method II 211: NMT 8 ppm acetic acid, and 20.0 mL of 0.1 N iodine VS and allow to
LEAD 251: NMT 20 ppm stand in a cool, dark place for 20 min. Add 25 mL of alcohol
to dissolve the precipitate, and titrate the excess iodine with
SPECIFIC TESTS 0.1 N sodium thiosulfate VS, using starch TS as the
LOSS ON DRYING 731: Dry it at 105 to constant weight: indicator. Each mL of 0.1 N iodine is equivalent to 9.412 mg
loses NMT 6.0% of its weight. of C11H12N2O.
ACIDITY OR ALKALINITY
Sample solution: 1.0 g in 50 mL of carbon dioxide-free
water
USP 32 Official Monographs / Antipyrine 261
Acceptance criteria: 99.0%100.5% Acceptance criteria: The IR absorption spectrum of this

solution exhibits maxima at the same wavelengths as that of
IMPURITIES a similar solution of USP Antipyrine RS, concomitantly
Inorganic Impurities measured.
RESIDUE ON IGNITION 281: NMT 0.15% C. The retention times of the major peaks of the Sample
HEAVY METALS, Method II 231: NMT 20 ppm solution correspond to those of the Standard solution, as
Sample solution: 1 g in 2 mL of 1 N acetic acid, and add obtained in the Assay.
water to make 25 mL
Organic Impurities ASSAY
PROCEDURE: ORDINARY IMPURITIES 466 PROCEDURE
Standard solution: Chloroform Solution A: 7.7 mg/mL of ammonium acetate in water
Sample solution: Chloroform Mobile phase: Acetonitrile and Solution A (1:3)
Eluant: Chloroform, acetone, butyl alcohol, and formic Standard solution: Transfer 15 mg of USP Benzocaine RS to
acid (60:15:15:15) a 100-mL volumetric flask. Add 15 J mg of USP Antipyrine
Visualization: 1 RS, accurately weighed, J being the ratio of the labeled
amount, in mg, of antipyrine to the labeled amount, in mg,
SPECIFIC TESTS of benzocaine per mL of Otic Solution. Dissolve in 50 mL of
MELTING RANGE OR TEMPERATURE 741: 110112.5 methanol, and dilute with methanol to volume. Transfer
LOSS ON DRYING 731: Dry it at 60 for 2 h: it loses NMT 10.0 mL of this solution to a 100-mL volumetric flask, dilute
1.0% of its weight. with Mobile phase to volume, and mix.
COMPLETENESS AND COLOR OF SOLUTION: It is completely Sample solution: Transfer an accurately measured volume
soluble in its own weight of cold water, the solution being of Otic Solution, equivalent to about 15 mg of benzocaine,
colorless or not more than slightly yellow when viewed to a 100-mL volumetric flask. Dissolve in 50 mL of
transversely in a tube having a diameter of 20 mm. methanol, dilute with methanol to volume, and mix.
Transfer 10.0 mL of this solution to a 100-mL volumetric
ADDITIONAL REQUIREMENTS flask, dilute with Mobile phase to volume, and mix
PACKAGING AND STORAGE: Preserve in tight containers Chromatographic system
USP Antipyrine RS Mode: LC
Detector: UV 280 nm
Column: 4-mm 15-cm; 5-m packing L15
Antipyrine and Benzocaine Otic Flow rate: 1 mL/min
Solution Injection size: 10 L
System suitability
(Comment on this Monograph)id=m5360=Antipyrine and Sample: Standard solution
Benzocaine Otic Solution=A-Monos.pdf) [NOTEThe relative retention times for antipyrine and
benzocaine are 0.35 and 1.0, respectively.]
Antipyrine and Benzocaine Otic Solution is a solution of Column efficiency: NLT 1500 theoretical plates,
Antipyrine and Benzocaine in Glycerin. It contains NLT 90.0% benzocaine peak
and NMT 110.0% of the labeled amounts of antipyrine Tailing factor: NMT 2.5, benzocaine peak
(C11H12N2O) and benzocaine (C9H11NO2). Relative standard deviation: NMT 2%
[NOTEIn the preparation of this Otic Solution, use Glycerin Analysis
that has a low water content, in order that the Otic Solution Samples: Standard solution and Sample solution
may comply with the limit for Water determination. This may Calculate the percentage of C9H11NO2 in each mL of the
be ensured by using Glycerin having a specific gravity of NLT Otic Solution taken:
1.2607, corresponding to a concentration of 99.5%.]
IDENTIFICATION Result = (rU/rS) (CS/CU) 100
A. PROCEDURE rU = peak response due to benzocaine in the Sample
Sample: 5 mL solution
Analysis: Transfer the Sample to a separator containing 25 rS = peak response due to benzocaine in the
mL of water, and extract the solution with two 25-mL Standard solution
portions of a mixture of equal volumes of ether and solvent CS = concentration of USP Benzocaine RS in the
hexane. Combine the extracts, and retain the water solution Standard solution (mg/mL)
for Identification test B. Extract the etherhexane solution CU = nominal concentration of benzocaine in the
with 50 mL of water, and discard the water layer. Evaporate Sample solution (mg/mL)
the etherhexane solution to dryness, dry the residue in a Calculate the percentage of C11H12N2O in each mL of the
vacuum at 40 to 50 for 1 h, and dissolve the residue in 1 Otic Solution taken by the same formula, changing the
mL of chloroform. terms to refer to antipyrine instead of benzocaine.
Acceptance criteria: The IR absorption spectrum of this Acceptance criteria: 90.0%110.0%
solution exhibits maxima at the same wavelengths as that of
a similar solution of USP Benzocaine RS, concomitantly SPECIFIC TESTS
measured. WATER DETERMINATION, Method I 921: NMT 1.0%
B. PROCEDURE
Analysis: Add 5 mL of 1 N sodium hydroxide solution to ADDITIONAL REQUIREMENTS
the water solution retained from Identification test A, and PACKAGING AND STORAGE: Preserve in tight, light-resistant
extract with two 25-mL portions of chloroform. Evaporate containers.
the combined extracts to dryness, dry the residue in a USP REFERENCE STANDARDS 11
vacuum at 4050 for 1 h, and dissolve the residue in 3 mL USP Antipyrine RS
of chloroform. USP Benzocaine RS
262 Antipyrine / Official Monographs USP 32
Antipyrine, Benzocaine, and Acceptance criteria: 90.0%110.0%

Phenylephrine Hydrochloride Otic ADDITIONAL REQUIREMENTS
Solution PACKAGING AND STORAGE: Preserve in tight, light-resistant
(Comment on this Monograph)id=m5365=Antipyrine, containers.
Benzocaine, and Phenylephrine Hydrochloride Otic Solution=A- USP REFERENCE STANDARDS 11
Monos.pdf) USP Antipyrine RS
USP Benzocaine RS
DEFINITION USP Phenylephrine Hydrochloride RS
Antipyrine, Benzocaine, and Phenylephrine Hydrochloride Otic
Solution is a solution of Antipyrine, Benzocaine, and
Phenylephrine Hydrochloride in a suitable nonaqueous solvent.
It contains NLT 90.0% and NMT 110.0% of the labeled Antithrombin III Human
amounts of antipyrine (C11H12N2O), benzocaine (C9H11NO2), (Comment on this Monograph)id=m5390=Antithrombin III
and phenylephrine hydrochloride (C9H13NO2 HCl). Human=A-Monos.pdf)
The retention times of the major peaks of the Sample solutions Antithrombin III Human is a glycoprotein, which is the major
correspond to those of the Standard solution, as obtained in inhibitor of thrombin and other activated clotting factors,
the Assay. including factors IX, X, XI, and XII, and the cofactor through
which heparin exerts its effect. It is obtained from human
ASSAY plasma of healthy donors who must, as far as can be
PROCEDURE ascertained, be free from detectable agents of infection
Mobile phase: Acetonitrile, 0.005 M solution of sodium 1- transmissible by transfusion of blood or blood derivatives. The
heptanesulfonate in water, and phosphoric acid (120:880:1) method of manufacturing includes steps that have been
Standard solution: 25 mg of USP Antipyrine RS, 25 mg of shown to remove or inactivate known agents of infection. If
USP Benzocaine RS, and 25 mg of USP Phenylephrine substances are used for inactivation of viruses during
Hydrochloride RS in a 250-mL volumetric flask. Add 5 mL of production, the subsequent purification procedure must be
a 0.5 mg/mL solution of p-aminobenzoic acid in Mobile validated to demonstrate that the concentration of these
phase. Add 150 mL of Mobile phase, and mix to dissolve, substances is reduced to an acceptable level and any residues
sonicating if necessary. Dilute with Mobile phase to volume. are such as not to compromise the safety of the preparation
Sample solution A: Nominally equivalent to 0.1 mg/mL of for patients. The antithrombin III concentrate is passed
antipyrine, from Otic Solution diluted in Mobile phase through a bacteria-retentive filter, filled aseptically into its final,
Sample solution B: Nominally equivalent to 0.1 mg/mL of sterile containers, and immediately frozen. It is then freeze-
benzocaine, from Otic Solution diluted in Mobile phase dried, and the containers are closed under vacuum. No
Sample solution C: Nominally equivalent to 0.1 mg/mL of antimicrobial preservative is added at any stage of production.
phenylephrine hydrochloride, from Otic Solution diluted in Antithrombin III Human complies with the requirements for
Mobile phase Biologics 1041. When reconstituted in the recommended
Chromatographic system volume of diluent, the potency is NLT 25 USP Antithrombin III
(See Chromatography 621, System Suitability.) Units/mL.
Mode: LC [NOTEOne USP Antithrombin III Unit is the amount of
Detector: UV 272 nm antithrombin III that forms a complex with one unit of
Column: 4.6-mm 30-cm; packing L11 thrombin at 25 in the presence of heparin at a pH of 8.4.]
Injection size: 20 or 25 L IDENTIFICATION
System suitability Meets the requirements of the Assay.
[NOTEThe relative retention times of p-aminobenzoic acid, ASSAY
phenylephrine, antipyrine, and benzocaine are about 0.19, PROCEDURE
0.26, 0.64, and 1.0, respectively.] Solution A: Mix Tris(hydroxymethyl)aminomethane, edetic
Suitability requirements acid, and sodium chloride in water containing 0.1%
Resolution: NLT 1.5 between phenylephrine and polyethylene glycol 6000 to obtain a solution having
aminobenzoic acid concentrations of 0.050 M, 0.0075 M, and 0.175 M,
Relative standard deviation: NMT 3.0% respectively. Adjust with hydrochloric acid or sodium
Analysis hydroxide solution to a pH of 8.4.
Samples: Standard solution and Sample solutions Solution B: 0.05% (w/v) of albumin human in Solution A
Calculate the percentage of C11H12N2O in each mL of the Solution C: 10 mg/mL of polybrene in Solution B
Otic Solution taken: Solution D: 15 USP Heparin Units/mL of USP Heparin
Sodium RS in Solution B
Result = (rU/rS) (CS/CU) 100 Solution E: Reconstitute thrombin bovine, and dilute with
Solution B to obtain a solution having a concentration of 2.0
rU = peak response from Sample solution A Thrombin Units/mL.
rS = peak response from the Standard solution Solution F: Prepare a solution of chromogenic substrate for
CS = concentration of the Standard solution (mg/mL) amidolytic test (see Reagents, Indicators, and Solutions
CU = nominal concentration of antipyrine in the Reagent Specifications) for factor IIa in water to obtain a
Sample solution (mg/mL) solution having a concentration of about 5.0 mM, and
Calculate the percentages of C9H11NO2 and C9H13NO2 HCl dilute the solution further with Solution C to 1.0 mM.
in each mL of the Otic Solution taken, changing the terms
to refer to benzocaine or phenylephrine hydrochloride
instead of antipyrine.
USP 32 Official Monographs / Antithrombin 263
Stopping solution: 20% of acetic acid Acceptance criteria: NMT 0.1 USP Heparin Unit/USP
Standard solution A: USP Antithrombin III Human RS in Antithrombin III Unit.
Solution D to obtain a solution containing 1.0 USP
Antithrombin III Unit SPECIFIC TESTS
Standard solutions B, C, D, and E: Dilute Standard solution STERILITY TESTS 71: Meets the requirements when tested as
A with Solution D 60-, 120-, 180-, and 300-fold. directed for Test for Sterility of the Product to be Examined,
Sample solution A: Dissolve a quantity of Antithrombin III Direct Inoculation of the Culture Medium
Human in Solution D to obtain a solution having the same WATER DETERMINATION, Method I 921: NMT 3.0%
concentration as Standard solution A. PYROGEN TEST 151: Inject 50 USP Antithrombin III Units/kg
Sample solutions B, C, D, and E: Dilute Sample solution A of the rabbits weight, calculated from the activity stated on
with Solution D 60-, 120-, 180-, and 300-fold. the label. Meets the requirements.
Analysis: Pipet 400 L each of Standard solutions B, C, D, GENERAL SAFETY: Meets the requirements for biologics as set
and E and Sample solutions B, C, D, and E into suitable tubes forth for under Biological Reactivity Tests, In Vivo 88, Safety
placed in a water bath set at 37. Add 200 L of Solution E, TestsBiologicals
prewarmed at 37 to each tube, mix, and incubate for 1 OSMOLALITY AND OSMOLARITY 785: Reconstitute with the
min. Add 200 L of Solution F prewarmed at 37 to each diluent according to the manufacturers instruction: NLT 240
tube, mix, and incubate for 60 s. Stop the reaction by mOsmol/kg for the solution.
adding 200 L Stopping solution. To prepare a blank, add PH 791: Reconstitute with the diluent according to the
the reagents in reverse order, starting with 200 L of manufacturers instruction: 6.07.5.
Stopping solution, followed by the addition of 200 L of MOLECULAR WEIGHT DISTRIBUTION
Solution F, then adding 200 L of Solution E, and ending Mobile phase: Solution containing 0.1 M sodium
with 400 L of Solution D. Record the absorbance at 405 phosphate, 0.15 M sodium chloride, and 0.05% sodium
nm against the blank. azide, having a pH of 6.5
For Standard solutions and Sample solutions, calculate the Solution A: 45 mg/mL of thyroglobulin in Mobile phase
regression of the absorbance against log concentrations, Sample solution: 810 mg/mL of Antithrombin III Human
and calculate the activity of Antithrombin III Human in USP System suitability solution: Dilute USP Albumin Human RS,
Antithrombin III Units, using a suitable statistical method if necessary, with water to obtain a solution containing 5%.
for parallel-line assays. The four independent relative Chromatographic system
activity estimates are then combined to obtain the final (See Chromatography 621, System Suitability.)
mean, and the confidence limits are calculated. Mode: LC
Acceptance criteria: 80%120%. The specific activity is Detector: UV 280 nm
NLT 6.0 USP Antithrombin III Units/mg of total protein. The Column: 7.5- 75-mm guard column and a 7.5- 300-
confidence interval (P = 0.95) is between 90% and 110%. mm analytical column, both containing packing L59
Temperature: Ambient
IMPURITIES Flow rate: 0.5 mL/min maintained constant to 1%
Organic Impurities Injection size: 10 L
PROCEDURE: HEPARIN CONTENT System suitability
Solution A: Mix Tris(hydroxymethyl)aminomethane, edetic Sample: System suitability solution
acid, and sodium chloride in water containing 0.1% Suitability requirements
polyethylene glycol 6000 to obtain a solution having Column efficiency: Greater than 1500 theoretical plates
concentrations of 0.050 M, 0.0075 M, and 0.175 M, Tailing factor: 0.52.5
respectively. Adjust with hydrochloric acid or sodium Analysis
hydroxide solution to a pH of 8.4. Samples: Solution A and Sample solution
Solution B: Solution of chromogenic substrate for Acceptance criteria: Note the retention times of the major
amidolytic test for factor Xa in water to obtain a solution of peak in the Solution A chromatogram. The relative peak
concentration of 2.5 mM area of the high molecular weight peak eluting at about
Solution C: Factor Xa in Solution A to obtain a solution the same retention time as the major peak in the Solution A
containing 20 nanokatalytic units (nkats) chromatogram, or earlier, is NMT 13%.
Solution D: 20% (v/v) of acetic acid in water TOTAL PROTEIN CONTENT
Standard solution: USP Antithrombin III Human RS in Solution A: 1000 mg/mL of trichloroacetic acid
Solution A to obtain a solution containing 1.0 USP Sample solution: 7.5 mg/mL of Antithrombin III Human in
Antithrombin III Unit 0.15 M sodium chloride solution
Sample solution: Antithrombin III Human in Solution A to Blank: 0.15 M solution of sodium chloride
obtain a solution containing 1.0 USP Antithrombin III Unit Analysis: To each of 2.0 mL of the Sample solution and the
Analysis: Pipet 250 L each of Solution A, the Standard Blank in suitable centrifuge tubes, add 1.5 mL of Solution A.
solution, and the Sample solution to suitable tubes placed in Mix, allow to stand for at least 10 min, centrifuge for 5 min,
a water bath set at 37. Add 250 L of Solution C and decant the supernatant. Resuspend the precipitates in
prewarmed at 37 to each tube, and incubate for 2 min. 1.5 mL of Solution A, centrifuge for 5 min, decant the
Add 250 L of Solution B prewarmed at 37 to each tube, supernatant, and hold the tubes inverted on a filter paper to
mix, and incubate for 120 s. Stop the reaction by adding drain. Transfer the residues with a minimum quantity of
250 L of Solution D. Record the absorbance at 405 nm, water to a micro-Kjeldahl flask, and determine the nitrogen
using Solution A as the blank. content using Method II (see Nitrogen Determination 461).
Calculate the USP Heparin Unit/USP Antithrombin III Unit: Multiply the result, corrected for the Blank, by 6.25 to
calculate the quantity of protein.
Result = PR (AF AU)/(AF AS)
PR = heparin content of USP Antithrombin III Human PACKAGING AND STORAGE: Use a Type I glass container with
RS in USP Heparin Unit/USP Antithrombin III an appropriate stopper and seal. Store protected from light
Unit between 2 and 8, excursions permitted up to 25.
AF = absorbance values from Solution A LABELING: The labeling should state the content of
AU = absorbance values from the Sample solution antithrombin III in USP Antithrombin III Units. The diluent
AS = Absorbance values from the Standard solution
264 Antithrombin / Official Monographs USP 32
and the volume to be used to reconstitute the preparation LABELING: Label it to indicate the species of spider against
are indicated. which the Antivenin is to be used, that it is not intended to
USP REFERENCE STANDARDS 11 protect against bites from other spider species, and to state
USP Albumin Human RS that it was prepared in the horse.
USP Antithrombin III Human RS EXPIRATION DATE: The expiration date for Antivenin
USP Heparin Sodium RS containing a 10% excess of potency is NMT 5 years after the
date of issue from manufacturers cold storage (5, 1 year;
0, 2 years).
Antivenin (Crotalidae) Polyvalent
(Comment on this Monograph)id=m5410=Antivenin
(Crotalidae) Polyvalent=A-Monos.pdf) Antivenin (Micrurus fulvius)
(Comment on this Monograph)id=m5440=Antivenin (Micrurus
DEFINITION fulvius)=A-Monos.pdf)
Antivenin (Crotalidae) Polyvalent conforms to the regulations of
the FDA concerning biologics (see Biologics 1041). It is a DEFINITION
sterile, non-pyrogenic preparation derived by drying a frozen Antivenin (Micrurus fulvius) conforms to the regulations of the
solution of specific venom-neutralizing globulins obtained from FDA concerning biologics (see Biologics 1041). It is the sterile,
the serum of healthy horses immunized against the venoms of non-pyrogenic preparation derived by drying a frozen solution
four species of pit vipers, Crotalus atrox, Crotalus adamanteus, of specific venom-neutralizing globulins obtained from the
Crotalus durissus terrificus, and Bothrops atrox (Fam. Crotalidae). serum of healthy horses immunized against venom of the
It is standardized by biological assay on mice, in terms of one Eastern Coral snake (Micrurus fulvius). It is standardized by
dose of antivenin neutralizing the venoms in NLT the number biological assay on mice, in terms of one dose of antivenin
of mouse LD50 stated, of Crotalus atrox (Western neutralizing the venom of Micrurus fulvius in NLT 250 mouse
diamondback), 180; Crotalus durissus terrificus (South American LD50. It may contain a suitable preservative. When constituted
rattlesnake), 1320; and Bothrops atrox (South American fer-de- as specified in the labeling, it is opalescent and contains NMT
lance), 780. It may contain a suitable preservative. When 20.0% of solids, determined by drying 1 mL at 105 to
constituted as specified in the labeling, it is opalescent and constant weight ( 1 mg).
contains NMT 20.0% of solids, determined by drying 1 mL at
105 to constant weight (1 mg). SPECIFIC TESTS
SAFETY: It meets the requirements for general safety (see
SPECIFIC TESTS Biological Reactivity Tests, In Vivo 88, Safety Tests
SAFETY: It meets the requirements for general safety (see Biologicals).
Biological Reactivity Tests, In Vivo 88, Safety Tests
Biologicals). ADDITIONAL REQUIREMENTS
ADDITIONAL REQUIREMENTS and avoid exposure to excessive heat.
PACKAGING AND STORAGE: Preserve in single-dose containers. LABELING: Label it to indicate the species of snake against
Avoid exposure to excessive heat. which the Antivenin is to be used, and to state that it was
LABELING: Label it to indicate the species of snakes against prepared in the horse.
which the Antivenin is to be used, and to state that it was EXPIRATION DATE: The expiration date for Antivenin
prepared from horse serum. containing a 10% excess of potency is NMT 5 years after the
EXPIRATION DATE: The expiration date for Antivenin date of issue from the manufacturers cold storage (5, 1
containing a 10% excess of potency is NMT 5 years after the year; or 0, 2 years).
date of issue from manufacturers cold storage (5, 1 year; or
0, 2 years).
Apomorphine Hydrochloride
(Comment on this Monograph)id=m5660=Apomorphine
Antivenin (Latrodectus mactans) Hydrochloride=A-Monos.pdf)
(Comment on this Monograph)id=m5420=Antivenin
(Latrodectus mactans)=A-Monos.pdf)
DEFINITION
Antivenin (Latrodectus mactans) conforms to the regulations of
the FDA concerning biologics (see Biologics 1041). It is the
sterile, non-pyrogenic preparation derived by drying a frozen
solution of specific venom-neutralizing globulins obtained from
the serum of healthy horses immunized against the venom of C17H17NO2 HCl 1/2H2O 312.79
black widow spiders (Latrodectus mactans). It is standardized C17H17NO2 HCl 303.79
by biological assay on mice, in terms of one dose of antivenin 4H-Dibenzo[de,g]quinoline-10,11-diol, 5,6,6a,7-tetrahydro-6-
neutralizing the venom of Latrodectus mactans in NLT 6000 methyl-, hydrochloride, hemihydrate, (R)-;
mouse LD50. Thimerosal 1:10,000 is added as a preservative. 6a-Aporphine-10,11-diol hydrochloride hemihydrate
When constituted as specified in the labeling, it is opalescent [41372-20-7].
and contains NMT 20.0% of solids. Anhydrous [314-19-2].
ADDITIONAL REQUIREMENTS DEFINITION
PACKAGING AND STORAGE: Preserve in single-dose containers. Apomorphine Hydrochloride contains NLT 98.5% and NMT
Avoid exposure to excessive heat. 100.5% of C17H17NO2 HCl, calculated on the dried basis.
USP 32 Official Monographs / Apomorphine 265
IDENTIFICATION sodium bicarbonate solution (1 in 20), and then add 0.50

A. INFRARED ABSORPTION 197K mL of iodine TS. Allow to stand for 30 s, add 0.60 mL of
B. PROCEDURE sodium thiosulfate solution (1 in 40), and dilute with water
Analysis: To 5 mL of a solution (1 in 100) add a slight to 10 mL.
excess of sodium bicarbonate solution (1 in 20): a white or
greenish-white precipitate is formed. Add 3 drops of iodine ADDITIONAL REQUIREMENTS
TS, and shake vigorously: an emerald-green color is PACKAGING AND STORAGE: Preserve in tight, light-resistant
produced. Add 5 mL of ether and, after vigorous shaking, containers.
allow the layers to separate: the ether is colored deep ruby- USP REFERENCE STANDARDS 11
red while the water layer retains its green color. USP Apomorphine Hydrochloride RS
C. PROCEDURE
Analysis: Dissolve it in nitric acid.
Acceptance criteria: A dark purple solution is produced. Apomorphine Hydrochloride Tablets
D. PROCEDURE
Analysis: To a solution of it add silver nitrate TS. (Comment on this Monograph)id=m5690=Apomorphine
Acceptance criteria: A white precipitate, which is insoluble Hydrochloride Tablets=A-Monos.pdf)
in nitric acid, is formed. This precipitate soon darkens by
reduction to metallic silver, the reduction being accelerated DEFINITION
by the addition of 6 N ammonium hydroxide. Apomorphine Hydrochloride Tablets contain NLT 90.0% and
NMT 110.0% of the labeled amount of C17H17NO2 HCl
ASSAY
1
/2H2O.
PROCEDURE
Sample solution: Dissolve 300 mg of Apomorphine IDENTIFICATION
Hydrochloride in 100 mL of glacial acetic acid with the aid PROCEDURE
of heat provided by a steam bath. Add 0.1 mL of acetic Analysis: To 5 mL of a filtered solution of Tablets,
anhydride to the hot solution, and stir for 5 min. Cool to containing about 10 mg of apomorphine hydrochloride, add
room temperature, and add 5 mL of mercuric acetate TS a slight excess of sodium bicarbonate solution (1 in 20): a
and 0.25 mL of crystal violet TS. white or greenish-white precipitate is formed. Add 3 drops
Analysis of iodine TS, and shake vigorously: an emerald-green color is
Sample: Sample solution produced. Add 5 mL of ether, and, after vigorous shaking,
Titrate with 0.1 N perchloric acid VS to a blue endpoint. allow the layers to separate: the ether is colored deep ruby-
Perform a blank determination and perform any necessary red while the water layer retains its green color.
correction (see Titrimetry 541). Each mL of 0.1 N ASSAY
perchloric acid is equivalent to 30.38 mg of C17H17NO2 PROCEDURE
HCl. Sample solution: Dissolve an equivalent to 50 mg of
Acceptance criteria: 98.5%100.5% apomorphine hydrochloride, from finely powdered tablets
IMPURITIES (NLT 20) in 25 mL of water in a separator. Add 500 mg of
Inorganic Impurities sodium bicarbonate, and completely extract with successive
RESIDUE ON IGNITION 281: NMT 0.1% small portions of ether. Combine the ether extracts in a
Organic Impurities separator, and wash them with three 5-mL portions of
PROCEDURE 1: ORDINARY IMPURITIES 466 water. Shake the combined water washings with 10 mL of
Solvent for sample solution: Methanol ether, and add this ether to the combined ether extracts.
Solvent for standard solution: Methanol Extract the ether solutions with 20.0 mL of 0.02 N sulfuric
Eluant: 1-Butanol, water, and formic acid (7:2:1) acid VS, and wash with three 5-mL portions of water.
Visualization: A freshly prepared mixture of 10% ferric Combine the acid extract and washings in a beaker, and
chloride solution and 5% potassium ferricyanide solution warm on a steam bath to expel any residual ether. Cool and
(2:1). Allow the chromatograms to develop until the add methyl red TS.
solvent front has moved about 8 cm. [NOTEThe Analysis: Titrate the excess acid with 0.02 N sodium
development time is 1.52 h.] Allow the plates to dry at hydroxide VS (see Titrimetry 541, Residual Titrations). Each
room temperature for 1 h prior to spraying. mL of 0.02 N sulfuric acid is equivalent to 6.256 mg of
PROCEDURE 2: DECOMPOSITION PRODUCTS C17H17NO2 HCl 1/2H2O.
Analysis: Shake 100 mg with 5 mL of ether. Acceptance criteria: 90.0%110.0%
Acceptance criteria: The latter acquires NMT a pale PERFORMANCE TESTS
reddish color. DISINTEGRATION 701
SPECIFIC TESTS Time: 15 min
OPTICAL ROTATION, Specific Rotation 781S: 60.5 to 63.0 UNIFORMITY OF DOSAGE UNITS 905
Sample solution: 15 mg/mL in dimethylsulfoxide Procedure for content uniformity
LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses Sample solution: Place 1 Tablet in a 500-mL volumetric
2.0%3.5% of its weight. flask containing 100 mL of 0.1 N hydrochloric acid, and
COLOR OF SOLUTION: Place 100 mg in a suitable test tube, shake for 15 min. Dilute with 0.1 N hydrochloric acid to
add 10 mL of cold, oxygen-free water, and agitate gently volume, mix, and filter, discarding the first 20 mL of
until dissolved: the color of the resulting solution, observed filtrate. Dilute a portion of the subsequent filtrate
promptly after the Apomorphine Hydrochloride has quantitatively and stepwise, if necessary, with 0.1 N
dissolved, is not more intense than that of a color standard hydrochloric acid to provide a solution containing
prepared as follows. Dissolve 5 mg of Apomorphine approximately 12 g/mL of apomorphine hydrochloride.
Hydrochloride in 100.0 mL of water. Transfer 1.0 mL of this Standard solution: 12 g/mL of anhydrous apomorphine
solution to a test tube of the same size as that used for the hydrochloride, from USP Apomorphine Hydrochloride RS in
Sample solution, dilute with 6 mL of water, add 1 mL of the same medium
266 Apomorphine / Official Monographs USP 32
Spectrometric conditions DEFINITION

Analytical wavelength: 273 nm Apraclonidine Hydrochloride contains NLT 98.0% and NMT
Cell: 1 cm 102.0% of C9H10Cl2N4 HCl, calculated on the dried basis.
Blank: 0.1 N hydrochloric acid
Samples: Sample solution and Standard solution A. INFRARED ABSORPTION 197K
Concomitantly determine the absorbances of the Sample B. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the
solution and the Standard solution. Calculate the requirements
percentage of C17H17NO2 HCl 1/2H2O in the Tablet
taken: ASSAY
PROCEDURE
Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 Sample solution: 125 mg of Apraclonidine Hydrochloride
in 40 mL of glacial acetic acid. Add 10 mL of mercuric
AU = absorbance of the Sample solution acetate TS.
AS = absorbance of the Standard solution Analysis: Titrate with 0.1 N perchloric acid VS, using a
CS = concentration of anhydrous apomorphine calomel-glass electrode system (see Titrimetry 541). Perform
hydrochloride in the Standard solution (g/mL) a blank determination. Each mL of 0.1 N perchloric acid is
CU = concentration of apomorphine hydrochloride in equivalent to 14.08 mg of C9H10Cl2N4 HCl.
the solution from the Tablet, based upon the Acceptance criteria: 98.0%102.0%
labeled quantity per Tablet and the extent of
dilution (g/mL) IMPURITIES
Mr1 = molecular weight of apomorphine hydrochloride Inorganic Impurities
hemihydrate, 312.80 RESIDUE ON IGNITION 281: NMT 0.1%
Mr2 = molecular weight of anhydrous apomorphine HEAVY METALS, Method II 231: NMT 20 ppm
hydrochloride, 303.79 Organic Impurities
Acceptance criteria: Meet the requirements PROCEDURE
Phosphate buffer: 6.8 mL of phosphoric acid in a 2000-
SPECIFIC TESTS mL volumetric flask. Add 1900 mL of water. Adjust with
COLOR OF SOLUTION: Dissolve a quantity of powdered sodium hydroxide solution (1 in 2) to a pH of 3.0, and
Tablets, equivalent to 5 mg of apomorphine hydrochloride, dilute with water to volume.
in water to make 100.0 mL. Transfer 1.0 mL of the solution Mobile phase: Acetonitrile, methanol, and Phosphate
to a test tube, dilute with 6 mL of water, and, if necessary, buffer (14:1:10)
filter through a small pledget of cotton. Add 1 mL of sodium System suitability solution: 0.8 mg/mL of USP
bicarbonate solution (1 in 20), then add 0.50 mL of iodine Apraclonidine Hydrochloride RS in Mobile phase
TS. Allow to stand for 30 s, then add 0.60 mL of sodium Sample solution: 0.8 mg/mL of Apraclonidine
thiosulfate solution (1 in 40), and dilute with water to 10 Hydrochloride in Mobile phase
mL. This solution represents the color standard. Chromatographic system
Place a quantity of powdered Tablets, equivalent to about (See Chromatography 621, System Suitability.)
50 mg of apomorphine hydrochloride, in a test tube of Mode: LC
suitably small size. Add 10.0 mL of cold, oxygen-free water, Detector: UV 220 nm
insert the stopper in the test tube, and agitate gently until Column: 8-mm 100-mm; packing L7
no more dissolves. If necessary, filter immediately through Flow rate: 3 mL/min
a small pledget of cotton. The color of the solution, Injection size: 20 L
observed promptly after preparation, is not more intense System suitability
than that of the color standard. Use closely matched test Sample: System suitability solution
tubes for the comparison. Suitability requirements
Tailing factor: NMT 2.2 for the apraclonidine peak
ADDITIONAL REQUIREMENTS Relative standard deviation: NMT 2.0%
PACKAGING AND STORAGE: Preserve in tight, light-resistant Analysis
containers. Sample: Sample solution
USP REFERENCE STANDARDS 11 [NOTEAllow about five times the elution time of
USP Apomorphine Hydrochloride RS apraclonidine before making the next injection.]
Calculate the percentage of each peak, other than the
solvent peak and the apraclonidine peak, in the portion
of Apraclonidine Hydrochloride taken:
Apraclonidine Hydrochloride
(Comment on this Monograph)id=m5700=Apraclonidine Result = (rU/rT) 100
Hydrochloride=A-Monos.pdf)
rU = response of each peak other than the principal
peak
rT = sum of the responses of all of the peaks,
excluding that of the solvent peak
Acceptance criteria
Individual impurities: NMT 1.0%
C9H10Cl2N4 HCl 281.57 SPECIFIC TESTS

1,4-Benzenediamine, 2,6-dichloro-N1-2-imidazolidinylidene-, PH 791: 5.06.6, in a solution (1 in 100)
monohydrochloride; LOSS ON DRYING 731: Dry it in a vacuum at 105 for 3 h: it
2-[(4-Amino-2,6-dichlorophenyl)imino]imidazolidine loses NMT 1.0% of its weight.
monohydrochloride [73218-79-8].
USP 32 Official Monographs / Apraclonidine 267
ADDITIONAL REQUIREMENTS Standard solution: 11.5 g/mL of USP Apraclonidine

PACKAGING AND STORAGE: Preserve in tight, light-resistant Hydrochloride RS (equivalent to about 10 g of
containers. apraclonidine per mL) diluted in Mobile phase
USP REFERENCE STANDARDS 11 System suitability solution: Transfer 1 mL of
USP Apraclonidine Hydrochloride RS propiophenone to a 100-mL volumetric flask, and dilute
with methanol to volume. Transfer 3.0 mL of this solution to
a 50-mL volumetric flask, and dilute with methanol to
volume. Transfer 1.0 mL of this solution and 5.0 mL of the
Apraclonidine Ophthalmic Solution Standard stock solution to a 100-mL volumetric flask, and
(Comment on this Monograph)id=m5705=Apraclonidine dilute with Mobile phase to volume.
Ophthalmic Solution=A-Monos.pdf) Sample stock solution: Equivalent to 0.2 mg/mL of
apraclonidine, from Ophthalmic Solution in water
DEFINITION Sample solution: 10 g/mL of Sample stock solution diluted
Apraclonidine Ophthalmic Solution is a sterile, aqueous solution in Mobile phase
of Apraclonidine Hydrochloride. It contains an amount of Chromatographic system
apraclonidine hydrochloride (C9H10Cl2N4 HCl) equivalent to (See Chromatography 621, System Suitability.)
NLT 90.0% and NMT 115.0% of the labeled amount of Mode: LC
apraclonidine (C9H10Cl2N4). Detector: UV 254 nm
Column: 8-mm 100-mm; packing L7
IDENTIFICATION Flow rate: 3 mL/min
A. The retention time of the major peak of the Sample Injection size: 20 L
solution corresponds to that of the major peak of the System suitability
Standard solution, as obtained in the Assay. Sample: System suitability solution
B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST [NOTEThe relative retention times for apraclonidine and
(see Chromatography 621.) propiophenone are 0.6 and 1.0, respectively.]
Adsorbent: 0.2-mm layer of chromatographic silica gel Suitability requirements
mixture, or equivalent Resolution: NLT 3.0 between the analyte and
Standard solution: 11.5 mg/mL of USP Apraclonidine propiophenone peaks
Hydrochloride RS in methanol Column efficiency: NLT 1000 theoretical plates for the
Sample solution: Apraclonidine Ophthalmic Solution analyte peak
Application volume: 2 L Tailing factor: NMT 2.2 for the analyte peak
Developing solvent system: Chloroform, methanol, and Relative standard deviation: NMT 2.0%
ammonium hydroxide (37:11:2) Analysis
Spray reagent: 1 mg/mL of fluorescamine in acetone Samples: Standard solution and Sample solution
Analysis Calculate the percentage of C9H10Cl2N4 in the portion of
Samples: Standard solution and Sample solution Solution taken:
Allow the applications to dry, and develop the
chromatogram in the developing solvent system, until the Result = (rU/rS) (CS/CU) (Mr1/Mr2) x 100
solvent front has moved about three-fourths of the length
of the plate. Locate the spots on the plate by viewing rU = peak response of apraclonidine from the Sample
under short-wavelength UV light. [NOTEThe solution
apraclonidine spot should appear as a blue spot.] Spray rS = peak response of apraclonidine from the
the plate with Spray reagent. [NOTEAvoid prolonged or Standard solution
repeated breathing of the aerosol from the fluorescamine CS = concentration of USP Apraclonidine
spray. Also avoid prolonged or repeated contact with skin. Hydrochloride RS in the Standard solution
Fluorescamine solution should be sprayed only in a hood.] (g/mL)
Examine the plate under normal light and long- CU = nominal concentration of apraclonidine in the
wavelength UV light. [NOTEThe apraclonidine spot Sample solution (g/mL)
should appear as a yellow spot under normal light and as Mr1 = molecular weight of apraclonidine, 245.11
a white spot under long-wavelength UV light.] Mr2 = molecular weight of apraclonidine hydrochloride,
Acceptance criteria: The RF value and appearance of the 281.57
principal spot of the Sample solution corresponds to that of Acceptance criteria: 90.0%115.0%
SPECIFIC TESTS
ASSAY STERILITY TESTS 71: It meets the requirements when tested
PROCEDURE as directed for Test for Sterility of the Product to Be Examined,
Phosphate buffer: 6.8 mL of phosphoric acid in a 2000-mL Membrane Filtration.
volumetric flask, and add 1900 mL of water. Adjust with PH 791: 4.47.8
sodium hydroxide solution (1 in 2) to a pH of 3.0, and
dilute with water to volume. ADDITIONAL REQUIREMENTS
Mobile phase: Acetonitrile, methanol, and Phosphate buffer PACKAGING AND STORAGE: Preserve in tight, light-resistant
(15:1:34) containers.
Standard stock solution: 0.23 mg/mL of USP Apraclonidine USP REFERENCE STANDARDS 11
Hydrochloride RS in water USP Apraclonidine Hydrochloride RS
268 Aprotinin / Official Monographs USP 32
Aprotinin immediately with 0.0015 M borate buffer to 40.0 mL. Allow

(Comment on this Monograph)id=m5740=Aprotinin=A- to stand at room temperature for 10 min, and then keep in
Monos.pdf) ice water. [NOTEUse within 6 h of preparation.]
Dilute trypsin solution: Trypsin solution and 0.0015 M
borate buffer (0.5:9.5)
Allow to stand at room temperature for 10 min, and then
keep in ice water.
Substrate solution: 6.9 mg/mL of N-benzoyl-L-arginine
ethyl ester hydrochloride [NOTEUse within 2 h.]
Analysis: Mix 9.0 mL of 0.0015 M borate buffer and 1.0 mL
of Substrate solution in a jacketed glass vessel with a capacity
C284H432N84O79S7 6511.44 of 30 mL and that contains a stirring device. The lid of the
Trypsin inhibitor, pancreatic basic; reaction vessel should contain five holes to accommodate
L-Arginyl-L-prolyl-L-aspartyl-L-phenylalanyl-L-cysteinyl-L-leucyl-L- the electrodes, the tip of a buret, a tube for the admission
glutamyl-L-prolyl-L-prolyl-L-tyrosyl-L-threonylglycyl-L-prolyl-L- of nitrogen, and the introduction of reactants. An
cysteinyl-L-lysyl-L-alanyl-L-arginyl-L-isoleucyl-L-isoleucyl-L- automated or manual titration apparatus may be used.
arginyl-L-tyrosyl-L-phenylalanyl-L-tyrosyl-L-asparaginyl-L-alanyl- Adjust to a pH of 8.0 by the addition of 0.1 N sodium
L-lysyl-L-alanylglycyl-L-leucyl-L-cysteinyl-L-glutaminyl-L-threonyl- hydroxide VS. Maintain an atmosphere of nitrogen within
L-phenylalanyl-L-valyl-L-tyrosylglycylglycyl-L-cysteinyl-L-arginyl- the vessel, and stir continuously. When the temperature has
L-alanyl-L-lysyl-L-arginyl-L-asparaginyl-L-asparaginyl-L- reached equilibrium at 25 0.1, add 1.0 mL of Trypsin and
phenylalanyl-L-lysyl-L-seryl-L-alanyl-L-glutamyl-L-aspartyl-L- aprotinin solution, and start a timer. Maintain at a pH of 8.0
cysteinyl-L-methionyl-L-arginyl-L-threonyl-L- by the addition of 0.1 N sodium hydroxide VS, and note the
cysteinylglycylglycyl-L-alanine cyclic (555), (1438), volume added every 30 s. Continue the reaction for 6 min.
(3051) tris(disulfide) [9087-70-1]. Determine the volume of 0.1 N sodium hydroxide added
per s, in mL (n1). Carry out a similar titration using 1.0 mL
DEFINITION of the Dilute trypsin solution. Determine the volume of 0.1 N
Aprotinin is a polypeptide consisting of a chain of 58 amino sodium hydroxide added per s, in mL (n2). For the
acid residues, which inhibits stoichiometrically the activity of lyophilized powder, calculate the aprotinin activity in USP
several proteolytic enzymes such as chymotrypsin, kallikrein, Aprotinin Units/mg:
plasmin, and trypsin. Aprotinin is obtained from bovine tissues
and purified by a suitable process, and is stored as a bulk Result = 4000 (2n2 n1)/m
solution or lyophilized powder. Its potency, calculated on the
dried basis, is NLT 3 USP Aprotinin Units/mg. In addition, the m = quantity of Aprotinin used to prepare 1 mL of
method of manufacture is validated to result in NMT 0.2 g of the Sample solution (mg)
histamine per 3 USP Aprotinin Units using validated methods. For the concentrated solution, calculate the USP Aprotinin
The origin and sourcing of bovine material must be specified Units/mL using the following formula:
in compliance with FDA requirements. The manufacturing
process is validated to demonstrate the clearance of potential Result = 4000 (2n2 n1) D
infectious agents (i.e., viruses, TSE agents). One USP Aprotinin
Unit is equivalent to 1800 Kallikrein Inhibition Units (K.I.U.). D = dilution factor of the concentrated solution used
to prepare the Sample solution
IDENTIFICATION Acceptance criteria: NLT 3 USP Aprotinin Units/mg
A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 potency
Sample solution: 15 USP Aprotinin Units/mL of Aprotinin
Developing solvent system: A mixture of glacial acetic acid IMPURITIES
and water (5:4) containing 100 g/L of sodium acetate Organic Impurities
Cupric chloride solution: 10 mg/mL of cupric chloride PROCEDURE 1: LIMIT OF DES-ALA-APROTININ AND DES-ALA-DES-
Spray reagent: 0.1 g of ninhydrin in a mixture of Cupric GLY-APROTININ
chloride solution, glacial acetic acid, and alcohol (6:21:70) Sample solution: 47 USP Aprotinin Units/mL of Aprotinin
Analysis: Proceed as directed in the chapter, except to [NOTEDilute a concentrated solution of Aprotinin with
spray the plate with the Spray reagent, and heat at 60 to water, or weigh out Aprotinin and dissolve in water.]
visualize the spots. Standard solution: Dilute USP Aprotinin RS with water to
B. The retention time of the major peak of the Sample obtain a solution having a concentration similar to that of
solution corresponds to that of the System suitability solution, the Sample solution.
as obtained in the test for Limit of N-pyroglutamyl-aprotinin Capillary zone electrophoresis buffer: 8.21 g of
and related compounds. monobasic potassium phosphate in 400 mL of water.
Adjust with phosphoric acid to a pH of 3.0, and dilute with
ASSAY water to 500 ml.
PROCEDURE Capillary zone electrophoresis system
0.0015 M borate buffer: Transfer 0.93 g of boric acid into (See Biotechnology-Derived ArticlesCapillary Electrophoresis
a 1000-mL volumetric flask, dissolve in 900 mL of water, 1053.)
adjust with 5 N sodium hydroxide to a pH of 8.0, and dilute The capillary electropherograph is equipped with a 214-nm
with water to volume. Transfer 100 mL of this solution into detector and a 45- to 60-cm uncoated fused silica
a 1000-mL volumetric flask, and dilute with water to capillary with an internal diameter of 75 m with the
volume. temperature controlled at 25. Apply a field strength of
Sample solution: 1.67 USP Aprotinin Units/mL (about 0.6 0.2 kV/cm for 30 min, using Capillary zone electrophoresis
mg/mL) of Aprotinin in 0.0015 M borate buffer buffer as the electrolyte in both buffer reservoirs.
Trypsin solution: 4300 USP Trypsin Units/mL of USP Electropherograph the Standard solution, and record the
Trypsin Crystallized RS in 0.001 N hydrochloric acid [NOTE peak responses as directed for Analysis: the relative
Use a freshly prepared solution, and keep in ice water.] migration times are 0.98 for des-Ala-des-Gly-aprotinin,
Trypsin and aprotinin solution: To 4.0 mL of the Trypsin 0.99 for des-Ala-aprotinin, and 1 for Aprotinin. The
solution, add 1.0 mL of the Sample solution. Dilute resolution, RS, between the des-Ala-des-Gly-aprotinin and
USP 32 Official Monographs / Aprotinin 269
des-Ala-aprotinin peaks is NLT 0.8, and the resolution ri = response of each impurity peak
between the des-Ala-aprotinin and aprotinin peaks is NLT rT = sum of the responses of all peaks of the Sample
0.5. The migration time for the Aprotinin peak is 1925 solution
min. The tailing factor, T, of the Aprotinin peak is NMT 3 Acceptance criteria
(see Chromatography 621 for calculation). The baseline is N-pyroglutamyl-aprotinin: NMT 1.0%
stable and shows little drift. Rinse the capillary for at least Any other impurity: NMT 0.5%
1 min with at least 10 total capillary volumes of 0.1 N Sum of all unknown impurities: NMT 1.0%
sodium hydroxide, followed by at least 10 total capillary PROCEDURE 3: LIMIT OF HIGH MOLECULAR WEIGHT PROTEINS
volumes of water, and by at least 20 capillary volumes of Mobile phase: Acetonitrile, glacial acetic acid, and water
Capillary zone electrophoresis buffer between injections. (1:1:3)
Analysis: Transfer a volume of the Sample solution, System suitability solution: Aprotinin solution that
approximately 15 mL, into the anodic end of the capillary contains 5 USP Aprotinin Units/mL with 2% aprotinin
(apply differential pressure of 3.5 kPa for 3 s either by oligomers [NOTEThis solution can be obtained by heating
vacuum or pressure), record an electropherogram, and lyophilized aprotinin at 112 for 2 h and dissolving the
measure the peak areas. solid at the specified concentration in water.]
Calculate the percentage contents of des-Ala-des-Gly- Sample solution: 5 USP Aprotinin Units/mL of Aprotinin in
aprotinin and des-Ala-aprotinin: water
Result = (ri/rT) 100 (See Chromatography 621, System Suitability.)
Mode: LC
ri = peak response corresponding to des-Ala-des-Gly- Detector: UV 280 nm
aprotinin or des-Ala-aprotinin Column: Series of three 7.8-mm 30-cm columns;
rT = sum of the responses of des-Ala-des-Gly- packing L33
aprotinin, des-Ala-aprotinin, and aprotinin Flow rate: 1 mL/min
peaks Injection size: 100 L
Acceptance criteria System suitability
des-Ala-des-Gly-aprotinin: NMT 8.0% Sample: System suitability solution
des-Ala-aprotinin: NMT 7.5% Suitability requirements
PROCEDURE 2: LIMIT OF N-PYROGLUTAMYL-APROTININ AND [NOTEThe relative retention times for the dimer and
RELATED COMPOUNDS aprotinin are 0.9 and 1.0, respectively.]
Solution A: 3.52 mg/mL of monobasic potassium Retention time: 24.525.5 min for aprotinin
phosphate and 7.26 mg/mL of dibasic sodium phosphate Resolution: NLT 1.3 between the dimer peak and the
Solution B: 3.52 mg/mL of monobasic potassium aprotinin peak
phosphate, 7.26 mg/mL of dibasic sodium phosphate, and Tailing factor: NMT 2.5 for the aprotinin peak
66.07 mg/mL of ammonium sulfate Analysis
System suitability solution: 5 USP Aprotinin Units/mL of Sample: Sample solution
USP Aprotinin System Suitability RS in Solution A Calculate the percentage of each oligomer peak:
Sample solution: 5 USP Aprotinin Units/mL of Aprotinin in
Solution A Result = (ri/rT) 100
(See Chromatography 621, System Suitability.) ri = response of each peak having a retention time
Mode: LC less than that of aprotinin monomer
Detector: UV 210 nm rT = sum of the responses of all peaks
Column: 7.5-mm 7.5-cm; packing L52 Acceptance criteria: NMT 1.0%
Temperature: 40 (constant temperature)
Flow rate: 1 mL/min SPECIFIC TESTS
ABSORBANCE
(See Spectrophotometry and Light-Scattering 851.) Prepare a
Time (min) Solution A Solution B
solution containing 3.0 USP Aprotinin Units/mL. The
0 92 8 solution shows an absorption maximum at 277 nm. The
21 64 36 absorbance at the maximum is NMT 0.80.
SAFETY: Prepare a solution of Aprotinin that contains 4 USP
30 0 100
Aprotinin Units/mL using a sufficient quantity of Water for
31 92 8 Injection. It meets the requirements when tested as directed
40 92 8 in Biological Reactivity Tests, In Vivo 88, Safety Tests
Biologicals.
Injection size: 40 L SPECIFIC ACTIVITY OF THE DRY RESIDUE
System suitability [NOTEThis test should only be performed when product is
Sample: System suitability solution a concentrated solution.]
[NOTEThe relative retention times for N-pyroglutamyl- Analysis: Evaporate 25.0 mL of Aprotinin concentrated
aprotinin and aprotinin are 0.9 and 1.0, respectively.] solution to dryness in a water bath, dry the residue at 110
Suitability requirements for 15 h, and weigh. From the weight of the residue and
Retention time: 1720 min for aprotinin the activity determined in the Assay, calculate the number of
Resolution: NLT 1.0 between N-pyroglutamyl-aprotinin USP Aprotinin Units/mg of dry residue.
and aprotinin Aceptance criteria: NLT 3.0 USP Aprotinin Units/mg of
Tailing factor: NMT 2.0 for the aprotinin peak dried residue is found.
Analysis LOSS ON DRYING 731
Sample: Sample solution [NOTEThis test should only be performed on the
Calculate the percentage of each impurity peak: lyophilized powder.]
Result = (ri/rT) 100
270 Aprotinin / Official Monographs USP 32
Dry 100 mg in a capillary-stoppered bottle in a vacuum at a reaction vessel should contain five holes to accommodate
pressure not exceeding 5 mm of mercury at 60 for 3 h: it the electrodes, the tip of a buret, a tube for the admission
loses NMT 6.0% of its weight. of nitrogen, and the introduction of reactants. An
BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.14 USP automated or manual titration apparatus may be used.
Endotoxin Unit/USP Aprotinin Unit. Use a solution that Adjust to a pH of 8.0 by the addition of 0.1 N sodium
contains 6 USP Aprotinin Units/mL. hydroxide VS. Maintain an atmosphere of nitrogen within
the vessel, and stir continuously. When the temperature has
ADDITIONAL REQUIREMENTS reached equilibrium at 25 0.1, add 1.0 mL of Trypsin and
PACKAGING AND STORAGE: For lyophilized powder, preserve aprotinin solution, and start a timer. Maintain at a pH of 8.0
in tight containers, and store in a cold place. Protect from by the addition of 0.1 N sodium hydroxide VS, and note the
light. For bulk solution, preserve in tight containers at a volume added every 30 s. Continue the reaction for 6 min.
temperature not exceeding 25. Avoid freezing. Determine the volume of 0.1 N sodium hydroxide added
LABELING: The labeling states the source of material and the per s, in mL (n1). Carry out a similar titration using 1.0 mL
number of Kallikrein Inhibition Units/mg or the number of of the Dilute trypsin solution. Determine the volume of 0.1 N
Kallikrein Inhibition Units/mL. sodium hydroxide added per s, in mL (n2). For the
USP REFERENCE STANDARDS 11 lyophilized powder, calculate the aprotinin activity in USP
USP Aprotinin RS Aprotinin Units/mg:
USP Aprotinin System Suitability RS
USP Endotoxin RS Result = 4000 (2n2 n1)/m
USP Trypsin Crystallized RS
m = quantity of Aprotinin used to prepare 1 mL of
the Sample solution (mg)
For the concentrated solution, calculate the USP Aprotinin
Aprotinin Injection Units/mL:
(Comment on this Monograph)id=m5745=Aprotinin
Injection=A-Monos.pdf) Result = 4000 (2n2 n1) D
DEFINITION D = dilution factor of the concentrated solution used

Aprotinin Injection is a sterile solution of Aprotinin in Water for to prepare the Sample solution
Injection that also contains sodium chloride. One USP Acceptance criteria: 90.0%110.0%
Aprotinin Unit is equivalent to 1800 Kallikrein Inhibition Units
(K.I.U.). It contains NLT 90.0% and NMT 110.0% of the OTHER COMPONENTS
potency stated on the label, expressed in Kallikrein Inhibition SODIUM CHLORIDE CONTENT
Units per mL. Sample: 5.0 mL of Injection
Analysis: Pipet Sample into 50 mL of water in a beaker. Add
IDENTIFICATION 10 mL of 25% nitric acid. Titrate with 0.1 N silver nitrate VS
A. The retention time of the major peak of the Sample to a potentiometric endpoint, using a silver combination
solution corresponds to that of the Standard solution, as electrode. Perform a blank determination (see Titrimetry
obtained in the test for Limit of N-pyroglutamyl-aprotinin and 541). Each mL of 0.1 N silver nitrate is equivalent to 5.844
related compounds. mg of sodium chloride.
B. The determination of activity by the Assay is based on Acceptance criteria: 42.547.5 mg
the specific inhibition of trypsin.
IMPURITIES
PROCEDURE PROCEDURE: LIMIT OF HIGH MOLECULAR WEIGHT PROTEINS
0.0015 M Borate buffer: Transfer 0.93 g of boric acid into Mobile phase: Acetonitrile, glacial acetic acid, and water
a 1000-mL volumetric flask, dissolve in 900 mL of water, (1:1:3)
adjust with 5 N sodium hydroxide to a pH of 8.0, and dilute System suitability solution: Aprotinin solution that
with water to volume. Transfer 100 mL of this solution into contains 5 USP Aprotinin Units/mL with 2% aprotinin
a 1000-mL volumetric flask, and dilute with water to oligomers [NOTEThis solution can be obtained by heating
volume. lyophilized aprotinin at 112 for 2 h and dissolving the
Sample solution: 1.67 USP Aprotinin Units/mL (about 0.6 solid at the specified concentration in water.]
mg/mL) of Aprotinin in 0.0015 M Borate buffer Sample solution: 5 USP Aprotinin Units/mL of Aprotinin
Trypsin solution: 4300 USP Trypsin Units/mL of USP Chromatographic system
Trypsin Crystallized RS in 0.001 N hydrochloric acid [NOTE (see Chromatography 621, System Suitability.)
Use a freshly prepared solution, and keep in ice water.] Mode: LC
Trypsin and aprotinin solution: To 4.0 mL of the Trypsin Detector: UV 280 nm
solution, add 1.0 mL of the Sample solution. Dilute Column: Series of three 7.8-mm 30-cm columns;
immediately with 0.0015 M Borate buffer to 40.0 mL. Allow packing L33
to stand at room temperature for 10 min, and then keep in Flow rate: 1 mL/min
ice water. [NOTEUse within 6 h of preparation.] Injection size: 100 L
Dilute trypsin solution: Trypsin solution and 0.0015 M System suitability
Borate buffer (0.5:9.5). Allow to stand at room temperature Sample: System suitability solution
for 10 min, and then keep in ice water. [NOTEThe relative retention times of dimer and
Substrate solution: 6.9 mg/mL of N-benzoyl-L-arginine aprotinin are 0.9 and 1.0, respectively.]
ethyl ester hydrochloride in water [NOTEUse within 2 h.] Suitability requirements
Analysis: Mix 9.0 mL of 0.0015 M Borate buffer and 1.0 mL Resolution: NLT 1.3 between the dimer peak and the
of Substrate solution in a jacketed-glass vessel with a capacity aprotinin peak
of 30 mL and that contains a stirring device. The lid of the
USP 32 Official Monographs / Arginine 271
Tailing factor: NMT 2.5 for the aprotinin peak Acceptance criteria
Retention time: 24.525.5 min for aprotinin N-pyroglutamyl-aprotinin: NMT 1.0%
Analysis Any other impurity: NMT 0.5%
Sample: Sample solution Sum of all unknown impurities: NMT 1.0%
Calculate the percentage of each oligomer peak in the BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.14 USP
chromatogram: Endotoxin Units/USP Aprotinin Unit.
Result = (ri/rT) 100 ADDITIONAL REQUIREMENTS
ri = response of each peak having a retention time store at up to 25, and avoid freezing.
less than that of aprotinin monomer USP REFERENCE STANDARDS 11
rT = sum of the responses of all peaks USP Aprotinin RS
Acceptance criteria: Sum of all oligomers is NMT 1.5% USP Aprotinin System Suitability RS
USP Endotoxin RS
SPECIFIC TESTS USP Trypsin Crystallized RS
as directed under Test for Sterility of the Product to Be
Examined, Membrane Filtration.
PARTICULATE MATTER IN INJECTIONS 788: Meets the Arginine
requirements (Comment on this Monograph)id=m5840=Arginine=A-
PH 791: 4.56.5 Monos.pdf)
INJECTIONS 1: Meets the requirements
LIMIT OF N-PYROGLUTAMYL-APROTININ AND RELATED
COMPOUNDS
phosphate and 7.26 mg/mL of dibasic sodium phosphate in
water
Solution B: 3.52 mg/mL of monobasic potassium
phosphate, 7.26 mg/mL of dibasic sodium phosphate, and
66.07 mg/mL of ammonium sulfate dissolved in water C6H14N4O2 174.20
System suitability solution: 5 USP Aprotinin Units/mL of L-Arginine [74-79-3].
USP Aprotinin System Suitability RS in Solution A
Sample solution: 5 USP Aprotinin Units/mL of Aprotinin in DEFINITION
Solution A Arginine contains NLT 98.5% and NMT 101.5% of (C6H14N4O2),
Chromatographic system as L-arginine, calculated on the dried basis.
(see Chromatography 621, System Suitability.)
Detector: UV 210 nm INFRARED ABSORPTION 197K
Column: 7.5-mm 7.5-cm; packing L52
Temperature: 40 (constant temperature) ASSAY
Flow rate: 1 mL/min PROCEDURE
Sample: 80 mg of Arginine
Analysis: Dissolve the Sample in a mixture of 3 mL of
Time (min) Solution A (%) Solution B (%) formic acid and 50 mL of glacial acetic acid in a 125-mL
0 92 8 flask. Titrate with 0.1 N perchloric acid VS. Perform a blank
21 64 36 determination (see Titrimetry 541). Each mL of 0.1 N
perchloric acid is equivalent to 8.710 mg of C6H14N4O2.
30 0 100 Acceptance criteria: 98.5%101.5%
31 92 8
IMPURITIES
40 92 8
Injection size: 40 L CHLORIDE AND SULFATE, Chloride 221: NMT 500 ppm. A
System suitability 1.0-g portion shows no more chloride than corresponds to
Sample: System suitability solution 0.70 mL of 0.020 N hydrochloric acid.
[NOTEThe relative retention times for N-pyroglutamyl- CHLORIDE AND SULFATE, Sulfate 221: NMT 300 ppm. A 1.0-
aprotinin and aprotinin are 0.9 and 1.0, respectively.] g portion shows no more sulfate than corresponds to 0.30
Suitability requirements mL of 0.020 N sulfuric acid.
Resolution: NLT 1.0 between N-pyroglutamyl-aprotinin IRON 241: NMT 30 ppm
and aprotinin HEAVY METALS 231, Method I: NMT 15 ppm
Tailing factor: NMT 2.0 for the aprotinin peak Organic Impurities
Retention time: 1720 min for aprotinin PROCEDURE
Sample: Sample solution mixture
Calculate the percentage of each impurity peak in the Standard solution: 0.05 mg/mL of USP L-Arginine RS in
chromatogram: 0.1 N hydrochloric acid [NOTEThis solution has a
Result = (ri/rT) 100 concentration equivalent to 0.5% of that of the Sample
solution.]
ri = response of each impurity peak Sample solution: 10 mg/mL of Arginine in 2 N
rT = sum of the responses of all peaks from the hydrochloric acid
Sample solution
272 Arginine / Official Monographs USP 32
System suitability solution: 0.4 mg/mL each of USP L- until the silver chloride flocculates and the mixture acquires
Arginine RS and USP L-Lysine Hydrochloride RS in 0.1 N a faint pink color, using 1 mL of dichlorofluorescein TS as
hydrochloric acid indicator. Each mL of 0.1 N silver nitrate is equivalent to
Application volume: 5 L 3.545 mg of chloride.
Spray reagent: 2 mg/mL of ninhydrin in a mixture of Acceptance criteria: 16.5%17.1%
butyl alcohol and 2 N acetic acid (19:1)
Application volume: 5 L IMPURITIES
Developing solvent system: Isopropyl alcohol and Inorganic Impurities
ammonium hydroxide (7:3) RESIDUE ON IGNITION 281: NMT 0.1%
Analysis CHLORIDE AND SULFATE, Sulfate 221: NMT 300 ppm. A 1.6-
Samples: Standard solution, Sample solution, and System g portion shows no more sulfate than corresponds to 0.50
suitability solution mL of 0.020 N sulfuric acid.
Proceed as directed under Chromatography 621, Thin- HEAVY METALS, Method I 231: NMT 20 ppm. Proceed as
Layer Chromatography. Dry the plate between 100 and directed except to dissolve 1.0 g in 20 mL of water, add 2
105 until the ammonia disappears completely. Spray mL of 1 N acetic acid, and dilute with water to 25 mL.
with Spray reagent, and heat between 100 and 105 for Organic Impurities
about 15 min. Examine the plate under white light. The PROCEDURE
chromatogram obtained from the System suitability Adsorbent: 0.25-mm layer of chromatographic silica gel
solution exhibits two clearly separated spots. Any mixture
secondary spot from the Sample solution is not larger or Standard solution: 0.05 mg/mL of USP Arginine
more intense than the principal spot from the Standard Hydrochloride RS in water. [NOTEThis solution has a
solution. concentration equivalent to 0.5% of that of the Sample
Acceptance criteria solution.]
Individual impurities: NMT 0.5% Sample solution: 10 mg/mL of Arginine Hydrochloride in
Total impurities: NMT 2.0% water
System suitability solution: 0.4 mg/mL each of USP
SPECIFIC TESTS Arginine Hydrochloride RS and USP L-Lysine Hydrochloride
OPTICAL ROTATION, Specific Rotation 781S: +26.3 to RS in water
+27.7 Spray reagent: 2 mg/mL of ninhydrin in butyl alcohol and
Sample solution: 80 mg/mL in 6 N hydrochloric acid 2 N acetic acid (19:1)
LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT Application volume: 5 L
0.5% of its weight. Developing solvent system: Isopropyl alcohol and
ammonium hydroxide (7:3)
PACKAGING AND STORAGE: Preserve in well-closed containers. Samples: Standard solution, Sample solution, and System
USP REFERENCE STANDARDS 11 suitability solution
USP L-Arginine RS Proceed as directed for Chromatography 621, Thin-Layer
USP L-Lysine Hydrochloride RS Chromatography. Dry the plate between 100 and105
until the ammonia disappears completely. Spray with
Spray reagent, and heat between 100 and 105 for 15
Arginine Hydrochloride min. Examine the plate under white light. The
chromatogram from the System suitability solution
(Comment on this Monograph)id=m5850=Arginine exhibits two clearly separated spots. Any secondary spot
Hydrochloride=A-Monos.pdf) from the Sample solution is not larger or more intense
C6H14N4O2 HCl 210.66 than the principal spot from the Standard solution.
L-Arginine monohydrochloride; Acceptance criteria
L-(+)-Arginine monohydrochloride [1119-34-2]. Individual impurities: NMT 0.5%
DEFINITION
Arginine Hydrochloride contains NLT 98.5% and NMT 101.5% SPECIFIC TESTS
of C6H14N4O2 HCl, calculated on the dried basis. OPTICAL ROTATION, Specific Rotation 781S: +21.4 to
+23.6 (t = 20)
IDENTIFICATION Sample solution: 80 mg/mL in 6 N hydrochloric acid
INFRARED ABSORPTION 197K LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT
0.2% of its weight.
ASSAY
Sample: 100 mg of Arginine Hydrochloride PACKAGING AND STORAGE: Preserve in well-closed containers.
Analysis: Dissolve Sample in 3 mL of 98% formic acid and USP REFERENCE STANDARDS 11
50 mL of glacial acetic acid. Add 6 mL of mercuric acetate USP Arginine Hydrochloride RS
TS and titrate with 0.1 N perchloric acid VS. Perform a blank USP L-Lysine Hydrochloride RS
determination (see Titrimetry 541). Each mL of 0.1 N
perchloric acid is equivalent to 10.53 mg of C6H14N4O2 HCl.
Acceptance criteria: 98.5%101.5% Arginine Hydrochloride Injection
OTHER COMPONENTS (Comment on this Monograph)id=m5880=Arginine
CHLORIDE CONTENT Hydrochloride Injection=A-Monos.pdf)
Sample: 350 mg
[NOTEUse a porcelain casserole.] DEFINITION
Analysis: Add 140 mL of water and 1 mL of Arginine Hydrochloride Injection is a sterile solution of Arginine
dichlorofluorescein TS. Titrate with 0.1 N silver nitrate VS Hydrochloride in Water for Injection. It contains NLT 9.5% and
USP 32 Official Monographs / Arsanilic 273
NMT 10.5% of C6H14N4O2 HCl. It contains no antimicrobial USP Endotoxin RS

agents.
[NOTEThe chloride ion content of Arginine Hydrochloride
Injection is approximately 475 mEq/L.]
Arsanilic Acid
IDENTIFICATION (Comment on this Monograph)id=m5940=Arsanilic Acid=A-
A. Transfer 1 mL of the Injection to a 200-mL volumetric Monos.pdf)
flask, and dilute with water to volume. To 1 mL of this
dilution add 2 mL of a solution of 0.02% 8-hydroxyquinoline
in 3 N sodium hydroxide, and add 1 mL of 0.1% N-
bromosuccinimide solution.
Acceptance criteria: An orange color is produced.
B. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the
requirements
ASSAY C6H8AsNO3 217.05
PROCEDURE p-Aminobenzenearsonic acid [98-50-0].
Color reagent: Dissolve 28.0 g of potassium hydroxide and
2.0 g of potassium sodium tartrate in 100 mL of water. DEFINITION
Cool, and add, in the order named, 100 mg of 2,4- Arsanilic Acid contains NLT 98.0% and NMT 102.0% of
dichloro-1-naphthol, 180 mL of alcohol, and 20.0 mL of C6H8AsNO3, calculated on the dried basis.
0.475% sodium hypochlorite solution. Mix by swirling, and
allow to stand at room temperature for 1 h before using. IDENTIFICATION
This Color reagent may be stored in a glass-stoppered bottle, INFRARED ABSORPTION 197K
in a refrigerator, for 2 months.
Standard solution: 40 g/mL of USP Arginine ASSAY
Hydrochloride RS in water PROCEDURE
Sample solution: Equivalent to 40 g/mL of arginine Sample solution: 125 mg of Arsanilic Acid transferred to a
hydrochloride, from a volume of Injection in water 50-mL conical flask
Analysis: Transfer 2.0-mL portions of the Sample solution Analysis: Add 10.0 mL of a mixture of sulfuric acid, nitric
and the Standard solution, respectively, to separate flasks, acid, and perchloric acid (1000:50:50) and several glass
and treat each as follows. Add 2.0 mL of potassium iodide beads. Digest on a hot plate for 1 h, increasing the
solution (3 in 1000), and allow to stand for 15 min. Add 6.0 temperature of the hot plate in steps until a ring of sulfuric
mL of Color reagent, and allow to stand for 15 min. Add 2.0 acid rises into the neck of the flask. Allow to cool, to the
mL of sodium hypochlorite solution (19 in 10,000), and colorless solution add 400 mg of hydrazine sulfate, and heat
allow to stand for 15 min. the flask vigorously on a hot plate until a ring of sulfuric
Spectrometric conditions acid rises into the neck of the flask. Allow to cool, and wash
Cell: 1 cm down the rim, neck, and insides of the flask with 1 mL of
Analytical wavelength: 520 nm water. Heat the flask again until a ring of sulfuric acid rises
Blank: Water into the neck of the flask. Allow to cool, and transfer the
Analysis colorless solution, with the aid of 80 mL of water, to a 125-
Samples: Standard solution and Sample solution mL conical flask. Add 10 mL of hydrochloric acid and several
Calculate the percentage of C6H14N4O2 HCl in each mL of drops of 0.002 M potassium iodide, cool to 05 and titrate
the Injection taken: with 0.1 N potassium permanganate VS to a pale pink
endpoint, maintaining a temperature of 05 during the
Result = (AU/AS) (CS/CU) 100 titration. Perform a blank determination (see Titrimetry
541). Each mL of 0.1 N potassium permanganate is
AU =
absorbances of the Sample solution equivalent to 10.852 mg of C6H8AsNO3.
AS =
absorbances of the Standard solution Acceptance criteria: 98.0%102.0%
CS =
concentration of the Standard solution (g/mL)
CU =
nominal concentration of the Sample solution IMPURITIES
(mg/mL) Organic Impurities
Acceptance criteria: 9.5%10.5% PROCEDURE 1: LIMIT OF O-ARSANILIC ACID
Mobile phase: 4.04 g of monobasic potassium phosphate
SPECIFIC TESTS in 985 mL of water. Add 2 mL of phosphoric acid. Add 10
PH 791: 5.06.5 mL of methanol.
OTHER REQUIREMENTS: Meets the requirements under Standard solution: Transfer 67 mg of o-arsanilic acid to a
Injections 1 100-mL volumetric flask, add 65 mg of warm (7080)
BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.01 USP water, and shake or sonicate to dissolve. Allow to cool, and
Endotoxin Unit/mg of arginine hydrochloride. dilute with water to volume. Dilute a portion of this
solution with water to obtain a solution having a known
ADDITIONAL REQUIREMENTS concentration of 0.0012 mg/mL of o-arsanilic acid.
PACKAGING AND STORAGE: Preserve in single-dose containers, Sample solution: Transfer 50 mg of Arsanilic Acid to a 50-
preferably of Type II glass. mL volumetric flask, add 30 mL of warm water, and shake
LABELING: The label states the total osmolar concentration in or sonicate to dissolve. Allow to cool, and dilute with water
mOsmol/L. Where the contents are less than 100 mL, or to volume.
where the label states that the Injection is not for direct Chromatographic system
injection but is to be diluted before use, the label (See Chromatography, 621 System Suitability.)
alternatively may state the total osmolar concentration in
mOsmol/mL.
USP Arginine Hydrochloride RS
274 Arsanilic / Official Monographs USP 32
Mode: LC W = weight of Arsanilic Acid in the Sample solution

Detector: UV 242nm (mg)
Column: 4.6-mm 15-cm; 5-m base-deactivated Acceptance criteria: NMT 0.045%
packing L1
Temperature: 30 SPECIFIC TESTS
Flow rate: 1.5 mL/min LOSS ON DRYING 731: Dry it in a vacuum at 80 for 4 h: it
Injection size: 20 L loses NMT 0.5% of its weight.
System suitability
Sample: Standard solution ADDITIONAL REQUIREMENTS
Suitability requirements PACKAGING AND STORAGE: Preserve in well-closed containers.
Relative standard deviation: NMT 2.5% LABELING: Label it to indicate that it is for veterinary use
Capacity factor: 2.83.8 for the o-arsanilic acid peak only.
Analysis USP REFERENCE STANDARDS 11
Samples: Standard solution and Sample solution USP Arsanilic Acid RS
Calculate the percentage of o-arsanilic acid in the portion
of C6H8AsNO3 taken:
Result = 5000 (CS/W) (rU/rS)
Ascorbic Acid
(Comment on this Monograph)id=m6030=Ascorbic Acid=A-
CS = concentration of o-arsanilic acid in the Standard Monos.pdf)
solution (mg/mL)
W = weight of Arsanilic Acid in the Sample solution
(mg)
rU = o-arsanilic acid peak response from the Sample
solution
rS = o-arsanilic acid peak response from the Standard
solution
PROCEDURE 2: LIMIT OF ANILINE C6H8O6 176.12
L-Ascorbic acid;
Mobile phase: 7.76 g of monobasic potassium phosphate
in 950 mL of water. Add 50 mL of methanol. 1-Deoxy-L-gulofuranose-2-ene-1-one [50-81-7].
Standard solution: Transfer 176 mg of aniline to a 25-mL DEFINITION
volumetric flask, add 1 mL of methanol, swirl, then add 15 Ascorbic Acid contains NLT 99.0% and NMT 100.5% of C6H8O6
mL of water, and shake to dissolve. Dilute with water to
volume. Dilute a portion of this solution to obtain a IDENTIFICATION
solution having a known concentration of 0.00045 mg/mL A. INFRARED ABSORPTION 197K
of aniline. B. A 20 mg/mL solution reduces alkaline cupric tartrate TS
Sample solution: Transfer 50 mg of Arsanilic Acid to a 50- slowly at room temperature but more readily upon heating.
mL volumetric flask, add 30 mL of warm (7080) water,
and shake or sonicate to dissolve. Allow to cool, and dilute ASSAY
with water to volume. PROCEDURE
Chromatographic system Sample solution: 3.2 mg/mL of Ascorbic Acid, in a mixture
(See Chromatography, 621 System Suitability.) of 2 N sulfuric acid and water (1:4)
Mode: LC Analysis: Titrate 125 mL of the Sample solution at once with
Detector: UV 235nm 0.1 N iodine VS, using 3 mL of starch TS as an indicator.
Column: 4.6-mm 15-cm; 5-m base-deactivated Each mL of 0.1 N iodine is equivalent to 8.806 mg of
packing L1 C 6 H8 O6 .
Temperature: 30 Acceptance criteria: 99.0%100.5%
Injection size: 50 L IMPURITIES
System suitability Inorganic Impurities
Sample: Standard solution RESIDUE ON IGNITION 281: NMT 0.1%
Suitability requirements HEAVY METALS 231: NMT 20 ppm
Capacity factor: 2.33.3 for the aniline peak Sample solution: 40 mg/mL in water
Relative standard deviation: NMT 3.0% SPECIFIC TESTS
Analysis OPTICAL ROTATION, Specific Rotation 781S: +20.5 to
Samples: Standard solution and Sample solution +21.5, the optical rotation being measured immediately,
Calculate the percentage of aniline in the portion of following the preparation of the solution
Arsanilic Acid taken: Sample solution: 100 mg/mL, in carbon dioxide-free water
Result = (rU/rS) (CS/W) 5000 ADDITIONAL REQUIREMENTS
rU = aniline peak response from the Sample solution containers.
rS = aniline peak response from the Standard solution USP REFERENCE STANDARDS 11
CS = concentration of aniline in the Standard solution USP Ascorbic Acid RS
(mg/mL)
USP 32 Official Monographs / Ascorbic 275
Ascorbic Acid Injection LIMIT OF OXALATE: Dilute a volume of Injection, equivalent

(Comment on this Monograph)id=m6050=Ascorbic Acid to 50 mg of ascorbic acid, with water to 5 mL. Add 0.2 mL
Injection=A-Monos.pdf) of acetic acid and 0.5 mL of calcium chloride TS: no
turbidity is produced in 1 min.
DEFINITION
Ascorbic Acid Injection is a sterile solution, in Water for ADDITIONAL REQUIREMENTS
Injection, of Ascorbic Acid prepared with the aid of Sodium PACKAGING AND STORAGE: Preserve in light-resistant, single-
Hydroxide, Sodium Carbonate, or Sodium Bicarbonate. It dose containers, preferably of Type I or Type II glass.
contains NLT 90.0% and NMT 110.0% of the labeled amount LABELING: In addition to meeting the requirements under
of ascorbic acid (C6H8O6). Injections 1, Labeling, fused-seal containers of the Injection
in concentrations of 250 mg/mL and greater are labeled to
IDENTIFICATION indicate that since pressure may develop on long storage,
A. To a volume of Injection, equivalent to 40 mg of precautions should be taken to wrap the container in a
ascorbic acid, add 4 mL of 0.1 N hydrochloric acid, then add protective covering while it is being opened.
4 drops of methylene blue TS, and warm to 40: the deep USP REFERENCE STANDARDS 11
blue color becomes appreciably lighter or is completely USP Ascorbic Acid RS
discharged within 3 min. USP Endotoxin RS
B. The retention time of the major peak of the Sample
solution corresponds to that of the Standard solution,
obtained as directed in the Assay.
C. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the Ascorbic Acid Oral Solution
requirements (Comment on this Monograph)id=m6070=Ascorbic Acid Oral
Solution=A-Monos.pdf)
ASSAY
Mobile phase: Dissolve 15.6 g of dibasic sodium phosphate Ascorbic Acid Oral Solution is a solution of Ascorbic Acid in a
and 12.2 g of monobasic potassium phosphate in 2000 mL hydroxylic organic solvent or an aqueous mixture thereof. It
of water, and adjust with phosphoric acid to a pH of 2.5 contains NLT 90.0% and NMT 110.0% of the labeled amount
0.05. of C6H8O6.
Standard solution: 0.5 mg/mL of USP Ascorbic Acid RS in
Mobile phase [NOTERefrigerate and store protected from IDENTIFICATION
light until use. The solution is stable for at least 24 h. Inject A. To a volume of Oral Solution equivalent to 40 mg of
within 3 h after removal from the refrigerator.] ascorbic acid add 4 mL of 0.1 N hydrochloric acid, then add
Sample solution: 0.5 mg/mL from Injection in Mobile phase 4 drops of methylene blue TS, and warm to 40: the deep
[NOTERefrigerate and store protected from light until use. blue color becomes appreciably lighter or is completely
The solution is stable for at least 24 h. Inject within 3 h after discharged within 3 min.
removal from the refrigerator.] B. To a volume of Oral Solution equivalent to 20 mg of
Chromatographic system ascorbic acid add 15 mL of trichloroacetic acid solution (1 in
(See Chromatography 621, System Suitability.) 20), add 200 mg of activated charcoal, shake the mixture
Mode: LC vigorously for 1 min, and pass through a small fluted filter,
Detector: UV 245 nm returning the filtrate, if necessary, until clear. To 5 mL of the
Column: 6-mm 150-cm; packing L39 filtrate add 1 drop of pyrrole, agitate gently until dissolved,
Flow rate: 0.6 mL/min then heat in a bath at 50: a blue color develops.
Sample: Standard solution Sample solution: Transfer an equivalent to 50 mg of
Suitability requirements ascorbic acid from a volume of the Oral Solution to a 100-
Column efficiency: NLT 3500 theoretical plates mL volumetric flask, previously diluted with water if
Tailing factor: NMT 1.6 necessary. Add 20 mL of metaphosphoric-acetic acids TS,
Relative standard deviation: NMT 1.5% and dilute with water to volume. Measure a volume of the
Analysis dilution, equivalent to 2 mg of ascorbic acid, into a 50 mL
Samples: Standard solution and Sample solution conical flask, and add 5 mL of metaphosphoric-acetic acids
Calculate the percentage of C6H8O6 in each mL of the TS.
Injection taken: Analysis: Titrate with standard dichlorophenolindophenol
Result = (rU/rS) (CS/CU) 100 solution until a rose-pink color persists for at least 5 s.
Correct for the volume of the dichlorophenolindophenol
rU = peak response from the Sample solution solution consumed by a mixture of 5.5 mL of
rS = peak response from the Standard solution metaphosphoric-acetic acids TS and 15 mL of water. From
CS = concentration of USP Ascorbic Acid RS in the the ascorbic acid equivalent of the standard
Standard solution (mg/mL) dichlorophenolindophenol solution, calculate the ascorbic
CU = nominal concentration of ascorbic acid in the acid content in each mL of the Oral Solution.
Sample solution (mg/mL) Acceptance criteria: If present, 90.0%110.0%
Acceptance criteria: 90.0%110.0% OTHER COMPONENTS
SPECIFIC TESTS ALCOHOL DETERMINATION, Method I 611: 90.0%110.0%
PH 791: 5.57.0 ADDITIONAL REQUIREMENTS
OTHER REQUIREMENTS: It meets the requirements under PACKAGING AND STORAGE: Preserve in tight, light-resistant
Injections 1. containers.
BACTERIAL ENDOTOXINS TEST 85: It contains NMT 1.2 USP
Endotoxin Units/mg of ascorbic acid.
276 Ascorbic / Official Monographs USP 32
LABELING: Label Oral Solution that contains alcohol to state Aspartic Acid
the alcohol content. (Comment on this Monograph)id=m6198=Aspartic Acid=A-
Monos.pdf)
Ascorbic Acid Tablets

(Comment on this Monograph)id=m6090=Ascorbic Acid
DEFINITION
Ascorbic Acid Tablets contain NLT 90.0% and NMT 110.0% of
the labeled amount of C6H8O6. C4H7NO4 133.10
L-Aspartic acid [56-84-8].
IDENTIFICATION
[NOTEThe Sample solution is prepared as follows.] DEFINITION
Sample solution: Triturate a quantity of finely powdered Aspartic Acid contains NLT 98.5% and NMT 101.5% of
Tablets with sufficient diluted alcohol to make approximately C4H7NO4, calculated on the dried basis.
the equivalent of a 1-in-50 solution of ascorbic acid, filter, and
proceed with the following tests. IDENTIFICATION
A. A portion of the filtrate reduces alkaline cupric tartrate TS INFRARED ABSORPTION 197K
slowly at room temperature but more readily upon heating.
B. To 2 mL of the filtrate add 4 drops of methylene blue TS, ASSAY
and warm to 40: the deep blue color becomes appreciably PROCEDURE
lighter or is completely discharged within 3 min. Sample: 0.1 g of Aspartic Acid
C. To 1 mL of the filtrate add 15 mL of a solution of Analysis: In a 125-mL flask, dissolve in 50 mL of carbon
trichloroacetic acid (1 in 20), add 200 mg of activated dioxide-free water, heating slightly if necessary. Cool, add
charcoal, shake the mixture vigorously for 1 min, and pass 0.1 mL of bromothymol blue TS, and titrate with 0.1 N
through a small fluted filter, returning the filtrate, if sodium hydroxide VS until the color changes from yellow to
necessary, until clear. To 5 mL of the filtrate add 1 drop of blue. Perform a blank determination (see Titrimetry 541).
pyrrole, and agitate gently until dissolved, then heat in a Each mL of 0.1 N sodium hydroxide is equivalent to 13.31
bath at 50: a blue color develops. mg of C4H7NO4.
ASSAY
Sample solution: Transfer NLT 20 Tablets to a 1000-mL Inorganic Impurities
volumetric flask containing 250 mL of metaphosphoric- RESIDUE ON IGNITION 281: NMT 0.1%
acetic acids TS. CHLORIDE AND SULFATE, Chloride 221: Dissolve 0.7 g in 10
Analysis: Insert the stopper in the flask, and shake by mL of diluted nitric acid, and dilute with water to make 15
mechanical means for 30 min or until the tablets have mL.
disintegrated completely. Dilute with water to volume. Acceptance criteria: The solution shows no more chloride
Transfer a portion of the solution to a centrifuge tube, and than corresponds to 0.20 mL of 0.020 N hydrochloric acid
centrifuge until a clear supernatant is obtained. (0.02%).
Quantitatively dilute the clear supernatant with water, if CHLORIDE AND SULFATE, Sulfate 221: Dissolve 0.8 g in 4 mL
necessary, to obtain a solution containing 500 g/mL of of hydrochloric acid, and dilute with water to make 15 mL.
ascorbic acid. Pipet 4 mL of the solution, equivalent to 2 mg Acceptance criteria: The solution shows no more sulfate
of ascorbic acid, into a 50-mL conical flask. Add 5 mL of than corresponds to 0.25 mL of 0.020 N sulfuric acid
metaphosphoric-acetic acids TS and titrate with standard (0.03%).
dichlorophenolindophenol solution until a rose-pink color IRON 241: NMT 10 ppm
persists for at least 5 s. Correct for the volume of the HEAVY METALS, Method II 231: NMT 10 ppm
dichlorophenolindophenol solution consumed by a mixture Organic Impurities
of 5.5 mL of metaphosphoric-acetic acids TS and 15 mL of PROCEDURE
water. From the ascorbic acid equivalent of the standard Adsorbent: 0.25-mm layer of chromatographic silica gel
dichlorophenolindophenol solution, calculate the content of System suitability solution: 10 mg each of USP Aspartic
ascorbic acid in each Tablet. Acid RS and glutamic acid in 2 mL of ammonia TS, dilute
Acceptance criteria: 90.0%110.0% with water to 25.0 mL
Sample solution: Transfer 0.1 g of Aspartic Acid to a 10-
PERFORMANCE TESTS mL volumetric flask, dissolve in 2 mL of 17% ammonia
DISSOLUTION, Procedure for a Pooled Sample 711 solution (prepared by diluting ammonium hydroxide, 6 in
Medium: Water; 900 mL 10), and dilute with water to volume.
Apparatus 2: 50 rpm Standard solution: Transfer 5 mg of USP Aspartic Acid RS
Time: 45 min to a 100-mL volumetric flask, dissolve in 2 mL of 17%
Analysis: Determine the amount of C6H8O6 dissolved, using ammonia solution (prepared by diluting ammonium
the Analysis set forth in the Assay and conducting the hydroxide, 6 in 10), and dilute with water to volume.
Analysis without delay, making any necessary modifications. Application volume: 5 L
Tolerances: NLT 75% (Q) of the labeled amount of C6H8O6 Developing solvent system: Butyl alcohol, glacial acetic
is dissolved. acid, and water (3:1:1)
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Spray reagent: 2 mg/mL of ninhydrin in a mixture of
ADDITIONAL REQUIREMENTS butyl alcohol and 2 N acetic acid (19:1)
containers.
USP 32 Official Monographs / Aspirin 277
Analysis Acceptance criteria: 99.5%100.5%

Samples: System suitability solution, Sample solution, and
Standard solution IMPURITIES
Analysis: Proceed as directed for Chromatography 621, Inorganic Impurities
Thin-Layer Chromatography, except to dry the plate at 80 RESIDUE ON IGNITION 281: NMT 0.05%
for 30 min, spray with Spray reagent, and heat at 80 for CHLORIDE AND SULFATE 221
30 min. Examine the plate under white light. The Sample: 1.5 g
chromatogram obtained from the System suitability Analysis: Boil Sample with 75 mL of water for 5 min, cool,
solution exhibits two clearly separated spots, and no add sufficient water to restore the original volume, and
secondary spot in the chromatogram of the Sample filter.
solution is larger or more intense than the principal spot in Acceptance criteria: A 25-mL portion of the filtrate shows
the chromatogram of the Standard solution. no more chloride than corresponds to 0.10 mL of 0.020 N
Acceptance criteria hydrochloric acid (140 ppm).
Individual impurities: NMT 0.5% SULFATE
Total impurities: NMT 2.0% Sample: 6.0 g
Analysis: Dissolve Sample in 37 mL of acetone, and add 3
SPECIFIC TESTS mL of water. Titrate potentiometrically with 0.02 M lead
OPTICAL ROTATION, Specific Rotation 781S: +24.0 to perchlorate, prepared by dissolving 9.20 g of lead
+26.0, at 20 perchlorate in water to make 1000 mL of solution, using a
Sample solution: 80 mg/mL, in 6 N hydrochloric acid pH meter capable of a minimum reproducibility of 0.1
LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT mV (see pH 791), and equipped with an electrode system
0.5% of its weight. consisting of a lead-specific electrode and a silversilver
chloride reference glass-sleeved electrode containing a
ADDITIONAL REQUIREMENTS solution of tetraethylammonium perchlorate in glacial
PACKAGING AND STORAGE: Preserve in well-closed containers, acetic acid (1 in 44) (see Titrimetry 541). [NOTEAfter
protected from light. use, rinse the lead-specific electrode with water, drain the
USP REFERENCE STANDARDS 11 reference electrode, flush with water, rinse with methanol,
USP Aspartic Acid RS and allow to dry.]
Acceptance criteria: NMT 1.25 mL of 0.02 M lead
perchlorate is consumed (NMT 400 ppm).
Aspirin HEAVY METALS
Sample: 2 g
(Comment on this Monograph)id=m6240=Aspirin=A- Analysis: Dissolve in 25 mL of acetone, and add 1 mL of
Monos.pdf) water. Add 1.2 mL of thioacetamideglycerin base TS and
2 mL of pH 3.5 Acetate Buffer (see Heavy Metals 231), and
allow to stand for 5 min.
Acceptance criteria: Any color produced is not darker
than that of a control made with 25 mL of acetone and 2
mL of Standard Lead Solution (see Heavy Metals 231),
treated in the same manner (NMT 1 ppm).
Organic Impurities
PROCEDURE 1: LIMIT OF FREE SALICYLIC ACID
C9H8O4 180.16 Sample: 2.5 g
Benzoic acid, 2-(acetyloxy)-; Analysis: Dissolve in sufficient alcohol to make 25.0 mL.
Salicylic acid acetate [50-78-2]. To each of two matched color-comparison tubes add 48
DEFINITION mL of water and 1 mL of a freshly prepared, diluted ferric
Aspirin contains NLT 99.5% and NMT 100.5% of C9H8O4, ammonium sulfate solution (prepared by adding 1 mL of 1
calculated on the dried basis. N hydrochloric acid to 2 mL of ferric ammonium sulfate TS,
and diluting with water to 100 mL). Into one tube pipet 1
IDENTIFICATION mL of a standard solution of salicylic acid in water,
A. Heat it with water for several min, cool, and add 1 or 2 containing 0.10 mg/mL of salicylic acid. Into the second
drops of ferric chloride TS: a violet-red color is produced. tube pipet 1 mL of the solution (1 in 10) of Aspirin. Mix
B. INFRARED ABSORPTION 197K the contents of each tube.
Acceptance criteria: After 30 s, the color in the second
ASSAY tube is not more intense than that in the tube containing
PROCEDURE the salicylic acid (NMT 0.1%).
Sample: 1.5 g of Aspirin in a flask PROCEDURE 2: READILY CARBONIZABLE SUBSTANCES TEST 271
Analysis: Add 50.0 mL of 0.5 N sodium hydroxide VS, and Sample solution: 100 mg/mL in sulfuric acid TS
boil the mixture gently for 10 min. Add phenolphthalein TS, Acceptance criteria: The solution has no more color than
and titrate the excess sodium hydroxide with 0.5 N sulfuric Matching Fluid Q.
acid VS. Perform a blank determination (see Titrimetry 541, PROCEDURE 3: SUBSTANCES INSOLUBLE IN SODIUM CARBONATE
Residual Titrations). Each mL of 0.5 N sodium hydroxide is TS: A solution of 500 mg in 10 mL of warm sodium
equivalent to 45.04 mg of C9H8O4. carbonate TS is clear.
278 Aspirin / Official Monographs USP 32

LOSS ON DRYING 731: Dry it over silica gel for 5 h: it loses
NMT 0.5% of its weight. PERFORMANCE TESTS
DISSOLUTION 711
ADDITIONAL REQUIREMENTS Medium: 0.5 M phosphate buffer, pH 7.4; 900 mL
PACKAGING AND STORAGE: Preserve in tight containers. Apparatus 2: 75 rpm
USP REFERENCE STANDARDS 11 Time: 45 min
USP Aspirin RS Detector: UV absorption at the wavelength of the isosbestic
point of aspirin and salicylic acid at 265 2 nm
Diluent: Acetonitrile and formic acid (99:1)
Aspirin Boluses Sample solutions: Filtered portions of the solution under
test, suitably diluted with Diluent
(Comment on this Monograph)id=m6245=Aspirin Boluses=A- Standard solution: USP Aspirin RS in Medium at a known
Monos.pdf) concentration [NOTEPrepare the solution at the time of
DEFINITION use.]
Aspirin Boluses contain NLT 90.0% and NMT 110.0% of the Analysis: Determine the amount of C9H8O4 dissolved by UV
labeled amount of aspirin (C9H8O4). absorption performed on the Sample solution, in comparison
to a Standard solution having a known concentration of USP
IDENTIFICATION Aspirin RS.
A. PROCEDURE Tolerances: NLT 80% (Q) of the labeled amount of C9H8O4
Analysis: Crush 1 Bolus. Boil a portion of the powder, is dissolved.
equivalent to 300 mg of aspirin, with 50 mL of water. Cool, UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
and add a drop of ferric chloride TS.
Acceptance criteria: A violet-red color is produced. IMPURITIES
B. The retention time of the aspirin peak of the Sample Organic Impurities
solution corresponds to that of the Standard solution, as PROCEDURE: LIMIT OF SALICYLIC ACID
obtained in the Assay. Analysis: Using the chromatograms of the Standard
solution and the Sample solution, obtained as directed in the
ASSAY Assay, calculate the percentage of C7H6O3 in the portion of
PROCEDURE Boluses taken:
Mobile phase: 2 mg/mL of sodium 1-heptanesulfonate in
acetonitrile and water (3:17). Adjust with glacial acetic acid Result = (rU/rS) (CS/CU) 100
to a pH of 3.4.
Diluent: Acetonitrile and formic acid (99:1) rU = peak response from the Sample solution
Standard solution: 0.4 mg/mL of USP Aspirin RS and 0.01 rS = peak response from the Standard solution
mg/mL of USP Salicylic Acid RS in Diluent CS = concentration of USP Salicylic Acid RS in the
Sample solution: Finely powder NLT 10 Boluses. Transfer a Standard solution (mg/mL)
portion of the powder to an appropriate volumetric flask CU = concentration of aspirin in the portion of Boluses
and dilute with Diluent to prepare a solution nominally taken for the Sample solution, as determined in
equivalent to 4 mg/mL. Stir the solution by mechanical the Assay (mg/mL)
means for 15 min. Pass a portion of this solution through a Acceptance criteria: NMT 0.3%
filter having a 0.5-m or finer porosity. ADDITIONAL REQUIREMENTS
Chromatographic system PACKAGING AND STORAGE: Preserve in tight containers.
(See Chromatography 621, System Suitability.) LABELING: Label Boluses to indicate that they are for
Mode: LC veterinary use only.
Detector: UV 254 nm USP REFERENCE STANDARDS 11
Column: 4.6-mm 25-cm; packing L1 USP Aspirin RS
Flow rate: 1 mL/min USP Salicylic Acid RS
System suitability
[NOTEThe relative retention times for salicylic acid and Aspirin Capsules
aspirin are 0.6 and 1.0, respectively.] (Comment on this Monograph)id=m6250=Aspirin Capsules=A-
Suitability requirements Monos.pdf)
Relative standard deviation: NMT 2.0% for the aspirin
peak DEFINITION
Analysis Aspirin Capsules contain NLT 93.0% and NMT 107.0% of the
Samples: Standard solution and Sample solution labeled amount of aspirin (C9H8O4).
Calculate the percentage of C9H8O4 in the portion of [NOTECapsules that are enteric-coated or the contents of
Boluses taken: which are enteric-coated meet the requirements for Aspirin
Delayed-Release Capsules.]
IDENTIFICATION
rU = peak response from the Sample solution A. PROCEDURE
rS = peak response from the Standard solution Sample: 1 Tablet
CS = concentration of USP Aspirin RS in the Standard Analysis: Crush and boil it with 50 mL of water for 5 min,
solution (mg/mL) cool, and add 1 or 2 drops of ferric chloride TS.
CU = nominal concentration of aspirin in the Sample
solution (mg/mL)
Acceptance criteria: A violet-red color is produced. Sample solution: Sample per Dissolution 711. Dilute with
B. INFRARED ABSORPTION 197K Medium, if necessary, and filter.
Sample: A quantity of the contents of Capsules, equivalent Analysis
to 500 mg of aspirin Samples: Standard solution and Sample solution
Analysis: Shake Sample with 10 mL of alcohol for several Determine the amount of C9H8O4 dissolved from UV
min. Centrifuge the mixture. Pour off the clear supernatant absorbances at the wavelength of the isosbestic point of
and evaporate it to dryness. Dry the residue in a vacuum at aspirin and salicylic acid at 265 2 nm of the Sample
60 for 1 h. solution in comparison with a Standard solution having a
Acceptance criteria: Meet the requirements known concentration.
Tolerances: NLT 80% (Q) of the labeled amount of C9H8O4
ASSAY is dissolved.
PROCEDURE UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
[NOTEUse chloroform recently saturated with water.]
Diluent: A solution (1 in 100) of glacial acetic acid in IMPURITIES
chloroform Organic Impurities
Standard stock solution: Transfer 50 mg of USP Aspirin RS PROCEDURE: LIMIT OF FREE SALICYLIC ACID
to a 50-mL volumetric flask, add 0.5 mL of glacial acetic Ferric chloride-urea reagent: Dissolve by swirling, without
acid, and add chloroform to volume. the aid of heat, 60 g of urea in a mixture of 8 mL of ferric
Standard solution: 50 g/mL of USP Aspirin RS from chloride solution (6 in 10) and 42 mL of 0.05 N
Standard stock solution diluted with Diluent hydrochloric acid. Adjust the resulting solution, if necessary,
Chromatographic column: Proceed as directed under with 6 N hydrochloric acid to a pH of 3.2.
Chromatography 621, Column Partition Chromatography, Standard solution: Transfer 75.0 mg of salicylic acid,
packing a chromatographic tube with a mixture of 3 g of previously dried over silica gel for 3 h, to a 100-mL
Solid Support and 2 mL of freshly prepared sodium volumetric flask, and add chloroform to volume. Transfer
bicarbonate solution (1 in 12). 10.0 mL of this solution to a second 100-mL volumetric
Sample solution: Remove, as completely as possible, the flask, and dilute with chloroform to volume. Transfer 10.0
contents of NLT 20 Capsules. Mix the combined contents, mL of this last solution to a 50-mL volumetric flask
and transfer a quantity of the powder, equivalent to 50 mg containing 10 mL of methanol, 2 drops of hydrochloric
of aspirin, to a 50-mL volumetric flask containing 1 mL of a acid, and 10 mL of a solution (1 in 10) of glacial acetic
solution (1 in 50) of hydrochloric acid in methanol, and add acid in ether, and dilute with chloroform to volume.
chloroform to volume. Transfer 5.0 mL of this solution to the Chromatographic column: Proceed as directed under
column, wash with 5 mL and then with 25 mL of Chromatography 621, Column Partition Chromatography,
chloroform, and discard the washings. Elute into a 100-mL packing a chromatographic tube with two segments of
volumetric flask with 10 mL of a solution (1 in 10) of glacial packing material. The lower segment is a mixture of 1 g of
acetic acid in chloroform and then with 85 mL of a solution Solid Support and 0.5 mL of 5 M phosphoric acid, and the
(1 in 100) of glacial acetic acid in chloroform, and dilute upper segment is a mixture of 3 g of Solid Support and 2
with the latter solvent to volume. mL of freshly prepared Ferric chloride-urea reagent.
Spectrometric conditions Sample solution: Weigh a portion of the contents of the
Mode: UV Capsules, as determined by the Assay, equivalent to 100
Analytical wavelength: 280 nm mg of aspirin, mix with 10 mL of chloroform by stirring for
Cell: 1 cm 3 min, and then transfer to the chromatographic column
Blank: Chloroform with the aid of a few mL of chloroform. Pass 50 mL of
Analysis chloroform through the column, rinse the tip of the
Samples: Standard solution and Sample solution chromatographic tube with chloroform, and discard the
Calculate the percentage of C9H8O4 in the portion of eluate. Prepare as a receiver a 50-mL volumetric flask
Capsules taken: containing 10 mL of methanol and 2 drops of hydrochloric
acid, and elute any salicylic acid from the column by
Result = (AU/AS) (CS/CU) 100 passing 10 mL of a solution (1 in 10) of glacial acetic acid
in ether that has been recently saturated with water,
AU = absorbances of the Sample solution followed by 30 mL of chloroform. Dilute the eluate with
AS = absorbances of the Standard solution chloroform to volume.
CS = concentration of USP Aspirin RS in the Standard Spectrometric conditions
solution (g/mL) Mode: UV
CU = nominal concentration of aspirin in the Sample Analytical wavelength: 306 nm
solution (mg/mL) Cell: 1 cm
Acceptance criteria: 93.0%107.0% Blank: A solvent mixture of the same composition as that
used for the Standard solution
PERFORMANCE TESTS Analysis
DISSOLUTION 711 Samples: Standard solution and Sample solution
0.05 M acetate buffer: Mix 2.99 g of sodium acetate Acceptance criteria: The absorbance of the Sample
trihydrate and 1.66 mL of glacial acetic acid with water to solution does not exceed that of the Standard solution
obtain 1000 mL of solution having a pH of 4.50 0.05. (NMT 0.75%, calculated on the labeled aspirin content).
Medium: 0.05 M acetate buffer; 500 mL
Apparatus 1: 100 rpm ADDITIONAL REQUIREMENTS
Time: 30 min PACKAGING AND STORAGE: Preserve in tight containers.
Standard solution: USP Aspirin RS in Medium USP REFERENCE STANDARDS 11
[NOTEPrepare the Standard solution at the time of use. An USP Aspirin RS
amount of alcohol not to exceed 1% of the total volume
of the Standard solution may be used to bring the
Reference Standard into solution prior to dilution with
Medium.]
Aspirin Delayed-Release Capsules hydrochloric acid or 2 N sodium hydroxide to a pH of 6.8

(Comment on this Monograph)id=m6260=Aspirin Delayed- 0.05.
Release Capsules=A-Monos.pdf) Standard solution: USP Aspirin RS of a known
concentration in the same medium
DEFINITION Sample solution: Filtered portion of the solution under test,
Aspirin Delayed-Release Capsules contain NLT 93.0% and NMT diluted, if necessary, with 0.1 N hydrochloric acid (analyzing
107.0% of the labeled amount of aspirin (C9H8O4). the Acid Stage) and with Diluent (analyzing the Buffer Stage)
Analysis
A. PROCEDURE Determine the amount of C9H8O4 dissolved by determining
Sample: 100 mg of Capsule contents UV absorbances at the wavelength of the isosbestic point
Analysis: Boil it with 10 mL of water for several min. Cool, of aspirin and salicylic acid (280 nm in the Acid Stage, and
and add 1 drop of ferric chloride TS. 265 nm in the Buffer Stage) of the Sample solution in
Acceptance criteria: A violet-red color is produced. comparison to the Standard solution.
B. INFRARED ABSORPTION 197K UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Sample: Shake a quantity of the contents of Capsules,
equivalent to 500 mg of aspirin, with 10 mL of alcohol for IMPURITIES
several min. Centrifuge the mixture. Pour off the clear Organic Impurities
supernatant and evaporate it to dryness. Dry the residue in a PROCEDURE: LIMIT OF FREE SALICYLIC ACID
vacuum at 60 for 1 h. Mobile phase and Diluent: Prepare as directed in the
Assay.
ASSAY Standard solution: 0.015 mg/mL of salicylic acid from
PROCEDURE USP Salicylic Acid RS dissolved in the Standard solution from
Mobile phase: Dissolve 2 g of sodium 1-heptanesulfonate the Assay.
in a mixture of 850 mL of water and 150 mL of acetonitrile, Sample solution: Use the Sample stock solution from the
and adjust with glacial acetic acid to a pH of 3.4. Assay.
Diluent: Acetonitrile and formic acid (99:1) Chromatographic system: Proceed as directed under
Standard solution: 0.5 mg/mL of USP Aspirin RS in Diluent Assay.
Sample stock solution: Remove, as completely as possible, System suitability
the contents of NLT 20 Capsules. Mix the combined Sample: Standard solution
contents, and transfer powder, equivalent to 100 mg of [NOTEThe relative retention times for salicylic acid and
aspirin, to a suitable container. Add 20.0 mL of Diluent and aspirin are about 0.7 and 1.0, respectively.]
10 glass beads. Shake vigorously for 10 min, and centrifuge. Suitability requirements
Sample solution: Dilute a volume of the Sample stock Resolution: NLT 2.0 between salicylic acid and aspirin
solution with nine volumes of Diluent. Relative standard deviation: NMT 4.0% of the salicylic
[NOTERetain the remaining portion of Sample stock acid peak responses
solution for the test for Procedure: Limit of free salicylic Analysis
acid.] Samples: Standard solution and Sample solution
Chromatographic system Calculate the percentage of C7H6O3 in the portion of
(See Chromatography 621, System Suitability.) Capsules taken:
Mode: LC
Detector: UV 280 nm Result = (rU/rS) (CS/CU) 100
Flow rate: 2 mL/min rU = peak response from the Sample solution
Injection size: 10 L rS = peak response from the Standard solution
System suitability CS = concentration of USP Salicylic Acid RS in the
Sample: Standard solution Standard solution (mg/mL)
Suitability requirements CU = nominal concentration of aspirin in the Sample
Tailing factor: NMT 2.0 solution as determined in the Assay (mg/mL)
Relative standard deviation: NMT 2.0% Acceptance criteria: NMT 3.0%
Analysis
Calculate the percentage of C9H8O4 in the portion of PACKAGING AND STORAGE: Preserve in tight containers.
Capsules taken: LABELING: The label indicates that the Capsules or the
contents thereof are enteric coated.
Result = (rU/rS) (CS/CU) 100 USP REFERENCE STANDARDS 11
USP Aspirin RS
rU = peak response from the Sample solution USP Salicylic Acid RS
CS = concentration of USP Aspirin RS in the Standard
solution (mg/mL)
CU = nominal concentration of aspirin in the Sample Aspirin Suppositories
solution (mg/mL) (Comment on this Monograph)id=m6280=Aspirin
Acceptance criteria: 93.0%107.0% Suppositories=A-Monos.pdf)
PERFORMANCE TESTS DEFINITION

DISSOLUTION 711: Proceed as directed under Apparatus 1 Aspirin Suppositories contain NLT 90.0% and NMT 110.0% of
and Apparatus 2, Delayed-Release Dosage Forms, Method B. the labeled amount of aspirin (C9H8O4).
Time: 90 min, for Buffer Stage
Diluent: 0.1 N hydrochloric acid and 0.20 M tribasic
sodium phosphate (3:1). Adjust, if necessary, with 2 N
IDENTIFICATION swirling and without the aid of heat, and adjust the
[NOTEThe Sample is prepared as follows.] resulting solution, if necessary, by the addition of 6 N
Sample: Transfer a portion of the melted Suppositories hydrochloric acid to a pH of 3.2.
obtained in the Assay, equivalent to 1 g of aspirin, to a 125- [NOTEPrepare on the day of use.]
mL conical flask. Add 20 mL of alcohol, and warm until Chromatographic column: Insert a small pledget of glass
completely disintegrated. Cool in an ice bath for 5 min, filter, wool above the stem constriction of a 20- 2.5-cm
and evaporate the filtrate to dryness: the residue meets the chromatographic tube, and uniformly pack with a mixture
requirements of the following tests. of 1 g of chromatographic siliceous earth and 0.5 mL of 5
A. Heat the residue with water for several min, cool, and M phosphoric acid. Directly above this layer, pack a similar
add 1 or 2 drops of ferric chloride TS: a violet-red color is mixture of 3 g of chromatographic siliceous earth, and 2
produced. mL of Ferric chloride-urea reagent.
B. INFRARED ABSORPTION 197K Standard stock solution: Dissolve a suitable quantity of
salicylic acid in chloroform to obtain a solution containing
ASSAY 150 g/mL of salicylic acid.
PROCEDURE Standard solution: Pipet 5 mL of the Standard stock
[NOTEIn this Assay, use chloroform that recently was solution into a 50-mL volumetric flask containing 10 mL of
saturated with water.] methanol, 0.1 mL of hydrochloric acid, and 10 mL of a
Chromatographic column: Uniformly pack a solution of glacial acetic acid in ether (1 in 10). Add
chromatographic tube, as described in Procedure: Limit of chloroform to volume.
free salicylic acid, with a mixture of 3 g of chromatographic Sample solution: Tare a small dish and glass rod, place in
siliceous earth and 2 mL of sodium bicarbonate solution (1 the dish NLT 5 Suppositories, heat gently on a steam bath
in 12) prepared on the day of use. until melted, then stir, and cool while stirring. Transfer a
Standard stock solution: Transfer 50 mg of USP Aspirin RS portion of the mass, equivalent to 50 mg of aspirin (see
to a 50-mL volumetric flask, add 0.5 mL of glacial acetic Sample solution in the Assay), to a small beaker. Add 10 mL
acid, and add chloroform to volume. of chloroform, warm slightly, and stir until dissolved. With
Standard solution: 0.05 mg/mL USP Aspirin RS from the aid of 5 mL of chloroform, transfer to the
Standard stock solution diluted with a solution of glacial chromatographic adsorption column. Pass 50 mL of
acetic acid in chloroform (1 in 100) chloroform in several portions through the column, rinse
Sample solution: Tare a small dish and glass rod, place in the tip of the chromatographic tube with chloroform, and
the dish NLT 5 Suppositories, heat gently on a steam bath discard the eluate. If the purple zone reaches the bottom
until melted, then stir, and cool while stirring. Transfer a of the tube, discard the column, and repeat the test with a
portion of the mass, equivalent to 50 mg of aspirin, to a 50- smaller quantity of melted Suppositories.
mL volumetric flask containing 1 mL of a solution of Elute the adsorbed salicylic acid into a 100-mL volumetric
hydrochloric acid in methanol (1 in 50), add 40 mL of flask containing 20 mL of methanol and 0.2 mL of
chloroform, and add chloroform to volume. hydrochloric acid by passing two 10-mL portions of a
Spectrometric conditions solution of glacial acetic acid in water-saturated ether (1
Mode: UV in 10), and then 30 mL of chloroform, through the
Analytical wavelength: 280 nm column, and dilute the eluate with chloroform to volume.
Cell: 1 cm Spectrometric conditions
Blank: Chloroform Mode: UV
Analysis: Pipet 5 mL of the Sample solution into the column, Analytical wavelength: 306 nm
wash with 5 mL of chloroform, then again with 25 mL of Cell: 1 cm
chloroform, and discard the washings. Without delay, elute Blank: A solvent mixture of the same composition as the
into a 100-mL volumetric flask with 10 mL of a solution of Standard solution
glacial acetic acid in chloroform (1 in 10), and then with 85 Analysis
mL of a solution of glacial acetic acid in chloroform (1 in Samples: Standard solution and Sample solution
100), and dilute with the latter solvent to volume. Without Concomitantly determine the absorbances.
delay, determine the absorbances of the eluted Sample Acceptance criteria: The absorbance of the Sample
solution and Standard solution. solution is NMT 3.0% that of the Standard solution.
Suppositories taken: ADDITIONAL REQUIREMENTS
PACKAGING AND STORAGE: Preserve in well-closed containers,
Result = (AU/AS) (CS/CU) 100 in a cool place.
AU = absorbance of the Sample solution USP Aspirin RS
solution (g/mL)
CU = nominal concentration of aspirin in the Sample Aspirin Tablets
solution (g/mL) (Comment on this Monograph)id=m6290=Aspirin Tablets=A-
Acceptance criteria: 90.0%110.0% Monos.pdf)
IMPURITIES DEFINITION
Organic Impurities Aspirin Tablets contain NLT 90.0% and NMT 110.0% of the
PROCEDURE: LIMIT OF FREE SALICYLIC ACID labeled amount of aspirin (C9H8O4). Tablets of larger than 81-
Ferric chloride-urea reagent: To a mixture of 8 mL of mg size contain no sweeteners or other flavors. [NOTETablets
ferric chloride solution (6 in 10) and 42 mL of 0.05 N that are enteric-coated meet the requirements for Aspirin
hydrochloric acid, add 60 g of urea. Dissolve the urea by Delayed-Release Tablets.]
IDENTIFICATION be used to bring the Reference Standard into solution before

A. PROCEDURE dilution with Medium.]
Sample: 1 Tablet Sample solution: Sample per Dissolution 711. Dilute with
Analysis: Crush and boil it with 50 mL of water for 5 min, Medium, if necessary, and filter.
cool, and add 1 or 2 drops of ferric chloride TS. Analysis
Acceptance criteria: A violet-red color is produced. Sample: Standard solution and Sample solution
B. INFRARED ABSORPTION 197K Determine the amount of C9H8O4 dissolved from UV
Sample: Shake a quantity of finely powdered Tablets, absorbances at the wavelength of the isosbestic point of
equivalent to 500 mg of aspirin, with 10 mL of alcohol for aspirin and salicylic acid at 265 2 nm of the Sample
several min. Centrifuge the mixture. Pour off the clear solution in comparison with the Standard solution.
supernatant, and evaporate it to dryness. Dry the residue in Tolerances: NLT 80% (Q) of the labeled amount of
a vacuum at 60 for 1 h. C9H8O4 is dissolved.
ASSAY
Mobile phase: Dissolve 2 g of sodium 1-heptanesulfonate Organic Impurities
in a mixture of 850 mL of water and 150 mL of acetonitrile, PROCEDURE: LIMIT OF FREE SALICYLIC ACID
and adjust with glacial acetic acid to a pH of 3.4. Mobile phase and Diluent: Prepare as directed in the
Diluent: Acetonitrile and formic acid (99:1) Assay.
Standard solution: 0.5 mg/mL of USP Aspirin RS in Diluent Standard solution: 0.015 mg/mL of salicylic acid in the
Sample stock solution: Transfer an equivalent to 100 mg of Standard solution prepared as directed in the Assay.
aspirin, from finely powdered Tablets (NLT 20), to a suitable Sample solution: Use the Sample stock solution from the
container. Add 20.0 mL of Diluent and 10 beads. Shake Assay.
vigorously for 10 min, and centrifuge. Chromatographic system: Proceed as directed in the
Sample solution: Dilute a volume of the Sample stock Assay.
solution with 9 volumes of Diluent. [NOTERetain the System suitability
remaining portion of Sample stock solution for the test for Sample: Standard solution [NOTEThe relative retention
Procedure: Limit of free salicylic acid.] times for salicylic acid and aspirin are about 0.7 and 1.0,
Chromatographic system respectively.]
Mode: LC Resolution: NLT 2.0, between salicylic acid and aspirin
Detector: UV 280 nm Relative standard deviation: NMT 4.0% for the salicylic
Column: 4.0-mm 30-cm; packing L1 acid peak responses
Flow rate: 2 mL/min Analysis
System suitability Calculate the percentage of salicylic acid (C7H6O3) in the
Sample: Standard solution portion of Tablets taken:
Tailing factor: NMT 2.0 Result = (rU/rS) (CS/CU) 100
Analysis rU = peak response of the salicylic acid from the
Samples: Standard solution and Sample solution Sample solution
Calculate the percentage of C9H8O4 in the portion of rS = peak response of the salicylic acid from the
Tablets taken: Standard solution
CS = concentration of USP Salicylic Acid RS in the
Result = (rU/rS) (CS/CU) 100 Standard solution (mg/mL)
CU = concentration of aspirin in the Sample solution as
rU = peak response of the aspirin from the Sample determined in the Assay (mg/mL)
solution Acceptance criteria: NMT 0.3%. For Tablets that are
rS = peak response of the aspirin from the Standard coated, NMT 3.0%.
solution
CS = concentration of USP Aspirin RS in the Standard ADDITIONAL REQUIREMENTS
solution (mg/mL) PACKAGING AND STORAGE: Preserve in tight containers.
CU = nominal concentration of the Sample solution Preserve flavored or sweetened Tablets of 81-mg size or
(mg/mL) smaller in containers holding NMT 36 Tablets each.
Acceptance criteria: 90.0%110.0% USP Aspirin RS
PERFORMANCE TESTS USP Salicylic Acid RS
DISSOLUTION 711
0.05 M acetate buffer: Mix 2.99 g of sodium acetate
trihydrate and 1.66 mL of glacial acetic acid with water to
obtain 1000 mL of solution having a pH of 4.50 0.05.
Buffered Aspirin Tablets
Medium: 0.05 M acetate buffer; 500 mL (Comment on this Monograph)id=m6291=Buffered Aspirin
Apparatus 1: 50 rpm Tablets=A-Monos.pdf)
Time: 30 min DEFINITION
Standard solution: USP Aspirin RS of a known Buffered Aspirin Tablets contain Aspirin and suitable buffering
concentration in Medium [NOTEPrepare the Standard agents. Tablets contain NLT 90.0% and NMT 110.0% of the
solution at the time of use. An amount of alcohol not to labeled amount of aspirin (C9H8O4).
exceed 1% of the total volume of the Standard solution may
IDENTIFICATION Standard solution may be used to bring the Reference

A. PROCEDURE Standard into solution prior to dilution with Medium.]
Sample: 1 Tablet Sample solution: Sample per Dissolution 711. Dilute with
Analysis: Crush and boil it with 50 mL of water for 5 min, Medium, if necessary, and filter.
cool, and add 1 or 2 drops of ferric chloride TS. Analysis
Acceptance criteria: A violet-red color is produced. Samples: Standard solution and Sample solution
B. INFRARED ABSORPTION 197K Determine the amount of C9H8O4 dissolved from UV
Sample: Shake a quantity of finely powdered Tablets, absorbances at the wavelength of the isosbestic point of
equivalent to 500 mg of aspirin, with 10 mL of alcohol for aspirin and salicylic acid at 265 2 nm of the Sample
several min. Centrifuge the mixture. Pour off the clear solution in comparison with a Standard solution having a
supernatant, and evaporate it to dryness. Dry the residue in known concentration.
a vacuum at 60 for 1 h. Tolerances: NLT 80% (Q) of the labeled amount of
C9H8O4 is dissolved.
ASSAY UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
PROCEDURE
Mobile phase: Dissolve 2 g of sodium 1-heptanesulfonate IMPURITIES
in a mixture of 850 mL of water and 150 mL of acetonitrile, Organic Impurities
and adjust with glacial acetic acid to a pH of 3.4. PROCEDURE: LIMIT OF FREE SALICYLIC ACID
Diluent: Acetonitrile and formic acid (99:1) Mobile phase and Diluent: Prepare as directed in the
Sample stock solution: Transfer an equivalent to 100 mg of Standard solution: 0.015 mg/mL of salicylic acid in
aspirin, from finely powdered Tablets (NLT 20), to a suitable Diluent
container. Add 20.0 mL of Diluent and 10 beads. Shake Sample solution: Use the Sample stock solution from the
vigorously for 10 min, and centrifuge. Assay.
Sample solution: Dilute a volume of the Sample stock Chromatographic system: Proceed as directed in the
solution with 9 volumes of Diluent. [NOTERetain the Assay.
remaining portion of Sample stock solution for the test for System suitability
Procedure: Limit of free salicylic acid.] Sample: Standard solution
Chromatographic system [NOTEThe relative retention times for salicylic acid and
(See Chromatography 621, System Suitability.) aspirin are about 0.7 and 1.0, respectively.]
Mode: LC Suitability requirements
Detector: UV 280 nm Resolution: NLT 2.0 between salicylic acid and aspirin
Column: 4.0-mm 30-cm; packing L1 Relative standard deviation: NMT 4.0% for the salicylic
Flow rate: 2 mL/min acid peak responses
Sample: Standard solution Calculate the percentage of C7H6O3 in the portion of
Suitability requirements Tablets taken:
Relative standard deviation: NMT 2.0% Result = (rU/rS) (CS/CU) 100
Analysis
Samples: Standard solution and Sample solution rU = peak response of the salicylic acid from the
Calculate the percentage of C9H8O4 in the portion of Sample solution
Tablets taken: rS = peak response of the salicylic acid from the
Standard solution
Result = (rU/rS) (CS/CU) 100 CS = concentration of USP Salicylic Acid RS in the
rU = peak response of the aspirin from the Sample CU = concentration of aspirin in the Sample solution as
solution determined in the Assay (mg/mL)
rS = peak response of the aspirin from the Standard Acceptance criteria: NMT 3.0%
solution
CS = concentration of USP Aspirin RS in the Standard SPECIFIC TESTS
solution (mg/mL) ACID-NEUTRALIZING CAPACITY 301: NLT 1.9 mEq of acid is
CU = nominal concentration of the Sample solution consumed for each 325 mg of aspirin in the Tablets.
(mg/mL)
PERFORMANCE TESTS USP REFERENCE STANDARDS 11
DISSOLUTION 711 USP Aspirin RS
0.05 M acetate buffer: Mix 2.99 g of sodium acetate USP Salicylic Acid RS
trihydrate and 1.66 mL of glacial acetic acid with water to
obtain 1000 mL of solution having a pH of 4.50 0.05.
Medium: 0.05 M acetate buffer; 500 mL Aspirin Delayed-Release Tablets
Apparatus 2: 75 rpm
[NOTEWhere the Tablet is composed of multiple layers, a (Comment on this Monograph)id=m6292=Aspirin Delayed-
stainless steel wire helix may be used, if needed, to hold Release Tablets=A-Monos.pdf)
the Tablet in proper orientation in the apparatus.]
Time: 30 min DEFINITION
Standard solution: USP Aspirin RS in Medium [NOTE Aspirin Delayed-Release Tablets contain NLT 95.0% and NMT
Prepare the Standard solution at the time of use. An amount 105.0% of the labeled amount of aspirin (C9H8O4).
of alcohol not to exceed 1% of the total volume of the
IDENTIFICATION Analysis
A. PROCEDURE Samples: Standard solution and Sample solution
Sample: 1 Tablet Determine the quantity of C9H8O4 dissolved by determining
Analysis: Crush and boil it with 50 mL of water for 5 min, UV absorbances at the wavelength of the isosbestic point
cool, and add 1 or 2 drops of ferric chloride TS. of aspirin and salicylic acid (280 nm in the Acid stage, and
Acceptance criteria: A violet-red color is produced. 265 nm in the Buffer stage) of the Sample solution in
B. INFRARED ABSORPTION 197K comparison to the Standard solution.
Sample: Shake a quantity of finely powdered Tablets, UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
equivalent to 500 mg of aspirin, with 10 mL of alcohol for
several min. Centrifuge the mixture. Pour off the clear IMPURITIES
supernatant, and evaporate it to dryness. Dry the residue in Organic Impurities
a vacuum at 60 for 1 h. PROCEDURE: LIMIT OF FREE SALICYLIC ACID
Mobile phase and Diluent: Prepare as directed in the
ASSAY Assay.
PROCEDURE Standard solution: 0.015 mg/mL of salicylic acid in
Mobile phase: 2 mg/mL of sodium 1-heptanesulfonate in Diluent
acetonitrile and water (3:17), and adjust with glacial acetic Sample solution: Use the Sample stock solution from the
acid to a pH of 3.4 Assay.
Diluent: Acetonitrile and formic acid (99:1) Chromatographic system: Proceed as directed under the
Sample stock solution: Transfer an equivalent to 100 mg of System suitability
aspirin, from finely powdered Tablets (NLT 20), to a suitable Sample: Standard solution
container. Add 20.0 mL of Diluent and 10 beads. Shake [NOTEThe relative retention times for salicylic acid and
vigorously for 10 min, and centrifuge. aspirin are about 0.7 and 1.0, respectively.]
Sample solution: Dilute a volume of the Sample stock Suitability requirements
solution with 9 volumes of Diluent. [NOTERetain the Resolution: NLT 2.0 between salicylic acid and aspirin
remaining portion of stock solution for the test for Procedure: Relative standard deviation: NMT 4.0% of the salicylic
Limit of free salicylic acid.] acid peak responses
Chromatographic system Analysis
(See Chromatography 621, System Suitability.) Samples: Standard solution and Sample solution
Mode: LC Calculate the percentage of C7H6O3 in the portion of
Detector: UV 280 nm Tablets taken:
Flow rate: 2 mL/min Result = (rU/rS) (CS/CU) 100
System suitability rU = peak response from the Sample solution
Sample: Standard solution rS = peak response from the Standard solution
Suitability requirements CS = concentration of USP Salicylic Acid RS in the
Tailing factor: NMT 2.0 Standard solution (mg/mL)
Relative standard deviation: NMT 2.0% CU = concentration of aspirin in the Sample solution as
Analysis determined in the Assay (mg/mL)
Samples: Standard solution and Sample solution Acceptance criteria: NMT 3.0%
Calculate the percentage of C9H8O4 in the Tablets taken:
Result = (rU/rS) (CS/CU) 100 PACKAGING AND STORAGE: Preserve in tight containers.
LABELING: The label indicates that the Tablets are enteric-
rU = peak response from the Sample solution coated.
rS = peak response from the Standard solution USP REFERENCE STANDARDS 11
CS = concentration of USP Aspirin RS in the Standard USP Aspirin RS
solution (mg/mL) USP Salicylic Acid RS
(mg/mL)
Acceptance criteria: 95.0%105.0% Aspirin Effervescent Tablets for Oral
PERFORMANCE TESTS Solution
DISSOLUTION 711: Proceed as directed for Procedure for (Comment on this Monograph)id=m6293=Aspirin Effervescent
Method B under Procedure, Apparatus 1 and Apparatus 2, Tablets for Oral Solution=A-Monos.pdf)
Delayed-Release Dosage Forms.
Apparatus 1: 100 rpm DEFINITION
Time: 90 min, for Buffer stage Aspirin Effervescent Tablets for Oral Solution contain Aspirin and
Diluent: 0.1 N hydrochloric acid and 0.20 M tribasic an effervescent mixture of a suitable organic acid and an alkali
sodium phosphate (3:1), and adjust, if necessary, with 2 N metal bicarbonate and/or carbonate. Tablets contain NLT
hydrochloric acid or 2 N sodium hydroxide to a pH of 6.8 90.0% and NMT 110.0% of the labeled amount of aspirin
0.05 (C9H8O4).
Standard solution: USP Aspirin RS of a known
concentration in Medium
Sample solution: Filtered portion of the solution under test,
diluted, if necessary, with 0.1 N hydrochloric acid (analyzing
the Acid stage) and with Diluent (analyzing the Buffer stage)
IDENTIFICATION Sample solution: Use the Sample stock solution prepared

A. PROCEDURE as directed in the Assay.
Analysis: Dissolve 1 Tablet in 50 mL of 1 N hydrochloric Chromatographic system: Proceed as directed in the
acid, boil for 5 min, and allow to cool. To 2 mL of the Assay.
resulting solution add 2 or 3 drops of ferric chloride TS. System suitability
Acceptance criteria: A violet-red color is produced. Sample: Standard solution
B. PROCEDURE [NOTEThe relative retention times for salicylic acid and
Analysis: Add 1/2 of a Tablet to 50 mL of water in a flask, aspirin are 0.7 and 1.0, respectively.]
and immediately stopper with a stopper fitted with tubing Suitability requirements
so that the evolved gas passes through calcium hydroxide Resolution: NLT 2.0 between salicylic acid and aspirin
TS. Relative standard deviation: NMT 4.0% of salicylic acid
Acceptance criteria: A white precipitate forms. Analysis: Calculate the percentage of salicylic acid
(C7H6O3) in the portion of Tablets taken:
ASSAY
PROCEDURE Result = (rU/rS) (CS/CU) 100
Mobile phase: 2 mg/mL of sodium 1-heptanesulfonate in
acetonitrile and water (3:17). Adjust with glacial acetic acid rU = peak response from the Sample solution
to a pH of 3.4. rS = peak response from the Standard solution
Diluent: Acetonitrile and formic acid (99:1) CS = concentration of USP Salicylic Acid RS in the
Standard solution: 0.5 mg/mL of USP Aspirin RS in Diluent Standard solution (mg/mL)
Sample stock solution: Transfer an equivalent to 100 mg of CU = concentration of aspirin in the portion of tablets
aspirin, from finely powdered Tablets (NLT 20), to a suitable taken for the Sample solution, as determined in
container. Add 20.0 mL of Diluent and 10 beads. Shake the Assay (mg/mL)
vigorously for 10 min, and centrifuge. Acceptance criteria: NMT 8.0%
Sample solution: Quantitatively dilute a measured volume
of the Sample stock solution with 9 volumes of Diluent. SPECIFIC TESTS
[NOTERetain the remaining portion of Sample stock ACID-NEUTRALIZING CAPACITY 301: NLT 5.0 mEq of acid is
solution for the test for Procedure: Limit of Free Salicylate.] consumed by 1 Tablet.
(See Chromatography 621, System Suitability.) ADDITIONAL REQUIREMENTS
Mode: LC PACKAGING AND STORAGE: Preserve in tight containers.
Detector: UV 280 nm USP REFERENCE STANDARDS 11
Column: 4.0-mm 30-cm; packing L1 USP Aspirin RS
Flow rate: 2 mL/min USP Salicylic Acid RS
System suitability
Sample: Standard solution Aspirin Extended-Release Tablets
Tailing factor: NMT 2.0 (Comment on this Monograph)id=m6295=Aspirin Extended-
Relative standard deviation: NMT 2.0% Release Tablets=A-Monos.pdf)
Analysis DEFINITION
Samples: Standard solution and Sample solution Aspirin Extended-Release Tablets contain NLT 95.0% and NMT
Calculate the percentage of C9H8O4 in the portion of 105.0% of the labeled amount of aspirin (C9H8O4).
Tablets taken:
IDENTIFICATION
Result = (rU/rS) (CS/CU) 100 A. PROCEDURE
Sample: 1 Tablet
rU = peak response from the Sample solution Analysis: Crush and boil it with 50 mL of water for 5 min,
rS = peak response from the Standard solution cool, and add 1 or 2 drops of ferric chloride TS.
CS = concentration of USP Aspirin RS in the Standard Acceptance criteria: A violet-red color is produced.
solution (mg/mL) B. INFRARED ABSORPTION 197K
CU = nominal concentration of aspirin in the Sample Sample: Shake a quantity of finely powdered Tablets,
solution (tablet/mL) equivalent to 500 mg of aspirin, with 10 mL of alcohol for
Acceptance criteria: 90.0%110.0% several min. Centrifuge the mixture, pour off the clear
PERFORMANCE TESTS supernatant, and evaporate it to dryness. Dry the residue in
SOLUTION TIME: 2 Tablets dissolve completely in 180 mL of a vacuum at 60 for 1 h.
water at 17.5 2.5 within 5 min. ASSAY
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements PROCEDURE
IMPURITIES Mobile phase: Dissolve 2 g of sodium 1-heptanesulfonate
Organic Impurities in a mixture of 850 mL of water and 150 mL of acetonitrile,
PROCEDURE: LIMIT OF FREE SALICYLATE and adjust with glacial acetic acid to a pH of 3.4.
Mobile phase and Diluent: Prepare as directed in the Diluent: Acetonitrile and formic acid (99:1)
Assay. Standard solution: 0.5 mg/mL of USP Aspirin RS in Diluent
Standard solution: Dissolve a quantity of USP Salicylic Sample stock solution: Transfer an equivalent to 100 mg
Acid RS in the Standard solution, prepared as directed in of aspirin, from finely powdered Tablets (NLT 20), to a
the Assay, to obtain a solution having a known suitable container. Add 20.0 mL of Diluent and 10 beads.
concentration of 0.015 mg/mL of salicylic acid. Shake vigorously for 10 min, and centrifuge.
Sample solution: Dilute a volume of the Sample stock Standard solution: USP Aspirin RS of a known
solution with 9 volumes of Diluent. concentration in Medium[NOTEPrepare the Standard
[NOTERetain the remaining portion of Sample stock solution at the time of use. An amount of alcohol not to
solution for the test for Procedure: Limit of Free Salicylic exceed 5% of the total volume of the Standard solution
Acid.] may be used to bring the USP Reference Standard into
Chromatographic system solution prior to dilution with Medium.]
(See Chromatography 621, System Suitability.) Sample solutions: Filtered portions of the solution under
Mode: LC test, suitably diluted with Medium, if necessary
Column: 4.0-mm 30-cm; packing L1 Samples: Standard solution and Sample solutions
Flow rate: 2 mL/min Determine the amount of C9H8O4 dissolved from UV
Injection size: 10 L absorbances at the wavelength of the isosbestic point of
System suitability aspirin at about 265 nm of the Sample solutions in
Sample: Standard solution comparison with the Standard solution.
Suitability requirements Tolerances: The percentages of the labeled amount of
Tailing factor: NMT 2.0 C9H8O4 dissolved at the times specified conform to
Relative standard deviation: NMT 2.0% Acceptance Table 2.
Analysis
Samples: Standard solution and Sample solution Time Amount Dissolved
Calculate the percentage of C9H8O4 in the portion of Tablets (h) (%)
taken:
1 1540
Result = (rU/rS) (CS/CU) 100 2 2560
4 3575
rU = peak response of the aspirin from the Sample
solution 8 NLT 70
rS = peak response of the aspirin from the Standard
solution UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
solution (mg/mL) IMPURITIES
CU = nominal concentration for the Sample solution Organic Impurities
(unit/mL) PROCEDURE: LIMIT OF FREE SALICYLIC ACID
Acceptance criteria: 95.0%105.0% Mobile phase and Diluent: Prepare as directed in the
Assay.
PERFORMANCE TESTS Standard solution: 0.015 mg/mL of salicylic acid in the
DISSOLUTION 711 Standard solution prepared as directed in the Assay
Test 1 Sample solution: Use the Sample stock solution from the
[NOTEIf the product complies with this test, the labeling Assay.
indicates that it meets USP Dissolution Test 1.] Chromatographic system: Proceed as directed in the
Medium: 0.1 N hydrochloric acid; 900 mL Assay.
Apparatus 2: 60 rpm System suitability
Time: 1 and 4 h Sample: Standard solution
Standard solution: USP Aspirin RS of a known [NOTEThe relative retention times for salicylic acid and
concentration in Medium aspirin are about 0.7 and 1.0, respectively.]
Sample solution: Filtered portions of the solution under Suitability requirements
test. Dilute with Medium, if necessary, and filter. Resolution: NLT 2.0 between salicylic acid and aspirin
Analysis Relative standard deviation: NMT 4.0% of the salicylic
Samples: Standard solution and Sample solution acid peak responses
Determine the amount of C9H8O4 dissolved from UV Analysis
absorbances at the wavelength of the isosbestic point of Samples: Standard solution and Sample solution
aspirin at 280 nm of the Sample solution in comparison Calculate the percentage of C7H6O3 in the portion of
with the Standard solution. Tablets taken:
Tolerances: The percentages of the labeled amount of
C9H8O4 dissolved at the times specified conform to Result = (rU/rS) (CS/CU) 100
Acceptance Table 1.
rU = peak response of the salicylic acid from the
Sample solution
Time Amount Dissolved rS = peak response of the salicylic acid from the
(h) (%) Standard solution
1 2055 CS = concentration of USP Salicylic Acid RS in the
4 NLT 80 Standard solution (mg/mL)
CU = concentration of aspirin in the Sample solution as
Test 2 determined in the Assay (mg/mL)
[NOTEIf the product complies with this test, the labeling Acceptance criteria: NMT 3.0%
indicates that it meets USP Dissolution Test 2.] ADDITIONAL REQUIREMENTS
Medium: Water; 1000 mL PACKAGING AND STORAGE: Preserve in tight containers.
Apparatus 2: 30 rpm LABELING: The labeling indicates the Dissolution Test with
Time: 1, 2, 4, and 8 h which the product complies.
Detector: UV absorbances at the isosbestic point at 265
nm
USP REFERENCE STANDARDS 11 Mode: LC

USP Aspirin RS Detector: UV 280 nm
USP Salicylic Acid RS Column: 4-mm 30-cm; 10-m packing L1
Flow rate: 2 mL/min
Injection size: 5 L
System suitability
Aspirin, Alumina, and Magnesia Tablets Sample: Standard solution
(Comment on this Monograph)id=m6300=Aspirin, Alumina, [NOTEThe relative retention times for salicylic acid,
and Magnesia Tablets=A-Monos.pdf) aspirin, phenacetin, and chloroform are about 0.3, 0.6,
1.0, and 1.8, respectively.]
DEFINITION [NOTERecord each chromatogram until the chloroform
Aspirin, Alumina, and Magnesia Tablets contain NLT 90.0% and peak.]
NMT 110.0% of the labeled amount of aspirin (C9H8O4), the Suitability requirements
equivalent of NLT 90.0% and NMT 110.0% of the labeled Resolution: NLT 2.0 between the salicylic acid, aspirin,
amount of aluminum hydroxide [Al(OH)3], and NLT 90.0% and internal standard peaks
and NMT 110.0% of the labeled amount of magnesium Tailing factor: NMT 2.0
hydroxide [Mg(OH)2]. Relative standard deviation: NMT 3.0%
Analysis
[NOTEThe Sample is prepared as follows.] Calculate the percentage of C9H8O4 in the portion of
Sample: To a 0.7-g portion of finely powdered Tablets, add Tablets taken:
20 mL of 3 N hydrochloric acid and 5 drops of methyl red TS,
heat to boiling, and add 6 N ammonium hydroxide until the Result = (RU/RS) (CS/CU) 100
color of the solution changes to deep yellow. Continue boiling
for 2 min, and filter. The filtrate is used in Identification test B RU = ratio of the peak responses of aspirin and
and the precipitate is used in Identification test C. phenacetin from the Sample solution
A. The retention time of the aspirin peak of the Sample RS = ratio of the peak responses of aspirin and
solution corresponds to that of the Standard solution, as phenacetin from the Standard solution
obtained in the Assay for Aspirin. CS = concentration of USP Aspirin RS in the Standard
B. IDENTIFICATION TESTSGENERAL, Magnesium 191 solution (mg/mL)
Sample solution: The Sample filtrate is used. CU = nominal concentration of the Sample solution
C. IDENTIFICATION TESTSGENERAL, Aluminum 191 (mg/mL)
Sample solution: Wash the Sample precipitate with a hot Acceptance criteria: 90.0%110.0%
solution of ammonium chloride (1 in 50), and dissolve the ALUMINUM HYDROXIDE
precipitate in hydrochloric acid. Edetate disodium titrant: Dissolve 18.6 g of edetate
Acceptance criteria: The solution so obtained meets the disodium in water to make 1000 mL, and standardize the
requirements. solution as follows. Weigh 2 g of aluminum wire, transfer to
a 1000-mL volumetric flask, and add 50 mL of a mixture of
ASSAY hydrochloric acid and water (1:1). Swirl the flask to ensure
ASPIRIN contact of the aluminum and the acid, and allow the
Mobile phase: Mix 225 mg of tetramethylammonium reaction to proceed until all of the aluminum has dissolved.
hydroxide pentahydrate and 200 mg of sodium 1- Dilute with water to volume. Pipet 10 mL of this solution
octanesulfonate in 700 mL of water. Add methanol, into a 250-mL beaker, add, in the order named and with
acetonitrile, and glacial acetic acid (150:150:1), and stir. continuous stirring, 25.0 mL of edetate disodium titrant, and
Diluent: To 2 g of anhydrous citric acid, add 990 mL of 20 mL of acetic acidammonium acetate buffer TS, and boil
acetonitrile, 990 mL of chloroform, and 20 mL of formic gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL
acid, and stir for 30 min. Allow to settle, and use the of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a
decanted clear solution. bright rose-pink color. Perform a blank determination,
Internal standard solution: 2 mg/mL of phenacetin in substituting 10 mL of water for the aluminum solution, and
Diluent make any necessary correction.
Standard stock solution: 1 mg/mL of USP Salicylic Acid RS Calculate the molarity of the solution taken:
in Diluent
Standard solution: Transfer about 325 mg of USP Aspirin Result = W/26.98V
RS to a 50-mL volumetric flask. Add 10.0 mL of Standard
stock solution and 5.0 mL of Internal standard solution, dilute W = weight of aluminum in the portion of solution
with Diluent to volume, and mix. taken (mg)
Sample solution: Transfer a portion of the powdered V = volume of edetate disodium titrant consumed
Tablets (NLT 20), equivalent to about 325 mg of aspirin, to (mL)
a screw-capped bottle, add 5.0 mL of Internal standard Sample solution: To a portion of the powdered Tablets
solution, and 45.0 mL of Diluent, cap the bottle, mix, and (NLT 20), equivalent to 250 mg of aluminum hydroxide,
sonicate for 25 min. Centrifuge, and use the resultant clear add 20 mL of water, stir, and slowly add 30 mL of 3 N
solution. hydrochloric acid. Heat gently, if necessary, to aid solution,
cool, and quantitatively transfer to a 200-mL volumetric flask Mode: LC

with water and dilute with water to volume. Detector: UV 280 nm
Analysis: To 50 mL of Sample solution, add, in the order Column: 4-mm 30-cm; 10-m packing L1
named and with continuous stirring, 25.0 mL of edetate Flow rate: 2 mL/min
disodium titrant and 20 mL of acetic acidammonium Injection size: 5 L
acetate buffer TS, and heat the solution near the boiling System suitability
temperature for 5 min. Cool, and add 50 mL of alcohol and Sample: Standard solution
2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS [NOTEThe relative retention times for salicylic acid,
until the color changes from green-violet to rose-pink. aspirin, phenacetin, and chloroform are about 0.3, 0.6,
Perform a blank determination, substituting 50 mL of water 1.0, and 1.8, respectively.]
for the Sample solution. Each mL of 0.05 M edetate disodium [NOTERecord each chromatogram until the chloroform
titrant consumed is equivalent to 3.900 mg of Al(OH)3. peak appears.]
Acceptance criteria: 90.0%110.0% Suitability requirements
MAGNESIUM HYDROXIDE Resolution: NLT 2.0 between the salicylic acid, aspirin,
Sample stock solution: Prepare as directed in the Assay for and internal standard peaks
Aluminum hydroxide. Tailing factor: NMT 2.0
Eriochrome black T indicator: Dissolve 200 mg of Relative standard deviation: NMT 3.0%
eriochrome black T in a mixture of 15 mL of triethanolamine Analysis
and 5 mL of dehydrated alcohol. Samples: Standard solution and Sample solution
Analysis: To a volume of Sample solution, equivalent to 80 Calculate the percentage of free salicylic acid in the
mg of magnesium hydroxide, add 200 mL of water, 20 mL Tablets taken:
of triethanolamine, 50 mL of ammoniaammonium chloride
buffer TS, and 2 drops of Eriochrome black T indicator. Cool Result = (RU/RS) (CS/CU) 100
the solution to between 3 and 4 by immersion of the
beaker in an ice bath, then remove, and titrate with 0.05 M RU = ratio of the peak responses of salicylic acid and
edetate disodium VS until the color changes to pure blue. phenacetin from the Sample solution
Perform a blank determination, substituting water for the RS = ratio of the peak responses of salicylic acid and
Sample solution, and make any necessary corrections. Each phenacetin from the Standard solution
mL of 0.05 M edetate disodium is equivalent to 2.916 mg of CS = concentration of USP Salicylic Acid RS in the
Mg(OH)2. Standard solution (mg/mL)
Acceptance criteria: 90.0%110.0% CU = concentration of aspirin in the portion of Tablets
taken for the Sample solution as determined in
PERFORMANCE TESTS the Assay (mg/mL)
DISSOLUTION 711 Acceptance criteria: NMT 3.0%
Medium: 0.05 M acetate buffer, prepared by mixing 2.99 g
of sodium acetate (trihydrate), and 1.66 mL of glacial acetic SPECIFIC TESTS
acid with water to obtain 1000 mL of solution having a pH ACID-NEUTRALIZING CAPACITY 301: NLT 1.9 mEq of acid is
of 4.50 0.05; 900 mL consumed for each 325 mg of aspirin in the Tablets.
Apparatus 2: 75 rpm
Time: 45 min ADDITIONAL REQUIREMENTS
Detector: From UV absorbances at the wavelength of the PACKAGING AND STORAGE: Preserve in tight containers.
isosbestic point of aspirin and salicylic acid at 265 2 nm USP REFERENCE STANDARDS 11
Sample solutions: Filtered portions of the solution under USP Aspirin RS
test, suitably diluted with Medium USP Salicylic Acid RS
Standard solution: USP Aspirin RS in Medium [NOTE
Prepare the Standard solution at the time of use. An amount
of methanol NMT 1% of the total volume of the Standard Aspirin, Alumina, and Magnesium
solution may be used to bring the Reference Standard into Oxide Tablets
solution prior to dilution with Medium.]
Tolerances: NLT 75% (Q) of the labeled amount of C9H8O4 (Comment on this Monograph)id=m6302=Aspirin, Alumina,
is dissolved. and Magnesium Oxide Tablets=A-Monos.pdf)
for Weight Variation with respect to aluminum hydroxide and DEFINITION
to magnesium hydroxide, and for Content Uniformity with Aspirin, Alumina, and Magnesium Oxide Tablets contain NLT
respect to aspirin. 90.0% and NMT 110.0% of the labeled amount of aspirin
(C9H8O4), the equivalent of NLT 90.0% and NMT 110.0% of
IMPURITIES the labeled amount of aluminum hydroxide [Al(OH)3], and
Organic Impurities NLT 90.0% and NMT 110.0% of the labeled amount of
PROCEDURE: LIMIT OF FREE SALICYLIC ACID magnesium oxide (MgO).
[NOTEThe results from the Assay for Aspirin may be used
for this test when calculated as described in the Analysis IDENTIFICATION
section of this test.] [NOTEThe Sample is prepared as follows.]
Mobile phase, Diluent, Internal standard solution, Sample: To a 0.7-g portion of finely powdered Tablets, add
Standard stock solution, Standard solution, and Sample 20 mL of 3 N hydrochloric acid and 5 drops of methyl red TS,
solution: Proceed as directed in Assay for Aspirin. heat to boiling, and add 6 N ammonium hydroxide until the
Chromatographic system color of the solution changes to deep yellow. Continue boiling
for 2 min, and filter. The filtrate is used in Identification B and Relative standard deviation: NMT 2.0% for salicylic acid
the precipitate is used in Identification C. and aspirin peaks, Aspirin standard solution and Salicylic
A. The retention time of the aspirin peak of the Sample acid standard solution
solution corresponds to that of the Standard solution, as Analysis
obtained in the Assay for Aspirin. Samples: Aspirin standard solution, Salicylic acid standard
B. IDENTIFICATION TESTSGENERAL, Magnesium 191 solution, and Sample solution
Sample solution: The Sample filtrate is used Calculate the percentage of C9H8O4 in each Tablet taken:
C. IDENTIFICATION TESTSGENERAL, Aluminum 191
Sample solution: Wash the Sample precipitate with a hot Result = (rU/rS) (CS/CU) 100
solution of ammonium chloride (1 in 50), and dissolve the
precipitate in hydrochloric acid. The solution so obtained rU = aspirin peak responses from the Sample solution
meets the requirements. rS = aspirin peak responses from the Standard solution
D. PROCEDURE CS = concentration of USP Aspirin RS in the Aspirin
Analysis: Where the Tablets are composed of two layers, standard solution (mg/mL)
scrape a small amount of each layer into separate test tubes. CU = nominal concentration of the Sample solution
Add 2 mL of water and 2 drops of methyl red TS to each (mg/mL)
tube, and shake for 15 s. Acceptance criteria: 90.0%110.0%
Acceptance criteria: The solution from the aspirin- ALUMINUM HYDROXIDE
containing layer is red, and the solution from the buffer- Edetate disodium titrant: Dissolve 18.6 g of edetate
containing layer is yellow. disodium in water to make 1000 mL, and standardize the
solution as follows. Weigh 2 g of aluminum wire, transfer to
ASSAY a 1000-mL volumetric flask, and add 50 mL of a mixture of
ASPIRIN hydrochloric acid and water (1:1). Swirl the flask to ensure
Mobile phase: Methanol, phosphoric acid, and water contact of the aluminum and the acid, and allow the
(30:3:70) reaction to proceed until all of the aluminum has dissolved.
Diluent: Hydrochloric acid and dehydrated alcohol (1:100) Dilute with water to volume. Pipet 10 mL of this solution
Aspirin standard stock solution: 5 mg/mL of USP Aspirin into a 250-mL beaker, add, in the order named and with
RS in Diluent, prepared by blending at high speed for 1.5 continuous stirring, 25.0 mL of edetate disodium titrant, and
min 20 mL of acetic acidammonium acetate buffer TS, and boil
Aspirin standard solution: 0.25 mg/mL of USP Aspirin RS gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL
from the Aspirin standard stock solution in dehydrated of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a
alcohol bright rose-pink color. Perform a blank determination,
[NOTEUse these solutions within 1 h.] substituting 10 mL of water for the aluminum solution, and
Salicylic acid standard stock solution: 5 mg/mL of USP make any necessary correction.
Salicylic Acid RS in dehydrated alcohol. Transfer 3.0 mL of Calculate the molarity of the solution taken:
this solution to a 100-mL volumetric flask, and dilute with
Diluent to volume. Result = W/Ar V
Salicylic acid standard solution: 7.5 g/mL of USP
Salicyclic Acid RS from Salicylic acid standard stock solution in W = weight of aluminum in the portion of solution
dehydrated alcohol taken (mg)
System suitability solution: Transfer 5.0 mL of the Aspirin Ar = atomic weight of aluminum, 26.98
standard stock solution to a 100-mL volumetric flask, add 5.0 V = volume of Edetate disodium titrant consumed
mL of the Salicylic acid standard stock solution, and dilute (mL)
with dehydrated alcohol to volume. Sample solution: To a portion of the powdered Tablets
Sample solution: Transfer a counted number of Tablets, (NLT 20), equivalent to 600 mg of aluminum hydroxide,
equivalent to 2500 mg of aspirin, to a 120-mL blender jar add 20 mL of water, stir, and slowly add 30 mL of 3 N
containing 100.0 mL of Diluent, and blend at high speed for hydrochloric acid. Heat gently, if necessary, to aid solution,
1.5 min. Immediately filter a portion of the mixture thus cool, and transfer to a 200-mL volumetric flask. Wash the
obtained, and transfer 1.0 mL of the filtrate to a 100-mL beaker with water, adding the washings to the flask, and
volumetric flask. Immediately dilute with dehydrated alcohol add water to volume.
to volume. [NOTEPromptly inject this Sample solution into Analysis: To 20 mL of the Sample solution, add 20 mL of
the chromatograph as directed for Analysis.] water, then add, in the order named and with continuous
Chromatographic system stirring, 25.0 mL of Edetate disodium titrant and 20 mL of
(See Chromatography 621, System Suitability.) acetic acid-ammonium acetate buffer TS, and heat the
Mode: LC solution near the boiling temperature for 5 min. Cool, and
Detector: UV 205 nm add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with
Column: 4.6-mm 3-cm; 5-m packing L7 0.05 M zinc sulfate VS until the color changes from green-
Flow rate: 3.5 mL/min violet to rose-pink. Perform a blank determination,
Injection size: 10 L substituting 10 mL of water for the Sample solution, and
System suitability make any necessary corrections. Each mL of 0.05 to 3.900
Samples: Aspirin standard solution, Salicylic acid standard mg of Al(OH)3.
solution, and System suitability solution Acceptance criteria: 90.0%110.0%
[NOTEThe relative retention times for aspirin and salicylic MAGNESIUM OXIDE
acid are 0.7 and 1.0, respectively.] Sample solution: Prepare as directed in the Assay for
Suitability requirements Aluminum hydroxide.
Resolution: NLT 2.0 between the aspirin peak and the Eriochrome black T indicator: Dissolve 200 mg of
salicylic acid peak, System suitablity solution eriochrome black T in a mixture of 15 mL of triethanolamine
Tailing factor: NMT 2.0 for salicylic acid and aspirin and 5 mL of dehydrated alcohol.
peaks, Aspirin standard solution and Salicylic acid standard Analysis: To a volume of Sample solution equivalent to 40
solution mg of magnesium oxide add, mixing, 20 mL of
triethanolamine and 200 mL of water. Cool the solution for Mobile phase: Methanol, phosphoric acid, and water
10 min, while stirring, by immersion in an ice bath. Remove (30:3:70)
from the ice bath, and add 15 mL of ammoniaammonium Diluent: Hydrochloric acid and dehydrated alcohol (1:100)
chloride buffer TS and 2 drops of eriochrome black T Aspirin standard stock solution: 5 mg/mL of USP Aspirin
indicator. Titrate with 0.05 M edetate disodium VS to a blue RS in Diluent, prepared by blending at high speed for 1.5
endpoint, allowing 60 s between drops of titrant as the min
endpoint is approached (after first color change is Aspirin standard solution: 0.25 mg/mL of USP Aspirin RS
observed). [NOTEThe titration should be completed within from the Aspirin standard stock solution in dehydrated
10 min after the addition of the buffer and indicator. If any alcohol
precipitate is observed prior to titration, the solution should [NOTEUse these solutions within 1 h.]
be discarded and a new solution prepared.] Perform a blank Salicylic acid standard stock solution: 5 mg/mL of USP
determination, substituting water for the Sample solution and Salicylic Acid RS in dehydrated alcohol. Transfer 3.0 mL of
make any necessary correction. Each mL of 0.05 M edetate this solution to a 100-mL volumetric flask, and dilute with
disodium consumed is equivalent to 2.015 mg of MgO. Diluent to volume.
Acceptance criteria: 90.0%110.0% Salicylic acid standard solution: 7.5 g/mL of USP
Salicylic Acid RS from Salicylic acid standard stock solution in
PERFORMANCE TESTS dehydrated alcohol
DISSOLUTION 711 System suitability solution: Transfer 5.0 mL of the Aspirin
Medium: 0.05 M acetate buffer, prepared by mixing 2.99 g standard stock solution to a 100-mL volumetric flask, add
of sodium acetate (trihydrate) and 1.66 mL of glacial acetic 5.0 mL of the Salicylic acid standard stock solution, and
acid with water to obtain 1000 mL of solution having a pH dilute with dehydrated alcohol to volume.
of 4.50 0.05; 900 mL Sample solution: Transfer a counted number of Tablets,
Apparatus 1 (10-mesh screen): 100 rpm equivalent to 2500 mg of aspirin, to a 120-mL blender jar
Time: 45 min containing 100.0 mL of Diluent, and blend at high speed
Detector: Determine the amount of C9H8O4 dissolved for 1.5 min. Immediately filter a portion of the mixture
employing the following method. thus obtained, and transfer 1.0 mL of the filtrate to a 100-
Alkaline detergent solution: 1 N sodium hydroxide and mL volumetric flask. Immediately dilute with dehydrated
30% solution of polyoxyethylene (23) lauryl ether (1000:0.5) alcohol to volume.[NOTEPromptly inject this Sample
pH 4.3 buffer detergent: 12.9 mg/mL of citric acid solution into the chromatograph as directed for Analysis.]
monohydrate and 20.6 mg/mL of dibasic sodium phosphate Chromatographic system
heptahydrate in water. Add 0.5 mL of a 30% solution of (See Chromatography 621, System Suitability.)
polyoxyethylene (23) lauryl ether. Mode: LC
Standard solution: 0.45 mg/mL of USP Aspirin RS in Detector: UV 205 nm
Medium Column: 4.6-mm 3-cm; 5-m packing L7
Analysis: Use an automatic analyzer consisting of (1) a Flow rate: 3.5 mL/min
liquid sampler, (2) a proportioning pump, (3) a suitable Injection size: 10 L
fluorometer equipped with a 0.4-cm flow cell and suitable System suitability
recording devices, and (4) a manifold consisting of the Samples: Aspirin standard solution, Salicylic acid standard
components illustrated in the diagram in Automated Methods solution, and System suitability solution
of Analysis 16. With the sample line pumping pH 4.3 buffer [NOTEThe relative retention times for aspirin and
detergent, the other lines pumping their respective reagents, salicylic acid are 0.7 and 1.0, respectively.]
the fluorometer set at an excitation wavelength of 298 nm Suitability requirements
and an emission wavelength of 425 nm, adjust the system Resolution: NLT 2.0 between the aspirin peak and the
until a steady fluorescence baseline has been achieved. Start salicylic acid peak, System suitablity solution
the sampler, and conduct determinations at a rate of 40/h, Tailing factor: NMT 2.0 for salicylic acid and aspirin
using a ratio of 5:1 for sample and wash time. Record the peaks, Aspirin standard solution and Salicylic acid standard
fluorescence values of the Standard solution and the solution solution
under test. Relative standard deviation: NMT 2.0% for salicylic
Calculate the percentage of C9H8O4 dissolved: acid and aspirin peaks, Aspirin standard solution and
Salicylic acid standard solution
Result = CS V (FU/FS) 100/L Analysis
Samples: Aspirin standard solution, Salicylic acid standard
CS = concentration of USP Aspirin RS in the Standard solution, and Sample solution
solution (mg/mL) Calculate the percentage of free salicylic acid in the
V = volume of medium (mL), 900 Tablets taken:
FU = fluorescence values of the solution under test
FS = fluorescence values of the Standard solution Result = (rU/rS) (CS/CU) 100
L = Tablet label claim (mg)
Tolerances: NLT 75% (Q) of the labeled amount of C9H8O4 rU = salicylic acid peak responses from the Sample
is dissolved. solution
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements rS = peak responses from the Salicylic acid standard
for Weight Variation with respect to aluminum hydroxide and solution
to magnesium oxide, and for Content Uniformity with respect CS = concentration of USP Salicylic Acid RS in the
to aspirin Salicylic acid standard solution (g/mL)
IMPURITIES (mg/mL)
Organic Impurities
PROCEDURE: LIMIT OF FREE SALICYLIC ACID
[NOTEThe results from the Assay for Aspirin may be used
for this test when calculated as described in the Analysis
section of this test.]
Acceptance criteria: NMT 3.0% System suitability

Samples: Standard solution and System suitability solution
ACID-NEUTRALIZING CAPACITY 301: NLT 1.9 mEq of acid is Resolution: NLT 2.5 between the caffeine and
consumed for each 325 mg of aspirin in the Tablets. dihydrocodeine peaks, NLT 1.0 between the
dihydrocodeine and aspirin peaks, and NLT 1.5 between
ADDITIONAL REQUIREMENTS the aspirin and salicylic acid peaks, System suitability
PACKAGING AND STORAGE: Preserve in tight containers. solution
USP REFERENCE STANDARDS 11 Relative standard deviation: NMT 2.0% for each
USP Aspirin RS analyte, Standard solution
USP Salicylic Acid RS Analysis
Samples: Standard solution, Salicylic acid standard solution,
and Sample solution
Aspirin, Caffeine, and Dihydrocodeine Calculate the percentage of C9H8O4, C8H10N4O2, and
Bitartrate Capsules C18H23NO3 C4H6O6 in each Capsule taken:
(Comment on this Monograph)id=m6312=Aspirin, Caffeine, Result = (rU/rS) (CS/CU) 100
and Dihydrocodeine Bitartrate Capsules=A-Monos.pdf)
rU = peak response of the relevant analyte from the
DEFINITION Sample solution
Aspirin, Caffeine, and Dihydrocodeine Bitartrate Capsules rS = peak response of the relevant analyte from the
contain NLT 90.0% and NMT 110.0% of the labeled amounts Standard solution
of aspirin (C9H8O4), caffeine (C8H10N4O2), and dihydrocodeine CS = concentration of the appropriate USP Reference
bitartrate (C18H23NO3 C4H6O6). Standard in the Standard solution (mg/mL)
CU = nominal concentration of the appropriate analyte
IDENTIFICATION Sample solution (mg/mL)
The retention times of the major peaks of the Sample solution Acceptance criteria: 90.0%110.0%
correspond to those of the Standard solution obtained as
directed in the Assay. PERFORMANCE TESTS
DISSOLUTION, Procedure for a Pooled Sample 711
ASSAY Medium: 0.05 M acetate buffer, prepared by mixing 2.99 g
PROCEDURE of sodium acetate trihydrate and 1.66 mL of glacial acetic
Mobile phase: Dissolve 1 g of sodium 1-pentanesulfonate acid with water to obtain 1000 mL of solution having a pH
and 2.3 g of monobasic ammonium phosphate in 850 mL of of 4.50 0.05; 500 mL
water. Add 150 mL of acetonitrile, degas, and adjust with Apparatus 1: 50 rpm
phosphoric acid to a pH of 2.5. Time: 45 min
Diluent: Mix acetonitrile and water (46:53), and adjust with Mobile phase and Chromatographic system: Prepare as
phosphoric acid to a pH of 2.5. directed in the Assay.
Standard solution: 0001A mg/mL of USP Aspirin RS, Standard solution: Prepare a solution in Medium containing
0.001C mg/mL of USP Caffeine RS, and 0.001D mg/mL of known concentrations of 0.002A mg of USP Aspirin RS,
USP Dihydrocodeine Bitartrate RS in Diluent (A, C, and D 0.002C mg of USP Caffeine RS, and 0.002D mg of USP
being the labeled amounts, in mg, of aspirin, caffeine, and Dihydrocodeine Bitartrate RS per mL, A, C, and D being the
dihydrocodeine bitartrate, respectively, in each Capsule) labeled amounts, in mg, of aspirin, caffeine, and
[NOTEUse this solution within 3 h.] dihydrocodeine bitartrate, respectively, in each Capsule.
Salicylic acid standard solution: 0.005A mg/mL of USP Sample solution: Filter a portion of the solution under test.
Salicylic Acid RS in Diluent (A being the labeled amount, in Analysis: Separately inject equal volumes (10 L) of the
mg, of aspirin per Capsule) [NOTEUse this solution within Standard solution and the Sample solution into the
3 h.] chromatograph, record the chromatograms, and measure
System suitability solution: 0.0001A mg/mL of USP the responses for the major peaks.
Salicylic Acid RS (A being the labeled amount, in mg, of Calculate the percentage of C9H8O4, C8H10N4O2, and
aspirin in each Capsule) [NOTEUse this solution within 3 C18H23NO3 C4H6O6 dissolved:
h.]
Sample solution: Transfer the contents of 10 Capsules to a Result = (rU/rS) CS V 100/L
500-mL volumetric flask. Dilute with Diluent to volume.
Transfer 5.0 mL of this mixture to a 100-mL volumetric flask, rU = peak response of the relevant analyte from the
and dilute with Diluent to volume. Centrifuge a portion of Sample solution
this mixture, and use the clear supernatant as the Sample rS = peak response of the relevant analyte from the
solution. [NOTEUse this solution within 3 h.] Standard solution
Chromatographic system CS = concentration of the appropriate USP Reference
(See Chromatography 621, System Suitability.) Standard in the Standard solution (mg/mL)
Mode: LC V = volume of the medium (mL), 500
Detector: UV 215 nm L = Tablet label claim of the appropriate analyte (mg)
Column: 4.6-mm 15-cm; packing L7 Tolerances: NLT 75% (Q) of the labeled amount of C9H8O4,
Flow rate: 2 mL/min C8H10N4O2, and C18H23NO3 C4H6O6 is dissolved.
[NOTEThe relative retention times for caffeine,
dihydrocodeine, aspirin, and salicylic acid are 0.2, 0.3,
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IDENTIFICATION

PROCEDURE
IMPURITIES Diluent, Mobile phase, Sample solution, Chromatographic
Organic Impurities System, and System suitability: Proceed as directed in the
PROCEDURE: LIMIT OF SALICYCLIC ACID Assay.
[NOTEThe results from the Assay may be used for this test Standard solution A: 3.3 mg/mL of USP Aspirin RS in
when calculated as described in the Analysis section of this Diluent
test.] Standard solution B: 1 mg/mL of USP Codeine Phosphate
Mobile phase, Diluent, Standard solution, Salicylic acid RS in Diluent
standard solution, System suitability solution, and Analysis
Sample solution: Proceed as directed in the Assay. Samples: Sample solution, Standard solution A, and
Chromatographic system Standard solution B
(See Chromatography 621, System Suitability.) Chromatograph these solutions as directed for the Assay.
Mode: LC Acceptance criteria: The retention times of the major peaks
Detector: UV 215 nm of the Sample solution correspond to those of Standard
Column: 4.6-mm 15-cm; packing L7 solution A and Standard solution B.
Flow rate: 2 mL/min
[NOTEThe relative retention times for caffeine, PROCEDURE
dihydrocodeine, aspirin, and salicylic acid are 0.2 , 0.3, Mobile phase: Dissolve 225 mg of tetramethylammonium
0.7, and 1.0, respectively.] hydroxide pentahydrate and 200 mg of sodium 1-
System suitability octanesulfonate in 700 mL of water. Add 150 mL of
Samples: Standard solution and System suitability solution methanol, 150 mL of acetonitrile, and 1.0 mL of glacial
Suitability requirements acetic acid, and stir.
Resolution: NLT 2.5 between the caffeine and Diluent: To 15 g of anhydrous citric acid add 200 mL of
dihydrocodeine peaks, NLT 1.0 between the methanol and 20 mL of glacial acetic acid, dilute with
dihydrocodeine and aspirin peaks, and NLT 1.5 between chloroform to 1000 mL, and mix until the citric acid is
the aspirin and salicylic acid peaks, System suitability dissolved.
solution Internal standard solution: 2 mg/mL of phenacetin in
Relative standard deviation: NMT 2.0% for each Diluent
analyte, Standard solution Salicylic acid standard stock solution: 1 mg/mL of USP
Analysis Salicylic Acid RS in Diluent
Samples: Standard solution, Salicylic acid standard solution, Salicylic acid standard solution: Salicylic acid standard stock
and Sample solution solution, Internal standard solution, and Diluent (1:1:8)
Calculate the percentage of salicylic acid in the Capsules Codeine phosphate standard stock solution: Transfer 325J
taken: mg of USP Codeine Phosphate RS, to a 25-mL volumetric
flask, J being the ratio of the labeled amount mg of codeine
Result = (rU/rS) (CS/CU) 100 phosphate to the labeled amount mg of aspirin/Tablet.
Dissolve in and dilute with Diluent to volume.
rU = peak response of salicylic acid from the Sample Standard solution: 6.5 mg/mL of USP Aspirin RS in a
solution solution of Codeine phosphate standard stock solution, Salicylic
rS = peak response of salicylic acid from the Standard acid standard stock solution, Internal standard solution, and
solution Diluent (5:1:1:3)
CS = concentration of USP Salicylic Acid RS in the Sample solution: Transfer an equivalent to 325 mg of
Standard salicylic acid solution (mg/mL) aspirin, from finely powdered Tablets (NLT 20), to a screw-
CU = nominal concentration of aspirin in the portion capped, 120-mL bottle, add 5.0 mL of Internal standard
of capsules taken of the Sample solution as solution and 45.0 mL of Diluent, and sonicate for 25 min.
determined in the Assay (mg/mL) Centrifuge, and use clear supernatant. [NOTEUse on the
Acceptance criteria: NMT 3.0% day prepared.]
ADDITIONAL REQUIREMENTS (See Chromatography 621, System Suitability.)
PACKAGING AND STORAGE: Preserve in tight containers. Mode: LC
USP Aspirin RS Column: 3.9-mm 30-cm; 10-m packing L1
USP Caffeine RS Flow rate: 2 mL/min
USP Dihydrocodeine Bitartrate RS Injection size: 5 L
USP Salicylic Acid RS System suitability
Samples: Salicylic acid standard solution and Standard
solution
Aspirin and Codeine Phosphate Tablets [NOTEThe relative retention times for salicylic acid,
aspirin, codeine, and phenacetin are about 0.3, 0.5, 0.8,
(Comment on this Monograph)id=m6313=Aspirin and Codeine and 1.0, respectively.]
Phosphate Tablets=A-Monos.pdf) Suitability requirements
DEFINITION Resolution: NLT 2.0 between salicylic acid and aspirin,
Aspirin and Codeine Phosphate Tablets contain NLT 90.0% and between aspirin and codeine, and between codeine and
NMT 110.0% of the labeled amounts of aspirin (C9H8O4) and phenacetin, Standard solution
codeine phosphate hemihydrate (C18H21NO3 H3PO4 1/2H2O).
Tailing factor: NMT 2.0 for each analyte peak Injection size: 10 L for System suitability and 50 L for
Relative standard deviation: NMT 3.0% of the ratios of Analysis
the peak responses of salicylic acid, aspirin, and codeine System suitability
to the peak response of phenacetin, Standard solution Sample: System suitability solution
Analysis Proceed as directed in the Assay.
Samples: Standard solution and Sample solution Analysis
Calculate the percentage of C9H8O4 in the portion of Samples: Standard solution A, Standard solution B, and
Tablets taken: Sample solution
Calculate the amount of codeine phosphate dissolved by
Result = (RU/RS) (CS/CU) 100 comparison of the relative peak response ratios for the
codeine phosphate peaks, obtained from Standard solution
RU = peak response ratio of aspirin to phenacetin B and the Sample solution.
from the Sample solution Calculate the percentage of aspirin dissolved:
RS = peak response ratio of aspirin to phenacetin
from the Standard solution Result = 100 [V C (RU/RS) + V C (RU/RS)
CS = concentration of USP Aspirin RS in the Standard (Mr1/Mr2)]/(L F)
solution (mg/mL)
CU = nominal concentration for the Sample solution V = volume of Medium (mL), 0.900
(mg aspirin/mL) C = concentration of USP Aspirin RS in Standard
Calculate the percentage of C18H21NO3 H3PO4 1/2H2O in solution A (g/mL)
the portion of Tablets taken: RU = peak response ratio for the aspirin component
from the Sample solution
Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 RS = peak response ratio for the aspirin component
from the Standard solution A
RU = peak response ratio of codeine phosphate to C = concentration of USP Salicylic Acid RS in
phenacetin from the Sample solution Standard solution B (g/mL)
RS = peak response ratio of codeine phosphate to RU = peak response ratio for the salicylic acid
phenacetin from the Standard solution component from the Sample solution
CS = concentration of USP Codeine Phosphate RS in RS = peak response ratio for the salicylic acid
the Standard solution mg/mL component from the Standard solution B
CU = nominal concentration for the Sample solution Mr1 = molecular weight of aspirin, 180.16
(mg/mL) Mr2 = molecular weight of salicylic acid, 138.12
Mr1 = molecular weight of codeine phosphate L = labeled amount of aspirin in 1 tablet (mg)
hemihydrate, 406.37 F = conversion factor, 1000 g/mg
Mr2 = molecular weight of anhydrous codeine Tolerances: NLT 75% (Q) of the labeled amounts of
phosphate, 397.37 C9H8O4 and C18H21NO3 H3PO4 1/2H2O is dissolved.
Acceptance criteria: 90.0%110.0% UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
for Content Uniformity with respect to aspirin and codeine
PERFORMANCE TESTS phosphate.
DISSOLUTION 711
0.05 M acetate buffer: Mix 2.99 g of sodium acetate IMPURITIES
trihydrate and 1.66 mL of glacial acetic acid with water to Organic Impurities
obtain 1000 mL of solution having a pH of 4.50 0.05. PROCEDURE: LIMIT OF FREE SALICYLIC ACID
Medium: 0.05 M acetate buffer, 900 mL Mobile phase, Diluent, Internal standard solution,
Apparatus 2: 75 rpm Salicylic acid standard stock solution, Salicylic acid
Time: 30 min standard solution, Codeine phosphate standard stock
Determine the amounts of C9H8O4 and C18H21NO3 H3PO4 solution, Standard solution, Sample solution,
1
/2H2O dissolved by employing the following method. Chromatographic system, and System suitability:
Mobile phase and Diluent: Proceed as directed in the Proceed as directed in the Assay.
Assay. Analysis
System suitability solution: Use the Standard solution from Samples: Salicylic acid standard solution and Sample
the Assay. solution
Internal standard solution: 0.07 mg/mL of phenacetin in Calculate the percentage of free salicylic acid in the
methanol Tablets taken:
Standard stock solution A: 0.36 mg/mL of USP Aspirin RS
in Diluent Result = (RU/RS) (CS/CU) 100
Standard stock solution B: Transfer 12 mg of USP Codeine
Phosphate RS and 25 mg of USP Salicylic Acid RS to a 50- RU = peak response ratio of salicylic acid to
mL volumetric flask, and add 2.5 mL of methanol. Add phenacetin from the Sample solution
Medium to volume. Pipet 10 mL of the resulting solution RS = peak response ratio of salicylic acid to
into a 100-mL volumetric flask, and add Medium to volume. phenacetin from the Salicylic acid standard
Standard solutions A and B: Pipet 10 mL of Standard stock solution
solution A and 10 mL of Standard stock solution B into CS = concentration of USP Salicylic Acid RS in the
separate containers, and add 3.0 mL of the Internal standard Salicylic acid standard solution (mg/mL)
solution to each container. CU = nominal concentration of aspirin in the Sample
Sample solution: Withdraw a portion of the solution under solution
test and filter, discarding the few mL of the filtrate. Pipet 10 Acceptance criteria: NMT 3.0%
mL of the filtrate and 3.0 mL of the Internal standard
solution into a suitable container. ADDITIONAL REQUIREMENTS
Chromatographic system: Proceed as directed under Assay PACKAGING AND STORAGE: Preserve in well-closed, light-
except to use the following injection size: resistant containers.
USP REFERENCE STANDARDS 11 the labeled amount, in mg, of codeine phosphate to the
USP Aspirin RS labeled amount, in mg, of aspirin per Tablet.) Dissolve in
USP Codeine Phosphate RS and dilute with Diluent to volume.
USP Salicylic Acid RS Standard solution: Transfer 65 mg of USP Aspirin RS to a
10-mL volumetric flask. Add 5.0 mL of Codeine phosphate
standard stock solution, 1.0 mL of Salicylic acid standard stock
solution, and 1.0 mL of Internal standard solution, and dilute
Aspirin, Codeine Phosphate, Alumina, with Diluent to volume.
and Magnesia Tablets Sample solution: Transfer powdered Tablets (NLT 20),
(Comment on this Monograph)id=m6314=Aspirin, Codeine equivalent to 325 mg of aspirin, to a suitable vessel, add 5.0
Phosphate, Alumina, and Magnesia Tablets=A-Monos.pdf) mL of Internal standard solution and 45.0 mL of Diluent, and
sonicate for 25 min. Centrifuge, and use the resultant clear
DEFINITION solution. [NOTEUse on the day prepared.]
Aspirin, Codeine Phosphate, Alumina, and Magnesia Tablets Chromatographic system
contain NLT 90.0% and NMT 110.0% of the labeled amounts (See Chromatography 621, System Suitability.)
of aspirin (C9H8O4), codeine phosphate hemihydrate Mode: LC
(C18H21NO3 H3PO4 1/2H2O), aluminum hydroxide [Al(OH)3], Detector: UV 280 nm
and magnesium hydroxide [Mg(OH)2]. Column: 3.9-mm 30-cm; 10-m packing L1
Flow rate: 2 mL/min
PROCEDURE System suitability
[NOTEThe Sample, Diluent, Aspirin standard solution, and Samples: Salicylic acid standard solution and Standard
Codeine phosphate standard solution are prepared as solution
follows.] [NOTEThe relative retention times for salicylic acid,
Sample: To a 0.7-g portion of finely powdered Tablets, add aspirin, codeine, and phenacetin are 0.3, 0.5, 0.8, and
10 mL of 3 N hydrochloric acid and 5 drops of methyl red 1.0, respectively.]
TS, heat to boiling, and add 6 N ammonium hydroxide until Suitability requirements
the color of the solution changes to deep yellow. Continue Resolution: NLT 2.0 between salicylic acid and aspirin;
boiling for 2 min, and filter. The filtrate is used in NLT 2.0 between aspirin and codeine; NLT 2.0 between
Identification B and the precipitate is used in Identification C. codeine and phenacetin, Standard solution
Diluent: To 15 g of anhydrous citric acid, add 200 mL of Tailing factor: NMT 2.0 for each analyte peak, Salicylic
methanol and 20 mL of glacial acetic acid, dilute with acid standard solution and Standard solution
chloroform to 1000 mL, and mix until the citric acid is Relative standard deviation: NMT 3.0% of the ratios of
dissolved. the peak responses of salicylic acid, aspirin, and codeine
Aspirin standard solution: 3.3 mg/mL of USP Aspirin RS in to the peak response of phenacetin, Salicylic acid standard
the Diluent solution and Standard solution
Codeine phosphate standard solution: 1 mg/mL of USP Analysis
Codeine Phosphate RS in the Diluent Samples: Salicylic acid standard solution, Standard solution,
A. The retention time of the aspirin and codeine peaks of and Sample solution
the Sample solution, obtained as in the Assay for Aspirin and Calculate the percentage of C9H8O4 in the portion of
Codeine Phosphate, corresponds to that of the Aspirin Tablets taken:
standard solution and Codeine phosphate standard solution.
B. IDENTIFICATION TESTSGENERAL, Magnesium 191 Result = (RU/RS) (CS/CU) 100
Sample solution: The Sample filtrate is used.
C. IDENTIFICATION TESTSGENERAL, Aluminum 191 RU = peak response ratio of aspirin to phenacetin
Sample solution: Wash the Sample precipitate with a hot from the Sample solution
solution of ammonium chloride (1 in 50), and dissolve the RS = peak response ratio of aspirin to phenacetin
precipitate in hydrochloric acid: the solution so obtained from the Standard solution
meets the requirements. CS = concentration of USP Aspirin RS in the Standard
solution (mg/mL)
ASSAY CU = nominal concentration of the Sample solution
ASPIRIN AND CODEINE PHOSPHATE (mg/mL)
Mobile phase: Dissolve 225 mg of tetramethylammonium Calculate the percentage of C18H21NO3 H3PO4 1/2H2O in
hydroxide pentahydrate and 200 mg of sodium 1- the portion of Tablets taken:
octanesulfonate in 700 mL of water. Add 150 mL of
methanol, 150 mL of acetonitrile, and 1.0 mL of glacial Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100
acetic acid, and stir.
Diluent: To 15 g of anhydrous citric acid, add 200 mL of RU = peak response ratio of codeine phosphate to
methanol and 20 mL of glacial acetic acid, dilute with phenacetin from the Sample solution
chloroform to 1000 mL, and mix until the citric acid is RS = peak response ratio of codeine phosphate to
dissolved. phenacetin from the Standard solution
Internal standard solution: 2 mg/mL of phenacetin in CS = concentration of USP Codeine Phosphate RS in
Diluent the Standard solution (mg/mL)
Salicylic acid standard stock solution: 1 mg/mL of USP CU = nominal concentration of the Sample solution
Salicylic Acid RS in Diluent (mg/mL)
Salicylic acid standard solution: Transfer 5.0 mL of Salicylic Mr1 = molecular weight of codeine phosphate
acid standard stock solution to a 50-mL volumetric flask, add hemihydrate, 406.37
5.0 mL of Internal standard solution, and dilute with Diluent Mr2 = molecular weight of anhydrous codeine
to volume. phosphate, 397.37
Codeine phosphate standard stock solution: 13J mg/mL Acceptance criteria: 90.0%110.0% of the labeled
of USP Codeine Phosphate RS in Diluent (J being the ratio of amounts of C9H8O4 and C18H21NO3 H3PO4 1/2H2O.
ALUMINUM HYDROXIDE Mobile phase, Diluent, and Aspirin and codeine phosphate
Edetate disodium titrant: Dissolve 18.6 g of edetate standard solution: Prepare as directed in the Assay for
disodium in water to make 1000 mL, and standardize the Aspirin and codeine phosphate and Procedure: Limit of free
solution as follows. Weigh 2 g of aluminum wire, transfer to salicylic acid.
a 1000-mL volumetric flask, and add 50 mL of a mixture of Internal standard solution: 0.07 mg/mL of phenacetin in
hydrochloric acid and water (1:1). Swirl the flask to ensure methanol
contact of the aluminum and the acid, and allow the Standard stock solution A: 0.36 mg/mL of USP Aspirin RS
reaction to proceed until all of the aluminum has dissolved. in Diluent
Dilute with water to volume. Pipet 10 mL of this solution Standard solution A: To 10 mL of Standard stock solution A,
into a 250-mL beaker, add, in the order named and with add 3.0 mL of the Internal standard solution.
continuous stirring, 25.0 mL of Edetate disodium titrant and Standard stock solution B: Transfer 12 mg of USP Codeine
20 mL of acetic acidammonium acetate buffer TS, and boil Phosphate RS and 25 mg of USP Salicylic Acid RS to a 50-
gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL mL volumetric flask, and add 2.5 mL of methanol. Add
of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a Medium to volume. Pipet 10 mL of the resulting solution
bright rose-pink color. Perform a blank determination, into a 100-mL volumetric flask, and add Medium to volume.
substituting 10 mL of water for the aluminum solution, and Standard solution B: To 10 mL of Standard stock solution B,
make any necessary correction. add 3.0 mL of the Internal standard solution.
Calculate the molarity of the solution taken: Sample solution: Withdraw a portion of the solution under
test and filter, discarding the few mL of the filtrate. Pipet 10
Result = W/26.98V mL of the filtrate and 3.0 mL of the Internal standard
solution into a suitable container.
W = weight of aluminum in the portion of solution Chromatographic system: Proceed as directed for
taken (mg) Chromatographic system in the Assay for Aspirin and codeine
V = volume of Edetate disodium titrant consumed phosphate except to use only the Standard solution for
(mL) evaluation of the suitability of the system.
Sample solution: Transfer an equivalent to 600 mg of Analysis: Proceed as directed in the Assay for Aspirin and
aluminum hydroxide, from finely powdered Tablets (NLT codeine phosphate except to inject 50 L of Standard solution
20), to a 150-mL beaker, add 20 mL of water, stir, and A, Standard solution B, and the Sample solution.
slowly add 30 mL of 3 N hydrochloric acid. Heat gently, if Calculate the amount of codeine phosphate dissolved by
necessary, to aid solution, cool, and filter into a 200-mL comparison of the relative peak response ratios for the
volumetric flask. Wash the filter with water into the flask, codeine phosphate peaks, of the Standard solution B, and
and add water to volume. the Sample solution.
Analysis: Pipet 10 mL of Sample solution into a 250-mL Calculate the percentage of aspirin dissolved:
beaker, add 20 mL of water, then add, in the order named
and with continuous stirring, 25.0 mL of Edetate disodium Result = [0.9 C (RU/RS) + 0.9C (RU/RS) (Mr1/Mr2)]/3.25
TS. Add 50 mL of alcohol and 2 mL of dithizone TS. Titrate C = concentration of USP Aspirin RS in Standard
with 0.05 M zinc sulfate VS until the color changes from solution A (g/mL)
green-violet to rose-pink. Perform a blank determination, RU = peak response ratio for the aspirin component
substituting 10 mL of water for the Sample solution, and from the Sample solution
make any necessary correction. Each mL of 0.05 M Edetate RS = peak response ratio for the aspirin component
disodium titrant is equivalent to 3.900 mg of Al(OH)3. from Standard solution A
Acceptance criteria: 90.0%110.0% C = concentration of USP Salicylic Acid RS in Standard
MAGNESIUM HYDROXIDE solution B (g/mL)
Sample solution: Prepare as directed in the Assay for RU = peak response ratios for the salicylic acid
Aluminum hydroxide. component from the Sample solution
Analysis: Pipet a volume of Sample solution, equivalent to RS = peak response ratio for the salicylic acid
40 mg of magnesium hydroxide, into a 400-mL beaker, add component from the Sample solution and
200 mL of water and 20 mL of triethanolamine, and stir. Standard solution B
Add 10 mL of ammoniaammonium chloride buffer TS and Mr1 = molecular weight of aspirin, 180.16
3 drops of an eriochrome black indicator solution (prepared Mr2 = molecular weight of salicylic acid, 138.12
by dissolving 200 mg of eriochrome black T in a mixture of Tolerances: NLT 75% (Q) of the labeled amounts of C9H8O4
15 mL of triethanolamine and 5 mL of dehydrated alcohol, and C18H21NO3 H3PO4 1/2H2O is dissolved.
and mixing). Cool the solution to between 3 and 4 by UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
immersion of the beaker in an ice bath, then remove, and for Content Uniformity with respect to aspirin and codeine
titrate with 0.05 M edetate disodium VS to a blue endpoint. phosphate and for Weight Variation with respect to
Perform a blank determination, substituting 10 mL of water aluminum hydroxide and magnesium hydroxide
for the Sample solution, and make any necessary correction.
Each mL of 0.05 M edetate disodium consumed is IMPURITIES
equivalent to 2.916 mg of Mg(OH)2. Organic Impurities
Acceptance criteria: 90.0%110.0% PROCEDURE: LIMIT OF FREE SALICYLIC ACID
[NOTEThe results from the Assay for Aspirin and codeine
PERFORMANCE TESTS phosphate may be used for this test when calculated as
DISSOLUTION 711 described in the Analysis section of this test.]
Medium: 0.05 M acetate buffer, prepared by mixing 2.99 g Mobile phase, Diluent, Salicylic acid standard stock
of sodium acetate trihydrate and 1.66 mL of glacial acetic solution, Salicylic acid standard solution, Codeine
acid with water to obtain 1000 mL of solution having a pH phosphate standard stock solution, Standard solution,
of 4.50 0.05; 900 mL and Sample solution: Proceed as directed in Assay for
Apparatus 2: 75 rpm Aspirin and codeine phosphate.
Time: 30 min Internal standard solution: 2 mg/mL of phenacetin in
Determine the amounts of C9H8O4 and C18H21NO3 H3PO4 Diluent
1
/2H2O dissolved by using the following method.

(See Chromatography 621, System Suitability.) INFRARED ABSORPTION 197K
Mode: LC
Detector: UV 280 nm ASSAY
Column: 3.9-mm 30-cm; 10-m packing L1 PROCEDURE
Flow rate: 2 mL/min Mobile phase: Acetonitrile, methanol, diethylamine, and
Injection size: 5 L 0.13 M ammonium acetate (230:470:1.0:300). Adjust with
System suitability glacial acetic acid to a pH of 7.5.
Samples: Salicylic acid standard solution and Standard Standard solution: 1 mg/mL of USP Astemizole RS in
solution Mobile phase
[NOTEThe relative retention times for salicylic acid, Sample solution: 1 mg/mL of Astemizole in Mobile phase
aspirin, codeine, and phenacetin are 0.3, 0.5, 0.8, and Chromatographic system
1.0, respectively.] (See Chromatography 621, System Suitability.)
Suitability requirements Mode: LC
Resolution: NLT 2.0 between salicylic acid and aspirin; Detector: UV 220 nm
NLT 2.0 between aspirin and codeine; NLT 2.0 between Column: 4.6-mm 25-cm; packing L1
codeine and phenacetin Flow rate: 2 mL/min
Tailing factor: NMT 2.0 for each analyte peak Injection size: 10 L
Relative standard deviation: NMT 3.0% of the ratios of System suitability
the peak responses of salicylic acid, aspirin, and codeine Sample: Standard solution
to the peak response of phenacetin Suitability requirements
Analysis: Calculate the percentage of free salicylic acid in Column efficiency: NLT 4000 theoretical plates
the Tablets taken: Tailing factor: NMT 1.8
Result = (RU/RS) (CS/CU) 100 Analysis
RU = ratio of the peak response of salicylic acid to Calculate the percentage of C28H31FN4O in the portion of
phenacetin from the Sample solution Astemizole taken:
RS = ratio of the peak response of salicylic acid to
phenacetin from the Salicylic acid standard Result = (rU/rS) (CS/CU) 100
solution
CS = concentration of USP Salicylic Acid RS in the rU = peak response from the Sample solution
Salicylic acid standard solution (mg/mL) rS = peak response from the Standard solution
CU = nominal concentration of aspirin in the portion CS = concentration of USP Astemizole RS in the
of tablets taken for the Sample solution as Standard solution (mg/mL)
determined in the Assay for Aspirin and codeine CU = concentration of the Sample solution (mg/mL)
phosphate (mg/mL) Acceptance criteria: 98.0%102.0%
IMPURITIES
ACID-NEUTRALIZING CAPACITY 301: NLT 1.9 mEq/Tablet RESIDUE ON IGNITION 281: NMT 0.1%
PACKAGING AND STORAGE: Preserve in well-closed, light- PROCEDURE
resistant containers. Solution A: 17 mg/mL of tetrabutylammonium hydrogen
USP REFERENCE STANDARDS 11 sulfate in water
USP Aspirin RS Solution B: Acetonitrile
USP Codeine Phosphate RS Standard solution: 25 g/mL of USP Astemizole RS in
USP Salicylic Acid RS methanol
Sample solution: 10 mg/mL of Astemizole in methanol
System suitability solution: 25 g/mL of USP Astemizole
Astemizole RS and 250 g/mL of ketoconazole in methanol
Mobile phase: See the gradient table below.
(Comment on this Monograph)id=m6328=Astemizole=A-
Monos.pdf)
(min) (%) (%)
Equilibrate 0 100
5 95 5
0 95 5
15 80 20
18 80 20
C28H31FN4O 458.58
1H-Benzimidazol-2-amine, 1-[(4-fluorophenyl)methyl]-N-[1-[2- 18.1 0 100
(4-methoxyphenyl)ethyl]-4-piperidinyl]-; 23 0 100
1-(p-Fluorobenzyl)-2-[[1-(p-methoxyphenethyl)-4-
piperidyl]amino]benzimidazole [68844-77-9]. Chromatographic system
DEFINITION
Astemizole contains NLT 98.0% and NMT 102.0% of
C28H31FN4O, calculated on the dried basis.
USP 32 Official Monographs / Astemizole 297
Mode: LC ASSAY
Detector: UV 278 nm PROCEDURE
Column: 4.6-mm 10-cm; contains base-deactivated 3- Mobile phase: Acetonitrile, methanol, diethylamine and
m packing L1 0.13 M ammonium acetate (230:470:1.0:300). Adjust with
Flow rate: 1 mL/min glacial acetic acid to a pH of 7.5.
Injection size: 10 L Standard solution: 1 mg/mL of USP Astemizole RS in
System suitability Mobile phase
Sample: System suitability solution Sample solution: Equivalent to 1 mg/mL of astemizole,
Suitability requirements from powdered Tablets in Mobile phase
Resolution: NLT 1.5 between the astemizole and [NOTEFinely powder NLT 20 Tablets. Mix a suitable
ketoconazole peaks quantity of powder with Mobile phase corresponding to
Analysis 50% of the final volume for 30 min. Dilute to volume and
Samples: Standard solution and Sample solution centrifuge. Use the supernatant.]
Calculate the percentage of each impurity in the portion Chromatographic system
of Astemizole taken: (See Chromatography 621, System Suitability.)
Mode: LC
Result = (ri/rS) (CS/CU) 100 Detector: UV 220 nm
ri = peak response for each impurity Flow rate: 2 mL/min
rS = peak response from the Standard solution Injection size: 10 L
CS = concentration of USP Astemizole RS in the System suitability
Standard solution (mg/mL) Sample: Standard solution
CU = concentration of the Sample solution (mg/mL) Suitability requirements
Acceptance criteria Column efficiency: NLT 4000 theoretical plates
Individual impurities: NMT 0.25% Tailing factor: NMT 1.8
Total impurities: NMT 0.5% Relative standard deviation: NMT 1.5%
Analysis
SPECIFIC TESTS Samples: Standard solution and Sample solution
LOSS ON DRYING 731: Dry it in a vacuum at 105 for 4 h: it Calculate the percentage of C28H31FN4O in the portion of
loses NMT 0.5% of its weight. Tablets taken:
PACKAGING AND STORAGE: Preserve in tight containers. rU = peak response from the Sample solution
USP Astemizole RS CS = concentration of USP Astemizole RS in the
CU = nominal concentration of astemizole in the
Astemizole Tablets Sample solution (mg/mL)
(Comment on this Monograph)id=m6329=Astemizole
Tablets=A-Monos.pdf) PERFORMANCE TESTS
DISSOLUTION 711
DEFINITION Medium: Simulated gastric fluid TS (without the enzyme);
Astemizole Tablets contain NLT 90.0% and NMT 110.0% of the 800 mL
labeled amount of astemizole (C28H31FN4O). Apparatus 2: 100 rpm
IDENTIFICATION Time: 45 min
THIN-LAYER CHROMATOGRAPHY621 Detector: UV 285 nm
Standard solution: 1 mg/mL of USP Astemizole RS in Sample solution: Sample per Dissolution 711. Dilute with
methanol Medium to a concentration that is similar to the Standard
Sample solution: Equivalent to 1 mg/mL of Astemizole, solution.
from finely ground Tablets in methanol [NOTEFilter before Standard solution: USP Astemizole RS in Medium
use.] Tolerances: NLT 80% (Q) of the labeled amount of
Chromatographic system C28H31FN4O is dissolved.
(See Chromatography 621, Thin-Layer Chromatography.) UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Mode: TLC IMPURITIES
Adsorbent: 0.25-mm layer of chromatographic silica gel Organic Impurities
mixture PROCEDURE
Application volume: 10 L Mobile phase, Standard solution, and Chromatographic
Developing solvent system: Toluene, dioxane, methanol, system: Proceed as directed in the Assay.
and ammonium hydroxide (60:30:10:1) Sample solution: Use the Sample solution from the Assay.
Analysis Analysis
Samples: Standard solution and Sample solution Sample: Sample solution
Develop the chromatogram in solvent system, until the Calculate the percentage of each impurity in the portion
solvent front has moved about three-fourths of the length of Tablets taken:
of the plate. Remove the plate from the developing
chamber, mark the solvent front, air-dry, and examine Result = (ri/rT) 100
under short-wavelength UV light.
Acceptance criteria: The RF value of the principal spot of ri = peak response for each impurity
the Sample solution corresponds to that of the Standard rT = sum of the responses of all of the peaks
solution.
298 Astemizole / Official Monographs USP 32
Acceptance criteria rS = peak response from the Standard solution

Individual impurities: NMT 0.25% CS = concentration of USP Atenolol RS in the Standard
Total impurities: NMT 1.0% solution (mg/mL)
CU = concentration of Atenolol in the Sample solution
PACKAGING AND STORAGE: Preserve in tight containers. Acceptance criteria: 98.0%102.0%
USP Astemizole RS IMPURITIES
CHLORIDE AND SULFATE, Chloride 221
Atenolol Sample solution: 10 mg/mL in 0.15 N nitric acid, made to
(Comment on this Monograph)id=m6330=Atenolol=A- 100 mL
Monos.pdf) Acceptance criteria: Shows no more turbidity with 1 mL
of silver nitrate TS than 1.4 mL of 0.020 N hydrochloric
acid in 100 mL of 0.15 N nitric acid (0.1%)
Organic Impurities
PROCEDURE
Mobile phase: Prepare as directed in the Assay.
Sample solution 1: 0.1 mg/mL of Atenolol in Mobile phase
Sample solution 2: 0.005 mg/mL Atenolol, made from
Sample stock solution diluted with Mobile phase
C14H22N2O3 266.34 Chromatographic system: Proceed as directed in the
Benzeneacetamide, 4-[2-hydroxy-3-[(1- Assay, except use the injection size listed below.
methylethyl)amino]propoxy]-; Injection size: 50 L
2-[p-[2-Hydroxy-3-(isopropylamino)propoxy]-phenyl]acetamide Analysis
[29122-68-7]. Samples: Sample solution 1 and Sample solution 2
[NOTEChromatograph Sample solution 1 for a period of
DEFINITION time that is 6 times the retention time of the atenolol
Atenolol contains NLT 98.0% and NMT 102.0% of C14H22N2O3, peak.]
calculated on the dried basis. Calculate the percentage of each impurity observed in the
IDENTIFICATION chromatogram obtained from the Sample stock solution:
B. ULTRAVIOLET ABSORPTION 197U Result = 0.5(ri/rA)
Sample solution: 50 g/mL in methanol ri = peak response of the Sample solution
ASSAY rA = response of the main atenolol peak of the
PROCEDURE Sample solution
Mobile phase: 1.1 g of sodium 1-heptanesulfonate and Acceptance criteria
0.71 g of anhydrous dibasic sodium phosphate in 700 mL of Individual impurities: NMT 0.25%
water. Add 2 mL of dibutylamine, and adjust with 0.8 M Total impurities: NMT 0.5%
phosphoric acid to a pH of 3.0. Add 300 mL of methanol, SPECIFIC TESTS
mix, and pass through a filter having a 0.5-m or finer MELTING RANGE OR TEMPERATURE, Class I 741: 152156.5
porosity. Degas this solution before use. LOSS ON DRYING 731: Dry it at 105 to constant weight: it
Standard solution: 0.01 mg/mL of USP Atenolol RS in loses NMT 0.5% of its weight.
Mobile phase
Sample solution: 0.01 mg/mL of Atenolol in Mobile phase ADDITIONAL REQUIREMENTS
Chromatographic system PACKAGING AND STORAGE: Preserve in well-closed containers.
(See Chromatography 621, System Suitability.) Store at room temperature.
Detector: UV 226 nm USP Atenolol RS
System suitability Atenolol Injection
Sample: Standard solution (Comment on this Monograph)id=m6332=Atenolol Injection=A-
Suitability requirements Monos.pdf)
Tailing factor: NMT 2.0 DEFINITION
Relative standard deviation: NMT 2.0% Atenolol Injection is a sterile solution of Atenolol in Water for
Analysis Injection. It contains a suitable buffering agent. It contains NLT
Samples: Standard solution and Sample solution 90.0% and NMT 110.0% of the labeled amount of atenolol
Calculate the percentage of C14H22N2O3 in the portion of (C14H22N2O3).
Atenolol taken:
IDENTIFICATION
Result = (rU/rS) (CS/CU) 100 A. The retention time of the main peak of the Sample
solution corresponds to that of the Standard solution,
rU = peak response from the Sample solution obtained in the Assay.
USP 32 Official Monographs / Atenolol 299
B. ULTRAVIOLET ABSORPTION 197U Atenolol Oral Solution

Solution: 10 g/mL of atenolol in methanol (Comment on this Monograph)id=m386=Atenolol Oral
ASSAY Solution=A-Monos.pdf)
Mobile phase: 930 mg of sodium octyl sulfate in 740 mL Atenolol Oral Solution contains NLT 90.0% and NMT 110.0%
of water. Add 8 mL of 3.6 N sulfuric acid and pass through of the labeled amount of C14H22N2O3. Prepare Atenolol Oral
a 1-m or finer porosity filter. To the filtrate add 250 mL of Solution at a 0.2% concentration, for example, as follows (see
acetonitrile. Pharmaceutical CompoundingNonsterile Preparations 795).
Citric acid buffer: 2.5 g of citric acid to a 500-mL
volumetric flask. Add 400 mL of water, and swirl to dissolve.
Adjust the solution with 2 N sodium hydroxide to a pH of Atenolol 200 mg
6.0, and dilute with water to volume. Glycerin 5 mL
Standard stock solution: 50 mg of USP Atenolol RS to a Vehicle for Oral Suspension 45 mL
100-mL volumetric flask. Add 80 mL of Citric acid buffer, and
sonicate for 30 s to achieve dissolution. Dilute with Citric Vehicle for Oral Solution, Sugar-Free A sufficient quantity
acid buffer to volume. To make 100 mL
Standard solution: 0.2 mg/mL of USP Atenolol RS from
Standard stock solution diluted with Citric acid buffer Calculate the quantity of each ingredient required for the total
Sample solution: Nominally equivalent to 0.2 mg/mL of volume and atenolol strength to be prepared. Mix the
atenolol from Injection diluted with Citric acid buffer Atenolol, previously pulverized, and Glycerin to form a smooth
Chromatographic system paste. Incorporate the Vehicle for Oral Suspension or an equal
(See Chromatography 621, System Suitability.) volume of Vehicle for Oral Solution, Sugar Free. [NOTEThe
Mode: LC Vehicle for Oral Suspension may be omitted.] Incorporate
Detector: UV 275 nm sufficient Vehicle for Oral Solution, Sugar Free in increments to
Column: 4.6-mm 25-cm; 5-m packing L1 bring to volume, and mix well. [NOTEDo not use a sucrose-
Flow rate: 1.7 mL/min containing vehicle for oral solution.] Package, and label.
System suitability ADDITIONAL REQUIREMENTS
Sample: Standard solution PACKAGING AND STORAGE: Package in amber, tight
Suitability requirements containers, and store at controlled room temperature.
Tailing factor: NMT 2 LABELING: Label it to state that it is to be shaken well before
Relative standard deviation: NMT 2.0% use, and discarded after 60 days. Label it to state that it is to
Analysis be kept out of reach of children. Label it to indicate the
Samples: Standard solution and Sample solution nominal atenolol concentration.
Calculate the percentage of C14H22N2O3 in each mL of the BEYOND-USE DATE: NMT 60 days after preparation.
Injection taken:
Atenolol Tablets
rU = peak response from the Sample solution (Comment on this Monograph)id=m6335=Atenolol Tablets=A-
rS = peak response from the Standard solution Monos.pdf)
CS = concentration of USP Atenolol RS in the Standard
solution (mg/mL) DEFINITION
CU = nominal concentration of atenolol in the Sample Atenolol Tablets contain NLT 90.0% and NMT 110.0% of the
solution (mg/mL) labeled amount of atenolol (C14H22N2O3).
IDENTIFICATION
SPECIFIC TESTS A. INFRARED ABSORPTION 197K
PARTICULATE MATTER IN INJECTIONS 788: Meets the Sample: Mix a quantity of powdered Tablets, equivalent to
requirements for small-volume injections 100 mg of atenolol, with 15 mL of methanol, heat the
PH 791: 5.56.5 mixture to 50, and shake for 5 min. Filter, and evaporate
STERILITY TESTS 71: It meets the requirements when tested the filtrate on a water bath to dryness. Add 10 mL of 0.1 N
as directed under Test for Sterility of the Product to Be hydrochloric acid to the residue, warm the solution, shake,
Examined, Membrane Filtration. and filter. To the filtrate add sufficient 1 N sodium hydroxide
BACTERIAL ENDOTOXINS TEST 85: It contains NMT 33.3 USP to make it alkaline, extract the solution with 10 mL of
Endotoxin Units/mg of atenolol. chloroform, drying the chloroform extract over anhydrous
sodium sulfate. Filter the dried chloroform solution,
ADDITIONAL REQUIREMENTS evaporate the filtrate on a water bath to dryness, and dry
PACKAGING AND STORAGE: Preserve in single-dose or in the residue at 105 for 1 h.
multiple-dose containers, preferably of Type I glass, in a cool B. The retention time of the atenolol peak of the Sample
place or at a controlled room temperature, protected from solution corresponds to that of the Standard solution, as
light. Avoid freezing. obtained in the Assay.
USP Atenolol RS
USP Endotoxin RS
300 Atenolol / Official Monographs USP 32
ASSAY CS = concentration of USP Atenolol RS in the Standard

PROCEDURE solution (mg/mL)
Mobile phase: 1.1 g of sodium 1-heptanesulfonate and D = dilution factor for the Sample solution
0.71 g of anhydrous dibasic sodium phosphate in 700 mL of L = label claim of Tablet (mg atenolol)
water. Add 2 mL of dibutylamine, and adjust with 0.8 M Tolerances: NLT 80% (Q) of the labeled amount of
phosphoric acid to a pH of 3.0. Add 300 mL of methanol C14H22N2O3 is dissolved.
and pass through a filter having a 0.5-m or finer porosity. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Degas this solution before use.
Standard solution: 0.01 mg/mL of USP Atenolol RS in ADDITIONAL REQUIREMENTS
Mobile phase PACKAGING AND STORAGE: Preserve in well-closed containers.
Sample stock solution: Transfer 10 Tablets to a 1000-mL USP REFERENCE STANDARDS 11
volumetric flask. Add 500 mL of Mobile phase, and sonicate USP Atenolol RS
for 15 min to disintegrate the Tablets. Dilute with Mobile
phase to volume.
Sample solution: Centrifuge a portion of the Sample stock Atenolol and Chlorthalidone Tablets
solution, and dilute a volume of the supernatant with Mobile
phase to obtain a solution nominally containing 0.01 mg/mL (Comment on this Monograph)id=m6338=Atenolol and
of atenolol. Chlorthalidone Tablets=A-Monos.pdf)
(See Chromatography 621, System Suitability.) DEFINITION
Mode: LC Atenolol and Chlorthalidone Tablets contain NLT 90.0% and
Detector: UV 226 nm NMT 110.0% of the labeled amounts of atenolol (C14H22N2O3)
Column: 3.9-mm 30-cm; packing L1 and chlorthalidone (C14H11ClN2O4S).
Injection size: 10 L A. THIN-LAYER CHROMATOGRAPHY
System suitability Standard solution A: 10 mg/mL of USP Chlorthalidone RS
Sample: Standard solution in methanol
Suitability requirements Standard solution B: 10J mg/mL of USP Atenolol RS in
Column efficiency: NLT 5000 theoretical plates methanol, J being the ratio of the labeled amount, in mg, of
Tailing factor: NMT 2.0 atenolol to the labeled amount, in mg, of
Relative standard deviation: NMT 2.0% chlorthalidone/Tablet
Analysis Sample solution: Equivalent to 10 mg/mL of
Samples: Standard solution and Sample solution chlorthalidone, from powdered Tablets in methanol. Shake a
Calculate the percentage of C14H22N2O3 in each Tablet quantity of powdered Tablets in methanol for 15 min, and
taken: filter.
Result = (rU/rS) (CS/CU) 100 (See Chromatography 621, Thin-Layer Chromatography.)
rU = peak response from the Sample solution Mode: TLC
rS = peak response from the Standard solution Adsorbent: 0.25-mm layer of chromatographic silica gel
CS = concentration of USP Atenolol RS in the Standard mixture
solution (mg/mL) Application volume: 10 L
CU = nominal concentration of atenolol in the Sample Developing solvent system: n-Butyl alcohol and 1 N
solution (mg/mL) ammonium hydroxide (5:1)
PERFORMANCE TESTS Sample solution
DISSOLUTION 711 Allow the spots to dry, and develop the chromatogram in a
Medium: 0.1 N acetate buffer, pH 4.6, prepared by mixing solvent system, until the solvent front has moved three-
44.9 parts of 0.1 N sodium acetate with 55.1 parts of 0.1 N fourths of the length of the plate. Locate the spots on the
acetic acid solution; 900 mL plate by viewing under short-wavelength UV light.
Apparatus 2: 50 rpm Acceptance criteria: The principal spots of the Sample
Time: 30 min solution correspond in RF value, size, and intensity to those
Determine the amount of C14H22N2O3 dissolved by using the of the respective Standard solutions.
following method. B. The retention times of the major peaks of the Sample
Mobile phase, Chromatographic system, and System solution correspond to those of the Standard solution, as
suitability: Proceed as directed in the Assay. obtained in the Assay.
Standard solution: 0.01 mg/mL of USP Atenolol RS in
Mobile phase ASSAY
Sample solution: Prepare a filtered portion of the solution PROCEDURE
under test. Quantitatively dilute an accurately measured Mobile phase: 250 mL of acetonitrile, 740 mL of water,
volume of the filtrate with Mobile phase to obtain a solution and 8 mL of 3.6 N sulfuric acid. Add 930 mg of sodium
estimated to contain 0.01 mg/mL of atenolol. octyl sulfate.
Analysis: Proceed as directed in the Assay. Standard solution: Dissolve quantities of USP Atenolol RS
Calculate the percentage of C14H22N2O3 dissolved: and USP Chlorthalidone RS in a mixture of water and
acetonitrile (3:1) to obtain a solution having concentrations
Result = V (rU/rS) CS D (100/L) of 0.25 mg of USP Chlorthalidone RS and 0.25J mg of USP
Atenolol RS/mL, J being the ratio of the labeled amount, in
V = volume of Medium, 900 mL mg, of atenolol to the labeled amount, in mg, of
rU = peak response from the Sample solution chlorthalidone/Tablet.
USP 32 Official Monographs / Atovaquone 301
Sample solution: Transfer 10 Tablets to a volumetric flask of V = volume of dissolution medium (mL), 900
such capacity that when filled to volume, a nominal D = dilution factor for the Sample solution
concentration of 0.5 mg of chlorthalidone/mL is obtained. rU = peak response from the Sample solution
Add a mixture of water and acetonitrile (1:1) to half the rS = peak response from the Standard solution
capacity of the flask, and shake by mechanical means for CS = nominal concentration of the appropriate USP
NLT 15 min to disintegrate the Tablets. Dilute with a Reference Standard in the Standard solution
mixture of water and acetonitrile (1:1) to volume. Pass a (mg/mL)
portion of this stock solution through a filter having a 0.5- Tolerances: NLT 80% (Q) of the labeled amount of
m or finer porosity. Transfer 25.0 mL of the clear filtrate to C14H22N2O3, and NLT 70% (Q) of the labeled amount of
a 50-mL volumetric flask, and dilute with water to volume. C14H11ClN2O4S is dissolved.
Chromatographic system UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
(See Chromatography 621, System Suitability.) Analysis for content uniformity: Proceed as directed in the
Mode: LC Assay, except prepare the Sample solution as follows. Transfer
Detector: UV 275 nm 1 Tablet to a volumetric flask of such capacity that when
Column: 4.6-mm 25-cm; packing L1 filled to volume, a nominal concentration of 0.25 mg of
Flow rate: 1.7 mL/min chlorthalidone/mL is obtained. Add a mixture of water and
Injection size: 10 L acetonitrile (1:1) to half the capacity of the flask, and shake
System suitability by mechanical means for NLT 15 min to disintegrate the
Sample: Standard solution Tablet. Dilute with water to volume. Pass a portion of this
[NOTEThe relative retention times for atenolol and solution through a filter having a 0.5-m or finer porosity,
chlorthalidone are 0.8 and 1.0, respectively.] and use the filtrate as the Sample solution.
Suitability requirements Calculate the percentage of C14H22N2O3 and C14H11ClN2O4S
Resolution: NLT 3.0 between the atenolol and in the Tablet taken:
chlorthalidone peaks
Relative standard deviation: NMT 2.0% Result = (rU/rS) (CS/CU) 100
Analysis
Samples: Standard solution and Sample solution rU = peak response from the Sample solution
Calculate the percentages of C14H22N2O3 and rS = peak response from the Standard solution
C14H11ClN2O4S in each Tablet taken: CS = concentration of the appropriate USP Reference
Result = (rU/rS) (CS/CU) 100 CU = nominal concentration of the Sample solution
(mg/mL)
CS = concentration of the appropriate USP Reference PACKAGING AND STORAGE: Preserve in well-closed containers.
Standard in the Standard solution (mg/mL) USP REFERENCE STANDARDS 11
CU = nominal concentration of atenolol or USP Atenolol RS
chlorthalidone in the Sample solution (mg/mL) USP Chlorthalidone RS
[NOTEIf a trailing peak or shoulder is observed on the
chlorthalidone peak with a relative retention time of NMT
1.1 in the chromatograms of both the Standard solution Atovaquone
and the Sample solution, sum the areas for the
chlorthalidone peak with the trailing peak or shoulder to (Comment on this Monograph)id=m6342=Atovaquone=A-
report the peak responses for chlorthalidone.] Monos.pdf)
PERFORMANCE TESTS
DISSOLUTION 711
Apparatus 2: 50 rpm
Time: 45 min
Determine the amounts of C14H22N2O3 and C14H11ClN2O4S C22H19ClO3 366.84
dissolved by using the following method. 1,4-Naphthalenedione, 2-[4-(4-chlorophenyl)cyclohexyl]-3-
Mobile phase, Chromatographic system, and System hydroxy-, trans-;
suitability: Proceed as directed in the Assay. 2-[trans-4-(p-Chlorophenyl)cyclohexyl]-3-hydroxy-1,4-
Diluent: Acetonitrile and 3.6 N sulfuric acid (125:4) naphthoquinone [95233-18-4].
Standard solvent: Diluent and water (3:10)
Standard solution: Dissolve USP Atenolol RS and USP DEFINITION
Chlorthalidone RS in Standard solvent to obtain a solution Atovaquone contains NLT 97.5% and NMT 101.5% of
having concentrations of 0.00085L mg of USP Atenolol RS C22H19ClO3, calculated on the anhydrous and organic solvent-
and 0.00085L mg of USP Chlorthalidone RS/mL, L and L free basis.
being the labeled amounts, in mg, of atenolol and
chlorthalidone, respectively, per Tablet. IDENTIFICATION
Sample solution: Filtered solution under test and Diluent A. INFRARED ABSORPTION 197M
(10:3) B. The retention time of the major peak in the Sample
Analysis solution corresponds to that of the Standard solution, as
Samples: Standard solution and Sample solution obtained in the Assay.
Calculate the percentage of C14H22N2O3 and C14H11ClN2O4S
dissolved:
Result = V D (rU/rS) CS 100
302 Atovaquone / Official Monographs USP 32
ASSAY Analysis
PROCEDURE Samples: Standard solution, Sample solution, and Blank
Mobile phase: Acetonitrile, methanol, water, and solution
phosphoric acid (525:175:300:5) Transfer 12.0 mL of the Sample solution to a 50-mL color-
Diluent: Acetonitrile and water (4:1) comparison tube, 10.0 mL of the Standard solution to
Standard solution: 0.25 mg/mL of USP Atovaquone RS in another, and 10.0 mL of the Blank solution to a third.
Diluent Then add 2.0 mL of the Sample solution to the Standard
System suitability solution: 0.25 mg/mL of USP solution as well as the Blank solution. Add 2 mL of pH 3.5
Atovaquone RS and 0.02 mg/mL of USP Atovaquone Related Acetate Buffer (see Heavy Metals 231) to each of the
Compound A RS in Diluent [NOTEStore in a low-actinic three tubes. Add 1.2 mL of thioacetamide-glycerin base
glass container.] TS. Allow to stand for 2 min, and view downward over a
Sample solution: 0.25 mg/mL of Atovaquone in Diluent white surface.
[NOTEUse a low-actinic volumetric flask.] Acceptance criteria: The solution from the Standard
Chromatographic system solution is slightly brown when compared with the solution
(See Chromatography 621, System Suitability.) from the Blank solution, and the color of the solution from
Mode: LC the Sample solution is not darker than that of the solution
Detector: UV 220 nm from the Standard solution (NMT 10 ppm).
Column: 4.6-mm 25-cm; packing L1 Organic Impurities
Flow rate: 3 mL/min PROCEDURE 1: RESIDUAL ORGANIC SOLVENTS
Injection size: 20 L Standard solution: 0.5 L of methanol, and 0.5 L of
System suitability glacial acetic acid per mL of dimethylformamide
Sample: Standard solution and System suitability solution Sample solution: 50 mg/mL of Atovaquone in
[NOTEThe relative retention times for atovaquone related dimethylformamide
compound A and atovaquone are about 0.85 and 1.0, Chromatographic system
respectively.] (See Chromatography 621, System Suitability.)
Suitability requirements Mode: GC
Resolution: NLT 5 between atovaquone related Detector: Flame ionization
compound A and atovaquone, System suitability solution Column: 4-mm 2.8-m; contains 10% liquid phase G16
Column efficiency: NLT 9000 theoretical plates, Standard on S2 support
solution Carrier gas: Nitrogen
Tailing factor: NMT 1.2, Standard solution Temperature
Relative standard deviation: NMT 2%, Standard solution Column: 180
Analysis Detector: 250
Samples: Standard solution and Sample solution Flow rate: 42.5 mL/min
Calculate the percentage of C22H19ClO3 in the portion of Injection size: 1 L
Atovaquone taken: System suitability
Result = (rU/rS) (CS/CU) 100 [NOTEThe relative retention times for methanol and
acetic acid are about 0.4 and 1.0, respectively.]
rU = peak response from the Sample solution Suitability requirements
rS = peak response from the Standard solution Resolution: NLT 14 between methanol and acetic acid
CS = concentration of USP Atovaquone RS in the Column efficiency: NLT 700 for the acetic acid peak
Standard solution (mg/mL) Tailing factor: NLT 0.8 for the acetic acid peak
CU = concentration of Atovaquone in the Sample Analysis
solution (mg/mL) Samples: Standard solution and Sample solution
Acceptance criteria: 97.5%101.5% Calculate the percentage of methanol and acetic acid in
the portion of Atovaquone taken:
IMPURITIES
Inorganic Impurities Result = (rU/rS) (CS/CU) F 100
HEAVY METALS 231 rU = peak area of methanol or acetic acid from the
Standard solution: Add 1.0 mL of Standard Lead Solution Sample solution
(see Heavy Metals 231, Special Reagents) to 0.5 g of rS = peak area of methanol or acetic acid from the
magnesium oxide, and dry between 100 and 105. Standard solution
Proceed as directed for Sample solution, starting with CS = concentration of the Standard solution (mg/mL)
Ignite to dull redness. CU = concentration of Atovaquone in the Sample
Sample solution: Mix 1.0 g of Atovaquone with 0.5 g of solution (mg/mL)
magnesium oxide thoroughly in a silica crucible. Ignite to F = specific gravity (0.79 for methanol; 1.05 for
dull redness until a homogeneous white or grayish white glacial acetic acid)
mass is obtained. If the mixture remains colored after 30 Acceptance criteria: NMT 0.2% methanol, NMT 0.2%
min, allow to cool, mix using a fine glass rod, and repeat acetic acid
the ignition. If necessary, repeat the operation. Heat the PROCEDURE 2
residue at 800 for about 1 h. Cool, take up the residue in Analysis: Using the chromatograms of the Sample solution
two 5-mL portions of 6 N hydrochloric acid, add 0.1 mL of and the System suitability solution obtained in the Assay,
phenolphthalein TS, and then add 13.5 N ammonium calculate the percentage of atovaquone related compounds
hydroxide until a pink color is obtained. Cool, add glacial in the portion of Atovaquone taken:
acetic acid until the solution is decolorized, and add 0.5
mL in excess. Filter, if necessary, and wash the filter with Result = (ri/rT) 100
water. Dilute with water to 20 mL.
Blank solution: Proceed as directed for Sample solution, ri = peak response of each impurity in the Sample
omitting the Atovaquone. solution
USP 32 Official Monographs / Atovaquone 303
rT = sum of all peak responses in the Sample solution with a mixture of methanol and water (1:1). [NOTE
Acceptance criteria Minimize exposure of this solution to light.]
Individual impurities: See Impurity Table 1. Sample stock solution: Transfer a volume of the Oral
Total impurities: NMT 1.5 % Suspension equivalent to 5.2 g of the formulation to a low-
actinic 250-mL volumetric flask. Add 50 mL of water, swirl
Impurity Table 1 for 5 min, add 150 mL of 0.1 M methanolic sodium
hydroxide, and sonicate for 15 min. Allow to cool, and
Relative Acceptance dilute with 0.1 M methanolic sodium hydroxide to volume.
Retention Criteria, Immediately filter a 20-mL portion, discarding the first 5 mL
Name Time NMT(%) of the filtrate.
Unnamed impurity 0.63 0.5 Sample solution: Transfer 3.0 mL of the clear filtrate of
Atovaquone related compound 0.85 1.0
Sample stock solution to a low-actinic 100-mL volumetric
A
flask, and dilute with a mixture of methanol and water (1:1)
to volume. [NOTEMinimize exposure of this solution to
Unnamed impurity 0.89 0.3 light.]
Unnamed impurity 1.8 0.5 Chromatographic system
Any other individual, 0.2
(See Chromatography 621, System suitability.)
unidentified impurity
Mode: LC
Detector: UV 220 nm
Sum of all other individual 1.0 Column: 4.6-mm 12.5-cm; packing L1
impurities Flow rate: 3 mL/min
System suitability
SPECIFIC TESTS Samples: System suitability solution and Standard solution
WATER DETERMINATION, Method I 921: NMT 1.0% [NOTEThe relative retention times for atovaquone related
ADDITIONAL REQUIREMENTS compound A and atovaquone are 0.86 and 1.0,
PACKAGING AND STORAGE: Preserve in tight, light-resistant respectively.]
containers. Suitability requirements
USP REFERENCE STANDARDS 11 Tailing factor: NMT 1, Standard solution
USP Atovaquone RS Relative standard deviation: NMT 2.0%, Standard
USP Atovaquone Related Compound A RS solution
Analysis
Calculate the percentage of C22H19ClO3 in each mL of the
Atovaquone Oral Suspension Oral Suspension taken:
(Comment on this Monograph)id=m6344=Atovaquone Oral
Suspension=A-Monos.pdf) Result = (rU/rS) (CS/V) D (100/L)
DEFINITION rU = atovaquone peak area from the Sample solution

Atovaquone Oral Suspension contains NLT 90.0% and NMT rS = atovaquone peak area from the Standard solution
110.0% of the labeled amount of C22H19ClO3. CS = concentration of USP Atovaquone RS in the
IDENTIFICATION V = volume of Oral Suspension taken to prepare the
A. ULTRAVIOLET ABSORPTION 197U Sample solution (mL)
Medium: Methanol and water (1:1) D = dilution factor of the Sample solution
Sample solution: Dilute 5 mL of Sample solution from the L = label claim (mg/mL)
Assay with Medium to 50 mL . Acceptance criteria: 90.0%110.0%
Standard solution: Dilute 5 mL of Standard solution from
Assay with Medium to 50 mL. PERFORMANCE TESTS
B. The retention time of the major peak of the Sample UNIFORMITY OF DOSAGE UNITS 905
solution corresponds to that of the Standard solution, as For oral suspension packaged in single-unit containers:
obtained in the Assay. Meets the requirements
DELIVERABLE VOLUME 698
ASSAY For Oral Suspension packaged in multiple-unit
PROCEDURE containers: Meets the requirements
Mobile phase: Acetonitrile, methanol, phosphoric acid, and
water (96:32:1:72) IMPURITIES
System suitability solution: 0.09 mg/mL of USP Organic Impurities
Atovaquone RS and 0.01 mg/mL of USP Atovaquone Related PROCEDURE
Compound A RS in 0.1 M methanolic sodium hydroxide Analysis
[NOTEStore in a low-actinic glass container.] Samples: System suitability solution, Standard solution, and
Standard stock solution: 30 mg of USP Atovaquone RS in the Sample solution obtained in the Assay
a low-actinic 10-mL volumetric flask, and add 2 mL of water Using the chromatograms of the Samples, calculate the
and 6 mL of 0.1 M methanolic sodium hydroxide. Sonicate percentage of atovaquone-related compounds, based on
for 5 min or until the material has dissolved. Allow to cool, the labeled strength of atovaquone:
and dilute with 0.1 M methanolic sodium hydroxide to
volume. Result = (rU/rS) (CS/CU) (1/F) D (100/L)
Standard solution: Transfer 3.0 mL of Standard stock
solution to a low-actinic 100-mL volumetric flask, and dilute
304 Atovaquone / Official Monographs USP 32
rU = individual peak response of an atovaquone Atracurium Besylate

related compound, if any, from the Sample (Comment on this Monograph)id=m6350=Atracurium
solution Besylate=A-Monos.pdf)
rS = peak response of atovaquone from the Standard
solution
CS = concentration of USP Atovaquone RS in the
CU = concentration of the Oral Suspension in the
Sample solution (g/mL)
F = relative response factor of an individual
atovaquone related compound relative to the
response of atovaquone: see Impurity Table 1 C65H82N2O18S2 1243.48
D = density of Oral Suspension (1.04 g/mL at Isoquinolinium, 2,2-[1,5-pentanediylbis[oxy(3-oxo-3,1-
2025) propanediyl)]]bis[1-[(3,4-dimethoxyphenyl)methyl]-1,2,3,4-
L = labeled amount of atovaquone in the Oral tetrahydro-6,7-dimethoxy-2-methyl-, dibenzenesulfonate;
Suspension (mg/mL) 2-(2-Carboxyethyl)-1,2,3,4-tetrahydro-6,7-dimethoxy-2-
[NOTEDisregard any peak having a relative retention methyl-1-veratrylisoquinolinium benzenesulfonate,
time of 0.3, which is due to photodegradation during pentamethylene ester [64228-81-5].
preparation of the Sample solution.]
Acceptance criteria: See Impurity Table 1 DEFINITION
Atracurium Besylate contains NLT 96.0% and NMT 102.0% of
Impurity Table 1
C65H82N2O18S2, calculated on the anhydrous basis. It contains
NLT 5.0% and NMT 6.5% of the trans-trans isomer, NLT
Relative Relative Acceptance 34.5% and NMT 38.5% of the cis-trans isomer, and NLT
Retention Response Criteria, 55.0% and NMT 60.0% of the cis-cis isomer.
Name Time Factor NMT(%)
Atovaquone related 0.65 1.08 0.5
IDENTIFICATION
compound
A. INFRARED ABSORPTION 197K: Meets the requirements
B. The retention times of the three main isomeric peaks of
Atovaquone related 0.86 0.85 1.0 the Sample solution correspond to those of the Standard
compound A solution, as obtained in the Assay.
Atovaquone related 0.88 1.0 0.3
compound ASSAY
PROCEDURE
Atovaquone 1.0 1.0 Buffer solution: 10.2 g of monobasic potassium phosphate
Any other atovaquone 1.0 0.2 in a 1000-mL volumetric flask. Dissolve in 950 mL of water.
related compound While stirring, adjust with phosphoric acid to a pH of 3.1,
Total related compound 2.0 and dilute with water to volume.
Solution A: Acetonitrile, methanol, and Buffer solution
(4:1:15)
SPECIFIC TESTS Solution B: Acetonitrile, methanol, and Buffer solution
PH 791: 3.57.0 (2:3:5)
SEDIMENTATION Mobile phase: See the gradient table below.
For Oral Suspension packaged in multiple-unit containers
Analysis: Transfer 50 mL of well-mixed Oral Suspension to Time Solution A Solution B
a glass-stoppered graduated cylinder, and allow to stand (min) (%) (%)
for 16 h. Measure the volume, if any, of clear liquid
0 80 20
observed in the cylinder.
Acceptance criteria: NMT 1 mL of clear liquid 5 80 20
15 40 60
PACKAGING AND STORAGE: Preserve in tight, light-resistant 25 40 60
containers. 30 0 100
USP Atovaquone RS
USP Atovaquone Related Compound A RS
USP 32 Official Monographs / Atracurium 305
Standard solution: 1 mg/mL of USP Atracurium Besylate RS CS = concentration of the cis-cis isomer in the
in Solution A Standard solution (mg/mL)
Sample solution: 1 mg/mL of Atracurium Besylate in CU = concentration of the Sample solution (mg/mL)
Solution A F = relative response factor of the impurity peak,
Chromatographic system which is 1.9 for laudanosine and 1.0 for all
(See Chromatography, 621 System Suitability.) other unidentified impurities
Mode: LC Acceptance criteria
Detector: UV 280 nm Laudanosine: NMT 0.5%
Column: 4.6-mm 25-cm; base-deactivated packing L1 Individual impurities: NMT 1.0%
Flow rate: 1 mL/min Total impurities: NMT 3.5%
Injection size: 20 L PROCEDURE 2: LIMIT OF METHYL BENZENESULFONATE
System suitability Buffer solution, Solution A, Solution B: Prepare as
Sample: Standard solution directed in the Assay.
[NOTEThe relative retention times for the trans-trans Standard stock solution: 0.2 mg/mL of methyl
isomer, the cis-trans isomer, and the cis-cis isomer are 0.8, benzenesulfonate in acetonitrile
0.9, and 1.0 respectively.] Standard solution: 1 g/mL of methyl benzenesulfonate
Suitability requirements from Standard stock solution diluted with Solution A
Resolution: NLT 1.1 between the trans-trans isomer and Sample solution: 10 mg/mL of Atracurium Besylate in
the cis-trans isomer and between the cis-trans isomer and Solution A
the cis-cis isomer System suitability solution: Transfer 1 mL of the Sample
Relative standard deviation: NMT 2.0% solution and 5 mL of Standard stock solution to a 100-mL
Analysis volumetric flask, and dilute with Solution A to volume.
Samples: Standard solution and Sample solution Mobile phase: See the gradient table below.
Calculate the percentage of C65H82N2O18S2 in the portion
taken: Time (min) Solution A (%) Solution B (%)
Result = (rT1/rT2) (CS/Cu) 100 0 80 20
5 80 20
rT1 = sum of the peak responses for the trans-trans
15 75 25
isomer, the trans-cis isomer, and the cis-cis
isomer from the Sample solution 25 75 25
rT2 = sum of the peak responses for the trans-trans 30 55 45
isomer, the trans-cis isomer, and the cis-cis
38 0 100
isomer from the Standard solution
CS = concentration of USP Atracurium Besylate RS in 45 0 100
CU = concentration of Atracurium Besylate in the Chromatographic system
Sample solution (mg/mL) Mode: LC
Acceptance criteria: 96.0%102.0%, calculated on the Detector: UV 217 nm
anhydrous basis [NOTEIt contains 5.0%6.5% of the trans- Column: 4.6-mm 25-cm; base-deactivated packing L1
trans isomer, 34.5%38.5% of the cis-trans isomer, and Flow rate: 1 mL/min
55.0%60.0% of the cis-cis isomer.] Injection size: 100 L
System suitability
IMPURITIES Samples: Standard solution and System suitability solution
Inorganic Impurities Suitability requirements
RESIDUE ON IGNITION 281: NMT 0.2% Resolution: NLT 12.0 between the trans-trans isomer
HEAVY METALS, Method II 231: NMT 20 ppm and methyl benzenesulfonate, System suitability solution
Organic Impurities Relative response: Responses for duplicate injections do
PROCEDURE 1 not differ from each other by NMT 12%
Buffer solution, Solution A, Solution B, and Mobile Analysis
phase: Proceed as directed in the Assay. Samples: Standard solution and Sample solution
Standard solution: 1.0 mL of the Standard solution, Measure the responses for the methyl benzenesulfonate
prepared as directed in the Assay, diluted in Solution A to peaks.
100 mL Acceptance criteria: NMT 0.01%, the peak response of
Sample solution: Prepare as directed in the Assay the Sample solution being NMT that of the Standard
Chromatographic system and System suitability: As solution
directed in the Assay [NOTEFor identification purposes, PROCEDURE 3: LIMIT OF TOLUENE
the relative retention time for laudanosine is 0.3.] Standard solution: 100 g/mL of toluene in organic-free
Analysis water (see Residual Solvents 467)
Samples: Standard solution and Sample solution Sample solution: 20 mg/mL of Atracurium Besylate in
Record the chromatograms, and measure all of the peak organic-free water (see Residual Solvents 467)
responses, except the three main isomeric peaks. Chromatographic system
Calculate the percentage of each impurity in the portion (See Chromatography 621, System Suitability.)
of C65H82N2O18S2 taken: Mode: GC
Result = (ri/rS) (CS/CU) (1/F) 100 Column: 0.53-mm 30-m fused silica analytical column
coated with a 5-m chemically cross-linked G27 stationary
ri = peak response for each impurity from the phase and a 0.53-mm 5-m silica guard column
Sample solution deactivated with phenylmethyl siloxane
rS = cis-cis isomer peak response from the Standard
solution
306 Atracurium / Official Monographs USP 32
Carrier gas: Helium with a linear velocity of 35 cm/s Solution B: Acetonitrile, methanol, and Buffer solution
[NOTEWhen a makeup gas is used, nitrogen is (2:3:5)
recommended.] Mobile phase: See the gradient table below.
Temperature: See the temperature program table below.
Time (min) Temperature (min) (%) (%)
Injection port 70 0 80 20
Detector port 260 5 80 20
Column 0 35 15 40 60
5 35 25 40 60
22.5 175 30 0 100
24.9 260
Standard solution: 1 mg/mL of USP Atracurium Besylate RS
40.9 260 in Solution A
Sample solution: Nominally equivalent to 1 mg/mL of
Injection size: 1 L atracurium besylate from Injection diluted with Solution A
System suitability Chromatographic system
Sample: Standard solution (See Chromatography 621, System Suitability.)
Suitability requirements Mode: LC
Relative standard deviation: NMT 15% of the toluene Detector: UV 280 nm
peak Column: 4.6-mm 25-cm; base-deactivated packing L1
Analysis Flow rate: 1 mL/min
Samples: Standard solution and Sample solution Injection size: 20 L
Acceptance criteria: NMT 0.5% of toluene is found; the System suitability
toluene peak from the Sample solution is NMT the toluene Sample: Standard solution
peak of the Standard solution. [NOTEThe relative retention times for atracurium besylate
trans-trans-isomer, cis-trans-isomer, and cis-cis-isomer are
SPECIFIC TESTS about 0.8, 0.9, and 1.0, respectively.]
WATER DETERMINATION, Method I 921: NMT 5.0% Suitability requirements
Resolution: NLT 2.0 between the atracurium besylate
ADDITIONAL REQUIREMENTS trans-trans-isomer and the cis-trans-isomer and between
PACKAGING AND STORAGE: Preserve in tight, light-resistant the atracurium besylate cis-trans-isomer and the cis-cis-
containers, in a cold place. [NOTEAtracurium Besylate is isomer
unstable at room temperature.] Relative standard deviation: NMT 2.0%
USP Atracurium Besylate RS Samples: Standard solution and Sample solution
Measure the responses for the three atracurium besylate
isomer peaks.
Atracurium Besylate Injection Calculate the percentage of C65H82N2O18S2 in each mL of
the Injection taken:
(Comment on this Monograph)id=m6356=Atracurium Besylate
Injection=A-Monos.pdf) Result = (rU/rS) (CS/CU) 100
DEFINITION rU = sum of the peak responses for the trans-trans
Atracurium Besylate Injection is a sterile solution containing NLT isomer, the trans-cis isomer, and the cis-cis
90.0% and NMT 115.0% of the labeled amount of atracurium isomer from the Sample solution
besylate (C65H82N2O18S2). It contains an amount of the trans- rS = sum of the peak responses for the trans-trans
trans-isomer equivalent to NLT 5.0% and NMT 6.5% of the isomer, the trans-cis isomer, and the cis-cis
labeled amount of atracurium besylate, an amount of the cis- isomer from the Standard solution
trans-isomer equivalent to NLT 34.5% and NMT 38.5% of the CS = concentration of USP Atracurium Besylate RS in
labeled amount of atracurium besylate, and an amount of the the Standard solution (mg/mL)
cis-cis-isomer equivalent to NLT 55.0% and NMT 60.0% of the CU = nominal concentration of atracurium besylate in
labeled amount of atracurium besylate. the Sample solution (mg/mL)
[NOTEThe Injection is unstable at room temperature. Store all Acceptance criteria: 90.0%115.0% of the labeled amount
samples in the refrigerator. Analyze all preparations as soon as of C65H82 N2O18S2. It contains an amount of the trans-trans-
possible, or use a refrigerated injector.] isomer equivalent to 5.0%6.5% of the labeled amount of
IDENTIFICATION atracurium besylate, an amount of the cis-trans-isomer
The retention times of the peaks of the three atracurium equivalent to 34.5%38.5% of the labeled amount of
besylate isomers of the Sample solution correspond to those atracurium besylate, and an amount of the cis-cis-isomer
of the Standard solution, as obtained in the Assay. equivalent to 55.0%60.0% of the labeled amount of
atracurium besylate.
ASSAY
Buffer solution: 10.2 g of monobasic potassium phosphate Organic Impurities
in a 1000-mL volumetric flask, and dissolve in 950 mL of PROCEDURE
water. While stirring, adjust with phosphoric acid to a pH of Buffer solution, Solution A, Solution B, Mobile phase, and
3.1, and dilute with water to volume. Standard stock solution: Use the Standard solution
Solution A: Acetonitrile, methanol, and Buffer solution prepared as directed in the Assay.
(4:1:15)
USP 32 Official Monographs / Atropine 307
System suitability solution: Heat a portion of the Atropine

Standard stock solution at 90 for 30 min, and immediately (Comment on this Monograph)id=m6390=Atropine=A-
chill to 5. Monos.pdf)
Standard solution: 0.02 mg/mL from Standard stock
solution diluted with Solution A
Sample solution and Chromatographic system: Proceed
as directed in the Assay.
System suitability
Sample: System suitability solution and Standard solution
[NOTEThe retention times relative to the atracurium
besylate cis-cis-isomer are 0.22 for the acidic compound;
0.29 for laudanosine; 0.44 and 0.50 for the trans- and C17H23NO3 289.37
cis-isomers, respectively, of the hydroxy compound; and Benzeneacetic acid, -(hydroxymethyl)-8-methyl-8-azabicyclo
1.28 and 1.33 for the trans- and cis-isomers, [3.2.1]oct-3-yl ester, endo-()-;
respectively, of the monoacrylate.] 1H,5H-Tropan-3-ol ()-tropate (ester) [51-55-8].
Analysis
Samples: Standard solution and Sample solution DEFINITION
Measure the peak responses, except the peak due to Atropine contains NLT 99.0% and NMT 100.5% of C17H23NO3,
benzenesulfonic acid occurring at a retention time of calculated on the anhydrous basis.
about 0.08 relative to the atracurium besylate cis-cis- [CAUTIONHandle Atropine with exceptional care, since it is
isomer. highly potent.]
of Sample solution taken: IDENTIFICATION
A. PROCEDURE
Result = (ri/rT) (CS/CU) 100 Standard solution: 36 mg of USP Atropine Sulfate RS
Sample solution: 30 mg
ri = peak response for each impurity from the Analysis: Dissolve Standard solution and Sample solution in
Sample solution individual 60-mL separators with the aid of 5-mL portions of
rT = sum of all the peak responses from the Standard water. To each separator, add 1.5 mL of 1 N sodium
solution hydroxide solution and 10 mL of chloroform. Shake for 1
CS = concentration of USP Atracurium Besylate RS in min, allow the layers to separate, and pass the chloroform
the Standard solution (mg/mL) extracts through separate filters of 2 g of anhydrous granular
CU = nominal concentration of atracurium besylate in sodium sulfate supported on pledgets of glass wool. Extract
the Sample solution (mg/mL) each aqueous layer with two additional 10-mL portions of
Acceptance criteria chloroform, filtering and combining with the respective
Acidic compound: NMT 6.0% main extracts. Evaporate the chloroform solutions under
Combined cis- and trans-isomers of the hydroxy reduced pressure to dryness, and dissolve each residue in 10
compound: NMT 6.0% mL of carbon disulfide.
Laudanosine: NMT 3.0% Acceptance criteria: The IR absorption spectrum,
Combined cis- and trans-isomers of the monoacrylate: determined in a 1-mm cell, of the solution of the Sample
NMT 3.0% exhibits maxima only at the same wavelengths as that of the
Other known synthetic impurities: NMT 2.0% solution of the Standard.
Other impurity: NMT 0.1 B. PROCEDURE
Total impurities: NMT 15.0% is found. Sample solution: A solution (1 in 50) in 3 N hydrochloric
acid
SPECIFIC TESTS Analysis: Add gold chloride TS to the Sample solution.
PH 791: 3.003.65 Acceptance criteria: A lusterless precipitate is formed
STERILITY TESTS 71: It meets the requirements when tested (distinction from hyoscyamine, which, similarly treated,
as directed for Test for Sterility of the Product to be Examined, yields a lustrous precipitate).
Membrane Filtration.
BACTERIAL ENDOTOXINS TEST 85: It contains NMT 5.56 USP ASSAY
Endotoxin Units/mg of atracurium besylate. PROCEDURE
INJECTIONS 1: Meets the requirements Sample: 400 mg of Atropine
Analysis: Dissolve in 50 mL of glacial acetic acid. Titrate
ADDITIONAL REQUIREMENTS with 0.1 N perchloric acid VS to a green endpoint, using 1
PACKAGING AND STORAGE: Preserve in single-dose or drop of crystal violet TS. Perform a blank determination (see
multiple-dose containers, preferably of Type I glass, in a Titrimetry 541). Each mL of 0.1 N perchloric acid is
refrigerator, and protect from freezing. Protect from light. equivalent to 28.94 mg of C17H23NO3.
USP Atracurium Besylate RS
308 Atropine / Official Monographs USP 32
Acceptance criteria: 99.0%100.5% DEFINITION

Atropine Sulfate contains NLT 98.5% and NMT 101.0% of
IMPURITIES (C17H23NO3)2 H2SO4, calculated on the anhydrous basis.
Inorganic Impurities [CAUTIONHandle Atropine Sulfate with exceptional care,
RESIDUE ON IGNITION 281: NMT 0.1% because it is highly potent.]
Organic Impurities
PROCEDURE 1: LIMIT OF FOREIGN ALKALOIDS AND OTHER IDENTIFICATION
IMPURITIES A. INFRARED ABSORPTION 197K
Standard solution: 24 mg/mL of USP Atropine Sulfate RS B. IDENTIFICATION TESTSGENERAL, Sulfate 191
in methanol Sample solution: 50 mg/mL
Sample solution A: 20 mg/mL of Atropine in methanol Acceptance criteria: Meets the requirements
Sample solution B: 1 mg/mL of Atropine from Sample
solution A diluted with methanol ASSAY
(See Chromatography 621, Thin-Layer Chromatography.) Sample: 1 g
Mode: TLC Analysis: Dissolve in 50 mL of glacial acetic acid and titrate
Adsorbent: 0.5-mm layer of chromatographic silica gel with 0.1 N perchloric acid VS. Perform a blank
Application volume determination (see Titrimetry 541). Each mL of 0.1 N
Standard solution: 5 L perchloric acid is equivalent to 67.68 mg of (C17H23 NO3)2
Sample solution A: 25 L H2SO4.
Sample solution B: 1 L Acceptance criteria: 98.5%101.0%
Developing solvent system: Chloroform, acetone, and
diethylamine (5:4:1) IMPURITIES
Spray reagent: Potassium iodoplatinate TS Inorganic Impurities
Analysis: Proceed as directed under General Chapter. RESIDUE ON IGNITION 281: NMT 0.2%
Allow the spots to dry, and develop the chromatogram in a Organic Impurities
solvent system, until the solvent front has moved three- OTHER ALKALOIDS
fourths of the length of the plate. Locate the spots on the Sample: 150 mg
plate by spraying with Spray reagent. Analysis: Dissolve in 10 mL of water. To 5 mL of the
Acceptance criteria: The RF value of the principal spot of solution, add a few drops of platinic chloride TS: no
each Sample solution corresponds to that of the Standard precipitate is formed. To the remaining 5 mL of the
solution; no secondary spot of the Sample solution A solution, add 2 mL of 6 N ammonium hydroxide, and
exhibits intensity equal to or greater than the principal spot shake vigorously.
of the Sample solution B (NMT 0.2%). Acceptance criteria: A slight opalescence may develop but
PROCEDURE 2: READILY CARBONIZABLE SUBSTANCES TEST 271 no turbidity is produced.
Sample solution: 200 mg in 5 mL of 2 N sulfuric acid SPECIFIC TESTS
Acceptance criteria: The solution has no more color than MELTING RANGE OR TEMPERATURE, Class Ia 741: Not lower
Matching Fluid A, and the solution is colored no more than than 187, determined after drying at 120 for 4 h
light yellow upon the addition of 0.2 mL of nitric acid. [NOTEBecause anhydrous Atropine Sulfate is hygroscopic,
SPECIFIC TESTS determine its melting temperature promptly on a specimen
OPTICAL ROTATION, Angular Rotation 781A: 0.70 to placed in the capillary tube immediately after drying.]
+0.05 (limit of hyoscyamine) OPTICAL ROTATION, Angular Rotation 781A: The observed
Sample solution: 1 g, previously dried at 105 for 1 h, in rotation, in degrees, multiplied by 200, and divided by the
sufficient 50% alcohol (w/w) to obtain a volume of 20 mL length, in mm, of the polarimeter tube used, is between
at 25 (200-mm tube being used) 0.60 and +0.05 (limit of hyoscyamine)
MELTING RANGE OR TEMPERATURE 741: 114118 Sample solution: 1 g, in water to make a volume of 20 mL
WATER DETERMINATION, Method I 921: NMT 0.2% at 25
ACIDITY
ADDITIONAL REQUIREMENTS Sample solution: 1.0 g
PACKAGING AND STORAGE: Preserve in tight, light-resistant Analysis: Dissolve in 20 mL of water. Titrate with 0.020 N
containers. sodium hydroxide.
USP REFERENCE STANDARDS 11 Acceptance criteria: NMT 0.30 mL is required to produce a
USP Atropine Sulfate RS yellow color.
Atropine Sulfate PACKAGING AND STORAGE: Preserve in tight containers.
(Comment on this Monograph)id=m6400=Atropine Sulfate=A- USP REFERENCE STANDARDS 11
Monos.pdf) USP Atropine Sulfate RS
(C17H23NO3)2 H2SO4 H2O 694.83

(C17H23NO3)2 H2SO4 676.83
Benzeneacetic acid, -(hydroxymethyl)-, 8-methyl-8-azabicyclo
[3.2.1]oct-3-yl ester, endo-()-, sulfate (2:1) (salt),
monohydrate;
1H,5H-Tropan-3--ol ()-tropate (ester), sulfate (2:1) (salt)
monohydrate [5908-99-6].
Anhydrous [55-48-1].
Atropine Sulfate Injection Acceptance criteria: 93.0%107.0%

(Comment on this Monograph)id=m6410=Atropine Sulfate SPECIFIC TESTS
Injection=A-Monos.pdf) PH 791: 3.06.5
DEFINITION BACTERIAL ENDOTOXINS TEST 85: It contains NMT 55.6 USP
Atropine Sulfate Injection is a sterile solution of Atropine Sulfate Endotoxin Units/mg of atropine sulfate.
in Water for Injection. It contains NLT 93.0% and NMT OTHER REQUIREMENTS: It meets the requirements under
107.0% of the labeled amount of (C17H23NO3)2 H2SO4 H2O. Injections 1.

THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 PACKAGING AND STORAGE: Preserve in single-dose or in
Adsorbent: Chromatographic silica gel multiple-dose containers, preferably of Type I glass.
Sample solution: Use undiluted USP REFERENCE STANDARDS 11
Application volume: 15 L USP Atropine Sulfate RS
Spray reagent: Potassium iodoplatinate TS USP Endotoxin RS
Developing solvent system: Chloroform and diethylamine
(9:1)
Analysis: Proceed as directed under Thin-Layer
Chromatographic Identification Test 201, the spots on the
Atropine Sulfate Ophthalmic Ointment
plate located by spraying with Spray reagent. (Comment on this Monograph)id=m6420=Atropine Sulfate
Acceptance criteria: Meets the requirements Ophthalmic Ointment=A-Monos.pdf)
ASSAY DEFINITION
PROCEDURE Atropine Sulfate Ophthalmic Ointment is Atropine Sulfate in a
Acetate buffer: 0.05 M sodium acetate, each L containing suitable ophthalmic ointment base. It contains NLT 90.0% and
2.9 mL of glacial acetic acid NMT 110.0% of the labeled amount of (C17H23NO3)2 H2SO4
Mobile phase: 5.1 g of tetrabutylammonium hydrogen H2O. It is sterile.
sulfate in a 1-L volumetric flask. IDENTIFICATION
Add 50 mL of acetonitrile, and dilute with Acetate buffer to A. IDENTIFICATIONORGANIC NITROGENOUS BASES 181
volume. Adjust with 5 N sodium hydroxide to a pH of 5.5 Sample solution: Transfer a portion of Ophthalmic
0.1. Ointment, equivalent to 50 mg of atropine sulfate, to a
Standard solution: 80 g/mL of USP Atropine Sulfate RS suitable separator, and dissolve in 25 mL of ether. Add 25
Sample solution: Nominally equivalent to 80 g/mL of mL of 0.01 N hydrochloric acid, shake vigorously, allow the
atropine from Injection diluted with water layers to separate, and discard the organic phase. Heat the
System suitability solution: 2.5 g/mL of p- aqueous phase gently on a steam bath while passing
hydroxybenzoic acid. nitrogen through the solution, to expel any residual ether.
Dilute one volume of this solution with four volumes of the Analysis: Proceed as directed under IdentificationOrganic
Standard solution. Nitrogenous Bases 181, beginning with In a second
Chromatographic system separator dissolve 50 mg.
(See Chromatography 621, System Suitability.) Acceptance critera: Meets the requirements
Mode: LC B. IDENTIFICATION TESTSGENERAL, Sulfate 191:
Detector: UV 254 nm Sample solution: Transfer 5 g of Ophthalmic Ointment to a
Column: 30-cm 3.9-mm; packing L1 separator, dissolve in 50 mL of ether, and extract with 20
Flow rate: 2 mL/min mL of water.
Injection size: 100 L Acceptance critera: Meets the requirements
System suitability
Sample: Standard solution and System suitability solution ASSAY
[NOTEThe retention time of p-hydroxybenzoic acid is 1.6 PROCEDURE
relative to that of atropine.] pH 9.0 Buffer: 34.8 g of dibasic potassium phosphate in
Suitability requirements 900 mL of water.
Resolution: NLT 2.2 between the p-hydroxybenzoic acid Adjust to a pH of 9.0 by the addition of 3 M hydrochloric
and atropine peaks, System suitability solution acid or 1 M sodium hydroxide, as necessary, with mixing.
Relative standard deviation: NMT 1.5%, Standard Internal standard solution: 0.5 mg/mL of homatropine
solution hydrobromide in water
Analysis [NOTEPrepare fresh daily.]
Samples: Sample solution and Standard solution Standard solution: 0.1 mg/mL of USP Atropine Sulfate RS
Calculate the percentage of (C17H23NO3)2 H2SO4 H2O in in water. Pipet 10 mL of this solution into a separator, add
each mL of the Injection taken: 2.0 mL of Internal standard solution and 5.0 mL of pH 9.0
Buffer, and adjust the solution in the separator with 1 M
Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 sodium hydroxide to a pH of 9.0. Extract with two 10-mL
portions of methylene chloride, filter the methylene chloride
rU = peak response from the Sample solution extracts through 1 g of anhydrous sodium sulfate supported
rS = peak response from the Standard solution by a small cotton plug in a funnel into a 50-mL beaker, and
CS = concentration of USP Atropine Sulfate RS in the evaporate under a stream of nitrogen to near-dryness.
Standard solution (mg/mL) Dissolve the residue in 2.0 mL of methylene chloride.
CU = nominal concentration of atropine in the Sample [NOTEPrepare fresh daily.]
solution (mg/mL) Sample solution: Transfer Ophthalmic Ointment, equivalent
Mr1 = molecular weight of atropine sulfate to 10 mg of atropine sulfate, to a separator containing 50
monohydrate, 694.85 mL of ether, shake to dissolve, extract with three 25-mL
Mr2 = molecular weight of anhydrous atropine sulfate, portions of 0.1 M sulfuric acid, collect the acid extracts in a
676.83
100-mL volumetric flask, dilute with 0.1 M sulfuric acid to 107.0% of the labeled amount of atropine sulfate
volume. Pipet 10 mL of this solution and treat as follows. [(C17H23NO3)2 H2SO4 H2O]. It may contain suitable stabilizers
Add 2.0 mL of Internal standard solution and 5.0 mL of pH and antimicrobial agents.
9.0 Buffer, and adjust the solution in the separator with 1 M
sodium hydroxide to a pH of 9.0. Extract with two 10-mL IDENTIFICATION
portions of methylene chloride, filter the methylene chloride A. INFRARED ABSORPTION 197M
extracts through 1 g of anhydrous sodium sulfate supported Sample: Ophthalmic Solution, equivalent to 30 mg,
by a small cotton plug in a funnel into a 50-mL beaker, and evaporated to dryness
evaporate under a stream of nitrogen to near-dryness. Standard: 36 mg USP Atropine Sulfate RS
Dissolve the residue in 2.0 mL of methylene chloride. Analysis: Dissolve Sample and Standard in individual 60-mL
Chromatographic system separators with the aid of 5-mL portions of water. To each
(See Chromatography 621, System Suitability.) separator, add 1.5 mL of 1 N sodium hydroxide solution and
Mode: GC 10 mL of chloroform. Shake for 1 min, allow the layers to
Detector: Flame ionization separate, and filter the chloroform extracts through separate
Column: 2-mm 1.8-m glass column packed with a 3% filters of 2 g of anhydrous granular sodium sulfate supported
phase G3 on support S1AB on pledgets of glass wool. Extract each aqueous layer with
Temperature: two additional 10-mL portions of chloroform, filtering and
Column: 225 combining with the respective main extracts. Evaporate the
Injection port: 250 chloroform solutions under reduced pressure to dryness, and
Detector: 250 dissolve each residue in 10 mL of carbon disulfide.
Flow rate: 25 mL/min Acceptance criteria: The IR absorption spectrum,
Carrier gas: Nitrogen determined in a 1-mm cell, of the solution of the Sample
Injection size: 1 L exhibits maxima only at the same wavelengths as that of the
System suitability solution of the Standard.
Sample: Standard solution B. IDENTIFICATION TESTSGENERAL, Sulfate 191: Meets the
Suitability requirements requirements
Resolution: NLT 4.0 Sample solution: Evaporate to dryness a quantity of
Tailing factor: NMT 2.0 Ophthalmic Solution. Prepare a solution from the residue
Relative standard deviation: NMT 2.0% that contains the equivalent of 50 mg of atropine
Analysis sulfate/mL.
Calculate the percentage of (C17H23NO3)2 H2SO4 H2O in ASSAY
the portion of Ophthalmic Ointment taken: PROCEDURE
pH 9.0 buffer: 34.8 g of dibasic potassium phosphate in
Result = (RU/RS) (CS/CU) Mr1/Mr2 100 900 mL of water.
Adjust to a pH of 9.0, determined electrometrically, by the
RU = peak area ratios of atropine sulfate to addition of 3 M hydrochloric acid or 1 M sodium
homatropine hydrobromide from the Sample hydroxide, as necessary, with mixing.
solution Internal standard solution: 0.5 mg/mL of homatropine
RS = peak area ratios of atropine sulfate to hydrobromide in water
homatropine hydrobromide from the Standard [NOTEPrepare fresh daily.]
solution Standard solution: 0.1 mg/mL of USP Atropine Sulfate RS
CS = concentration of USP Atropine Sulfate RS in the in water.
Standard solution (mg/mL) [NOTEPrepare fresh daily.]
CU = concentration of the Sample solution (unit/mL) Pipet 10 mL of this solution into a separator, add 2.0 mL of
Mr1 = molecular weight of atropine sulfate Internal standard solution and 5.0 mL of pH 9.0 buffer, and
monohydrate, 694.85 adjust the solution in the separator with 1 M sodium
Mr2 = molecular weight of anhydrous atropine sulfate, hydroxide to a pH of 9.0. Extract with two 10-mL portions
676.83 of methylene chloride, filter the methylene chloride extracts
Acceptance criteria: 90.0%110.0% through 1 g of anhydrous sodium sulfate supported by a
small cotton plug in a funnel into a 50-mL beaker, and
SPECIFIC TESTS evaporate under a stream of nitrogen to near-dryness.
STERILITY TESTS 71: Meets the requirements Dissolve the residue in 2.0 mL of methylene chloride.
METAL PARTICLES IN OPHTHALMIC OINTMENTS 751: Meets Sample solution: Ophthalmic Solution, equivalent to 10
the requirements mg of atropine sulfate, in a 100-mL volumetric flask.
ADDITIONAL REQUIREMENTS and treat as follows. Add 2.0 mL of Internal standard
PACKAGING AND STORAGE: Preserve in collapsible ophthalmic solution and 5.0 mL of pH 9.0 buffer, and adjust the
ointment tubes. solution in the separator with 1 M sodium hydroxide to a
USP REFERENCE STANDARDS 11 pH of 9.0. Extract with two 10-mL portions of methylene
USP Atropine Sulfate RS chloride, filter the methylene chloride extracts through 1 g
of anhydrous sodium sulfate supported by a small cotton
plug in a funnel into a 50-mL beaker, and evaporate under
Atropine Sulfate Ophthalmic Solution a stream of nitrogen to near-dryness. Dissolve the residue
in 2.0 mL of methylene chloride.
(Comment on this Monograph)id=m6430=Atropine Sulfate Chromatographic system
Ophthalmic Solution=A-Monos.pdf) Mode: GC
DEFINITION Column: 2-mm 1.8-m glass column packed with a 3%
Atropine Sulfate Ophthalmic Solution is a sterile, aqueous phase G3 on support S1AB
solution of Atropine Sulfate. It contains NLT 93.0% and NMT
Temperature Adjust to a pH of 9.0, determined electrometrically, by the

Column: 225 addition of 3 M hydrochloric acid or 1 M sodium
Injection port and detector: 250 hydroxide, as necessary, with mixing.
Flow rate: 25 mL/min Internal standard solution: 0.5 mg/mL of homatropine
Carrier gas: Nitrogen hydrobromide in water
Injection size: 1 L [NOTEPrepare fresh daily.]
System suitability Standard solution: 0.1 mg/mL of USP Atropine Sulfate RS
Sample: Standard solution in water
Suitability requirements Pipet 10 mL of this solution into a separator, add 2.0 mL of
Resolution: NLT 4.0 Internal standard solution and 5.0 mL of pH 9.0 Buffer, and
Tailing factor: NMT 2.0 adjust the solution in the separator with 1 M sodium
Relative standard deviation: NMT 2.0% hydroxide to a pH of 9.0. Extract with two 10-mL portions
Analysis of methylene chloride, filter the methylene chloride extracts
Samples: Standard solution and Sample solution through 1 g of anhydrous sodium sulfate supported by a
Calculate the percentage of (C17H23NO3)2 H2SO4 H2O in small cotton plug in a funnel into a 50-mL beaker, and
each mL of Ophthalmic Solution taken: evaporate under a stream of nitrogen to near-dryness.
Dissolve the residue in 2.0 mL of methylene chloride.
Result = (RU/RS) (CS/CU) Mr1/Mr2 100 [NOTEPrepare fresh daily.]
Sample solution: Finely powder NLT 20 Tablets. Transfer a
RU = peak area ratios of atropine sulfate to portion of the powder, equivalent to 1 mg of atropine
homatropine hydrobromide from the Sample sulfate, to a separator. Add 2.0 mL of Internal standard
solution solution and 5.0 mL of pH 9.0 Buffer, and adjust the solution
RS = peak area ratios of atropine sulfate to in the separator with 1 M sodium hydroxide to a pH of 9.0.
homatropine hydrobromide from the Standard Extract with two 10-mL portions of methylene chloride, filter
solution the methylene chloride extracts through 1 g of anhydrous
CS = concentration of USP Atropine Sulfate RS in the sodium sulfate supported by a small cotton plug in a funnel
Standard solution (mg/mL) into a 50-mL beaker, and evaporate under a stream of
CU = nominal concentration of the Sample solution nitrogen to near-dryness. Dissolve the residue in 2.0 mL of
(mg/mL) methylene chloride.
Mr1 = molecular weight of atropine sulfate Chromatographic system
monohydrate, 694.85 (See Chromatography 621, System Suitability.)
Mr2 = molecular weight of anhydrous atropine sulfate, Mode: GC
676.83 Detector: Flame ionization
Acceptance criteria: 90.0%110.0% Column: 2-mm 1.8-m glass column, packed with a 3%
phase G3 on support S1AB
SPECIFIC TESTS Temperature
PH 791: 3.56.0 Column: 225
STERILITY TESTS 71: Meets the requirements Injection port and detector: 250
ADDITIONAL REQUIREMENTS Flow rate: 25 mL/min
PACKAGING AND STORAGE: Preserve in tight containers. Carrier gas: Nitrogen
USP REFERENCE STANDARDS 11 Injection size: 1 L
USP Atropine Sulfate RS System suitability
Resolution: NLT 4.0
Atropine Sulfate Tablets Tailing factor: NMT 2.0
(Comment on this Monograph)id=m6440=Atropine Sulfate Relative standard deviation: NMT 2.0%
Tablets=A-Monos.pdf) Analysis
DEFINITION Calculate the percentage of (C17H23NO3)2 H2SO4 H2O in
Atropine Sulfate Tablets contain NLT 90.0% and NMT 110.0% the portion of Tablets taken:
of the labeled amount of atropine sulfate [(C17H23NO3)2 H2SO4
H2O]. Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100
IDENTIFICATION RU = peak area ratios of atropine sulfate to

A. IDENTIFICATIONORGANIC NITROGENOUS BASES 181: homatropine hydrobromide from the Sample
Sample: A quantity of Tablets, equivalent to 5 mg of solution
atropine sulfate RS = peak area ratios of atropine sulfate to
Analysis: Triturate with 10 mL of water for a few minutes, homatropine hydrobromide from the Standard
and filter into a small separator. Render the solution alkaline solution
with 6 N ammonium hydroxide, and extract with 50 mL of CS = concentration of USP Atropine Sulfate RS in the
chloroform. Filter the chloroform layer, and evaporate to Standard solution (mg/mL)
dryness. CU = nominal concentration of atropine sulfate in the
Acceptance criteria: The residue so obtained meets the Sample solution (mg/mL)
requirements. Mr1 = molecular weight of atropine sulfate
B. IDENTIFICATION TESTSGENERAL, Sulfate 191: A filtered monohydrate, 694.85
solution of Tablets meets the requirements of the tests. Mr2 = molecular weight of anhydrous atropine sulfate,
676.83
ASSAY
PROCEDURE
pH 9.0 Buffer: 34.8 g of dibasic potassium phosphate in
900 mL of water
Acceptance criteria: 90.0%110.0% SPECIFIC TESTS

LOSS ON DRYING 731: Dry at 105 to constant weight: it
PERFORMANCE TESTS loses NMT 4.0% of its weight.
DISINTEGRATION 701 POWDER FINENESS: Add 50 g to 450 mL of water containing
Time: 15 min 5 g of sodium pyrophosphate, and stir for 10 min. Pour the
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements resulting dispersion slowly through a No. 325 standard sieve
(see Particle Size Distribution Estimation by Analytical Sieving
ADDITIONAL REQUIREMENTS 786), and carefully wash the residue until clean. Dry the
PACKAGING AND STORAGE: Preserve in well-closed containers. residue at 105 to constant weight: the dry weight of the
USP REFERENCE STANDARDS 11 residue so obtained is NMT 0.30% of the weight of the
USP Atropine Sulfate RS sample taken.
Acceptance criteria: The dry weight of the residue is NMT
0.10% of the weight of the sample taken.
Activated Attapulgite MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED
MICROORGANISMS 62: It meets the requirements of the test
(Comment on this Monograph)id=m6470=Activated for absence of Escherichia coli.
Attapulgite=A-Monos.pdf) PH 791: Disperse 1.0 g in 10 mL of carbon dioxide-free
DEFINITION water: the pH of the mixed dispersion so obtained is
Activated Attapulgite is a highly heat-treated, processed, native 7.09.5.
magnesium aluminum silicate. ADSORPTIVE CAPACITY: To 10 mL of a suspension (1 in 10) of
the sample in water, add 80 mL of methylene blue solution
IDENTIFICATION (1 in 1000), and shake. Add 10 mL of barium chloride
PROCEDURE solution (1 in 50), and shake. Allow to stand for 15 min.
Analysis: Add 2 g in small portions to 100 mL of water, Transfer 40 mL of the supernatant to a 50-mL centrifuge
with vigorous agitation. Allow to stand for at least 12 h to tube, and centrifuge. To 5 mL of the clear supernatant add
ensure complete hydration. Place 2 mL of the resulting 495 mL of water, and mix.
mixture on a suitable glass slide, and allow to air-dry at Acceptance criteria: The color of the solution so obtained
room temperature to produce a uniform film. Place the slide is not deeper than that of a solution containing 1.5 g of
in a vacuum desiccator over a free surface of ethylene methylene blue per mL.
glycol. Evacuate the desiccator, and close the stopcock so
that the ethylene glycol saturates the desiccator chamber. ADDITIONAL REQUIREMENTS
Allow to stand for 12 h. Record the X-Ray diffraction pattern PACKAGING AND STORAGE: Preserve in well-closed containers.
(see X-Ray Diffraction 941), and calculate the d values.
Acceptance criteria: Several peaks are observed; the
characteristic peak corresponds to a d value between 10.3
and 10.7 Angstroms.
Colloidal Activated Attapulgite
(Comment on this Monograph)id=m6472=Colloidal Activated
IMPURITIES Attapulgite=A-Monos.pdf)
LOSS ON IGNITION 733: When ignited at 1000 for 1 h, it DEFINITION
loses 4.0%12.0% of its weight. Colloidal Activated Attapulgite is a purified native magnesium
ARSENIC AND LEAD: To 5.0 g add 50 mL of 1 N nitric acid, aluminum silicate.
and boil for 30 min, adding 1 N nitric acid at times to IDENTIFICATION
maintain the volume. Filter into a 100-mL volumetric flask, PROCEDURE
wash the filter with water, and dilute the combined filtrate Analysis: Add 2 g in small portions to 100 mL of water,
and washings with water to volume. with vigorous agitation. Allow to stand for at least 12 h to
Arsenic: Determine the arsenic in the solution by atomic ensure complete hydration. Place 2 mL of the resulting
absorption spectrometry (see Spectrophotometry and Light- mixture on a suitable glass slide, and allow to air-dry at
Scattering 851), using a graphite furnace to volatilize the room temperature to produce a uniform film. Place the slide
arsenic, as directed by the manufacturer of the instrument in a vacuum desiccator over a free surface of ethylene
used, and measuring the absorbance at 189.0 nm against a glycol. Evacuate the desiccator, and close the stopcock so
standard. that the ethylene glycol saturates the desiccator chamber.
Acceptance criteria: NMT 2 ppm Allow to stand for 12 h. Record the X-Ray diffraction pattern
Lead: Determine the lead in the solution by atomic (see X-Ray Diffraction 941), and calculate the d values:
absorption spectrometry (see Spectrophotometry and Light- several peaks are observed.
Scattering 851), using a graphite furnace to volatilize the Acceptance criteria: The characteristic peak corresponds to
lead, as directed by the manufacturer of the instrument a d value between 10.3 and 10.7 Angstroms.
used, and measuring the absorbance at 283.3 nm against a
standard. IMPURITIES
Acceptance criteria: NMT 0.001% Inorganic Impurities
CARBONATE: Mix 1.0 g with 15 mL of 0.5 N sulfuric acid. LOSS ON IGNITION 733: When ignited at 1000 for 1 h, it
Acceptance criteria: No effervescence occurs. loses 17.0%27.0% of its weight.
ACID-SOLUBLE MATTER: Boil 2.0 g with 100 mL of 0.2 N ARSENIC AND LEAD: To 5.0 g, add 50 mL of 1 N nitric acid,
hydrochloric acid for 5 min, and cool. Add water to adjust and boil for 30 min, adding 1 N nitric acid at times to
the volume to 100 mL, and filter. Evaporate 50 mL of the maintain the volume. Filter into a 100-mL volumetric flask,
filtrate so obtained to dryness, and ignite the residue at wash the filter with water, and dilute the combined filtrate
600. and washings with water to volume.
Acceptance criteria: NMT 0.25 g (25%). Arsenic: Determine the arsenic in the solution by atomic
VOLATILE MATTER: Ignite at 600 for 1 h. absorption spectrometry (see Spectrophotometry and Light-
Acceptance criteria: It loses 3.0%7.5% of its weight on Scattering 851), using a graphite furnace to volatilize the
the dried basis.
USP 32 Official Monographs / Aurothioglucose 313
arsenic, as directed by the manufacturer of the instrument DEFINITION

used, and measuring the absorbance at 189.0 nm against a Aurothioglucose contains NLT 95.0% and NMT 105.0% of
standard. C6H11AuO5S, calculated on the dried basis. It is stabilized by
Acceptance criteria: NMT 2 ppm the addition of a small amount of Sodium Acetate.
Lead: Determine the lead in the solution by atomic
absorption spectrometry (see Spectrophotometry and Light- IDENTIFICATION
Scattering 851), using a graphite furnace to volatilize the A. PROCEDURE
lead, as directed by the manufacturer of the instrument Standard solution: 4 mg/mL of USP Aurothioglucose RS
used, and measuring the absorbance at 283.3 nm against a Sample solution: 4 mg/mL
standard. Application volume: 10 L
Acceptance criteria: NMT 0.001% Developing solvent system: n-propyl alcohol, ethyl
ACID-SOLUBLE MATTER: Boil 2.0 g with 100 mL of 0.2 N acetate, and water (3:1:3)
hydrochloric acid for 5 min, and cool. Add water to adjust Analysis
the volume to 100 mL, and filter. Evaporate 50 mL of the Samples: Sample solution and Standard solution
filtrate so obtained to dryness, and ignite the residue at Apply Samples to a suitable thin-layer chromatographic
600. glass microfilament sheet (see Chromatography 621)
Acceptance criteria: NMT 0.15 g (15%) impregnated with silicic acid and a suitable fluorescing
CARBONATE: Mix 1.0 g with 15 mL of 0.5 N sulfuric acid. substance. Allow the spots to dry, and develop the
Acceptance criteria: No effervescence occurs. chromatogram in a solvent system, until the solvent front
VOLATILE MATTER: When ignited at 600 for 1 h, it loses has moved three-fourths of the length of the plate.
7.5%12.5% of its weight on the dried basis. Remove the sheet from the developing chamber, mark the
solvent front, and allow the solvent to evaporate. Locate
SPECIFIC TESTS the spots on the plate by examination under short-
MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED wavelength UV light.
MICROORGANISMS 62: It meets the requirements of the test Acceptance criteria: The RF value of the principal spot
for absence of Escherichia coli. obtained from the solution under test corresponds to that
PH 791: Disperse 1.0 g in 10 mL of carbon dioxide-free obtained from the Standard solution.
water: the pH of the mixed dispersion so obtained is B. PROCEDURE
7.09.5. Analysis: To a portion of the filtrate obtained in the Assay,
LOSS ON DRYING 731: Dry at 105 to constant weight: it add barium chloride TS.
loses 5.0%17.0% of its weight. Acceptance criteria: A heavy, white precipitate is formed.
POWDER FINENESS: Add 50 g to 450 mL of water containing
5 g of sodium pyrophosphate, and stir for 10 min. Pour the ASSAY
resulting dispersion slowly through a No. 325 standard sieve PROCEDURE
(see Particle Size Distribution Estimation by Analytical Sieving Analysis
786), and carefully wash the residue until clean. Dry the Sample: 1 g of Aurothioglucose
residue at 105 to constant weight. Dissolve the Sample in 100 mL of water in a 300-mL
Acceptance criteria: The dry weight of the residue so Kjeldahl flask. Slowly add 10 mL of nitric acid, and when
obtained is NMT 0.30% of the weight of the sample taken. the reaction has subsided, boil the mixture for 5 min.
ADSORPTIVE CAPACITY: To 10 mL of a suspension (1 in 10) of Filter, wash well the separated gold with hot water, dry,
the specimen in water, add 80 mL of methylene blue and ignite to constant weight. The weight of the gold so
solution (1 in 1000), and shake. Add 10 mL of barium obtained, multiplied by 1.991, represents the weight of
chloride solution (1 in 50), and shake. Allow to stand for 15 C6H11AuO5S in the portion of Aurothioglucose taken.
min. Transfer 40 mL of the supernatant to a 50-mL Acceptance criteria: 95.0%105.0%
centrifuge tube, and centrifuge. To 5 mL of the clear
supernatant, add 495 mL of water, and mix. SPECIFIC TESTS
Acceptance criteria: The color of the solution so obtained OPTICAL ROTATION, Specific Rotation 781S: +65 to +75
is not deeper than that of a solution containing 1.5 g of Sample solution: 10 mg/mL
methylene blue per mL. LOSS ON DRYING 731: Dry over phosphorus pentoxide for
24 h: it loses NMT 1.0% of its weight.
PACKAGING AND STORAGE: Preserve in well-closed containers. ADDITIONAL REQUIREMENTS
containers. Store at room temperature.
Aurothioglucose USP Aurothioglucose RS
(Comment on this Monograph)id=m6520=Aurothioglucose=A-
Monos.pdf)
Aurothioglucose Injectable Suspension
(Comment on this Monograph)id=m6545=Aurothioglucose
Injectable Suspension=A-Monos.pdf)
DEFINITION
Aurothioglucose Injectable Suspension is a sterile suspension of
Aurothioglucose in a suitable vegetable oil. It contains NLT
C6H11AuO5S 392.18 90.0% and NMT 110.0% of the labeled amount of
Gold, (1-thio-D-glucopyranosato)-; C6H11AuO5S. It may contain suitable thickening agents.
(1-Thio-D-glucopyranosato)gold [12192-57-3].
314 Aurothioglucose / Official Monographs USP 32
IDENTIFICATION Avobenzone
PROCEDURE (Comment on this Monograph)id=m6560=Avobenzone=A-
Standard solution: 4 mg/mL of USP Aurothioglucose RS Monos.pdf)
Sample solution: Injectable Suspension, equivalent to 200
mg of aurothioglucose, in a centrifuge separator containing
20 mL of ethyl acetate and 50 mL of water
Shake the mixture thoroughly, and centrifuge until the
liquid phases have been clearly separated. Withdraw the
lower, aqueous phase, and filter, discarding the first 10 mL
of the filtrate. Collect the filtrate in a glass-stoppered vessel.
Developing solvent system: n-propyl alcohol, ethyl C20H22O3 310.39
acetate, and water (3:1:3) 1,3-Propanedione, 1-[4-(1,1-dimethylethyl)phenyl]-3-(4-
Analysis methoxyphenyl)-;
Samples: Sample solution and Standard solution 1-(p-tert-Butylphenyl)-3-(p-methoxyphenyl)-1,3-propanedione.
Apply the Sample to a suitable thin-layer chromatographic [70356-09-1].
glass microfilament sheet (see Chromatography 621)
impregnated with silicic acid and a suitable fluorescing DEFINITION
substance. Allow the spots to dry, and develop the Avobenzone contains NLT 95.0% and NMT 105.0% of
chromatogram in a solvent system, until the solvent front C20H22O3, calculated on the dried basis.
has moved about three-fourths of the length of the plate.
Remove the sheet from the developing chamber, mark the IDENTIFICATION
solvent front, and allow the solvent to evaporate. Locate A. INFRARED ABSORPTION 197K
the spots on the plate by examination under short- B. ULTRAVIOLET ABSORPTION 197U
wavelength UV light. Analytical wavelength: 360 nm
Acceptance criteria: The RF value of the principal spot from Solution: 5 g/mL in alcohol
the solution under test corresponds to that from the Absorptivities: Do not differ by more than 3.0%
Standard solution. ASSAY
ASSAY PROCEDURE
PROCEDURE Standard solution: 50 mg/mL of USP Avobenzone RS in
Sample solution: Transfer with a pipet, calibrated to acetone
contain rather than to deliver, a measured volume of Sample solution: 50 mg/mL of Avobenzone in acetone
Injectable Suspension, equivalent to 200 mg of Chromatographic system
aurothioglucose, to a beaker containing 400 mL of acetone. (See Chromatography 621, System Suitability.)
Analysis: Wash the pipet into the beaker with a small Mode: GC
quantity of acetone, allow the solids to settle, and decant Detector: Flame ionization
the supernatant through a filter. Wash the solids with Column: 0.32-mm 25-m fused silica capillary column
another 400-mL portion of acetone, and repeat the coated with phase G1
decantation. Transfer the solids to the filter with the aid of Temperature: See the temperature program table below.
acetone, then transfer the filter and its contents to a short-
necked, 300-mL Kjeldahl flask, and add 5 mL of water and Time (min) Temperature
20 mL of nitric acid. To this solution, add 15 mL of sulfuric Injection port 200
acid slowly. Heat over a low flame, gently at first, and then
increase the heat until fumes of sulfur trioxide are evolved. Detector port 280
Allow the flask and contents to cool to room temperature, Column 0 200
add 30 mL of water slowly, and 20 mL of hydrogen (4/min) 20 280
peroxide TS, again heat to fumes of sulfur trioxide, cool, and
dilute with 30 mL of water. Pass the mixture through an Carrier gas: Helium
ignited, tared filtering crucible, wash with water, heat the Injection size: 1 L
crucible and contents over a low flame to dry the System suitability
precipitate, and ignite at 650 50 to constant weight. The Sample: Standard solution
weight of gold so obtained, multiplied by 1.991, represents Suitability requirements
the weight of C6H11AuO5S in the portion of Injectable Resolution: NLT 1.0 between avobenzone and any
Suspension taken. adjacent peak
Acceptance criteria: 90.0%110.0% Relative standard deviation: NMT 2.0%
SPECIFIC TESTS Analysis
OTHER REQUIREMENTS: It meets the requirements under Samples: Standard solution and Sample solution
Injections 1. Calculate the percentage of C20H22O3 in the portion taken:

PACKAGING AND STORAGE: Preserve in single-dose or in
multiple-dose containers, preferably of Type I glass. Protect rU = peak response of the Sample solution
from light. rS = peak response of the Standard solution
USP REFERENCE STANDARDS 11 CS = concentration of USP Avobenzone RS in the
USP Aurothioglucose RS Standard solution (mg per mL)
USP 32 Official Monographs / Azaperone 315
CU = concentration of avobenzone in the Sample Acceptance criteria: 98.0%102.0%

solution (mg/mL)
IMPURITIES RESIDUE ON IGNITION 281: NMT 0.1%
Organic Impurities Organic Impurities
PROCEDURE PROCEDURE
Sample solution: Proceed as directed for the Sample Adsorbent: 0.2-mm layer of chromatographic silica gel
solution in the Assay. mixture with chemically bonded amino groups
Chromatographic system: Proceed as directed in the Allow the spots to dry.
Assay. Standard solution A: 0.50 mg/mL of USP Azaperone RS in
Analysis acetone and methylene chloride (5:1)
Sample: Sample solution Standard solution B: Quantitatively dilute Standard
Calculate the percentage of each impurity in the portion of solution A with a (5:1) mixture of acetone and methylene
C20H22O3 taken: chloride to obtain a 0.25 mg/mL solution of USP
Azaperone RS.
Result = (rU/rT) 100 Sample solution: 50 mg/mL of Azaperone in acetone and
methylene chloride (5:1)
rU = peak response for each impurity, other than the Application volume: 1 L
avobenzone peak, of the Sample solution Developing solvent system: Cyclohexane, acetone, and
rT = sum of all of the peak responses of the Sample methanol (13:6:1)
solution Analysis
Acceptance criteria Samples: Standard solution A, Standard solution B, and
Individual impurities: NMT 3.0% Sample solution
Sum of all of the impurities: NMT 4.5% Analysis: Proceed as directed for Chromatography 621,
Thin-Layer Chromatography. Develop the chromatograms
SPECIFIC TESTS in the Developing solvent system, until the solvent front has
MELTING RANGE OR TEMPERATURE, Class I 741: 8186 moved three-fourths of the length of the plate. Remove
LOSS ON DRYING 731: Dry in vacuum at 70 for 4 h: it the plate from the chromatographic chamber, and allow
loses NMT 0.5% of its weight. the plate to air-dry. Examine the plate under short-
ADDITIONAL REQUIREMENTS wavelength UV light, and compare the intensities of any
PACKAGING AND STORAGE: Preserve in tight, light-resistant secondary spots, other than any spot at the origin,
containers. observed in the chromatogram of the Sample solution with
USP REFERENCE STANDARDS 11 those of the principal spots in the chromatograms of
USP Avobenzone RS Standard solution A and Standard solution B.
Acceptance criteria: The sum of the intensities of the
secondary spots of the Sample solution corresponds to not
more than the intensity of the principal spot of Standard
Azaperone solution A (1.0%).
(Comment on this Monograph)id=m6570=Azaperone=A- SPECIFIC TESTS
Monos.pdf) MELTING RANGE OR TEMPERATURE 741: 9295
LOSS ON DRYING 731: Dry in vacuum at 60 for 4 h: it
protected from light. Store at room temperature.
only.
C19H22FN3O 327.40 USP REFERENCE STANDARDS 11
1-Butanone, 1-(4-fluorophenyl)-4-[4-(2-pyridinyl)-1-piperazinyl]-; USP Azaperone RS
4-Fluoro-4-[4-(2-pyridyl)-1-piperazinyl]butyrophenone
[1649-18-9].
DEFINITION
Azaperone contains NLT 98.0% and NMT 102.0% of
Azaperone Injection
C19H22FN3O, calculated on the dried basis. (Comment on this Monograph)id=m6575=Azaperone
IDENTIFICATION
INFRARED ABSORPTION 197K: Previously dried DEFINITION
Azaperone Injection is a sterile solution of Azaperone in Water
ASSAY for Injection, prepared with the aid of Tartaric Acid. It may
PROCEDURE contain a suitable preservative and a stabilizing agent. It
Sample: 120 mg of Azaperone contains NLT 90.0% and NMT 110.0% of the labeled amount
Analysis: Dissolve in 50 mL of a mixture of methyl ethyl of C19H22FN3O.
ketone and glacial acetic acid (7:1). Add 3 drops of p-
naphtholbenzein TS, and titrate with 0.1 N perchloric acid IDENTIFICATION
VS. Perform a blank determination, and make any necessary The chromatogram of the Sample solution exhibits a major
correction. Each mL of 0.1 N perchloric acid is equivalent to peak for azaperone, the retention time of which corresponds
16.37 mg of C19H22FN3O. to that of the Standard solution, as obtained in the Assay.
316 Azaperone / Official Monographs USP 32
ASSAY Azatadine Maleate

PROCEDURE (Comment on this Monograph)id=m6600=Azatadine
Mobile phase: Acetonitrile and 0.01 M dibasic potassium Maleate=A-Monos.pdf)
phosphate mixture (3:2)
Adjust by the addition of dilute phosphoric acid (1 in 10) to
a pH of 7.8 0.1.
Internal standard solution: 0.5 mg/mL of benzophenone
in methanol
Standard stock solution: 0.5 mg/mL of USP Azaperone RS
in methanol
Standard solution: Combine 2.5 mL of Standard stock
solution and 2.5 mL of Internal standard solution, and dilute C20H22N2 2C4H4O4 522.55
quantitatively with methanol to 10.0 mL. 5H-Benzo[5,6]cyclohepta[1,2-b]pyridine, 6,11-dihydro-11-(1-
Sample stock solution: 0.5 mg/mL of azaperone from methyl-4-piperidinylidene)-, (Z)-2-butenedioate (1:2);
Injection in methanol 6,11-Dihydro-11-(1-methyl-4-piperidylidene)-5H-benzo
Sample solution: Combine 2.5 mL of Sample stock solution [5,6]cyclohepta[1,2-b]pyridine maleate (1:2) [3978-86-7].
and 2.5 mL of Internal standard solution, and dilute
quantitatively with methanol to 10.0 mL. DEFINITION
Chromatographic system Azatadine Maleate contains NLT 98.0% and NMT 102.0% of
(See Chromatography 621, System Suitability.) C20H22N2 2C4H4O4, calculated on the dried basis.
Mode: LC
Detector: UV 243 nm IDENTIFICATION
Column: 4.6-mm 25-cm; packing L1 A. INFRARED ABSORPTION 197M
Flow rate: 2 mL/min B. ULTRAVIOLET ABSORPTION 197U
Injection size: 10 L Solution: 40 g/mL
System suitability Medium: 0.25 N hydrochloric acid in methanol
Sample: Standard solution ASSAY
Suitability requirements PROCEDURE
Resolution: NLT 2.7 between the azaperone and the Sample solution: Dissolve about 650 mg of Azatadine
internal standard peaks, Standard solution Maleate in 50 mL of glacial acetic acid, and add 2 drops of
Relative standard deviation: NMT 2.0%, Standard crystal violet TS.
solution Analysis: Titrate with 0.1 N perchloric acid VS. Perform a
Analysis blank determination (see Titrimetry 541). Each mL of 0.1 N
Samples: Standard solution and Sample solution perchloric acid is equivalent to 26.13 mg of C20H22N2
Calculate the percentage of C19H22FN3O in each mL of the 2C4H4O4.
Injection taken: Acceptance criteria: 98.0%102.0%
Result = (RU/RS) (CS/CU) 100 IMPURITIES
RU = ratio of the azaperone peak to the RESIDUE ON IGNITION 281: NMT 0.1%
benzophenone peak of the Sample solution Organic Impurities
RS = ratio of the azaperone peak to the PROCEDURE
benzophenone peak of the Standard solution Adsorbent: 0.25-mm layer of chromatographic silica gel
CS = concentration of USP Azaperone RS in the mixture
Standard solution (mg/mL) Sample solution: 7 mg/mL of the Azatadine Maleate in a
CU = concentration of azaperone in the Sample mixture of toluene and methanol (1:1)
solution (mg/mL) Standard solution: 7 mg/mL of USP Azatadine Maleate RS
Acceptance criteria: 90.0%110.0% in a mixture of toluene and methanol (1:1)
SPECIFIC TESTS Application volume: 100 L
PH 791: 4.05.6 Developing solvent system: Toluene, isopropyl alcohol,
OTHER REQUIREMENTS: Meets the requirements under and diethylamine (10:10:1)
Injections 1 Analysis 1
ADDITIONAL REQUIREMENTS Proceed as directed for Chromatography 621, Thin-Layer
PACKAGING AND STORAGE: Preserve in single-dose or in Chromatography. Allow the spots to dry, and develop the
multiple-dose containers, preferably of Type I glass, chromatogram in the Developing solvent system until the
protected from light. solvent front has moved about three-fourths of the
LABELING: Label it to indicate that it is for veterinary use length of the plate. Remove the plate from the
only. developing chamber, mark the solvent front, and allow
USP REFERENCE STANDARDS 11 the solvent to evaporate. Locate the spots on the plate
USP Azaperone RS by visualization under short-wavelength UV light.
USP 32 Official Monographs / Azatadine 317
Separately transfer the silica gel mixture containing the solvent hexane extracts on a steam bath under a stream
principal spot from each track to suitable stoppered of nitrogen to dryness, pipet 1 mL of solvent hexane into
centrifuge tubes. each flask, insert the stoppers, and mix by use of a vortex
Similarly transfer an equal amount of silica gel from a mixer (or equivalent) until the residues have dissolved. Use
blank section of the plate to a separate, suitable these solutions as the Standard solution and the Sample
stoppered centrifuge tube. To each of the three tubes, solution, respectively.
add 15.0 mL of a solvent mixture consisting of methanol Apply 100 L of each of the Sample and Standard solutions
and 0.5 N hydrochloric acid (4:1), shake vigorously for separately to a suitable thin-layer chromatographic plate.
about 15 min, centrifuge, and use the supernatants for Allow the spots to dry, and develop the chromatogram in
the next spectrometric Analysis. the Developing solvent system, until the solvent front has
[NOTETake care to separate the principal spots from moved three-fourths of the length of the plate. Remove
any adjacent spots.] the plate from the developing chamber, mark the solvent
Spectrometric conditions front, and allow the plate to air-dry. Examine the plate
Cell: 1 cm under short-wavelength UV light.
Analytical wavelength: At 284 nm Acceptance criteria: The RF value and intensity of the
Blank: Solution obtained from the blank section of the principal spot in the chromatogram of the Sample solution
plate correspond to those from the Standard solution.
Analysis 2
Samples: Sample solution and the Standard solution ASSAY
Concomitantly determine the absorbances. PROCEDURE
Calculate the chromatographic purity in the portion of Standard solution: 0.06 mg/mL of USP Azatadine Maleate
Azatadine Maleate taken: RS in 0.1 N hydrochloric acid
(AU/AS) (CS/CU) 100 Transfer a portion of the powder, equivalent to 1.5 mg of
azatadine maleate, to a 50-mL flask fitted with a glass
AU = absorbance of the Sample solution stopper. Add 25.0 mL of 0.1 N hydrochloric acid, insert the
AS = absorbance of the Standard solution stopper, and shake the mixture by mechanical means for 30
CS = concentration of USP Azatadine Maleate RS in min. Filter the mixture into a suitable glass-stoppered vessel,
the Standard solution (mg/mL) discarding the first 5 mL of the filtrate (0.06 mg/mL of
CU = nominal concentration of azatadine maleate in azatadine maleate).
the Sample solution (mg/mL) Analysis: Separately transfer 15.0 mL of the Standard
Acceptance criteria: NLT 98.0% solution, 15.0 mL of the Sample solution, and 15.0 mL of 0.1
N hydrochloric acid to provide the reagent blank to three
SPECIFIC TESTS 50-mL centrifuge tubes fitted with glass stoppers. To each
LOSS ON DRYING 731: Dry it in vacuum at 60 for 3 h: it centrifuge tube, add 10.0 mL of 1.0 N sodium hydroxide
loses NMT 1.0% of its weight. and 20 mL of solvent hexane, insert the stoppers, rotate the
centrifuge tubes for 15 min, and centrifuge until the
ADDITIONAL REQUIREMENTS supernatants (solvent hexane phase) are clear. With the aid
PACKAGING AND STORAGE: Preserve in well-closed containers. of separate syringes, transfer the supernatants to separate
USP REFERENCE STANDARDS 11 50-mL centrifuge tubes fitted with glass stoppers. Rinse each
USP Azatadine Maleate RS syringe with 10 mL of solvent hexane, and add the rinse to
the aqueous phase from which the respective supernatant
was removed. Insert the stoppers, rotate each tube for 10
Azatadine Maleate Tablets min, and centrifuge. Transfer each supernatant to the
respective supernatant previously collected. Pipet 15 mL of
(Comment on this Monograph)id=m6630=Azatadine Maleate 0.1 N hydrochloric acid into each centrifuge tube containing
Tablets=A-Monos.pdf) the combined supernatants, insert the stoppers, rotate each
DEFINITION tube for 15 min, and centrifuge. Remove and discard the
Azatadine Maleate Tablets contain NLT 90.0% and NMT supernatants. Concomitantly determine the absorbances of
110.0% of the labeled amount of C20H22N2 2C4H4O4. the solutions.
IDENTIFICATION Cell: 1 cm
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Analytical wavelength: At 283 nm
Adsorbent: 0.25-mm layer of chromatographic silica gel Analysis
mixture Samples: Standard solution, Sample solution, and blank
Application volume: 100 L [NOTEUsing the prepared reagent blank, zero the
Developing solvent system: Toluene, diethylamine, and spectrophotometer with 0.1 N hydrochloric acid.]
isopropyl alcohol (10:1:10) Calculate the percentage of C20H22N2 2C4H4O4 in the
Analysis portion of Tablets taken:
Transfer 15.0 mL of the Standard solution and 15.0 mL of Result = (AU/AS) (CS/CU) 100
the Sample solution, respectively, prepared as directed in
the Assay, to separate 50-mL centrifuge tubes fitted with AU = absorbance of the Sample solution
glass stoppers. To each centrifuge tube, add 10.0 mL of AS = absorbance of the Standard solution
1.0 N sodium hydroxide and 20 mL of solvent hexane, CS = concentration of USP Azatadine Maleate RS in
insert the stoppers, rotate the centrifuge tubes for 15 min, the Standard solution (mg/mL)
and centrifuge. Transfer the solvent hexane extracts CU = nominal concentration of azatadine maleate in
(upper phase) from each centrifuge tube to separate 50- the Sample solution (mg/mL)
mL conical flasks fitted with glass stoppers. Evaporate the
318 Azatadine / Official Monographs USP 32
Acceptance criteria: 90.0%110.0% Application volume: 5 L

Analysis
PERFORMANCE TESTS Samples: Sample solution, Standard solution A, and
DISSOLUTION 711 Standard solution B
Medium: 0.01 N hydrochloric acid; 500 mL Proceed as directed for Chromatography 621, Thin-Layer
Apparatus 2: 50 rpm Chromatography. Apply the Samples at points 2 cm from
Time: 30 min the bottom edge of a thin-layer chromatographic plate.
Detector: UV 283 nm Allow the spots to dry, and develop the chromatogram
Standard solution: USP Azatadine Maleate RS in Medium in a suitable chamber, using butyl alcohol, previously
Sample solutions: Sample per Dissolution 711 saturated with 6 N ammonium hydroxide, as the solvent,
Dilute with Medium to a concentration that is similar to that until the solvent front has moved 15 cm from the point
of the Standard solution. of application. Remove the plate, air-dry, and locate the
Tolerances: NLT 80% (Q) of the labeled amount of spots by viewing under short- and long-wavelength UV
C20H22N2 2C4H4O4 light.
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Acceptance criteria: Any spot from Azathioprine, other
than the principal spot, is not more intense than the spot
ADDITIONAL REQUIREMENTS from USP Mercaptopurine RS (1.0%).
USP REFERENCE STANDARDS 11 SPECIFIC TESTS
USP Azatadine Maleate RS ACIDITY OR ALKALINITY: Shake 2.0 g with 100 mL of water
for 15 min, and filter: 20.0 mL of the filtrate requires for
neutralization NMT 0.10 mL of 0.020 N hydrochloric acid or
Azathioprine NMT 0.10 mL of 0.020 N sodium hydroxide, methyl red TS
being used as the indicator.
(Comment on this Monograph)id=m6690=Azathioprine=A- LOSS ON DRYING 731: Dry it in vacuum at 105 for 5 h: it
Monos.pdf) loses NMT 1.0% of its weight.
containers.
USP Azathioprine RS
USP Mercaptopurine RS
C9H7N7O2S 277.26
1H-Purine, 6-[(1-methyl-4-nitro-1H-imidazol-5-yl)thio]-;
6-[(1-Methyl-4-nitroimidazol-5-yl)thio]purine [446-86-6]. Azathioprine Oral Suspension
(Comment on this Monograph)id=m1378=Azathioprine Oral
DEFINITION Suspension=A-Monos.pdf)
Azathioprine contains NLT 98.0% and NMT 101.5% of
C9H7N7O2S, calculated on the dried basis. DEFINITION
Azathioprine Oral Suspension contains NLT 90.0 %and NMT
IDENTIFICATION 110.0 %of the labeled amount of azathioprine (C9 H7 N7 O2
A. INFRARED ABSORPTION 197K S).
B. The principal spot from the Sample solution in the test for Prepare Azathioprine Oral Suspension 50 mg/mL as follows (See
Limit of mercaptopurine shows the same RF value as that Pharmaceutical CompoundingNonsterile Solutions 795):
obtained from the solution of USP Azathioprine RS.
ASSAY Azathioprine 5g
PROCEDURE Vehicle: a mixture of Vehicle for Oral A sufficient quantity
Sample: 300 mg of Azathioprine Solution, (regular or sugar-free), NF and
Analysis: Dissolve in 80 mL of dimethylformamide. Add 5 Vehicle for Oral Suspension, NF (1:1)
drops of a 1 in 100 solution of thymol blue in
dimethylformamide. Titrate with 0.1 N tetrabutylammonium To make 100 mL
hydroxide VS, using a magnetic stirrer, and taking
precautions to prevent absorption of atmospheric carbon If using Tablets, comminute them to a fine powder in a suitable
dioxide. Perform a blank determination (see Titrimetry mortar, or add Azathioprine powder to the mortar. Add about
541). Each mL of 0.1 N tetrabutylammonium hydroxide is 10 mL of the Vehicle, and mix to a uniform paste. Add the
equivalent to 27.73 mg of C9H7N7O2S. Vehicle in small portions almost to volume, and mix
Acceptance criteria: 98.0%101.5% thoroughly after each addition. Transfer the contents of the
mortar, stepwise and quantitatively, to a calibrated bottle. Add
IMPURITIES sufficient Vehicle to bring to final volume, and mix well.
Inorganic Impurities [CAUTIONAvoid skin contact or inhalation of azathioprine by
RESIDUE ON IGNITION 281: NMT 0.1% using protective gloves and a fume hood or surgical mask.]
Organic Impurities
LIMIT OF MERCAPTOPURINE ASSAY
Adsorbent: 0.25-mm layer of microcrystalline cellulose PROCEDURE
Standard solution A: 20 mg/mL of USP Azathioprine RS in Mobile phase: Dissolve 1.1 g of sodium-1-heptanesulfonate
6 N ammonium hydroxide in 700 ml water, add 30 mL of methanol. Adjust with 1 N
Standard solution B: 200 g/mL of USP Mercaptopurine hydrochloric acid to a pH of 3.5.
RS, on the anhydrous basis in 6 N ammonium hydroxide Standard solution: 25 mg of USP Azathioprine RS, in a 50-
Sample solution: 20 mg/mL of Azathioprine in 6 N mL volumetric flask
ammonium hydroxide
USP 32 Official Monographs / Azathioprine 319
Add 15 mL of methanol and 0.5 mL of ammonium Filter the solution.

hydroxide to the flask, swirl and sonicate for 2 mins. Dilute Chromatographic system
with methanol to volume. Transfer 10 mL of this solution Adsorbent: 0.25-mm layer of microcrystalline cellulose
to a 50-mL volumetric flask, and dilute with water to Application volume: 5 L
volume. Developing solvent system: Butyl alcohol, previously
Sample solution: Agitate the container of Oral Suspension saturated with 6 N ammonium hydroxide
for 30 mins on a rotating mixer, remove a 5-mL sample, Analysis
and store in a clear glass vial at 70 until analyzed. At the Samples: Standard solution and Sample solution
time of analysis, remove the sample from the freezer, allow Proceed as directed in the chapter.
it to reach room temperature, and mix with a vortex mixer
for 30 s. Pipet 1.0 mL of the Sample solution into a 100-mL ASSAY
volumetric flask, and dilute with Mobile phase to volume. PROCEDURE
Chromatographic system Mobile phase: To 1.1 g of sodium 1-heptanesulfonate in
(See Chromatography 621, System Suitability.) 700 mL of water, add 300 mL of methanol. Adjust the
Mode: LC solution with 1 N hydrochloric acid to a pH of 3.5. Filter the
Detector: UV 254 nm solution through a 0.8-m solvent-resistant membrane, and
Column: 4.6-mm 25-cm; 5-m packing L1 degas.
Flow rate: 2 mL/min Standard stock solution: Transfer 25 mg of USP
Injection size: 20 L Azathioprine RS to a 50-mL volumetric flask. Add 15 mL of
System suitability methanol and 0.5 mL of ammonium hydroxide to the flask,
Sample: Standard solution swirl, and sonicate for 2 min. Dilute with methanol to
Suitability requirements volume.
Relative standard deviation: NMT 1.3% Standard solution: Transfer 10.0 mL of Standard stock
Retention time: 4 min solution to a 50-mL volumetric flask, and dilute with water
Analysis to volume.
Samples: Sample solution and Standard solution Sample stock solution: Finely powder NLT 20 Tablets.
Calculate the percentage of C9H7N7O2S in the volume of Weigh a portion of the powder, equivalent to 50 mg of
Oral Suspension : azathioprine, and transfer to a 100-mL volumetric flask. Add
25 mL of methanol and 1.0 mL of ammonium hydroxide to
Result = (rU/rS) (CS/CU) 100 the flask, swirl, and sonicate for 2 min. Dilute with methanol
to volume. Allow the excipients to settle.
rU = peak response from the Sample solution Sample solution: Transfer 10.0 mL of the Sample stock
rS = peak response from the Standard solution solution to a 50-mL volumetric flask, and dilute with water
CS = concentration of USP Azathioprine RS in the to volume.
Standard solution (g/mL) Chromatographic system
CU = nominal concentration of the Sample solution (See Chromatography 621, System Suitability.)
(g/mL) Mode: LC
Acceptance criteria: 90.0%110.0% Detector: UV 254 nm
SPECIFIC TESTS Flow rate: 2 mL/min
PH 791: 3.84.8 Injection size: 10 L
System suitability
PACKAGING AND STORAGE: Preserve in tight, light-resistant Suitability requirements
containers. Store at room temperature, or in a cold place. Column efficiency: NLT 800 theoretical plates, Standard
LABELING: Label it to state that it is to be well shaken before solution
use, and to state the beyond-use date. Tailing factor: NMT 1.5 for the azathioprine peak,
Beyond-use date: 60 days after the day on which it was Standard solution
compounded Relative standard deviation: NMT 2.0%, Standard
USP REFERENCE STANDARDS 11 solution
USP Azathioprine RS Analysis
Calculate the percentage of C9H7N7O2S in the portion of
Tablets taken:
Azathioprine Tablets
(Comment on this Monograph)id=m6720=Azathioprine Result = (rU/rS) (CS/CU) 100
rU = peak response for azathioprine from the Sample
DEFINITION solution
Azathioprine Tablets contain NLT 93.0% and NMT 107.0% of rS = peak response for azathioprine from the
the labeled amount of azathioprine (C9H7N7O2S). Standard solution
CS = concentration of USP Azathioprine RS in the
IDENTIFICATION Standard solution (mg/mL)
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 CU = concentration of azathioprine in the Sample
Standard solution: 20 mg/mL of USP Azathioprine RS in 6 solution (mg/mL)
N ammonium hydroxide
Sample solution: Equivalent to 20 mg/mL of azathioprine,
from Powdered Tablets in 6 N ammonium hydroxide
320 Azathioprine / Official Monographs USP 32
Acceptance criteria: 93.0%107.0% CS = concentration of USP Azathioprine RS in the

PERFORMANCE TESTS CU = concentration of azathioprine in the Sample
DISSOLUTION 711 solution (unit/mL)
Medium: Water; 900 mL L = label claim (mg/unit)
Apparatus 2: 50 rpm Acceptance criteria: 93.0%107.0%
Time: 30 min
Detector: UV 280 nm PERFORMANCE TESTS
Sample solutions: Sample per Dissolution 711 UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Filter and dilute, if necessary, with Medium to a
concentration that is similar to that of the Standard IMPURITIES
solution. Organic Impurities
Standard solution: USP Azathioprine RS in Medium LIMIT OF MERCAPTOPURINE
Tolerances: NLT 75% (Q) of the labeled amount of Standard solution A: 10 mg/mL of USP Azathioprine RS in
C9H7N7O2S dimethylformamide
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Standard solution B: 100 g/mL of USP Mercaptopurine
RS in dimethylformamide
ADDITIONAL REQUIREMENTS Sample solution: 10 mg/mL of Azathioprine Sodium for
PACKAGING AND STORAGE: Protect from light. Injection in dimethylformamide
USP Azathioprine RS (See Chromatography 621, Thin-Layer Chromatography.)
Adsorbent: 250-m layer of microcrystalline cellulose
Application volumes
Standard solution B: 15 L
Azathioprine Sodium for Injection Sample solution and Standard solution A: 5 L
(Comment on this Monograph)id=m6730=Azathioprine Sodium Developing solvent system: Butyl alcohol, previously
for Injection=A-Monos.pdf) saturated with 5 N ammonium hydroxide
Analysis
DEFINITION Samples: Sample solution, Standard solution A, and
Azathioprine Sodium for Injection is a sterile solid prepared by Standard solution B
the freeze-drying of an aqueous solution of Azathioprine and Proceed as directed for Chromatography 621, Thin-Layer
Sodium Hydroxide. It contains NLT 93.0% and NMT 107.0% Chromatography. Apply Samples, at points 2 cm from the
of the labeled amount of azathioprine (C9H7N7O2S). bottom edge of a thin-layer chromatographic plate.
Allow the spots to dry, and develop the chromatogram
IDENTIFICATION until the solvent front has moved 15 cm from the point
The principal spot of the Sample solution obtained in the test of application. Remove the plate, air-dry, and locate the
for Limit of mercaptopurine shows the same RF value as that spots by viewing under short- and long-wavelength UV
obtained with Standard solution A. light.
ASSAY Acceptance criteria: Any spot from azathioprine, other
PROCEDURE than the principal spot, is not more intense than the spot
Standard solution: Transfer 25 mg of USP Azathioprine RS obtained with USP Mercaptopurine RS (3.0%).
to a 50-mL volumetric flask, dissolve in 2.5 mL of 0.1 N SPECIFIC TESTS
sodium hydroxide, and dilute with water to volume. Pipet COMPLETENESS OF SOLUTION 641: The contents of 1
10.0 mL of this solution into a 50-mL volumetric flask, and container are soluble in 10 mL of water, to give a clear,
dilute with 0.1 N sulfuric acid to volume. bright yellow solution, essentially free from foreign matter.
Sample solution: Transfer the contents of 1 vial of PH 791
Azathioprine Sodium for Injection with the aid of water to a Sample solution: The contents of 1 container dissolved in
100-mL volumetric flask, and dilute with water to volume. 10 mL of water
Pipet 10 mL of this solution into another 100-mL volumetric Acceptance criteria: 9.811.0
flask, and dilute with 0.1 N sulfuric acid to volume. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 1.0 USP
Analysis: Transfer 20 mL each of the Standard solution and Endotoxin Unit/mg of azathioprine.
the Sample solution, separately, to polarographic cells, and WATER DETERMINATION, Method I 921: NMT 7.0%, the
deaerate for 10 min with nitrogen that previously has been Sample solution being prepared as directed for a hygroscopic
saturated with 0.1 N sulfuric acid. Blanket the solution with specimen.
saturated nitrogen, insert the dropping mercury electrode of OTHER REQUIREMENTS: It meets the requirements under
a suitable polarograph, and record the polarogram from Injections 1.
0.60 to 1.00 V, using a saturated calomel electrode as the
reference electrode. Determine the height of the diffusion ADDITIONAL REQUIREMENTS
current as the difference between the residual current and PACKAGING AND STORAGE: Preserve as described under
diffusion current plateau. Injections 1, Containers for Sterile Solids, at controlled room
Calculate the percentage of C9H7N7O2S in the volume of temperature.
solution taken from the vial used for the Sample solution: USP REFERENCE STANDARDS 11
USP Azathioprine RS
Result = (id)U/(id)S (CS/CU) (100/L) USP Endotoxin RS
USP Mercaptopurine RS
(id)U = diffusion currents of the Sample solution
(id)S = diffusion currents of the Standard solution
USP 32 Official Monographs / Azithromycin 321
Azithromycin Chromatographic system

(Comment on this Monograph)id=m6740=Azithromycin=A- (See Chromatography 621, System Suitability.)
Monos.pdf) Mode: LC
Detector: Amperometric electrochemical detector
Detector type: Dual glassy carbon electrodes
Detector mode: Oxidative screen mode
Detector settings: Electrode 1 set at +0.70 0.05 V,
electrode 2 set at +0.82 0.05 V, background current
optimized to 85 15 nanoamperes
Column
Guard column: 4.6-mm 5-cm; 5-m packing L29
C38H72N2O12 xH2O (anhydrous) 749.00 Analytical column: 4.6-mm 15-cm; 5-m packing L29
[83905-01-5]. or 3-m packing L49 without the guard column
1-Oxa-6-azacyclopentadecan-15-one, 13-[(2,6-dideoxy-3-C- Flow rate: 1.5 mL/min
methyl-3-O-methyl--L-ribo-hexopyranosyl)oxy]-2- Injection size: 50 L
ethyl-3,4,10-trihydroxy-3,5,6,8,10,12,14-heptamethyl-11-[ System suitability
[3,4,6-trideoxy-3-(dimethylamino)--D-xylo- Samples: Standard solution and System suitability solution
hexopyranosyl]oxy]- [2R(2R*,3S*,4R*,5R*,8R*,10R*,11R*,12S*, [NOTEThe relative retention times for azaerythromycin A
13S*,14R*)]; and azithromycin with the L29 column are 0.7 and 1.0,
(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-13-[(2,6-Dideoxy-3-C- respectively; the relative retention times for
methyl-3-O-methyl--L-ribo-hexopyranosyl)oxy]-2- azaerythromycin A and azithromycin with the L49 column
ethyl-3,4,10-trihydroxy-3,5,6,8,10,12,14-heptamethyl-11-[ are 0.8 and 1.0, respectively.]
[3,4,6-trideoxy-3-(dimethylamino)--D-xylo- Suitability requirements
hexopyranosyl]oxy]-1-oxa-6-azacyclopentadecan-15-one; Resolution: NLT 2.5 between azaerythromycin A and
9-Deoxo-9a-aza-9a-methyl-9a-homoerythromycin azithromycin, System suitability solution
A Monohydrate 767.02 Column efficiency: NLT 1000 theoretical plates, Standard
[121479-24-4]. solution
Dihydrate 785.02 Tailing factor range: 0.91.5 for azithromycin, Standard
[117772-70-0]. solution
DEFINITION solution
Azithromycin is anhydrous or contains one or two molecules of Analysis
water of hydration. It contains the equivalent of NLT 945 g Samples: Standard solution and Sample solution
and NMT 1030 g of azithromycin (C38H72N2O12) per mg, Calculate the quantity, in g, of C38H72N2O12 in each mg of
calculated on the anhydrous basis. Azithromycin taken:
[NOTEUse water that has a resistivity of NLT 18 Mohm-cm
(0.055S-cm) for all tests in this monograph.] Result = (rU/rS) (CS/CU) P
IDENTIFICATION rU = peak response from the Sample solution
A. INFRARED ABSORPTION 197K: If a difference appears in rS = peak response from the Standard solution
the IR spectra of the analyte and the standard, dissolve equal CS = concentration of USP Azithromycin RS in the
portions of the test specimen and the Reference Standard in Standard solution
equal volumes of methanol. Evaporate the solutions to CU = concentration of Azithromycin in the Sample
dryness on a water bath, and dry at 80 for 30 min under solution
vacuum. Perform the test on the residues. P = potency of USP Azithromycin RS (g/mg of
B. The retention time of the azithromycin peak in the azithromycin)
Sample solution corresponds to that in the Standard solution, Acceptance criteria: Equivalent of 9451030 g
as obtained in the Assay.
IMPURITIES
ASSAY Inorganic Impurities
PROCEDURE RESIDUE ON IGNITION 281: NMT 0.3%, the charred residue
Mobile phase: Dissolve 5.8 g of monobasic potassium being moistened with 2 mL of nitric acid and 5 drops of
phosphate in 2130 mL of water, add 870 mL of acetonitrile. sulfuric acid
Adjust with 6 mL of 10 N potassium hydroxide to a pH of HEAVY METALS, Method II 231: NMT 25 ppm
11.0 0.1 and filter. Organic Impurities
Standard stock solution: 0.165 mg/mL of USP PROCEDURE 1
Azithromycin RS in acetonitrile [NOTEPerform either Procedure 1 or Procedure 2, depending
Standard solution: 3.3 g/mL of USP Azithromycin RS, on the manufacturing process used.]
from the Standard stock solution in Mobile phase [NOTEUse water that has a resistivity of NLT 18 Mohm-
Sample stock solution: 0.165 mg/mL of Azithromycin in cm.]
acetonitrile Mobile phase: Proceed as directed in the Assay.
Sample solution: 3.3 g/mL of Sample stock solution in Solution A: 2.7 mg/mL of monobasic potassium
Mobile phase phosphate in water
System suitability stock solution: 0.16 mg/mL of USP Adjust with 10 N potassium hydroxide to a pH of 7.5 0.1.
Azaerythromycin A RS in acetonitrile and Mobile phase (1:9) Diluent: Acetonitrile and Solution A (1:3)
[NOTEDissolve in acetonitrile by swirling and with the aid Standard stock solution: 45 g/mL of USP
of brief sonication, and then dilute with Mobile phase to Desosaminylazithromycin RS, 105 g/mL of USP N-
volume.] Demethylazithromycin RS, and 160 g/mL of USP
System suitability solution: Transfer 2.0 mL of the System Azithromycin RS in acetonitrile
suitability stock solution and 2.0 mL of the Standard stock Standard solution: 0.9 g/mL of USP
solution to a 100-mL volumetric flask, and dilute with Mobile Desosaminylazithromycin RS, 2.1 g/mL of USP N-
phase to volume.
322 Azithromycin / Official Monographs USP 32
Demethylazithromycin RS, and 3.2 g/mL of USP Acceptance criteria: See Impurity Table 1.
Azithromycin RS from Standard stock solution in Diluent
Sample solution: 33 mg of Azithromycin in a 100-mL Impurity Table 1
volumetric flask
Add 5 mL of acetonitrile, and sonicate for about 20 s to Acceptance
dissolve. Dilute with Diluent to volume. Relative Criteria
[NOTEUse this solution within 6 h.] Name Retention Time (NMT %)
Chromatographic system Desosaminylazithromycin 0.20 0.3
(See Chromatography 621, System Suitability.) N-demethylazithromycin 0.26 0.7
Mode: LC
Detector: Amperometric electrochemical detector Any other individual 0.34 1.0
electrodes impurity
Detector type: Dual glassy carbon Sum of all impurities 0.37 3.0
Detector mode: Oxidative screen mode
Detector settings: Electrode 1 set at +0.70 0.05 V, PROCEDURE 2
electrode 2 set at +0.85 0.05 V, background current [NOTEPerform either Procedure 1 or Procedure 2, depending
optimized to 95 25 nanoamperes on the manufacturing process used.]
[NOTEIn general, maintain electrode 1 at 0.12 V less Solution A: 8.7 mg/mL of dibasic potassium phosphate in
than electrode 2, and maintain the electrodes at a water
constant temperature of about 26.] Adjust with 20% phosphoric acid to a pH of 8.2.
Column Mobile phase: Acetonitrile and Solution A (3:2)
Guard column: 4.6-mm 5-cm; 5-m packing L29 Standard solution A: 35 g/mL of USP Azithromycin RS in
Analytical column: 4.6-mm 15-cm; 5-m packing L29 Mobile phase
or 3-m packing L49 without the guard column. Standard solution B: 7 mg/mL of USP Azithromycin
Flow rate: 0.4 mL/min Identity RS in Mobile phase
Injection size: 50 L Standard solution C: 14 g/mL of USP Azithromycin-N-
System suitability Oxide RS in Mobile phase
Sample: Standard solution Sample solution: 7 mg/mL of Azithromycin in Mobile
[NOTEThe relative retention times for phase
desosaminylazithromycin, N-demethylazithromycin, and System suitability solution: 0.07 mg/mL of USP
azithromycin are 0.38, 0.54, and 1.0.] Azaerythromycin A RS and 7 mg/mL of USP Azithromycin
Suitability requirements RS in Mobile phase
Column efficiency: NLT 1500 theoretical plates for the Chromatographic system
azithromycin peak (See Chromatography 621, System Suitability.)
Tailing factor: NMT 1.5 for each peak Mode: LC
Relative standard deviation: NMT 5% for each of these Detector: UV 210 nm
compounds Column: 4.6-mm 15-cm; 5-m packing L1
Analysis Temperature: 30
Samples: Standard solution and Sample solution Flow rate: 0.9 mL/min
[NOTERecord chromatograms for NLT 3.3 times the Injection size: 50 L
elution time of the azithromycin peak.] System suitability
Calculate the percentages of desosaminylazithromycin and Sample: System suitability solution
N-demethylazithromycin in the Azithromycin taken: [NOTEFor the purpose of identification, the relative
retention times for azaerythromycin A and azithromycin
Result = (rU/rS) (CS/CU) 100 are 0.47 and 1.00, respectively.]
rU = peak area for the relevant analyte from the Resolution: NLT 8.0 between azaerythromycin A and
Sample solution azithromycin
rS = peak area for the relevant analyte from the Tailing factor: NMT 2.5 for the azithromycin
Standard solution Analysis
CS = concentration of the appropriate USP Reference Samples: Mobile phase, Standard solution A, Standard
Standard in the Standard solution (g/mL) solution B, Standard solution C, and Sample solution
CU = concentration of the Sample solution [NOTEDisregard any peak due to the solvent front and
Calculate the percentages of other related substances in any peak corresponding to those obtained from the
the Azithromycin taken: Mobile phase.]
Result = (rU/rS) (CS/CU) 100 of Azithromycin taken:
rU = peak area of each additional impurity from the Result = (rU/rS) (CS/CU) 100/RRF
Sample solution
rS = peak area of the azithromycin peak from the rU = peak area for each impurity from the Sample
Standard solution solution
CS = concentration of USP Azithromycin RS in the rS = peak area for each impurity from the Standard
Standard solution (g/mL) solution A
CS = concentration of USP Azithromycin RS in about 70, and 1.8%2.6% between the inflection point at
Standard solution A (mg/mL) about 70 and the inflection point at about 130.
CU = concentration of Sample solution (mg/mL)
RRF = relative response factor (see Impurity Table 2) ADDITIONAL REQUIREMENTS
LABELING: Label it to indicate whether it is anhydrous, or the
Impurity Table 2
monohydrate or the dihydrate. The amorphous form is so
Relative Relative Acceptance labeled. Where the quantity of azithromycin is indicated in
Retention Response Criteria, the labeling of any preparation containing Azithromycin, this
Name Time Factor (NMT %) shall be understood to be in terms of anhydrous
Azithromycin-N-oxide 0.20 0.45 0.40 azithromycin (C38H72N2O12). The labeling states with which
Organic Impurities test the article complies, if other than
3-(N,N-didemethyl)-3-N- 0.26 1.8 0.30 Procedure 1.
formylazithromycin USP REFERENCE STANDARDS 11
3-N-demethyl-3-N- 0.34 4.1 0.15 USP Azaerythromycin A RS
formylazithromycin USP Azithromycin RS
(rotamer 1) USP Azithromycin Identity RS
3-N-demethyl-3-N- 0.37 4.1 0.15 USP Azithromycin-N-Oxide RS
formylazithromycin USP Desosaminylazithromycin RS
(rotamer 2) USP N-Demethylazithromycin RS
6-Demethylazithromycin 0.47 0.67 0.50
(azaerythromycin A)
3-De(dimethylamino)-3- 0.80 1.9 0.25 Azithromycin Capsules
oxoazithromycin (Comment on this Monograph)id=m6745=Azithromycin
2-Desethyl-2- 1.52 1.0 0.50 Capsules=A-Monos.pdf)
propylazithromycin
DEFINITION
3-Deoxyazithromycin 1.60 1.0 0.50 Azithromycin Capsules contain the equivalent of NLT 90.0%
(azithromycin B) and NMT 110.0% of the labeled amount of azithromycin
3-N-demethyl-3-N-[(4- 2.14 7.0 0.50 (C38H72N2O12).
methylphenyl)sulfonyl]-
azithromycin
IDENTIFICATION
The retention time for the azithromycin peak of the Sample
Individual unknown 1.0 0.20 solution corresponds to that of the Standard solution, as
impurity obtained in the Assay.
Total impurities 2.0
ASSAY
PROCEDURE
SPECIFIC TESTS [NOTEUse water that has a resistivity of NLT 18 Mohm-
OPTICAL ROTATION, Specific Rotation 781S: 45 to 49, at cm.]
20 Mobile phase: Dissolve 5.8 g of monobasic potassium
Sample solution: 20 mg/mL, in dehydrated alcohol phosphate in 2130 mL of water, and add 870 mL of
CRYSTALLINITY 695: Meets the requirements except, where acetonitrile. Adjust with 6 mL of 10 N potassium hydroxide
it is labeled as amorphous, most of the particles do not to a pH of 11.0 0.1, and filter.
exhibit birefringence and extinction positions Standard stock solution: 0.165 mg/mL of USP
PH 791: 9.011.0, in a mixture of methanol and water Azithromycin RS in acetonitrile
(1:1) containing 2 mg/mL, prepared by diluting a solution in [NOTEFirst dissolve in a small quantity of acetonitrile by
methanol containing 4 mg/mL with an equal volume of swirling and with the aid of brief sonication, and then
water dilute it to volume.]
WATER DETERMINATION, Method I 921 Standard solution: 0.0033 mg/mL of USP Azithromycin RS,
Where it is labeled as anhydrous: NMT 2.0% from Standard stock solution in Mobile phase
Where it is labeled as the dehydrate: 4.0%5.0% Sample stock solution: Remove, as completely as possible,
Where it is labeled as the monohydrate: 1.8%4.0%, the contents of NLT 20 Capsules. Mix the combined
except that it may be 4.0%6.5% when the requirements of contents, and transfer a quantity of the powder, equivalent
the Loss on Drying test are met to 250 mg of anhydrous azithromycin, to a 250-mL
LOSS ON DRYING 731 (where it is labeled as Azithromycin volumetric flask. Add 175 mL of acetonitrile, and shake by
monohydrate and has a water content of 4.0%6.5%) mechanical means for 30 min. Dilute with acetonitrile to
(See Thermal Analysis 891.) volume. Place 40 mL of the resulting suspension in a
[NOTEThe quantity taken for the determination may be centrifuge tube, and centrifuge. Transfer 2.0 mL of the clear
adjusted, if necessary, for instrument sensitivity.] supernatant to a 50-mL volumetric flask, and dilute with
Procedure: Determine the percentage of volatile substances Mobile phase to volume.
by thermogravimetric analysis in an appropriately calibrated Sample solution: Transfer 2.0 mL of Sample stock solution
instrument, using about 10 mg of Azithromycin. Heat the to a 25-mL volumetric flask, and dilute with Mobile phase to
specimen at the rate of 10/min between ambient volume.
temperature and 150 in an atmosphere of nitrogen at a System suitability stock solution: 0.16 mg/mL of USP
constant flow rate of about 35 mL/min. From the Azaerythromycin A RS in acetonitrile and Mobile phase (1:9)
thermogram plot the derivatives of the loss on drying [NOTEDissolve in acetonitrile by swirling and with the aid
(percent loss per minute), identify the inflection points of of brief sonication, and then dilute with Mobile phase to
the two weight loss steps at about 70 and 130. volume.]
Acceptance criteria: It loses NMT 4.5% of its weight System suitability solution: Transfer 2.0 mL of the System
between ambient temperature and the inflection point at suitability stock solution and 2.0 mL of Standard stock solution
324 Azithromycin / Official Monographs USP 32
to a 100-mL volumetric flask, and dilute with Mobile phase solution to a second 25-mL volumetric flask, and dilute with
to volume. Mobile phase to volume.
Chromatographic system Analysis: Determine the amount of C38H72N2O12 dissolved,
(See Chromatography 621, System Suitability.) using filtered portions of the Sample solution and using the
Mode: LC Analysis in the Assay, making any necessary modifications.
Detector: Amperometric electrochemical detector with Calculate the quantity, in mg, of C38H72N2O12 dissolved:
dual glassy carbon electrodes operated in the oxidative
screen mode with electrode 1 set at +0.70 0.05 V and Result = 70.31 (rU/rS) (CS P)
electrode 2 set at +0.82 0.05 V, and the background
current optimized to 85 15 nanoamperes rU = Azithromycin peak response from the Sample
Columns solution
Guard column: 4.6-mm 5-cm; 5-m packing L29 rS = Azithromycin peak response from the Standard
Analytical column: 4.6-mm 15-cm; 5-m packing L29 solution
or 3-m packing L49 without the guard column CS = concentration of USP Azithromycin RS in the
Flow rate: 1.5 mL/min Standard solution (mg/mL)
Injection size: 50 L P = potency of USP Azithromycin RS (g/mg)
System suitability Tolerances: NLT 75% (Q) of the labeled amount of
Samples: Standard solution and System suitability solution azithromycin is dissolved in 45 min.
[NOTEThe relative retention times for azaerythromycin A UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
and azithromycin with the L29 column are 0.7 and 1.0,
respectively; the relative retention times for SPECIFIC TESTS
azaerythromycin A and azithromycin with the L49 column WATER DETERMINATION, Method I 921: NMT 5.0%
are 0.8 and 1.0, respectively.)] ADDITIONAL REQUIREMENTS
Suitability requirements PACKAGING AND STORAGE: Preserve in well-closed containers.
Resolution: NLT 2.5 between azaerythromycin A and Where packaged in unit-of-use containers, each container
azithromycin, System suitability solution contains six 250-mg Capsules, and the label indicates the
Column efficiency: NLT 1000 theoretical plates, Standard intended sequential day of use for each Capsule.
solution USP REFERENCE STANDARDS 11
Tailing factor: NLT 0.9 for the azithromycin peak and USP Azaerythromycin A RS
NMT 1.5, Standard solution USP Azithromycin RS
solution
Analysis
Samples: Standard solution and Sample solution Azithromycin for Oral Suspension
Calculate the percentage of C38H72N2O12 in the portion of (Comment on this Monograph)id=m6750=Azithromycin for
Capsules taken: Oral Suspension=A-Monos.pdf)
Result = (rU/rS) (CS P/CU) 100 DEFINITION
Azithromycin for Oral Suspension is a dry mixture of
rU = Azithromycin area from the Sample solution Azithromycin and one or more buffers, sweeteners, diluents,
rS = Azithromycin area response from the Standard anticaking agents, and flavors. It contains NLT 90.0% and
solution NMT 110.0% of the labeled amount of azithromycin
CS = concentration of USP Azithromycin RS in (C38H72N2O12).
P = potency of USP Azithromycin RS (g/mg) IDENTIFICATION
CU = concentration of Azithromycin in Sample solution The Sample solution, obtained as directed in the Assay,
(g/mL) exhibits a major peak for azithromycin, the retention time of
Acceptance criteria: Equivalent of 90.0%110.0% which corresponds to that exhibited in the chromatogram of
the Standard solution, obtained as directed in the Assay.
PERFORMANCE TESTS
DISSOLUTION 711 ASSAY
[NOTEUse water that has a resistivity of NLT 18 Mohm- PROCEDURE
cm.] [NOTEUse water that has a resistivity of NLT 18 Mohm-
Solution A: Prepare 6 L of 0.1 M dibasic sodium phosphate, cm.]
adjust with 40 mL of hydrochloric acid to a pH of 6.0 Mobile phase: Dissolve 5.8 g of monobasic potassium
0.05, and add 600 mg of trypsin. phosphate in 2130 mL of water, add 870 mL of acetonitrile.
Medium: Solution A: 900 mL Adjust with 6 mL of 10 N potassium hydroxide to a pH of
Apparatus 2: 100 rpm 11.0 0.1, and filter.
Time: 45 min Standard stock solution: 0.165 mg/mL of USP
Standard solution: Transfer 15 mg of USP Azithromycin RS Azithromycin RS in acetonitrile
to a 50-mL volumetric flask. Add 25 mL of Medium, and [NOTEFirst dissolve in a small quantity of acetonitrile by
sonicate briefly to dissolve. Dilute with Medium to volume. swirling and with the aid of brief sonication, and then
Transfer 2.0 mL of this solution to a 25-mL volumetric flask, dilute it to volume.]
and dilute with Mobile phase (prepared as directed in the Standard solution: 0.0033 mg/mL of USP Azithromycin RS,
Assay) to volume. Transfer 4.0 mL of this solution to a from Standard stock solution in Mobile phase
second 25-mL volumetric flask, and dilute with Mobile phase Solvent: Dissolve 2.2 g of monobasic potassium phosphate
to volume. in 1590 mL of water, add 600 mL of isopropyl alcohol, 480
Sample solution: Filter a portion of the Sample solution mL of alcohol, and 330 mL of acetonitrile. Adjust with 10 N
through a filter having a porosity of 0.5-m or less. Transfer potassium hydroxide to a pH of 8.4 0.1, and shake by
2.0 mL of the filtrate to a 25-mL volumetric flask, and dilute mechanical means for 30 min.
with Mobile phase to volume. Transfer 4.0 mL of this
Sample solution A (where packaged in a single-unit Suitability requirements

container): Transfer the contents of a container of Resolution: NLT 2.5 between azaerythromycin A and
Azithromycin for Oral Suspension to a volumetric flask of azithromycin, System suitability solution
such capacity, V, in mL, that when diluted to volume as Column efficiency: NLT 1000 theoretical plates, Standard
directed in the following two sentences the solution will solution
contain 2 mg of azithromycin/mL. Add a volume of Solvent Tailing factor: NLT 0.9 for the azithromycin peak and
equal to 70% of the volume of the flask, and shake by NMT 1.5, Standard solution
mechanical means for 30 min. Dilute with Solvent to Relative standard deviation: NMT 2.0%, Standard
volume. Transfer 40 mL of this suspension to a stoppered solution
centrifuge tube, and centrifuge for 20 min. Transfer 2.0 mL Analysis
of the clear supernatant to a 50-mL volumetric flask, and Samples: Standard solution and Sample solution
dilute with Mobile phase to volume. Transfer 2.0 mL of this Calculate the quantity, in mg, of C38H72N2O12 in the single-
solution to a second 50-mL volumetric flask, and dilute with unit container of Azithromycin for Oral Suspension taken:
Mobile phase to volume.
Sample solution B (where packaged in a multiple-unit Result = (rU/rS) (125V/200) CP
container): Constitute Azithromycin for Oral Suspension as
directed in the labeling. Transfer 5.0 mL of the suspension rU = Azithromycin peak response from the Sample
so obtained, freshly mixed and free from air bubbles, to a solution A
volumetric flask of such capacity, Vm, in mL, that when rS = Azithromycin peak response from the Standard
diluted to volume as directed below, the solution will solution
contain 0.4 mg of azithromycin/mL. Add a volume of V = volume, 2 mg of azithromycin/mL
Solvent equal to 70% of the volume of the flask, and shake C = concentration of USP Azithromycin RS in the
by mechanical means for 30 min. Dilute with Solvent to Standard solution (mg/mL)
volume. Transfer 40 mL of the suspension so obtained to a P = potency of USP Azithromycin RS (g/mg)
stoppered centrifuge tube, and centrifuge for 20 min. Calculate the percentage of C38H72N2O12 in each 5 mL of
Transfer 5.0 mL of the clear supernatant to a 50-mL suspension taken from a container of Azithromycin for
volumetric flask, and dilute with Mobile phase to volume. Oral Suspension where packaged in a multiple-unit
Transfer 5.0 mL of this solution to a second 50-mL container:
volumetric flask, and dilute with Mobile phase to volume.
System suitability stock solution: 0.16 mg/mL of USP Result = (rU/rS) (Vm/10) CP
Azaerythromycin A RS in acetonitrile and Mobile phase (1:9)
[NOTEDissolve in acetonitrile by swirling and with the aid rU = Azithromycin peak response from Sample solution
of brief sonication, and then dilute with Mobile phase to B
volume.] rS = Azithromycin peak response from the Standard
System suitability solution: Transfer 2.0 mL of System solution
suitability stock solution and 2.0 mL of Standard stock Vm = volume, 0.4 mg of azithromycin/mL
solutionto a 100 mL volumetric flask, and dilute with Mobile C = concentration of USP Azithromycin RS in the
phase to volume. Standard solution (mg/mL)
Chromatographic system P = potency of USP Azithromycin RS (g/mg)
(See Chromatography 621, System Suitability.) Acceptance criteria: 90.0%110.0%
Mode: LC SPECIFIC TESTS
Detector: Amperometric electrochemical detector with PH 791
dual glassy carbon electrodes operated in the oxidative For solid packaged in single-unit containers: 9.011.0, in
screen mode with electrode 1 set at +0.70 0.05 V and the suspension constituted as directed in the labeling
electrode 2 set at +0.82 0.05 V, and the background For solid packaged in multiple-unit containers: 8.511.0,
current optimized to 85 15 nanoamperes in the suspension constituted as directed in the labeling
Column WATER DETERMINATION, Method I 921: NMT 1.5%
Guard column: 4.6-mm 5-cm; 5-m packing L29
Analytical column: 4.6-mm 15-cm; 5-m packing L29 PERFORMANCE TESTS
or 3-m packing L49 without the guard column DELIVERABLE VOLUME 698: Meets the requirements
Flow rate: 1.5 mL/min UNIFORMITY OF DOSAGE UNITS 905: It meets the
Injection size: 50 L requirements for solid packaged in single-unit containers.
System suitability
Samples: System suitability solution and Standard solution ADDITIONAL REQUIREMENTS
[NOTEThe relative retention times for azaerythromycin A PACKAGING AND STORAGE: Preserve in tight containers.
and azithromycin with the L29 column are 0.7 and 1.0, USP REFERENCE STANDARDS 11
respectively; the relative retention times for USP Azithromycin RS
azaerythromycin A and azithromycin with the L49 column
are 0.8 and 1.0, respectively.]
326 Aztreonam / Official Monographs USP 32
Aztreonam Tailing factor: NMT 2.0 for aztreonam

(Comment on this Monograph)id=m6772=Aztreonam=A- Relative standard deviation: NMT 2.0%
Monos.pdf) Analysis
Aztreonam taken:
C13H17N5 O8S2 435.43 CS = concentration of USP Aztreonam RS in the
Propanoic acid, 2-[[[1-(2-amino-4-thiazolyl)-2-[(2-methyl-4- Standard solution (mg/mL)
oxo-1-sulfo-3-azetidinyl)amino]-2- CU = concentration of aztreonam in the Sample
oxoethylidene]amino]oxy]-2-methyl-, [2S-[2,3(Z)]]-; solution (mg/mL)
(Z)-2-[[[(2-Amino-4-thiazolyl)[[(2S,3S)-2-methyl-4-oxo-1-sulfo-3- Acceptance criteria: 90.0%105.0%
azetidinyl]carbamoyl]methylene]amino]oxy]-2- IMPURITIES
methylpropionic acid [78110-38-0]. Inorganic Impurities
DEFINITION RESIDUE ON IGNITION 281: NMT 0.1%, the charred residue
Aztreonam is anhydrous or hydrated. The anhydrous form being moistened with 2 mL of nitric acid and 5 drops of
contains NLT 90.0% and NMT 105.0% of C13H17N5 O8S2, sulfuric acid
calculated on the as-is basis. The hydrated form contains NLT HEAVY METALS, Method II 231: NMT 30 ppm
92.0% and NMT 105.0% of C13H17N5O8S2, calculated on the SPECIFIC TESTS
anhydrous basis. STERILITY TESTS 71: Where the label states that Aztreonam
IDENTIFICATION is sterile, it meets the requirements for Test for Sterility of the
A. INFRARED ABSORPTION 197K: If a difference appears in Product to Be ExaminedMembrane Filtration, using Fluid A,
the IR spectra of the analyte and the standard, dissolve equal to which 23.4 g of sterile arginine/1000 mL has been added.
portions of the test specimen and the Reference Standard in WATER DETERMINATION, Method I 921: NMT 2.0%; if
equal volumes of methanol. labeled as the hydrated form: 12.0%18.0%
[NOTETo achieve a complete dissolution, it is suggested to [NOTEThe term hydrated form refers to the -form of
use about 25 mL of methanol for each 50 mg of material, Aztreonam, which is not a stoichiometric hydrate.]
and stir the mixture for 40 min at room temperature.] BACTERIAL ENDOTOXINS TEST 85: Where the label states that
Evaporate the solutions to dryness under vacuum, and dry aztreonam is sterile or must be subjected to further
at 40 for 4 h under vacuum. Perform the test on the processing during the preparation of injectable dosage
residues. forms, it contains NMT 0.17 USP Endotoxin Unit/mg of
aztreonam.
ASSAY
Buffer: 6.8 mg/mL of monobasic potassium phosphate in PACKAGING AND STORAGE: Preserve in tight containers.
water; adjust with 1 M phosphoric acid to a pH of 3.0 0.1. LABELING: Where it is intended for use in preparing
Mobile phase: Methanol and Buffer (1:4) injectable dosage forms, the label states that it is sterile or
Standard solution: 1 mg/mL of USP Aztreonam RS in must be subjected to further processing during the
Mobile phase preparation of injectable dosage forms. Where it is the
System suitability solution: 0.2 mg/mL of USP Aztreonam hydrated form, the label so indicates.
RS and 0.2 mg/mL of USP Aztreonam E-Isomer RS in Mobile USP REFERENCE STANDARDS 11
phase USP Aztreonam RS
Sample solution: 1 mg/mL of Aztreonam in Mobile phase USP Aztreonam E-Isomer RS
Chromatographic system USP Endotoxin RS
Mode: LC
Detector: UV 270 nm
Column
Aztreonam Injection
Guard: 2-mm 10-cm; packing L2 (Comment on this Monograph)id=m6773=Aztreonam
Analytical: 4.6-mm 30-cm; packing L1 Injection=A-Monos.pdf)
Flow rate: 1.5 mL/min DEFINITION
Injection size: 20 L Aztreonam Injection is a sterile solution of Aztreonam and
System suitability Arginine and a suitable osmolality adjusting substance in Water
Sample: System suitability solution for Injection. It contains NLT 90.0% and NMT 120.0% of the
[NOTEThe relative retention times for aztreonam and labeled amount of aztreonam (C13H17N5O8S2).
aztreonam E-isomer are 0.6 and 1.0, respectively.]
Resolution: NLT 2.0 between aztreonam and aztreonam The retention times of the major peaks of the Sample solution
E-isomer correspond to those of the Standard solution, as obtained in
Column efficiency: NLT 1000 theoretical plates for the Assay.
aztreonam
USP 32 Official Monographs / Aztreonam 327
ASSAY USP REFERENCE STANDARDS 11

PROCEDURE USP L-Arginine RS
Mobile phase: 1.15 g of monobasic ammonium phosphate USP Aztreonam RS
in 800 mL of water USP Open Ring Aztreonam RS
Adjust with phosphoric acid to a pH of 2.0 0.1, and dilute
with water to 1000 mL. Prepare a suitable mixture of
acetonitrile and this solution (3:1).
Standard solution: 0.2 mg/mL each of USP Aztreonam RS Aztreonam for Injection
and USP L-Arginine RS, in Mobile phase (Comment on this Monograph)id=m6781=Aztreonam for
System suitability solution: 0.2 mg/mL of each, USP Injection=A-Monos.pdf)
Aztreonam RS and USP Open Ring Aztreonam RS, in Mobile
phase DEFINITION
Sample solution: 0.2 mg/mL of aztreonam, from volume of Aztreonam for Injection is a dry mixture of sterile Aztreonam
Injection in Mobile phase and Arginine. It contains NLT 90.0% and NMT 105.0% of
Chromatographic system aztreonam (C13H17N5O8S2), calculated on the anhydrous and
(See Chromatography 621, System Suitability.) arginine-free basis. Each container contains NLT 90.0% and
Mode: LC NMT 120.0% of the labeled amount of aztreonam
Detector: UV 206 nm (C13H17N5O8S2).
Column: 4.6-mm 50-cm saturator precolumn containing
packing L27, and a 4-mm 25-cm analytical column IDENTIFICATION
containing packing L20 The retention times of the major peaks of the Sample solution
Flow rate: 1 mL/min correspond to those of the Standard solution, as obtained in
Injection size: 20 L the Assay.
Sample: System suitability solution PROCEDURE
[NOTEThe relative retention times for aztreonam and Mobile phase: 1.15 g of monobasic ammonium phosphate
open ring aztreonam are 0.8 and 1.0, respectively.] in 800 mL of water
Suitability requirements Adjust with phosphoric acid to a pH of 2.0 0.1, and dilute
Resolution: NLT 2.0 between aztreonam and open ring with water to 1000 mL. Prepare a suitable mixture of
aztreonam in the System suitability solution acetonitrile and this solution (3:1).
Column efficiency: NLT 1000 theoretical plates Standard solution: 0.2 mg/mL each of USP Aztreonam RS
determined from the aztreonam peak, System suitability and USP L-Arginine RS, in Mobile phase
solution System suitability solution: 0.2 mg/mL each of USP
Tailing factor: NMT 2.0 for the aztreonam peak, System Aztreonam RS and USP Open Ring Aztreonam RS in Mobile
suitability solution phase
Relative standard deviation: NMT 2.0%, System Sample solution A: Weigh one container of Aztreonam for
suitability solution Injection. Transfer the contents of the container to a 100-mL
Analysis volumetric flask. Weigh the empty container, and calculate
Samples: Standard solution and Sample solution the weight, in mg, of Aztreonam for Injection removed.
Calculate the percentage of C13H17N5O8S2 in the volume of Dissolve the powder in the volumetric flask in Mobile phase,
Injection taken: and dilute with Mobile phase to volume. Dilute a volume of
this solution with Mobile phase to obtain a solution having a
Result = (rU/rS) (CS/CU) 100 concentration of 0.2 mg/mL of aztreonam.
rU = peak response of the Sample solution Sample solution B: Constitute one container of Aztreonam
rS = peak response of the Standard solution for Injection with a volume of water, corresponding to the
CS = concentration of USP Aztreonam RS in the volume of solvent specified in the labeling, except where the
Standard solution (mg/mL) capacity of the container is 100 mL or greater to constitute
CU = concentration of aztreonam in the Sample with 10 mL of water. Withdraw the total withdrawable
solution (mg/mL) contents of the container, and dilute with Mobile phase to
Acceptance criteria: 90.0%120.0% obtain a solution containing 0.2 mg/mL of aztreonam.
SPECIFIC TESTS (See Chromatography 621, System Suitability.)
PYROGEN TEST 151: It meets the requirements, the test Mode: LC
dose being an accurately measured volume of Injection Detector: UV 206 nm
sufficient to provide 50 mg of aztreonam per kg. Column: 4.6-mm 50-cm saturator precolumn containing
STERILITY TESTS 71: It meets the requirements when tested packing L27, and a 4-mm 25-cm analytical column
as directed for Test for Sterility of the Product to Be Examined, containing packing L20
Membrane Filtration. Flow rate: 1 mL/min
PH 791: 4.57.5 Injection size: 20 L
PARTICULATE MATTER IN INJECTIONS 788: It meets the System suitability
requirements for small-volume injections. Sample: System suitability solution
[NOTEThe relative retention times for aztreonam and
ADDITIONAL REQUIREMENTS open ring aztreonam are about 0.8 and 1.0, respectively.]
PACKAGING AND STORAGE: Preserve as described under Suitability requirements
Injections 1, Containers for Sterile Solids. Maintain in the Resolution: NLT 2.0 between aztreonam and open ring
frozen state. aztreonam, System suitability solution
LABELING: It meets the requirements for Injections 1, Labels Column efficiency: NLT 1000 theoretical plates
and Labeling. The label states that it is to be thawed just determined from the aztreonam peak, System suitability
prior to use, describes conditions for proper storage of the solution
resultant solution, and directs that the solution is not to be
refrozen.
328 Aztreonam / Official Monographs USP 32
Tailing factor: NMT 2.0 for the aztreonam peak, System Sample solution: Use Sample solution A, prepared as
suitability solution directed in the Assay.
Relative standard deviation: NMT 2.0%, System Analysis: Proceed as directed in the Assay.
suitability solution Calculate the percentage of C6H14N4O2 in the Aztreonam for
Analysis Injection taken:
Samples: Standard solution, Sample solution A, and Sample
solution B Result = (rU/rS) (CS/CU) 100
[NOTEThe relative retention times for aztreonam and
arginine are 0.3 and 1.0, respectively.] rU = peak response from the Sample solution
Calculate the percentage of C13H17N5O8S2 in the Aztreonam rS = peak response from the Standard solution
for Injection taken: CS = concentration of USP L-Arginine RS in the
Result = 0.1(rU/rS) (CS PS/CU) CU = concentration of Aztreonam for Injection in
Sample solution A, based on the weight of
rU = peak response from Sample solution A Aztreonam for Injection removed from the
rS = peak response from the Standard solution container (mg) and the extent of dilution
CS = concentration of USP Aztreonam RS in the (mg/mL)
Standard solution (mg/mL) Use this percentage to calculate, on the anhydrous and
PS = assigned purity of USP Aztreonam RS (g/mg) arginine-free basis, the Assay result from Sample solution A,
CU = concentration of Aztreonam for Injection in obtained as directed in the Assay.
Sample solution A (mg/mL), based on the
weight of Aztreonam for Injection removed SPECIFIC TESTS
from the container (mg) and the extent of CONSTITUTED SOLUTION: At the time of use, it meets the
dilution requirements for Injections 1, Constituted Solutions.
Calculate the quantity, in mg, of C13H17N5O8S2 in the BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.17 USP
container of Aztreonam for Injection used to prepare Endotoxin Unit/mg of aztreonam.
Sample solution B: STERILITY TESTS 71: It meets the requirements when tested
as directed for Test for Sterility of the Product to be Examined,
Result = (rU/rS) (CS PS L/1000 CU) Membrane Filtration.
PH 791
rU = peak response from Sample solution B Sample solution: 100 mg/mL
rS = peak response from the Standard solution Acceptance criteria: 4.57.5
CS = concentration of USP Aztreonam RS in the WATER DETERMINATION, Method I 921: NMT 2.0%
Standard solution (mg/mL) PARTICULATE MATTER IN INJECTIONS 788: Meets the
PS = assigned purity of USP Aztreonam RS (g/mg) requirements for small-volume injections
L = labeled quantity of aztreonam in the container OTHER REQUIREMENTS: It meets the requirements for
of Aztreonam for Injection (mg) Injections 1, Labeling.
CU = concentration of aztreonam in Sample solution B
(mg/mL), on the basis of the labeled quantity PERFORMANCE TESTS
of aztreonam in the container (mg) and the UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
extent of dilution
Each container contains 90.0%120.0% of the labeled PACKAGING AND STORAGE: Preserve as described in Injections
amount of C13H17N5O8S2. 1, Containers for Sterile Solids.
OTHER COMPONENTS USP L-Arginine RS
CONTENT OF ARGININE USP Aztreonam RS
Mobile phase, Standard solution, System suitability USP Endotoxin RS
solution, and Chromatographic system: Proceed as USP Open Ring Aztreonam RS
Bacampicillin Hydrochloride Benzethonium Chloride Tincture

Bacampicillin Hydrochloride for Oral Suspension Benzocaine
Bacampicillin Hydrochloride Tablets Benzocaine Topical Aerosol
Bacitracin Benzocaine Cream
Bacitracin for Injection Benzocaine Gel
Bacitracin Ointment Benzocaine Lozenges
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Bacitracin and Polymyxin B Sulfate Topical Aerosol Benzocaine Otic Solution
Soluble Bacitracin Methylene Disalicylate Benzocaine Topical Solution
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cal Aerosol
Bacitracin Zinc
Benzocaine, Butamben, and Tetracaine Hydrochloride Gel
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Barium Sulfate Paste Benzoyl Peroxide Gel
Barium Sulfate Suspension Benzoyl Peroxide Lotion
Barium Sulfate for Suspension Benztropine Mesylate
Barium Sulfate Tablets Benztropine Mesylate Injection
BCG Live Benztropine Mesylate Tablets
BCG Vaccine Benzyl Benzoate
Beclomethasone Dipropionate Benzyl Benzoate Lotion
Belladonna Leaf Benzylpenicilloyl Polylysine Concentrate
Belladonna Extract Benzylpenicilloyl Polylysine Injection
Belladonna Extract Tablets Beta Carotene
Belladonna Tincture Beta Carotene Capsules
Benazepril Hydrochloride Betahistine Hydrochloride
Benazepril Hydrochloride Tablets Betaine Hydrochloride
Bendroflumethiazide Betamethasone
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Benzethonium Chloride Betamethasone Acetate
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Bisoprolol Fumarate and Hydrochlorothiazide Tablets
Betamethasone Valerate
Bleomycin Sulfate
Betamethasone Valerate Cream
Bleomycin for Injection
Betamethasone Valerate Lotion
Anti-A Blood Grouping Serum
Betamethasone Valerate Ointment
Anti-B Blood Grouping Serum
Betaxolol Hydrochloride
Blood Grouping Serums
Betaxolol Ophthalmic Solution
Blood Grouping Serums Anti-D, Anti-C, Anti-E, Anti-c, Anti-e
Betaxolol Tablets
Whole Blood
Bethanechol Chloride
Red Blood Cells
Bethanechol Chloride Injection
Platelets
Bethanechol Chloride Oral Solution
Botulism Antitoxin
Bethanechol Chloride Oral Suspension
Bovine Acellular Dermal Matrix
Bethanechol Chloride Tablets
Bretylium Tosylate
Bicalutamide
Bretylium Tosylate Injection
Bicalutamide Tablets
Bretylium Tosylate in Dextrose Injection
Biological Indicator for Dry-Heat Sterilization, Paper Carrier
Brinzolamide
Biological Indicator for Ethylene Oxide Sterilization, Paper
Carrier Brinzolamide Ophthalmic Suspension
Biological Indicators for Moist Heat, Dry Heat, and Gaseous Bromocriptine Mesylate
Modes of Sterilization, Liquid Spore Suspensions Bromocriptine Mesylate Capsules
Biological Indicators for Moist Heat, Dry Heat, and Gaseous Bromocriptine Mesylate Tablets
Modes of Sterilization, Nonpaper Carriers
Bromodiphenhydramine Hydrochloride
Biological Indicator for Steam Sterilization, Paper Carrier
Bromodiphenhydramine Hydrochloride Oral Solution
Biological Indicator for Steam Sterilization, Self-Contained
Bromodiphenhydramine Hydrochloride and Codeine Phos-
Biotin phate Oral Solution
Biperiden Brompheniramine Maleate
Biperiden Hydrochloride Brompheniramine Maleate Injection
Biperiden Hydrochloride Tablets Brompheniramine Maleate Oral Solution
Biperiden Lactate Injection Brompheniramine Maleate Tablets
Bisacodyl Brompheniramine Maleate and Pseudoephedrine Sulfate
Bisacodyl Suppositories Oral Solution
Bisacodyl Rectal Suspension Budesonide
Bisacodyl Delayed-Release Tablets Bumetanide
Milk of Bismuth Bumetanide Injection
Bismuth Citrate Bumetanide Tablets
Bismuth Subcarbonate Bupivacaine Hydrochloride
Bismuth Subgallate Bupivacaine Hydrochloride Injection
Bupivacaine Hydrochloride in Dextrose Injection Butalbital

Bupivacaine Hydrochloride and Epinephrine Injection Butalbital, Acetaminophen, and Caffeine Capsules
Buprenorphine Hydrochloride Butalbital, Acetaminophen, and Caffeine Tablets
Bupropion Hydrochloride Butalbital and Aspirin Tablets
Bupropion Hydrochloride Tablets Butalbital, Aspirin, and Caffeine Capsules
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Buspirone Hydrochloride Butalbital, Aspirin, Caffeine, and Codeine Phosphate
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Buspirone Hydrochloride Tablets
Butamben
Busulfan
Butoconazole Nitrate
Busulfan Tablets
Butoconazole Nitrate Vaginal Cream
Butabarbital
Butorphanol Tartrate
Butabarbital Sodium
Butorphanol Tartrate Injection
Butabarbital Sodium Oral Solution
Butorphanol Tartrate Nasal Solution
Butabarbital Sodium Tablets
USP 32 Official Monographs / Bacampicillin 1
Bacampicillin Hydrochloride
132270
Mode: LC
(Comment on this Monograph)id=m6805=Bacampicillin Detector: UV 254 nm
Hydrochloride=B-Monos.pdf) Column: 3.9-mm 15-cm; packing L1
Flow rate: 1 mL/min
System suitability
Analysis
C21H27N3O7S HCl 501.98 Samples: Standard solution and Sample solution
4-Thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, 6- Calculate the quantity, in g/mg, of the ampicillin
[(aminophenylacetyl)amino]-3,3-dimethyl-7-oxo,1- (C16H19N3O4S) equivalent in the portion of Bacampicillin
[(ethoxycarbonyl)oxyethyl ester, monohydrochloride, [2S- Hydrochloride taken:
[2,5,6(S*)]]-;
(2S,5R,6R)-6-[(R)-(2-Amino-2-phenylacetamido)]-3,3-dimethyl-7- Result = (rU/rS) (CS/CU) P (Mr1/Mr2)
oxo-4-thia-1-azabicyclo[3.2.0] heptane-2-carboxylic acid ester
with ethyl 1-hydroxyethyl carbonate, monohydrochloride rU = peak area from the Sample solution
[37661-08-8]. rS = peak area from the Standard solution
CS = concentration of USP Bacampicillin RS in the
DEFINITION Standard solution (mg/mL)
Bacampicillin Hydrochloride has a potency of NLT 623 g and CU = concentration of the Sample solution (mg/mL)
NMT 727 g of ampicillin (C16H19N3O4S) per mg. P = potency of USP Bacampicillin Hydrochloride RS
(g of ampicillin/mg of RS)
IDENTIFICATION Mr1 = molecular weight of anhydrous ampicillin,
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 349.41
Adsorbent: 0.25-mm layer of chromatographic silica gel Mr2 = molecular weight of bacampicillin hydrochloride,
mixture 501.98
Standard solution: 2 mg/mL of USP Bacampicillin Acceptance criteria: 623727 g ampicillin in each mg of
Hydrochloride RS in alcohol Bacampicillin Hydrochloride
Sample solution: 2 mg/mL in alcohol
Mode: TLC IMPURITIES
Application volume: 5 L Organic impurities
Developing solvent system: Methylene chloride, DIMETHYLANILINE 223: Meets the requirements
chloroform, and alcohol (10:1:1) Sample solution: Transfer 1.0 g of the sample to a suitable
Analysis: Apply two 5-L portions of the Sample solution 4.0 centrifuge tube, add 5.0 mL of 2 N sodium hydroxide, swirl
cm apart. After the spots dry, apply two 5-L portions of the to dissolve the specimen, add 1.0 mL of Internal Standard
Standard solution, one midway between the Sample solution Solution prepared as directed in the chapter, shake
spots and the other to one of the Sample solution spots. vigorously for 1 min, and centrifuge. Use the clear
When the solvent front has moved three-fourths of the supernatant.
length of the plate, remove the plate from the chamber,
and allow to dry. Spray the plate with a spray reagent SPECIFIC TESTS
containing 1 g of ninhydrin and 1 mL of pyridine in each PH 791: 3.04.5, in a solution 20 mg/mL
100 mL of solution in butyl alcohol, and heat at 100 for 10 WATER DETERMINATION, Method I 921: NMT 1.0%
min. ADDITIONAL REQUIREMENTS
Acceptance criteria: Bacampicillin appears as a purple spot, PACKAGING AND STORAGE: Preserve in tight containers.
and the RF values of the spots from the Sample solution and USP REFERENCE STANDARDS 11
from the combined Sample solution and Standard solution, USP Bacampicillin Hydrochloride RS
respectively, correspond to the RF value of the spot obtained
from the Standard solution.
ASSAY
PROCEDURE
Bacampicillin Hydrochloride for Oral
Solution A: 0.02 M dibasic sodium phosphate. Suspension
Adjust with 0.02 M monobasic sodium phosphate to a pH of (Comment on this Monograph)id=m6810=Bacampicillin
6.8 0.05. Hydrochloride for Oral Suspension=B-Monos.pdf)
Mobile phase: Acetonitrile and Solution A (1:1), filtered
Standard solution: 0.8 mg/mL of USP Bacampicillin DEFINITION
Hydrochloride RS Bacampicillin Hydrochloride for Oral Suspension contains an
Sample solution: 0.8 mg/mL of Bacampicillin amount of Bacampicillin Hydrochloride equivalent to NLT
Hydrochloride 90.0% and NMT 125.0% of the labeled amount of ampicillin
[NOTESonicate the Standard solution and the Sample (C16H19N3O4S) when constituted as directed. It contains one or
solution for about 20 min to achieve complete dissolution, more suitable buffers, colors, flavors, suspending agents, and
and pass through a filter of 0.5 m or finer porosity.] sweetening ingredients.
2 Bacampicillin / Official Monographs USP 32
IDENTIFICATION V = volume of constituted Sample solution taken (mL)

THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Acceptance criteria: 90.0%125.0%
mixture PERFORMANCE TESTS
Standard solution: 2 mg/mL of USP Bacampicillin UNIFORMITY OF DOSAGE UNITS 905
Hydrochloride RS in alcohol Meets the requirements for Solids packaged in single-unit
Sample solution: Equivalent to 1.4 mg/mL of ampicillin containers
from Bacampicillin Hydrochloride for Oral Suspension in DELIVERABLE VOLUME 698: Meets the requirements
alcohol
Constitute Bacampicillin Hydrochloride for Oral Suspension SPECIFIC TESTS
as directed in the labeling. Transfer a portion of the PH 791: 6.58.0, in the suspension constituted as directed
resulting suspension, equivalent to 140 mg of ampicillin, to in the labeling
a 100-mL volumetric flask, add 70 mL of alcohol, shake by LOSS ON DRYING 731: Dry about 100 mg in a capillary-
mechanical means for 30 min, and dilute with alcohol to stoppered bottle in vacuum at 60 for 3 h: it loses NMT
volume. 2.0% of its weight.
Developing solvent system: Methylene chloride, ADDITIONAL REQUIREMENTS
chloroform, and alcohol (10:1:1) PACKAGING AND STORAGE: Preserve in tight containers.
Samples: Sample solution and Standard solution USP Ampicillin RS
Apply two 5-L portions of the Sample solution 4.0 cm USP Bacampicillin Hydrochloride RS
apart. After the spots dry, apply two 5-L portions of the
Standard solution, one midway between the Sample
solution spots and the other to one of the Sample solution
spots, allow spots to dry and place the plate in the
Bacampicillin Hydrochloride Tablets
chamber. When the solvent front has moved three-fourths (Comment on this Monograph)id=m6815=Bacampicillin
of the length of the plate, remove the plate from the Hydrochloride Tablets=B-Monos.pdf)
chamber, and allow to dry. DEFINITION
Spray the plate with a spray reagent containing 1 g of Bacampicillin Hydrochloride Tablets contain the equivalent of
ninhydrin and 1 mL of pyridine in each 100 mL of NLT 90.0% and NMT 125.0% of the labeled amount of
solution in butyl alcohol, and heat at 100 for 10 min. ampicillin (C16H19N3O4S).
Acceptance criteria: Bacampicillin appears as a purple spot,
and the RF values of the spots from the Sample solution and IDENTIFICATION
from the combined Sample solution and Standard solution, THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
respectively, correspond to the RF value of the spot obtained Standard solution: 2 mg/mL of USP Bacampicillin
from the Standard solution. Hydrochloride RS in alcohol
Sample solution: 2 mg/mL of ampicillin from powdered
ASSAY Tablets diluted in alcohol
PROCEDURE Developing solvent system: Methylene chloride,
Standard solution: Using USP Ampicillin RS, prepare as chloroform, and alcohol (10:1:1)
directed for the Standard preparation under Iodometric Spray reagent: 10 mg/mL of ninhydrin in butyl alcohol
AssayAntibiotics 425. with 1 mL of pyridine in each 100 mL of solution
Sample solution: Equivalent to 0.35 mg/mL of ampicillin Chromatographic system
from Bacampicillin Hydrochloride for Oral Suspension in (See Chromatography 621.)
alcohol and 0.1 M phosphoric acid (4:1) Mode: TLC
[NOTETransfer a volume of Bacampicillin Hydrochloride Adsorbent: 0.25-mm layer of chromatographic silica gel
for Oral Suspension, constituted as directed in the labeling mixture
and free from bubbles, equivalent to about 87.5 mg of Application volume: 5 L
ampicillin, to a 250-mL volumetric flask. Add 200 mL of a Analysis
solvent mixture consisting of alcohol and 0.1 M Samples: Sample solution and Standard solution
phosphoric acid (4:1). Shake by mechanical means for 30 Apply two 5-L portions of the Sample solution 4.0 cm
min. Centrifuge a portion of the resulting suspension. apart on a suitable thin-layer chromatographic plate. After
Pipet 4.0 mL of the clear solution so obtained into each of the spots dry, apply two 5-L portions of the Standard
two glass-stoppered, 125-mL conical flasks.] solution, one midway between the Sample solution spots
Analysis: Proceed as directed under Iodometric Assay and the other to one of the Sample solution spots.
Antibiotics 425, Procedure. Develop the chromatogram in the Developing solvent
Calculate the quantity, in mg, of C16H19N3O4S in each mL of system until the solvent front has moved three-fourths the
the constituted Bacampicillin Hydrochloride for Oral length of the plate. Spray the plate with the Spray
Suspension taken: reagent, and heat at 100 for 10 min.
Result = (0.0625 F)(B I)/V Acceptance criteria: Bacampicillin appears as a purple
spot, and the RF values of the spots from the Sample
F = microgram (or unit) equivalent solution and from the combined Sample solution and
B = volume of 0.01 N sodium thiosulfate consumed Standard solution, respectively, correspond to the RF value
in the Blank determination (mL) of the spot obtained from the Standard solution.
I = volume of 0.01 N sodium thiosulfate consumed
in the Inactivation and Titration (mL)
USP 32 Official Monographs / Bacitracin 3
ASSAY Bacitracin
PROCEDURE (Comment on this Monograph)id=m6830=Bacitracin=B-
Solution A: To 0.02 M dibasic sodium phosphate, add Monos.pdf)
portions of 0.02 M monobasic sodium phosphate, until a pH
6.8 0.05 is reached.
Mobile phase: Acetonitrile and Solution A (1:1)
Standard solution: 0.8 mg/mL of USP Bacampicillin
Hydrochloride RS. [NOTESonicate for 20 min to achieve
complete dissolution. Pass through a filter of 0.5-m or finer
porosity.]
Sample solution: Transfer a portion of finely powdered
Tablets (NLT 20 Tablets), equivalent to 56 mg of Bacitracin [1405-87-4].
C16H19N3O4S to a 100-mL volumetric flask, add 90 mL of
water, and sonicate for 20 min. Dilute to volume with DEFINITION
water, and pass through a filter of 0.5-m or finer porosity. Bacitracin is a polypeptide produced by the growth of an
Chromatographic system organism of the licheniformis group of Bacillus subtilis (Fam.
(See Chromatography 621, System Suitability.) Bacillacaea). It has a potency of NLT 40 Bacitracin Units/mg.
Mode: LC
Column: 3.9-mm 15-cm; packing L1 THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
Flow rate: 1 mL/min 201BNP: Meets the requirements
System suitability ANTIBIOTICSMICROBIAL ASSAYS 81: Proceed as directed in
Sample: Standard solution the chapter for Bacitracin.
Suitability requirements Acceptance criteria: NLT 40 Bacitracin Units/mg
Relative standard deviation: NMT 2.0% SPECIFIC TESTS
Analysis PH 791: 5.57.5, in a solution containing 10,000
Samples: Standard solution and Sample solution Bacitracin Units/mL
Calculate the percentage of C16H19N3O4S in the portion of LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered
Tablets taken: bottle in vacuum at a pressure not exceeding 5 mm of
mercury at 60 for 3 h: it loses NMT 5.0% of its weight.
Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 STERILITY TESTS 71: Where the label states that the
Bacitracin is sterile, it meets the requirements.
rU = peak response from the Sample solution BACTERIAL ENDOTOXINS TEST 85: Where the label states that
rS = peak response from the Standard solution Bacitracin is sterile or must be subjected to further
CS = concentration of USP Bacampicillin processing during the preparation of injectable dosage
Hydrochloride RS in the Standard solution forms, it meets the requirements.
(mg/mL)
CU = nominal concentration of the Sample solution ADDITIONAL REQUIREMENTS
(mg/mL) PACKAGING AND STORAGE: Preserve in tight containers, and
Mr1 = molecular weight of anhydrous ampicillin, store in a cool place.
349.41 LABELING: Where it is packaged for prescription
Mr2 = molecular weight of bacampicillin hydrochloride, compounding, label it to indicate that it is not sterile and
501.99 that the potency cannot be assured for longer than 60 days
Acceptance criteria: 90.0%125.0% after opening, and to state the number of Bacitracin Units
per milligram. Where it is intended for use in preparing
PERFORMANCE TESTS injectable or other sterile dosage forms, the label states that
DISSOLUTION 711 it is sterile or must be subjected to further processing during
Medium: Water; 900 mL the preparation of injectable or other sterile dosage forms.
Time: 30 min USP Bacitracin Zinc RS
Standard solution: 0.3 mg/mL of USP Ampicillin RS in USP Endotoxin RS
water
Analysis: Determine the amounts of C16H19N3O4S dissolved
as directed in AntibioticsHydroxylamine Assay, Automated
Methods of Analysis 16, Procedure. Bacitracin for Injection
(Comment on this Monograph)id=m6840=Bacitracin for
C16H19N3O4S is dissolved.
Injection=B-Monos.pdf)
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
DEFINITION
SPECIFIC TESTS
Bacitracin for Injection has a potency of NLT 50 Bacitracin Units
/mg. It contains NLT 90.0% and NMT 115.0% of the labeled
ADDITIONAL REQUIREMENTS amount of bacitracin.
USP Ampicillin RS
USP Bacampicillin Hydrochloride RS
4 Bacitracin / Official Monographs USP 32
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
201BNP: Meets the requirements 201BNP: Meets the requirements
ASSAY ASSAY
ANTIBIOTICSMICROBIAL ASSAYS 81 ANTIBIOTICSMICROBIAL ASSAYS 81
Sample solution 1: Constitute one container of Bacitracin Sample: Use a portion of Ointment shaken with about 50
for Injection as directed in the labeling. Using a suitable mL of ether in a separator, and extracted with four 20-mL
hypodermic needle and syringe, withdraw the contents of portions of Buffer No. 1.
the container, and dilute quantitatively with Buffer No. 1 to Combine the buffer extracts, and dilute with Buffer No. 1 to
obtain a solution containing about 100 Bacitracin Units/mL. an appropriate volume to obtain a stock solution. Add
Sample solution 2 (where the label states the number of sufficient 0.01 N hydrochloric acid to a measured portion
Bacitracin Units in a given volume of constituted solution): of the stock solution so that the amount of hydrochloric
Constitute one container of Bacitracin for Injection as acid in the Test Dilution will be the same as in the median
directed in the labeling. Dilute a volume of the constituted dose level of the Standard, and quantitatively dilute with
solution with Buffer No. 1 to obtain a solution containing Test Dilution having a bacitracin concentration assumed to
100 Bacitracin Units/mL. be equal to the median dose level of the Standard.
Samples: Sample solution 1 and Sample solution 2
Proceed as directed. Add sufficient 0.01 N hydrochloric acid SPECIFIC TESTS
to the Sample solution so that the amount of hydrochloric WATER DETERMINATION, Method I 921: NMT 0.5%
acid in the Test Dilution will be the same as in the median [NOTEUse 20 mL of a mixture of toluene and methanol
dose level of the Standard, and dilute with Buffer No. 1 to (7:3) in place of methanol in the titration vessel.]
obtain a Test Dilution having a bacitracin concentration MINIMUM FILL 755: Meets the requirements
assumed to be equal to the median dose level of the
Standard. ADDITIONAL REQUIREMENTS
Acceptance criteria: 90.0%115.0% PACKAGING AND STORAGE: Preserve in well-closed containers
containing NMT 60 g, unless labeled solely for hospital use,
PERFORMANCE TESTS preferably at controlled room temperature.
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements USP REFERENCE STANDARDS 11
USP Bacitracin Zinc RS
IMPURITIES
being moistened with 2 mL of nitric acid and 5 drops of Bacitracin Ophthalmic Ointment
sulfuric acid (Comment on this Monograph)id=m6870=Bacitracin
HEAVY METALS, Method II 231: NMT 30 ppm Ophthalmic Ointment=B-Monos.pdf)
SPECIFIC TESTS DEFINITION
CONSTITUTED SOLUTION: At the time of use, it meets the Bacitracin Ophthalmic Ointment is a sterile preparation of
requirements under Injections 1. Bacitracin in an anhydrous ointment base. It contains NLT
PH 791: 5.57.5, 10,000 Bacitracin Units/mL 90.0% and NMT 140.0% of the labeled amount of bacitracin.
LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered
bottle in vacuum at a pressure of NMT 5 mm of mercury at IDENTIFICATION
60 for 3 h: it loses NMT 5.0% of its weight. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
INJECTIONS 1: Meets the requirements 201BNP: Meets the requirements
as directed under Test for Sterility of the Product to Be ASSAY
Examined, Membrane Filtration. ANTIBIOTICSMICROBIAL ASSAYS 81
BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.01 USP Sample: Use a portion of Ophthalmic Ointment shaken
Endotoxin Unit/Bacitracin Unit. with 50 mL of ether in a separator, and extracted with four
20-mL portions of Buffer No. 1. Combine the buffer
ADDITIONAL REQUIREMENTS extracts, and dilute with Buffer No. 1 to an appropriate
PACKAGING AND STORAGE: Preserve as described under volume to obtain a stock solution. Add sufficient 0.01 N
Injections 1, Containers for Sterile Solids, and store in a cool hydrochloric acid to a portion of the stock solution so that
place. the amount of hydrochloric acid in the Test Dilution will be
USP REFERENCE STANDARDS 11 the same as in the median dose level of the Standard, and
USP Bacitracin Zinc RS quantitatively dilute with Test Dilution having a bacitracin
USP Endotoxin RS concentration assumed to be equal to the median dose level
of the Standard.
Bacitracin Ointment SPECIFIC TESTS
(Comment on this Monograph)id=m6860=Bacitracin WATER DETERMINATION, Method I 921: NMT 0.5%
Ointment=B-Monos.pdf) [NOTEUse 20 mL of a mixture of toluene and methanol
(7:3) in place of methanol in the titration vessel.]
DEFINITION METAL PARTICLES IN OPTHALMIC OINTMENTS 751: Meets the
Bacitracin Ointment is Bacitracin in an anhydrous ointment requirements
base. It contains NLT 90.0% and NMT 140.0% of the labeled
amount of bacitracin. It may contain a suitable anesthetic.
STERILITY TESTS 71: Meets the requirements toluene and methanol (7:3) in place of methanol in the
titration vessel.]
ADDITIONAL REQUIREMENTS OTHER REQUIREMENTS
PACKAGING AND STORAGE: Preserve in collapsible ophthalmic It meets the requirements for Aerosols, Nasal Sprays,
ointment tubes. Metered-Dose Inhalers, and Dry Powder Inhalers 601,
USP REFERENCE STANDARDS 11 Pressure Test, Minimum Fill, and Leakage Test.
PACKAGING AND STORAGE: Preserve in pressurized containers,
and avoid exposure to excessive heat.
Bacitracin and Polymyxin B Sulfate USP REFERENCE STANDARDS 11
Topical Aerosol USP Bacitracin Zinc RS
(Comment on this Monograph)id=m6885=Bacitracin and USP Polymyxin B Sulfate RS
Polymyxin B Sulfate Topical Aerosol=B-Monos.pdf)
DEFINITION
Bacitracin and Polymyxin B Sulfate Topical Aerosol is a Soluble Bacitracin Methylene
suspension of Bacitracin and Polymyxin B Sulfate in a suitable Disalicylate
vehicle, packaged in a pressurized container with a suitable (Comment on this Monograph)id=m6890=Soluble Bacitracin
inert propellant. It contains NLT 90.0% and NMT 130.0% of Methylene Disalicylate=B-Monos.pdf)
the labeled amounts of bacitracin and polymyxin B. It may
contain a suitable local anesthetic. DEFINITION
[NOTEPrepare the specimen for the following tests and assays Soluble Bacitracin Methylene Disalicylate is a mixture of
as follows. Maintain the container in the inverted position Bacitracin Methylene Disalicylate and Sodium Bicarbonate. It
throughout this procedure. Store the container in a freezer at has a potency of NLT 8 Bacitracin Units/mg, calculated on the
70 for 16 to 24 h. Remove the container from the freezer, dried basis.
promptly puncture the container, and allow the propellant to
volatilize. Open the container, and mix the contents.] ASSAY
ANTIBIOTICSMICROBIAL ASSAYS 81
IDENTIFICATION Sample stock solution: Transfer a sample to a high-speed
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST glass blender jar, add 99.0 mL of a 20 mg/mL sodium
201BNP: A portion of the contents of one container bicarbonate solution and 1.0 mL of polysorbate 80, and
prepared as directed above and tested as directed under For blend for 3 min. Add a sufficient volume of 0.01 N
Creams, Lotions, and Ointments meets the requirements. hydrochloric acid so that the amount of hydrochloric acid
will be the same as in the median dose level of the
ASSAY Standard.
BACITRACIN Sample solution: Dilute the Sample stock solution with
(See AntibioticsMicrobial Assays 81.) Buffer No. 1 (see Phosphate Buffers and Other Solutions) to
Sample: A portion of the contents of one container, obtain a concentration of bacitracin assumed to be equal to
prepared as directed, equivalent to about 500 USP Bacitracin the median dose level of the Standard.
Units Analysis: Proceed as directed for Bacitracin in Antibiotics
Analysis: Transfer to a suitable separator containing 50 mL Microbial Assays 81.
of ether, and extract with three 25-mL portions of Buffer No. Acceptance criteria: NLT 8 Bacitracin Units/mg
1. Combine the buffer extracts in a 100-mL volumetric flask,
dilute with Buffer No. 1 to volume, and mix. Add sufficient SPECIFIC TESTS
0.01 N hydrochloric acid to this solution so that the amount PH 791: 8.09.5, in a solution of 25 mg/mL
of hydrochloric acid in the Test solution will be the same as LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered
in the median dose level of the Standard, and quantitatively bottle in vacuum at a pressure not exceeding 5 mm of
dilute with Buffer No. 1 to obtain a Test Dilution having a mercury at 60 for 3 h: it loses NMT 8.5% of its weight.
bacitracin concentration assumed to be equal to the median
dose level of the Standard. ADDITIONAL REQUIREMENTS
Acceptance criteria: 90.0%130.0% PACKAGING AND STORAGE: Preserve in well-closed containers.
POLYMYXIN B LABELING: Label it to indicate that it is for veterinary use
(See AntibioticsMicrobial Assays 81.) only.
Sample: A portion of the contents of one container, USP REFERENCE STANDARDS 11
prepared as directed, equivalent to 5000 USP Polymyxin B USP Bacitracin Zinc RS
Units
Analysis: Transfer to a suitable separator containing 50 mL
of ether, and extract with three 25-mL portions of Buffer No. Bacitracin Methylene Disalicylate
6. Combine the buffer extracts in a 100-mL volumetric flask,
dilute with Buffer No. 6 to volume, and mix. Dilute this Soluble Powder
solution, quantitatively and stepwise, with Buffer No. 6 to (Comment on this Monograph)id=m6894=Bacitracin Methylene
obtain a Test Dilution having a polymyxin B concentration Disalicylate Soluble Powder=B-Monos.pdf)
assumed to be equal to the median dose level of the
Standard. DEFINITION
Acceptance criteria: 90.0%130.0% Bacitracin Methylene Disalicylate Soluble Powder contains NLT
90.0% and NMT 120.0% of the labeled amount of bacitracin.
SPECIFIC TESTS
[NOTEUse a portion of the contents of one container,
prepared as directed above, and 20 mL of a mixture of
ASSAY Standard stock solution: Equivalent to 10 mg/mL of zinc

ANTIBIOTICSMICROBIAL ASSAYS 81 (from zinc oxide) in 1 N hydrochloric acid [NOTEWarm to
Sample stock solution: Transfer a sample to a high-speed dissolve then allow to cool before diluting to volume.]
glass blender jar. Add 99.0 mL of sodium bicarbonate Standard solutions: 0.5, 1.5, and 2.5 g/mL of zinc in
solution (1 in 50) and 1.0 mL of polysorbate 80, and blend 0.001 N hydrochloric acid, from Standard stock solution
for about 3 min. Add a sufficient volume of 0.01 N Sample stock solution: 2 mg/mL of Bacitracin Zinc in 0.01
hydrochloric acid to a measured volume of Sample solution N hydrochloric acid
so that the amount of hydrochloric acid will be the same as Sample solution: 0.02 mg/mL in 0.001 N hydrochloric
in the median dose level of the Standard. acid, from Sample stock solution
Sample solution: Dilute the Sample stock solution with Blank: 0.001 N hydrochloric acid
Buffer No. 1 (see Phosphate Buffers and Other Solutions) to Spectrometric conditions
obtain a concentration of bacitracin assumed to be equal to (See Spectrophotometry and Light-Scattering 851).
the median dose level of the Standard. Mode: Atomic absorption spectrophotometry
Detector: Zinc hollow-cathode lamp and an airacetylene
SPECIFIC TESTS flame
PH 791: 8.09.5, 50 mg/mL Analytical wavelength: Zinc resonance line of 213.8 nm
LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered Analysis
bottle in vacuum at a pressure not exceeding 5 mm of Samples: Standard solutions, Sample solution, and Blank.
mercury at 60 for 3 h: it loses NMT 8.5% of its weight. Plot the absorbances of the Standard solutions versus
concentration, in g/mL, of zinc, and draw the straight
ADDITIONAL REQUIREMENTS line best fitting the three plotted points and determine
PACKAGING AND STORAGE: Preserve in tight containers. the concentration, in g/mL, of zinc in the Sample
LABELING: Label it to indicate that it is for veterinary use solution.
only. Label it to state the content of bacitracin in terms of Calculate the content of zinc, as a percentage, in the
grams per pound, each gram of bacitracin being equivalent portion of Bacitracin Zinc taken:
to 42,000 Bacitracin Units.
USP REFERENCE STANDARDS 11 Result = C/Cu 100
C = concentration of zinc in the Sample solution
obtained from the curve (g/mL)
Bacitracin Zinc Cu = concentration of Bacitracin Zinc in the Sample
solution (g/mL)
(Comment on this Monograph)id=m6910=Bacitracin Zinc=B- Acceptance criteria: 4.0%-6.0% of Zinc, calculated on the
Monos.pdf) dried basis
Bacitracins zinc complex [1405-89-6]. COMPOSITION
Solution A: Dissolve 34.8 g of potassium phosphate,
DEFINITION dibasic, in 1 L of water. Adjust with 27.2 g of potassium
Bacitracin Zinc is the zinc complex of bacitracin, which consists phosphate, monobasic, dissolved in 1 L of water, to a pH of
of a mixture of antimicrobial polypeptides, the main 6.0.
components being bacitracins A, B1, B2, and B3. It has a Mobile phase: Mixture of methanol, acetonitrile, Solution A,
potency of NLT 65 Bacitracin Units/mg, calculated on the and water (26:2:5:15)
dried basis. It contains NLT 4.0% and NMT 6.0% of zinc (Zn), Mix well, and degas.
calculated on the dried basis. Diluent: Dissolve 40 g of sodium edentate in 1 L of water.
Adjust with dilute sodium hydroxide to a pH of 7.0.
IDENTIFICATION System suitability solution: 2.0 mg/mL of USP Bacitracin
A. Thin-Layer Chromatographic Identification Test Zinc RS in Diluent
201BNP: Meets the requirements Reporting threshold solution: Dilute quantitatively, with
B. Meets the requirements of the liquid chromatographic water, a suitable volume of the System suitability solution to
procedure in the test for Composition obtain a solution with a known concentration of 0.01
mg/mL.
ASSAY Peak identification solution: Dissolve a quantity of USP
ANTIBIOTICSMICROBIAL ASSAYS 81 : NLT 65 Bacitracin Bacitracin Zinc RS in a suitable volume of Diluent to obtain a
Units/mg, calculated on the dried basis 2.0 mg/mL solution. Heat in a boiling water bath for 30
SPECIFIC TESTS min. Cool to room temperature.
PH 791: 6.07.5, in a (saturated) solution 100 mg/mL Sample solution: 2.0 mg/mL of Bacitracin Zinc in Diluent
LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered Chromatographic system
bottle in vacuum at 60 for 3 h: it loses NMT 5.0% of its (See Chromatography 621, System Suitability.)
weight. Mode: LC
STERILITY TESTS 71: Where the label states that it is sterile, Detector: UV 254 nm
it meets the requirements when tested as directed under Test Column: 4.6-mm 250-mm; 5-m packing L1
for Sterility of the Product to Be Examined, Membrane Filtration, Flow rate: 1 mL/min
except to add 20g of edetate disodium to Fluid A. Injection size: 100 L
ZINC CONTENT System suitability
[NOTEThe Standard solutions and the Sample solution may Sample: Peak identification solution and System suitability
be quantitatively diluted with 0.001 N hydrochloric acid, if solution
necessary, to obtain solutions of suitable concentrations,
adaptable to the linear or working range of the
instrument.]
Suitability requirements Acceptance criteria: The sum of bacitracin A, B1, B2,

Peak-to-valley ratio: NLT 1.2 and B3 is NLT 70.0% of the Total area.
Limit of Early Eluting Peptides
Impurity Table 1 Calculate the percentage of all peaks eluting before the
peak due to bacitracin B1:
Relative
Retention Result = (rPreB1/Total area) 100
Time
Name (approximate) rPreB1 = sum of the responses of all peaks eluting before the
Bacitracin C1 0.5 peak for bacitracin B1
Acceptance criteria: The limit of early eluting peptides is
Bacitracin C2 0.6 NMT 20.0%.
Bacitracin C3 0.6 Limit of Bacitracin F
Bacitracin B1 0.7 Calculate the percentage of bacitracin F:
Bacitracin B2 0.7 Result = (rF/rA) 100
Bacitracin B3 0.8
rF = response of bacitracin F from the Sample solution
Bacitracin A 1.0
rA = response of bacitracin A from the Sample solution
Bacitracin F 2.4 Acceptance criteria: The limit of bacitracin F, a known
impurity, is NMT 6.0%.
Chromatograph the Peak identification solution (detector set
at 300 nm) and then the System suitability solution ADDITIONAL REQUIREMENTS
(detector set at 254 nm). Identify the location of bacitracin PACKAGING AND STORAGE: Preserve in tight containers, and
F, which is a known impurity, using the relative retention store in a cool place.
time shown in Impurity Table 1. Identify the peaks of the LABELING: Label it to indicate that it is to be used in the
most active components of bacitracin (bacitracins A, B1, manufacture of nonparenteral drugs only. Where it is
B2, and B3), early eluting peptides (those eluting before packaged for prescription compounding, label it to indicate
the peak due to bacitracin B1), and the impurity, bacitracin that it is not sterile and that the potency cannot be assured
F, using the relative retention values in Impurity Table 1. for longer than 60 days after opening, and to state the
Calculate the peak-to-valley ratio: number of Bacitracin Units/milligram. Where it is intended
for use in preparing sterile dosage forms, the label states
Result = HP/HV that it is sterile or must be subjected to further processing
during the preparation of sterile dosage forms.
HP = height above the baseline of the peak due to USP REFERENCE STANDARDS 11
bacitracin B1 USP Bacitracin Zinc RS
HV = height above the baseline of the lowest point of
the curve separating the bacitracin B1 peak
from the peak due to bacitracin B2
Analysis Bacitracin Zinc Ointment
Sample: Diluent, Sample solution, and Reporting threshold (Comment on this Monograph)id=m6915=Bacitracin Zinc
solution. Ointment=B-Monos.pdf)
Record the chromatograms for about three times the
retention time of bacitracin A. Identify the peaks using the DEFINITION
relative retention times shown in Impurity Table 1. Bacitracin Zinc Ointment is Bacitracin Zinc in an anhydrous
[NOTEDisregard any peak in the Sample solution having ointment base. It contains NLT 90.0% and NMT 140.0% of
an area less than the area of the bacitracin A peak in the the labeled amount of bacitracin.
Reporting threshold solution; disregard any peak observed
in the Diluent.] IDENTIFICATION
[NOTEThe total area in the following calculations is THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
defined as the area of all peaks except the reporting 201BNP: Meets the requirements
threshold.] ASSAY
Content of Bacitracin A PROCEDURE
Calculate the percentage of bacitracin A: (See AntibioticsMicrobial Assays 81.)
Result = (rA/Total area) 100 Standard solution: For each test dilution of the Standard,
add additional hydrochloric acid to obtain the same
rA = area response from bacitracin A concentration of hydrochloric acid as in the Sample solution.
Acceptance criteria: Bacitracin A content is NLT 40.0% Sample stock solution: Weigh a portion of Ointment and
of the Total area. shake with about 50 mL of ether in a separator. Extract with
Content of Active Bacitracin four 20-mL portions of 0.01 N hydrochloric acid. Combine
Calculate the percentage of active bacitracin (bacitracin A, the acid extracts, and dilute with 0.01 N hydrochloric acid
B1, B2, and B3): to an appropriate volume.
Sample solution: Dilute the Sample stock solution
Result = (rS/Total area) 100 quantitatively and stepwise with Buffer No. 1 to obtain a
concentration assumed to be equal to the median dose level
rS = rA, rB1, rB2, and rB3 area responses from bacitracin of the Standard solution.
A, B1, B2, and B3, respectively
Analysis: Proceed as directed. best fitting the three plotted points. From the graph so
Acceptance criteria: NLT 90.0% and NMT 140.0% obtained, determine the concentration, in g/mL, of zinc in
the Sample solution.
SPECIFIC TESTS Calculate the zinc content, in g, in relation to each 42,000
WATER DETERMINATION, Method I 921 Bacitracin Units in the specimen taken:
Analysis: Proceed as directed, except use 20 mL of a
mixture of toluene and methanol (7:3) in place of methanol Result = 280,000C/(W A)
in the titration vessel.
Acceptance criteria: NMT 0.5% C = concentration of zinc in the Sample solution
MINIMUM FILL 755: Meets the requirements (g/mL)
W = weight of the Powder taken (mg)
ADDITIONAL REQUIREMENTS A = bacitracin content in the Powder (Bacitracin
PACKAGING AND STORAGE: Preserve in well-closed containers Units/g)
containing NMT 60 g, unless labeled solely for hospital use, Acceptance criteria: NMT 2.0 g for each 42,000 Bacitracin
preferably at controlled room temperature. Units
USP Bacitracin Zinc RS SPECIFIC TESTS
LOSS ON DRYING 731
Sample: 100 mg
Analysis: Dry the Sample in a vacuum at a pressure not
Bacitracin Zinc Soluble Powder exceeding 5 mm of mercury at 60 for 3 h.
(Comment on this Monograph)id=m6917=Bacitracin Zinc Acceptance criteria: The sample loses NMT 5.0% of its
Soluble Powder=B-Monos.pdf) weight.
DEFINITION ADDITIONAL REQUIREMENTS
Bacitracin Zinc Soluble Powder is a mixture of Bacitracin Zinc PACKAGING AND STORAGE: Preserve in tight containers.
and zinc proteinates. It contains NLT 90.0% and NMT 120.0% LABELING: Label it to indicate that it is for veterinary use
of the labeled amount of bacitracin. only. Label it to state the content of bacitracin in terms of
g/lb, each g of bacitracin being equivalent to 42,000
ASSAY Bacitracin Units.
(See AntibioticsMicrobial Assays 81.) USP Bacitracin Zinc RS
Standard solution: For each test dilution of the Standard,
add additional hydrochloric acid to each to obtain the same
concentration of hydrochloric acid as in the Sample solution.
Sample stock solution: Equivalent to 100 Bacitracin Bacitracin Zinc and Polymyxin B
Units/mL in 0.01 N hydrochloric acid Sulfate Ointment
Sample solution: Dilute the Sample stock solution stepwise (Comment on this Monograph)id=m6926=Bacitracin Zinc and
with Buffer No. 1 to obtain a concentration assumed to be Polymyxin B Sulfate Ointment=B-Monos.pdf)
equal to the median dose level of the Standard solution.
Analysis: Proceed as directed. DEFINITION
Acceptance criteria: NLT 90.0% and NMT 120.0% Bacitracin Zinc and Polymyxin B Sulfate Ointment contains the
equivalent of NLT 90.0% and NMT 130.0% of the labeled
OTHER COMPONENTS amounts of bacitracin and polymyxin B. It may contain a
ZINC CONTENT suitable local anesthetic.
[NOTEThe Standard solutions and the Sample solution may
be diluted with 0.001 N hydrochloric acid, if necessary, to IDENTIFICATION
obtain solutions of suitable concentrations, adaptable to THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
the linear or working range of the instrument.] 201BNP: Meets the requirements
Standard stock solution: Transfer 3.11 g of zinc oxide to a
250-mL volumetric flask, add 80 mL of 1 N hydrochloric ASSAY
acid, warm to dissolve, cool, and dilute with water to BACITRACIN
volume. This solution contains 10 mg/mL of zinc. (See AntibioticsMicrobial Assays 81.)
Standard solutions: 0.5 g/mL, 1.5 g/mL, and 2.5 g/mL Analysis: Shake a portion of Ointment with about 50 mL of
of zinc in 0.001 N hydrochloric acid: from the Standard stock ether in a separator, and extract with four 20-mL portions of
solution 0.01 N hydrochloric acid. Combine the acid extracts, and
Sample solution: Transfer an amount of Powder equivalent dilute with 0.01 N hydrochloric acid to an appropriate
to 200 mg of Bacitracin Zinc to a 100-mL volumetric flask. volume to obtain a Sample solution. Dilute this Sample
Dissolve in and dilute with 0.01 N hydrochloric acid to solution quantitatively and stepwise with Buffer No. 1 to
volume. Dilute 2 mL of this solution with 0.001 N obtain a Test Dilution having a concentration assumed to be
hydrochloric acid to 200 mL. equal to the median dose level of the Standard, adding
Blank: 0.001 N hydrochloric acid additional hydrochloric acid to each Dilute solution of the
Spectrometric conditions Standard to obtain the same concentration of hydrochloric
(See Spectrophotometry and Light-Scattering 851.) acid as in the Test Dilution.
Mode: Atomic absorption spectrophotometry POLYMYXIN B
Detector: Zinc hollow-cathode lamp and an air-acetylene (See AntibioticsMicrobial Assays 81.)
flame Analysis: Transfer a portion of Ointment to a suitable
Analytical wavelength: Zinc resonance line, 213.8 nm separator containing 50 mL of ether, and extract with four
Analysis: 20-mL portions of Buffer No. 6. Combine the aqueous
Sample: Standard solutions, Sample solution, and Blank. extracts, and dilute with Buffer No. 6 to an appropriate
Plot the absorbances of the Standard solutions versus volume to obtain a Sample solution. Dilute this Sample
concentration, in g/mL of zinc, and draw the straight line solution quantitatively and stepwise with Buffer No. 6 to
USP 32 Official Monographs / Baclofen 9
obtain a Test Dilution having a concentration assumed to be STERILITY TESTS 71

equal to the median dose level of the Standard. Analysis: Test as directed for Test for Sterility of the Product
Acceptance criteria: 90.0%130.0% to Be Examined, Membrane Filtration.
Acceptance criteria: Meets the requirements
SPECIFIC TESTS
WATER DETERMINATION, Method I 921: NMT 0.5%, 20 mL ADDITIONAL REQUIREMENTS
of a mixture of toluene and methanol (7:3) being used in PACKAGING AND STORAGE: Preserve in collapsible ophthalmic
place of methanol in the titration vessel ointment tubes.
MINIMUM FILL 755: Meets the requirements USP REFERENCE STANDARDS 11
ADDITIONAL REQUIREMENTS USP Polymyxin B Sulfate RS
USP Bacitracin Zinc RS Baclofen
USP Polymyxin B Sulfate RS (Comment on this Monograph)id=m6940=Baclofen=B-
Monos.pdf)
Bacitracin Zinc and Polymyxin B

Sulfate Ophthalmic Ointment
(Comment on this Monograph)id=m6927=Bacitracin Zinc and
Polymyxin B Sulfate Ophthalmic Ointment=B-Monos.pdf)
DEFINITION C10H12ClNO2 213.66
Bacitracin Zinc and Polymyxin B Sulfate Ophthalmic Ointment Butanoic acid, 4-amino-3-(4-chlorophenyl)-;
contains the equivalent of NLT 90.0% and NMT 130.0% of -(Aminomethyl)-p-chlorohydrocinnamic acid [1134-47-0].
the labeled amounts of bacitracin and polymyxin B.
DEFINITION
IDENTIFICATION Baclofen contains NLT 99.0% and NMT 101.0% of
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST C10H12ClNO2, calculated on the anhydrous basis.
201BNP: Meets the requirements
IDENTIFICATION
ASSAY
BACITRACIN INFRARED ABSORPTION 197M
(See AntibioticsMicrobial Assays 81.) ASSAY
Standard solution: For each test dilution of the Standard, PROCEDURE
add additional hydrochloric acid to each to obtain the same Sample solution: Dissolve 200 mg of Baclofen in a suitable
concentration of hydrochloric acid as in the Sample solution. volume of glacial acetic acid, sufficient to immerse the
Sample stock solution: Weigh a portion of Ophthalmic electrodes.
Ointment and shake with about 50 mL of ether in a Analysis: Titrate the Sample solution with 0.1 N perchloric
separator. Extract with four 20-mL portions of 0.01 N acid VS, using a calomel electrode containing a saturated
hydrochloric acid. Combine the acid extracts, and dilute solution of lithium chloride in glacial acetic acid. Perform a
with 0.01 N hydrochloric acid to an appropriate volume. blank determination, and make any necessary correction
Sample solution: Dilute the Sample stock solution with (see Titrimetry 541). Each mL of 0.1 N perchloric acid is
Buffer No. 1 to obtain a concentration assumed to be equal equivalent to 21.37 mg of C10H12ClNO2.
to the median dose level of the Standard solution. Acceptance criteria: 99.0%101.0%
Analysis: Proceed as directed.
Acceptance criteria: NLT 90.0% and NMT 130.0% IMPURITIES
POLYMYXIN B Inorganic Impurities
(See AntibioticsMicrobial Assays 81.) RESIDUE ON IGNITION 281: NMT 0.3%
Sample stock solution: Weigh a portion of Ophthalmic HEAVY METALS, Method II 231: NMT 10 ppm
Ointment and shake with 50 mL of ether in a separator. Organic Impurities
Extract with four 20-mL portions of Buffer No. 6. Combine PROCEDURE
the aqueous extracts, and dilute with Buffer No. 6 to an [NOTEAvoid contact with o-tolidine when performing this
appropriate volume. test, and conduct the test in a well-ventilated hood.]
Sample solution: Dilute the Sample stock solution with Diluent: Alcohol and glacial acetic acid (4:1)
Buffer No. 6 to obtain a concentration assumed to be equal Standard stock solution: 0.1 mg/mL of USP Baclofen RS
to the median dose level of the Standard. in Diluent
Analysis: Proceed as directed. Standard solution A: 0.05 mg/mL of USP Baclofen RS in
Acceptance criteria: NLT 90.0% and NMT 130.0% Diluent, from the Standard stock solution
Standard solution B: 0.03 mg/mL of USP Baclofen RS in
SPECIFIC TESTS Diluent, from the Standard stock solution
WATER DETERMINATION, Method I 921 Identification solution: 0.1 mg/mL USP Baclofen Related
Analysis: Proceed as directed, except use 20 mL of a Compound A RS in Diluent
mixture of toluene and methanol (7:3) in place of methanol Sample solution: 10 mg/mL of Baclofen in Diluent
in the titration vessel. Chromatographic system
Acceptance criteria: NMT 0.5% (See Chromatography 621, Thin-Layer Chromatography.)
MINIMUM FILL 755: Meets the requirements Adsorbent: 0.25-mm layer of chromatographic silica gel
METAL PARTICLES IN OPHTHALMIC OINTMENTS 751: Meets mixture
the requirements
10 Baclofen / Official Monographs USP 32
Application volume: 10 L Adjust with phosphoric acid to a pH of 3.5.

Developing solvent system: Butyl alcohol, glacial acetic Standard stock solution: 1.0 mg/mL of USP Baclofen RS in
acid, and water (4:1:1) water
Spray reagent: Dissolve 200 mg of o-tolidine in 2 mL of Standard solution: 5 g/mL of USP Baclofen RS in water,
glacial acetic acid with the aid of a hot water bath. Dilute from the Standard stock solution
to 100 mL with water. Mix equal volumes of o-tolidine Sample solution: Shake thoroughly by hand each bottle of
solution and 0.83% potassium iodide solution (1:1). Oral Suspension. Pipet 0.5 mL of Oral Suspension from each
Analysis bottle to a 500-mL volumetric flask, dilute with water to
Samples: Standard solution A, Standard solution B, volume to obtain a concentration of 5 g/mL, and pass
Identification solution, and Sample solution through a 0.22-m polyvinylidenefluoride (PVDF) filter.
Proceed as directed. Remove the plate from the Chromatographic system
developing chamber, and dry under a current of warm (See Chromatography 621, System Suitability.)
air. Transfer the dry plate to another chromatographic Mode: LC
chamber containing a beaker with 1 g of potassium Detector: UV 220 nm
permanganate. Add 10 mL of 40% hydrochloric acid to Column: 4.6-mm 25-cm; 5-m packing L1
the beaker. Cover the chamber, and allow the chamber Flow rate: 1 mL/min
to become saturated with chlorine gas. Expose the plate Injection size: 15 L
to the chlorine gas for 8 min. Remove the plate and System suitability
expose to the air for 2 min, then spray with freshly Sample: Standard solution
prepared Spray reagent. [NOTEThe retention time of baclofen is about 5.5 min.]
Acceptance criteria: The intensity of the secondary spot Suitability requirements
from the Sample solution, corresponding in RF value to the Relative standard deviation: NMT 2.0%
principal spot from the Identification solution, is NMT that Analysis
of the principal spot from the Identification solution (1.0%), Samples: Standard solution and Sample solution
and no other secondary spot from the Sample solution is Calculate the percentage of C10H12ClNO2 in the volume of
greater in intensity than the principal spot from Standard Oral Suspension:
solution A (0.5%). The sum of all secondary spots from the
Sample solution is NMT 2.0%. Result = (rU/rS) (CS/CU) 100
WATER DETERMINATION, Method I 921: NMT 3.0% rS = peak response from the Standard solution
CS = concentration of USP Baclofen RS in the
ADDITIONAL REQUIREMENTS Standard solution (g/mL)
PACKAGING AND STORAGE: Preserve in tight containers. CU = nominal concentration of baclofen in the Sample
USP REFERENCE STANDARDS 11 solution (g/mL)
USP Baclofen RS Acceptance criteria: 90.0%110.0%
USP Baclofen Related Compound A RS
SPECIFIC TESTS
PH 791: 4.25.2
Baclofen Oral Suspension ADDITIONAL REQUIREMENTS
(Comment on this Monograph)id=m1722=Baclofen Oral PACKAGING AND STORAGE: Preserve in tight, light-resistant
Suspension=B-Monos.pdf) containers. Store in a cold place.
LABELING: Label it to state that it is to be well shaken, and
DEFINITION to state the beyond-use date.
Baclofen Oral Suspension contains NLT 90.0% and NMT BEYOND-USE DATE: 35 days after the day on which it was
110.0% of the labeled amount of baclofen (C10 H12 ClNO2). compounded
Prepare Baclofen Oral Suspension 5 mg/mL as follows (see USP REFERENCE STANDARDS 11
Pharmaceutical CompoundingNonsterile Solutions 795): USP Baclofen RS
Baclofen 500 mg
Vehicle: a mixture of Vehicle for Oral A sufficient quantity Baclofen Tablets
Solution(regular or sugar-free), NF, and (Comment on this Monograph)id=m6970=Baclofen Tablets=B-
Vehicle for Oral Suspension, NF (1:1) Monos.pdf)
To make 100 mL
DEFINITION
If using Baclofen Tablets, place the Tablets in a suitable mortar Baclofen Tablets contain NLT 90.0% and NMT 110.0% of the
and comminute the Tablets to a fine powder, or add Baclofen labeled amount of C10H12ClNO2.
powder. Add 5 mL of the Vehicle to wet the powder, and
triturate the powder to form a fine paste. Add the Vehicle in IDENTIFICATION
small portions almost to volume, and mix thoroughly after A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
each addition. Transfer, stepwise and quantitatively, the Sample solution: Equivalent to 5 mg/mL of baclofen from
contents of the mortar to a calibrated bottle. Add sufficient powdered Tablets in dehydrated alcohol and glacial acetic
Vehicle to bring to final volume, and mix well. acid (4:1)
[NOTEShake for 30 min and centrifuge.]
ASSAY Standard solution: 5 mg/mL of USP Baclofen RS in
PROCEDURE dehydrated alcohol and glacial acetic acid (4:1)
Mobile phase: Acetonitrile and 0.05 M monobasic sodium Chromatographic system
phosphate (1:4) (See Chromatography 621, Thin-Layer Chromatography.)
USP 32 Official Monographs / Bandage 11
Mode: TLC Mode: LC

Adsorbent: 0.25-mm layer of chromatographic silica gel Detector: UV 265 nm
mixture Column: 3.9-mm 30-cm; packing L1
Application volume: 20 L Flow rate: 0.6 mL/min
Developing solvent system: Butyl alcohol, glacial acetic Injection size: 190 L
acid, and water (4:1:1) Analysis
Spray reagent: 0.4 g of ninhydrin in 95 mL of butyl Samples: Sample solution and Standard solution
alcohol mixed with 5 mL of dilute glacial acetic acid (1 in Calculate the quantity of C10H12ClNO2 dissolved.
10) Tolerances: NLT 75% (Q) of the labeled amount of
Analysis: Proceed as directed. Develop the chromatogram C10H12ClNO2
until the solvent front has moved about three-fourths the UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
length of the plate. Remove the plate from the developing
chamber, and dry under a current of warm air. Spray the IMPURITIES
plate with the Spray reagent until the plate is slightly wet. Organic Impurities
Place the plate in an oven maintained at 100 for 10 min. PROCEDURE
Acceptance criteria: The RF value of the principal orange- Diluent: Proceed as directed in the Assay.
red spot of the Sample solution corresponds to that of the Mobile phase: Proceed as directed in the Assay.
Standard solution. Standard stock solution: 1 mg/mL of USP Baclofen
B. The retention time of the Sample solution corresponds to Related Compound A RS in methanol
that of the Standard solution, as obtained in the Assay. Standard solution: 0.16 mg/mL of USP Baclofen Related
Compound A in Diluent from the Standard stock solution
ASSAY Sample solution: Proceed as directed in the Assay.
PROCEDURE Chromatographic system and System suitability:
Diluent: Methanol, glacial acetic acid, and water (15:2:33) Proceed as directed in the Assay.
Mobile phase: 0.3 N acetic acid, Methanol, and 0.36 N Analysis
sodium 1-pentanesulfonate (55:44:2) Samples: Standard solution and Sample solution
Standard solution: 4 mg/mL of USP Baclofen RS in Diluent Calculate the percentage of baclofen related compound A
Sample solution: Weigh and finely powder NLT 20 Tablets. in the portion of the Tablets taken:
Transfer an amount equivalent to 40 mg to a 50-mL flask.
Add 10.0 mL of Diluent to the flask. Sonicate to disperse, Result = (rU/rS) (CS/CU) 100
and shake by mechanical means for 30 min. Centrifuge a
portion of this solution for 5 min, and filter. rU = peak area response of the Sample solution
Chromatographic system rS = peak area response of the Standard solution
(See Chromatography 621, System Suitability.) CS = concentration of USP Baclofen Related
Mode: LC Compound A RS in the Standard solution
Detector: UV 265 nm (mg/mL)
Column: 3.9-mm 30-cm; packing L1 CU = nominal concentration of baclofen in the Sample
Flow rate: 0.6 mL/min solution (mg/mL)
Injection size: 10 L Acceptance criteria: NMT 4.0% of baclofen related
System suitability compound A
Relative standard deviation: NMT 2.0% PACKAGING AND STORAGE: Preserve in well-closed containers.
Samples: Standard solution and Sample Solution USP Baclofen RS
[NOTESample solution to elute at NLT three times the USP Baclofen Related Compound A RS
retention time of baclofen.]
Result = (rU/rS) (CS/CU) 100 Adhesive Bandage
rU = peak area response of the Sample solution (Comment on this Monograph)id=m7020=Adhesive
rS = peak area response of the Standard solution Bandage=B-Monos.pdf)
Cs = concentration of USP Baclofen RS in the
Standard solution (mg/mL) DEFINITION
Cu = nominal concentration of the Sample solution Adhesive Bandage consists of a compress of four layers of Type
(mg/mL) I Absorbent Gauze or other suitable material affixed to a film
Acceptance criteria: 90.0%110.0% or fabric coated with a pressure-sensitive adhesive substance. It
is sterile. The compress may contain a suitable antimicrobial
PERFORMANCE TESTS agent and may contain one or more suitable colors. The
DISSOLUTION, Procedure for a Pooled Sample 711 adhesive surface is protected by a suitable removable covering.
Medium: 0.01 N hydrochloric acid; for NMT 10 mg of
drug, 500 mL for more than 10 mg of drug, 1000 mL SPECIFIC TESTS
Apparatus 2: 50 rpm STERILITY TESTS 71: Meets the requirements
Time: 30 min ADDITIONAL REQUIREMENTS
Mobile phase: Proceed as directed in the Assay. PACKAGING AND STORAGE: Package Adhesive Bandage that
Standard solution: USP Baclofen RS in Medium does not exceed 15 cm (6 inches) in width individually in
Sample solution: Sample per Dissolution 711 such manner that sterility is maintained until the individual
12 Bandage / Official Monographs USP 32
package is opened. Package individual packages in a second WIDTH: Measure width at each of the 5 points selected for
protective container. the determination of the thread count: the average of 5
LABELING: The label of the second protective container bears measurements is NMT 1.6 mm (1/16 inch) less than the
a statement that the contents may not be sterile if the labeled width of the Bandage.
individual package has been damaged or previously opened, LENGTH: Measure the length of the unrolled Gauze Bandage,
and it bears the names of any added antimicrobial agents. smoothed without tension, along the center line of the
Each individual package is labeled to indicate the dimensions Gauze Bandage: the length is NLT 98.0% of the labeled
of the compress and the name of the manufacturer, packer, length of the Bandage.
or distributor, and each protective container indicates also WEIGHT: Weigh the entire Bandage: the calculated weight in
the address of the manufacturer, packer, or distributor. g/0.894 m2 (1 linear yard Type I gauze), using the
measurements obtained as described under Width and
Length is NLT 39.2 g.
ABSORBENCY: Hold a rolled Gauze Bandage horizontal to and
Gauze Bandage almost in contact with the surface of water at 25, and allow
(Comment on this Monograph)id=m7030=Gauze Bandage=B- to drop lightly upon the water: complete submersion takes
Monos.pdf) place in NMT 30 s.
IGNITED RESIDUE, ACID OR ALKALI, AND DEXTRIN OR STARCH, IN
DEFINITION WATER EXTRACT
Gauze Bandage is Type I Absorbent Gauze. Its length is NLT Sample solution: Place 20 0.1 g of Gauze Bandage in
98.0% of that declared on the label, and its average width is 500 mL of water, and boil the mixture for 15 min, adding
NMT 1.6 mm less than the declared width. It contains no dye boiling water as necessary to maintain the original volume.
or other additives. Pour the water through a funnel into a 1000-mL volumetric
flask, transfer the Absorbent Gauze to the funnel, press out
IMPURITIES the excess water with a glass rod, and wash it with two
Inorganic Impurities 250-mL portions of boiling water, pressing the gauze after
RESIDUE ON IGNITION each washing. Cool the combined washings, dilute to
Sample: Place 5 g in a suitable dish, and moisten with 2 N volume, and mix.
sulfuric acid. Analysis 1 (Ignited Residue): Evaporate 400 mL of the
Analysis: Gently heat the Sample mixture until it is Sample solution, filtering if necessary, in a suitable dish on a
charred, then ignite more strongly until the carbon is steam bath, and dry the residue at 105 to constant
completely consumed. weight. Ignite the dried residue in a muffle furnace at a
Acceptance criteria: The weight of the residue dull-red heat to constant weight.
corresponds to NMT the percentage of the weight of the Acceptance criteria 1: The weight of the ignited residue
Gauze, calculated as follows: does not exceed an amount, in mg, calculated as follows:
Result = 0.002C + 0.015(100 C) Result = 20 0.14C
C = corrected percentage of cotton (0.89% C = corrected percentage of cotton (13 mg
maximum) maximum, or 0.16%)
Organic Impurities Analysis 2 (Acid or Alkali): To separate 200-mL portions
FATTY MATTER of the Sample solution, add 3 drops of phenolphthalein TS
Sample solution: Pack 10 0.01 g of Gauze Bandage in a and 1 drop of methyl orange TS, respectively.
continuous-extraction thimble with a tared flask, and Acceptance criteria 2: No pink color develops in either
extract with ether for 5 h, adjusting the rate so that the portion.
ether siphons NLT four times/h. Analysis 3 (Dextrin or Starch): To a 200-mL portion of
[NOTEThe ether extract in the flask shows no trace of the Sample solution, add 1 drop of iodine TS.
blue, green, or brownish color.] Acceptance criteria 3: No red, violet, or blue color
Analysis: Evaporate the Sample solution extract to dryness, develops.
and dry at 105 to constant weight. ALCOHOL-SOLUBLE DYES: Pack 10 g of Gauze Bandage in a
Acceptance criteria: The weight of the residue does not narrow percolator, and extract slowly with alcohol until the
exceed an amount, in mg, calculated as follows: percolate measures 50 mL: when observed downward in a
Result = 0.4C + 30 column 20 cm in depth, the percolate may show a yellowish
color, but neither a blue nor a green tint.
C = corrected percentage of cotton (70 mg ADDITIONAL REQUIREMENTS
maximum, or 0.7%) PACKAGING AND STORAGE: Gauze Bandage that has been
SPECIFIC TESTS rendered sterile is so packaged that the sterility of the
STERILITY TESTS 71: Meets the requirements contents is maintained until the package is opened for use.
THREAD COUNT: Count the number of warp and filling LABELING: The width and length of the Bandage, the
threads in areas of 1.27 cm (1/2 inch) square at 5 points number of pieces contained, and the name of the
evenly spread along the center line of the Bandage, no point manufacturer, packer, or distributor, are stated on the
being within 30.5 cm (12 inches) of either end of the package. The designation non-sterilized or not sterilized
Bandage, and calculate the average number of threads/2.54 appears prominently on the package unless the Gauze
cm (1 inch) in each direction. A variation of NMT 3 Bandage has been rendered sterile, in which case it may be
threads/inch is allowed in either warp or filling, provided labeled to indicate that it is sterile and that the contents may
that the combined variations do not exceed 5 not be sterile if the package bears evidence of damage or if
threads/square inch. [NOTEBefore determining the thread the package has been previously opened.
count, dimensions, and weight, hold the Bandage, unrolled,
for NLT 4 h in a standard atmosphere of 65 2% relative
humidity at 21 1.1C (70 2F).]
USP 32 Official Monographs / Barium 13
Barium Hydroxide Lime Acceptance criteria: The increase in weight is NLT 19.0%
(Comment on this Monograph)id=m7080=Barium Hydroxide of the weight of Barium Hydroxide Lime used for the test.
Lime=B-Monos.pdf) ADDITIONAL REQUIREMENTS
DEFINITION PACKAGING AND STORAGE: Preserve in tight containers.
Barium Hydroxide Lime is a mixture of barium hydroxide LABELING: If an indicator has been added, the name and
octahydrate and Calcium Hydroxide. It may also contain color change of such indicator are stated on the container
Potassium Hydroxide and may contain an indicator that is label. The container label indicates also the mesh size in
inert toward anesthetic gases such as Ether, Cyclopropane, terms of standard-mesh sieve sizes (see Powder Fineness
and Nitrous Oxide and that changes color when the Barium 811).
Hydroxide Lime no longer can absorb carbon dioxide.
[CAUTIONBecause Barium Hydroxide Lime contains a soluble

form of barium, it is toxic if swallowed.]
Barium Sulfate
(Comment on this Monograph)id=m7110=Barium Sulfate=B-
IDENTIFICATION Monos.pdf)
A. PROCEDURE BaSO4 233.39
Place a granule of it on a piece of moistened red litmus Sulfuric acid, barium salt (1:1);
paper: the paper turns blue immediately. Barium sulfate (1:1) [7727-43-7].
B. IDENTIFICATION TESTSGENERAL, Barium, Calcium, and
Potassium 191: A 100 mg/mL solution in 6 N acetic acid DEFINITION
responds to the tests for Barium and for Calcium, and it may Barium Sulfate contains NLT 97.5% and NMT 100.5% of
respond also to the flame test for Potassium. BaSO4.
SPECIFIC TESTS IDENTIFICATION
PARTICLE SIZE DISTRIBUTION ESTIMATION BY ANALYTICAL SIEVING A. IDENTIFICATION TESTSGENERAL 191, Sulfate
786: Screen 100 g for 5 min as directed in the chapter, Sample solution: Mix 0.5 g of Barium Sulfate with 2 g each
using a mechanical shaker. It passes completely through a of anhydrous sodium carbonate and anhydrous potassium
No. 2 standard-mesh sieve, and NMT 2.0% passes through a carbonate, heat the mixture in a crucible until fusion is
No. 40 standard-mesh sieve. NMT 7.0% is retained on the complete, treat the resulting fused mass with hot water, and
coarse-mesh sieve, and NMT 15.0% passes through the fine- filter.
mesh sieve designated on the label. Acceptance criteria: The filtrate, acidified with hydrochloric
LOSS ON DRYING 731: Weigh about 10 g in a tared acid, meets the requirements.
weighing bottle, and dry at 105 for 2 h: it loses between B. IDENTIFICATION TESTSGENERAL 191, Barium
11.0% and 16.0% of its weight. Sample solution: Dissolve a portion of the well-washed
HARDNESS residue from Identification test A in 6 N acetic acid.
Sample: 200 g Acceptance criteria: The solution meets the requirements.
Analysis: Screen the Sample on a mechanical sieve shaker
(see Particle Size Distribution Estimation by Analytical Sieving ASSAY
786) having a frequency of oscillation of 285 3 PROCEDURE
cycles/min, for 3 min, to remove granules coarser than 4- Sample: 0.58 g0.62 g, weighed in a tared platinum
mesh and finer than 8-mesh. Weigh 50 g of the granules crucible
retained on the screen, and place them in a hardness pan of Analysis: Add 10 g of anhydrous sodium carbonate to the
the following description: crucible, and mix by rotating the crucible. Fuse over a blast
the hardness pan has a diameter of 200 mm and a concave burner until a clear melt is obtained, and heat for an
brass bottom, and the bottom of the pan is 7.9 mm thick additional 30 min. Cool, place the crucible in a 400-mL
at the circumference and 3.2 mm thick at the center and beaker, add 250 mL of water, stir with a glass rod, and heat
has an inside spherical radius of curvature of 109 cm. Add to dislodge the melt. Remove the crucible from the beaker,
15 steel balls of 7.9-mm diameter, and shake on a and wash with water, collecting the washings in the beaker.
mechanical sieve shaker for 30 min. Remove the steel balls, Rinse the inside of the crucible with 2 mL of 6 N acetic acid
brush the contents of the hardness pan onto a sieve of the and then with water, again collecting the washings in the
fine-mesh size designated on the label, shake for 3 min on beaker, and continue heating and stirring until the melt is
the mechanical sieve shaker, and weigh. disintegrated. Cool the beaker in an ice bath until the
Acceptance criteria: The percentage of Barium Hydroxide precipitate settles, decant the clear liquid through filter
Lime retained on the screen is NLT 75.0%, and represents paper (Whatman No. 40, or equivalent), taking care to
the hardness. transfer as little precipitate as possible to the paper. Wash
CARBON DIOXIDE ABSORBENCY twice by decantation as follows. Wash down the inside of
Analysis: Fill the lower transverse section of a U-shaped the beaker with 10 mL of cold sodium carbonate solution (1
drying tube of about 15-mm internal diameter and 15-cm in 50), swirl the contents of the beaker, allow the precipitate
height with loosely packed glass wool. Place in one arm of to settle, and decant the supernatant through the same filter
the tube, about 5 g of anhydrous calcium chloride, and paper as before, transferring as little precipitate as possible.
weigh the tube and the contents. Into the other arm of the Place the beaker containing the bulk of the barium
tube, place 9.5 g10.5 g of Barium Hydroxide Lime, and carbonate precipitate under the funnel, wash the filter paper
again weigh. Insert stoppers in the open arms of the U-tube, with five 1-mL portions of 3 N hydrochloric acid, and wash
and connect the side tube of the arm filled with Barium the paper with water.
Hydroxide Lime to a calcium chloride drying tube, which in [NOTEThe solution may be slightly hazy.]
turn is connected to a suitable source of supply of carbon Add 100 mL of water, 5.0 mL of hydrochloric acid, 10.0 mL
dioxide. Pass the carbon dioxide through the U-tube at a of ammonium acetate solution (2 in 5), 25 mL of
rate of 75 mL/min for 20 min, timed. Disconnect the U- potassium dichromate solution (1 in 10), and 10.0 g of
tube, cool to room temperature, remove the stoppers, and urea. Cover the beaker with a watch glass, and digest at
weigh. 80 to 85 for NLT 16 h. Filter while hot through a tared,
14 Barium / Official Monographs USP 32
fine-porosity, sintered-glass crucible, transferring all of the 90.0% and NMT 110.0% of the labeled amount of barium
precipitate with the aid of a rubber-tipped stirring rod. sulfate (BaSO4). It may contain one or more suitable colors,
Wash the precipitate with potassium dichromate solution (1 flavors, suspending or dispersing agents, and preservatives.
in 200), and finally with 20 mL of water. Dry at 105 for 2
h, cool, and weigh. The weight of the barium chromate so IDENTIFICATION
obtained, multiplied by 0.9213, represents the weight of A. IDENTIFICATION TESTSGENERAL, Sulfate 191
BaSO4. Sample: Ignite a quantity of Paste equivalent to 0.5 g of
Acceptance criteria: 97.5%100.5% of BaSO4 barium sulfate to constant weight.
Analysis: Mix 0.5 g of the ignited Sample with 2 g each of
IMPURITIES anhydrous sodium carbonate and anhydrous potassium
Inorganic Impurities carbonate, heat the mixture in a crucible until fusion is
HEAVY METALS 231: NMT 10 ppm complete, treat the resulting fused mass with hot water, and
Sample solution: Boil 4.0 g with a mixture of 2 mL of filter. Proceed as directed.
glacial acetic acid and 48 mL of water for 10 min. Dilute Acceptance criteria: The filtrate, acidified with hydrochloric
with water to 50 mL, filter, and use 25 mL of the filtrate. acid, meets the requirements.
LIMIT OF SULFIDE B. IDENTIFICATION TESTSGENERAL, Barium 191
Sample solution: Transfer 10 g to a 500-mL conical flask. Sample solution: Dissolve a portion of the well-washed
Add 100 mL of 0.3 N hydrochloric acid. residue from Identification test A in 6 N acetic acid.
Control: 100 mL of 0.3 N hydrochloric acid containing 5 Acceptance criteria: The solution meets the requirements.
g of sulfide in a 500-mL conical flask
Analysis: Cover the mouth of each conical flask with a ASSAY
circle of filter paper that has been moistened at the area PROCEDURE
over the mouth of the flask with 0.15 mL of lead acetate Sample: Barium Sulfate Paste, equivalent to 0.60 g of
TS, the paper being held in place with a string tied around barium sulfate, weighed in a tared platinum crucible
the neck of the flask. Boil each mixture gently for 10 min, Analysis: Ignite the Sample over a low flame until any
taking care to avoid spattering the paper. organic matter is thoroughly carbonized. Cool, cautiously
Acceptance criteria: Any darkening of the paper is not add 0.5 mL of nitric acid and 0.5 mL of sulfuric acid, and
greater than that produced by the similarly treated Control continue the ignition over a low flame until the residue
(NMT 0.5 ppm). becomes gray in color, then ignite over the full heat of a
LIMIT OF ACID-SOLUBLE SUBSTANCES blast burner. Allow the contents of the crucible to cool to
Sample solution: Cool the mixture obtained in the test for room temperature.
Limit of Sulfide, add water to restore approximately the [NOTEIf the specimen contains a silicate, such as
original volume, and filter it through paper that previously bentonite, proceed as follows. Add 10 mL of water and 1
has been washed with a mixture of 10 mL of 3 N mL of sulfuric acid to the residue in the crucible, mix, and
hydrochloric acid and 90 mL of water, returning the first add 10 mL of hydrofluoric acid. Heat gently over a low
portions, if necessary, to obtain a clear filtrate. flame until fumes of sulfur trioxide appear. Add 5 mL
Analysis: Evaporate 50 mL of the filtrate on a steam bath more of hydrofluoric acid, heat again over a low flame to
to dryness, and add 2 drops of hydrochloric acid and 10 the appearance of dense fumes, and continue heating
mL of hot water. Filter again through acid-washed paper, until the sulfuric acid has been completely volatilized.
prepared as directed above, wash the filter with 10 mL of Allow the contents of the crucible to cool.]
hot water, and evaporate the combined filtrate and [NOTEIf the specimen does not contain a silicate, omit
washings in a tared dish on a steam bath to dryness. the treatment of the specimen with hydrofluoric and
Acceptance criteria: The residue, when dried at 105 for 1 sulfuric acids.]
h, weighs NMT 15 mg (NMT 0.3%). Add to the treated or untreated specimen in the platinum
LIMIT OF SOLUBLE BARIUM SALTS crucible, 10 g of anhydrous sodium carbonate, fuse over a
Sample: Residue obtained in the test for Limit of Acid- blast burner until a clear melt is obtained, and heat for an
Soluble Substances additional 30 min. Cool, place the crucible in a 400-mL
Control: 10 mL of water containing 0.5 mL of 2 N sulfuric beaker, add 250 mL of water, stir with a glass rod, and
acid and 50 g of barium heat to dislodge the melt. Remove the crucible from the
Analysis: Treat the Sample with 10 mL of water, pass the beaker, and wash with water, collecting the washings in
solution through a filter previously washed with 100 mL of the beaker. Rinse the inside of the crucible with 2 mL of 6
0.3 N hydrochloric acid, and add 0.5 mL of 2 N sulfuric N acetic acid and then with water, again collecting the
acid. washings in the beaker, and continue heating and stirring
Acceptance criteria: Any turbidity formed within 30 min is until the melt is disintegrated. Cool the beaker in an ice
NMT that produced in the similarly treated Control (NMT bath until the precipitate settles, decant the clear liquid
0.001%). through filter paper (Whatman No. 40, or equivalent),
taking care to transfer as little precipitate as possible to the
SPECIFIC TESTS paper.
PH 791: 3.510.0, in a 10% (w/w) aqueous suspension Wash twice by decantation as follows. Wash down the inside
of the beaker with 10 mL of cold sodium carbonate
ADDITIONAL REQUIREMENTS solution (1 in 50), swirl the contents of the beaker, allow
PACKAGING AND STORAGE: Preserve in well-closed containers. the precipitate to settle, and decant the supernatant
through the same filter paper as before, transferring as little
precipitate as possible. Place the beaker containing the bulk
Barium Sulfate Paste of the barium carbonate precipitate under the funnel, wash
the filter paper with five 1-mL portions of 3 N hydrochloric
(Comment on this Monograph)id=m7120=Barium Sulfate acid, and wash the paper with water.
Paste=B-Monos.pdf) [NOTEThe solution may be slightly hazy.]
Add 100 mL of water, 5.0 mL of hydrochloric acid, 10.0 mL
DEFINITION of ammonium acetate solution (2 in 5), 25 mL of
Barium Sulfate Paste is a semisolid formulation of finely divided potassium dichromate solution (1 in 10), and 10.0 g of
particles of Barium Sulfate in a suitable base. It contains NLT
USP 32 Official Monographs / Barium 15
urea. Cover the beaker with a watch glass, and digest at acid and 0.5 mL of sulfuric acid, and continue the ignition
8085 for NLT 16 h. Filter while hot through a tared, over a low flame until the residue becomes gray in color,
fine-porosity, sintered-glass crucible, transferring all of the then ignite over the full heat of a blast burner. Allow the
precipitate with the aid of a rubber-tipped stirring rod. contents of the crucible to cool to room temperature.
Wash the precipitate with potassium dichromate solution (1 [NOTEIf the specimen contains a silicate, such as
in 200), and finally with 20 mL of water. Dry at 105 for 2 bentonite, proceed as follows. Add 10 mL of water and 1
h, cool, and weigh. The weight of the barium chromate so mL of sulfuric acid to the residue in the crucible, mix, and
obtained, multiplied by 0.9213, represents the weight of add 10 mL of hydrofluoric acid. Heat gently over a low
BaSO4. flame until fumes of sulfur trioxide appear. Add 5 mL
Acceptance criteria: 90.0%110.0% more of hydrofluoric acid, heat again over a low flame to
the appearance of dense fumes, and continue heating
SPECIFIC TESTS until the sulfuric acid has been completely volatilized.
MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED Allow the contents of the crucible to cool.]
MICROORGANISMS 62: For products labeled for oral [NOTEIf the specimen does not contain a silicate, omit
administration or labeled for oral and rectal administration, the treatment of the specimen with hydrofluoric and
the total aerobic microbial count does not exceed 100 cfu/g, sulfuric acids.]
and the total combined molds and yeasts count does not Add to the treated or untreated specimen in the platinum
exceed 10 cfu/g. For products labeled for rectal crucible, 10 g of anhydrous sodium carbonate, fuse over a
administration, the total aerobic microbial count does not blast burner until a clear melt is obtained, and heat for an
exceed 1000 cfu/g, and the total combined molds and additional 30 min. Cool, place the crucible in a 400-mL
yeasts count does not exceed 100 cfu/g. For all products, it beaker, add 250 mL of water, stir with a glass rod, and
meets the requirements of the tests for absence of heat to dislodge the melt. Remove the crucible from the
Salmonella species, Escherichia coli, Staphylococcus aureus, and beaker, and wash with water, collecting the washings in
Pseudomonas aeruginosa, and the total enterobacterial count the beaker. Rinse the inside of the crucible with 2 mL of 6
does not exceed 10 cfu/g. N acetic acid and then with water, again collecting the
PH 791: 3.010.0 washings in the beaker, and continue heating and stirring
until the melt is disintegrated. Cool the beaker in an ice
ADDITIONAL REQUIREMENTS bath until the precipitate settles, decant the clear liquid
PACKAGING AND STORAGE: Preserve in tight containers, through filter paper (Whatman No. 40, or equivalent),
protected from freezing and from excessive heat. taking care to transfer as little precipitate as possible to the
paper. Wash twice by decantation as follows. Wash down
the inside of the beaker with 10 mL of cold sodium
Barium Sulfate Suspension carbonate solution (1 in 50), swirl the contents of the
beaker, allow the precipitate to settle, and decant the
(Comment on this Monograph)id=m7135=Barium Sulfate supernatant through the same filter paper as before,
Suspension=B-Monos.pdf) transferring as little precipitate as possible. Place the beaker
DEFINITION containing the bulk of the barium carbonate precipitate
Barium Sulfate Suspension contains NLT 90.0% and NMT under the funnel, wash the filter paper with five 1-mL
110.0% of the labeled amount of barium sulfate (BaSO4). It portions of 3 N hydrochloric acid, and wash the paper with
contains suitable dispersing and/or suspending agents so that water.
when mixed as directed in the labeling, it yields a uniformly [NOTEThe solution may be slightly hazy.]
dispersed suspension. It may contain one or more suitable Add 100 mL of water, 5.0 mL of hydrochloric acid, 10.0 mL
colors, flavors, fluidizing agents, and preservatives. of ammonium acetate solution (2 in 5), 25 mL of
potassium dichromate solution (1 in 10), and 10.0 g of
IDENTIFICATION urea. Cover the beaker with a watch glass, and digest at
A. IDENTIFICATION TESTSGENERAL 191, Sulfate 80 to 85 for NLT 16 h. Filter while hot through a tared,
Sample: Shake the Suspension and transfer a volume fine-porosity, sintered-glass crucible, transferring all of the
equivalent to 0.5 g of barium sulfate to a suitable container. precipitate with the aid of a rubber-tipped stirring rod.
Ignite to constant weight. Wash the precipitate with potassium dichromate solution (1
Analysis: Mix 0.5 g of the ignited Sample with 2 g each of in 200), and finally with 20 mL of water. Dry at 105 for 2
anhydrous sodium carbonate and anhydrous potassium h, cool, and weigh. The weight of the barium chromate so
carbonate, heat the mixture in a crucible until fusion is obtained, multiplied by 0.9213, represents the weight of
complete, treat the resulting fused mass with hot water, and BaSO4.
filter. Proceed as directed. Acceptance criteria: 90.0%110.0%
Acceptance criteria: The filtrate, acidified with hydrochloric
acid, meets the requirements. SPECIFIC TESTS
B. IDENTIFICATION TESTSGENERAL 191, Barium MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED
Sample solution: Dissolve a portion of the well-washed MICROORGANISMS 62: The total bacterial count does not
residue from Identification test A in 6 N acetic acid. exceed 100 cfu/mL; the total combined molds and yeasts
Acceptance criteria: The solution meets the requirements. count does not exceed 10 cfu/mL; and it meets the
requirements of the tests for absence of Salmonella species,
ASSAY Staphylococcus aureus, and Pseudomonas aeruginosa.
PROCEDURE PH 791: 3.510.0
Sample: A volume of Suspension, previously well shaken in
its original container, equivalent to 0.60 g of barium sulfate, ADDITIONAL REQUIREMENTS
in a tared platinum crucible PACKAGING AND STORAGE: Preserve in tight containers, and
Analysis: Ignite over a low flame until any organic matter is avoid freezing.
thoroughly carbonized. Cool, cautiously add 0.5 mL of nitric
16 Barium / Official Monographs USP 32
Barium Sulfate for Suspension portions of 3 N hydrochloric acid, and wash the paper with
(Comment on this Monograph)id=m7140=Barium Sulfate for water.
Suspension=B-Monos.pdf) [NOTEThe solution may be slightly hazy.]
Add 100 mL of water, 5.0 mL of hydrochloric acid, 10.0 mL
DEFINITION of ammonium acetate solution (2 in 5), 25 mL of
Barium Sulfate for Suspension is a dry mixture of Barium Sulfate potassium dichromate solution (1 in 10), and 10.0 g of
and one or more suitable dispersing and/or suspending urea. Cover the beaker with a watch glass, and digest at
agents. It contains NLT 90.0% and NMT 110.0% of the 80 to 85 for NLT 16 h. Filter while hot through a tared,
labeled amount of barium sulfate (BaSO4). It may contain one fine-porosity, sintered-glass crucible, transferring all of the
or more suitable colors, flavors, fluidizing agents, and precipitate with the aid of a rubber-tipped stirring rod.
preservatives. Wash the precipitate with potassium dichromate solution (1
in 200), and finally with 20 mL of water. Dry at 105 for 2
IDENTIFICATION h, cool, and weigh. The weight of the barium chromate so
A. IDENTIFICATION TESTSGENERAL 191, Sulfate obtained, multiplied by 0.9213, represents the weight of
Sample: Ignite 1g to constant weight BaSO4.
Analysis: Mix 0.5 g of the ignited Sample with 2 g each of Acceptance criteria: 90.0%110.0%
anhydrous sodium carbonate and anhydrous potassium
carbonate, heat the mixture in a crucible until fusion is SPECIFIC TESTS
complete, treat the resulting fused mass with hot water, and LOSS ON DRYING 731: Dry at 105 for 4 h: it loses NMT
filter. 1.0% of its weight.
Acceptance criteria: The filtrate, acidified with hydrochloric PH 791: 3.510.0, in a 60% (w/w) aqueous suspension, or
acid, meets the requirements. constituted for its intended use as directed in the labeling
B. IDENTIFICATION TESTSGENERAL191, Barium
Sample solution: Dissolve a portion of the well-washed ADDITIONAL REQUIREMENTS
residue from Identification test A in 6 N acetic acid. PACKAGING AND STORAGE: Preserve in well-closed containers.
Acceptance criteria: The solution meets the requirements.
ASSAY
PROCEDURE Barium Sulfate Tablets
Sample: Barium Sulfate for Suspension, equivalent to 0.60 g (Comment on this Monograph)id=m7155=Barium Sulfate
of barium sulfate, weighed in a tared platinum crucible Tablets=B-Monos.pdf)
Analysis: Ignite over a low flame until any organic matter is
thoroughly carbonized. Cool, cautiously add 0.5 mL of nitric DEFINITION
acid and 0.5 mL of sulfuric acid, and continue the ignition Barium Sulfate Tablets are flat-sided disks between 11.5 mm
over a low flame until the residue becomes gray in color, and 13.5 mm in diameter and contain NLT 90.0% and NMT
then ignite over the full heat of a blast burner. Allow the 110.0% of the labeled amount of BaSO4.
contents of the crucible to cool to room temperature. IDENTIFICATION
[NOTEIf the specimen contains a silicate, such as A. IDENTIFICATION TESTSGENERAL, Sulfate 191
bentonite, proceed as follows. Add 10 mL of water and 1 Sample: A portion of powdered Tablets equivalent to 0.6 g
mL of sulfuric acid to the residue in the crucible, mix, and of barium sulfate
add 10 mL of hydrofluoric acid. Heat gently over a low Analysis: Mix the Sample with 2 g each of anhydrous
flame until fumes of sulfur trioxide appear. Add 5 mL sodium carbonate and anhydrous potassium carbonate, heat
more of hydrofluoric acid, heat again over a low flame to the mixture in a crucible until fusion is complete, treat the
the appearance of dense fumes, and continue heating resulting fused mass with hot water, and filter. Proceed as
until the sulfuric acid has been completely volatilized. directed.
Allow the contents of the crucible to cool.] Acceptance criteria: The filtrate, acidified with hydrochloric
[NOTEIf the specimen does not contain a silicate, omit acid, meets the requirements.
the treatment of the specimen with hydrofluoric and B. IDENTIFICATION TESTSGENERAL, Barium 191
sulfuric acids.] Sample solution: Dissolve a portion of the well-washed
Add to the treated or untreated specimen in the platinum residue from Identification test A in 6 N acetic acid.
crucible, 10 g of anhydrous sodium carbonate, fuse over a Acceptance criteria: The solution meets the requirements.
blast burner until a clear melt is obtained, and heat for an
additional 30 min. Cool, place the crucible in a 400-mL ASSAY
beaker, add 250 mL of water, stir with a glass rod, and PROCEDURE
heat to dislodge the melt. Remove the crucible from the Sample: A portion of powdered Tablets, equivalent to 0.6 g
beaker, and wash with water, collecting the washings in of barium sulfate, weighed in a tared platinum crucible
the beaker. Rinse the inside of the crucible with 2 mL of 6 Analysis: Add 10 g of anhydrous sodium carbonate to the
N acetic acid and then with water, again collecting the crucible, and mix by rotating the crucible. Fuse over a blast
washings in the beaker, and continue heating and stirring burner until a clear melt is obtained, and heat for an
until the melt is disintegrated. Cool the beaker in an ice additional 30 min. Cool, place the crucible in a 400-mL
bath until the precipitate settles, decant the clear liquid beaker, add 250 mL of water, stir with a glass rod, and heat
through filter paper (Whatman No. 40, or equivalent), to dislodge the melt. Remove the crucible from the beaker,
taking care to transfer as little precipitate as possible to the and wash with water, collecting the washings in the beaker.
paper. Wash twice by decantation as follows. Wash down Rinse the inside of the crucible with 2 mL of 6 N acetic acid
the inside of the beaker with 10 mL of cold sodium and then with water, again collecting the washings in the
carbonate solution (1 in 50), swirl the contents of the beaker, and continue heating and stirring until the melt is
beaker, allow the precipitate to settle, and decant the disintegrated. Cool the beaker in an ice bath until the
supernatant through the same filter paper as before, precipitate settles, decant the clear liquid through filter
transferring as little precipitate as possible. Place the beaker paper (Whatman No. 40, or equivalent), taking care to
containing the bulk of the barium carbonate precipitate transfer as little precipitate as possible to the paper. Wash
under the funnel, wash the filter paper with five 1-mL
USP 32 Official Monographs / BCG 17
twice by decantation as follows. Wash down the inside of total of at least 4 mg of the Sample solution intramuscularly
the beaker with 10 mL of cold sodium carbonate solution (1 or subcutaneously in the rear left internal thigh, and observe
in 50), swirl the contents of the beaker, allow the precipitate them for a period of 6 weeks. Note the number of animals
to settle, and decant the supernatant through the same filter that survive at the end of the observation period, and then
paper as before, transferring as little precipitate as possible. sacrifice them. Perform autopsies of all animals postmortem
Place the beaker containing the bulk of the barium to examine them for evidence of tuberculous infections,
carbonate precipitate under the funnel, wash the filter paper particularly at the popliteal and inguinal lymph nodes, liver,
with five 1-mL portions of 3 N hydrochloric acid, and wash spleen, pancreas, and lungs, as well as at the injection site. If
the paper with water. [NOTEThe solution may be slightly any abnormalities are found, perform a histological
hazy.] Add 100 mL of water, 5.0 mL of hydrochloric acid, examination using standard and Acid-Fast staining
10.0 mL of ammonium acetate solution (2 in 5), 25 mL of techniques to detect Acid-Fast organisms.
potassium dichromate solution (1 in 10), and 10.0 g of urea. Acceptance criteria: The product complies with the test if
Cover the beaker with a watch glass, and digest at 8085 none of the animals show signs of tuberculosis and NMT
for NLT 16 h. Filter while hot through a tared, fine-porosity, one-third of the animals die during the observation period.
sintered-glass crucible, transferring all of the precipitate with SKIN REACTIVITY
the aid of a rubber-tipped stirring rod. Wash the precipitate Sample solutions: Using the same diluent and the Sample
with potassium dichromate solution (1 in 200), and finally solution, prepared as directed in the test for Virulent
with 20 mL of water. Dry at 105 for 2 h, cool, and weigh. Mycobacteria, further dilute aseptically by making three serial
The weight of the barium chromate so obtained, multiplied tenfold dilutions.
by 0.9213, represents the weight of BaSO4. Analysis: Randomly select two guinea pigs (male or
Acceptance criteria: 90.0%110.0% female), each weighing 250300 g. Inject 0.1 mL of each of
the four suspensions intradermally at different sites on the
PERFORMANCE TESTS back of each animal. After 4 weeks, the animals are shaved
DISINTEGRATION 701 so that the injection sites and any reactions are made clearly
Time: NLT 10 min and NMT 30 min visible. The diameters of the reactions are measured, and the
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements presence of necrosis or nodules are noted.
Acceptance criteria: The reaction for the largest dose is
ADDITIONAL REQUIREMENTS between 410 mm and the smallest dose induces a nodule
PACKAGING AND STORAGE: Preserve in well-closed containers. less than or equal to 4 mm. Each animal gains weight
during the observation period.
TUBERCULIN SENSITIVITY
BCG Live Tuberculin solution: Use tuberculin, purified protein
derivative, to prepare a solution containing 25 U.S.
(Comment on this Monograph)id=m703=BCG Live=B- Tuberculin Units/0.1 mL. Dilute aseptically, if necessary, with
Monos.pdf) sterile 0.9% sodium chloride solution.
DEFINITION Analysis: Use the same animals on which the Skin Reactivity
BCG Live (intravesical) for immunotherapy is a freeze-dried test is performed. After the Skin Reactivity test is completed,
solution of attenuated live bacteria derived from a culture of inject each animal intradermally on the back with 0.1 mL of
Bacillus Calmette-Guerin (Mycobacterium bovis, var. BCG) and the Tuberculin solution, and observe after 1824 h.
used intravesically in the treatment of carcinoma in situ and Acceptance criteria: An erythematous reaction of NLT 10
papilloma tumors of the urinary bladder. The bacteria are mm in diameter is measured on each animal.
grown in a medium that does not contain substances known RESIDUAL MOISTURE: NMT the limit approved for the
to cause toxic or allergic reactions in human beings or to particular product, determined by a suitable validated
cause the bacteria to become virulent for guinea pigs. The method
culture is harvested and formulated to contain one or more Limits vary in accordance with the method.
excipients. The freeze-dried solution is reconstituted and VIABILITY: Determine the potencies of BCG Live using NLT 5
further diluted aseptically with a sterile diluent for use. A containers before freeze-drying, and an equal number of
reconstituted dose contains 1.0-19.2 108 colony-forming containers after freeze-drying, following the procedure under
units (cfu). BCG Live does not contain a preservative. Potency, except use the suspension before freeze-drying as is.
The loss in viability due to freeze-drying is NMT 90%.
IDENTIFICATION POTENCY: Determine the number of viable units/mL by
BCG Live is identified by microscopic examination of the viable count on solid medium using a method suitable for
bacilli in stained smears demonstrating their acid-fast the product to be examined. Alternatively, a validated
property. Alternatively, validated molecular biology biochemical method may be used.
techniques may be used.
SPECIFIC TESTS PACKAGING AND STORAGE: BCG Live is sensitive to light and
STERILITY TESTS 71: It meets the requirements when tested therefore must be preserved and stored in a glass container
as directed for Test for Sterility of the Product to be Examined, where it is protected from direct light at a temperature
Direct Inoculation of the Culture Medium. between 2 and 8.
GENERAL SAFETY: It meets the requirements as set forth for EXPIRATION DATE: The product is stable for 3 years when
under Biological Reactivity Tests, In Vivo 88, Safety Tests stored between 2 and 8.
Biologicals, modified as follows. Guinea pigs are injected LABELING: Label it to indicate the dry weight of bacteria in a
intraperitoneally with 3.0 mL of the reconstituted product. vial, cfu/dose, the storage conditions, the expiration date,
VIRULENT MYCOBACTERIA and that it is not to be used after the expiration date given
Sample solution: Reconstitute the freeze-dried BCG Live as on the package. Label it to state that it should be protected
per the manufacturers instructions for human use with the from light and that it should be used immediately after
diluent recommended by the manufacturer, and dilute reconstitution/dilution. Label it to indicate that it is for
aseptically to about 2 mg/mL with sterile BCG diluents. intravesical use.
Analysis: Randomly select NLT six guinea pigs of the same
sex, each weighing 250300 g. Inject each animal with a
18 BCG / Official Monographs USP 32
BCG Vaccine Sample solution: Transfer 4.0 mL of Sample stock solution

(Comment on this Monograph)id=m7200=BCG Vaccine=B- to a suitable vial, and add 4.0 mL of Internal standard
Monos.pdf) solution.
DEFINITION (See Chromatography 621, System Suitability.)
BCG Vaccine conforms to the regulations of the FDA Mode: LC
concerning biologics (see Biologics 1041). It is a dried, living Detector: UV 254 nm
culture of the bacillus Calmette-Guerin strain of Mycobacterium Column: 4-mm 30-cm; packing L1 and a pump capable
tuberculosis var. bovis, grown in a suitable medium from a of operating at a column pressure of up to 3500 psi
seed strain of known history that has been maintained to [NOTEAdjust the specimen size and other operating
preserve its capacity for conferring immunity. It contains an parameters such that the peak from the internal standard
amount of viable bacteria such that inoculation, in the in the Standard solution is about 0.60.9 full-scale.]
recommended dose, of tuberculin-negative persons results in Injection size: 525 L
an acceptable tuberculin conversion rate. It is free from other System suitability
organisms, and contains a suitable stabilizer. It contains no Sample: Standard solution
antimicrobial agent. [NOTEThe retention time of beclomethasone dipropionate
[NOTEUse the Vaccine immediately after its constitution, and is about 6 min, and that of testosterone propionate is
discard any unused portion after 2 h.] about 10 min.]
ADDITIONAL REQUIREMENTS Relative standard deviation: NMT 3.0% for five replicate
PACKAGING AND STORAGE: Preserve in hermetic containers, injections
preferably of Type I glass, at a temperature between 2 and Analysis:
8. Samples: Sample solution and Standard solution
EXPIRATION DATE: The expiration date is not later than 6 Calculate the percentage of C28H37ClO7 in the portion of
months after date of issue, or not later than 1 year after date Beclomethasone Dipropionate taken:
of issue if stored at a temperature below 5.
RU = peak height ratio of beclomethasone
Beclomethasone Dipropionate dipropionate to the internal standard from the
(Comment on this Monograph)id=m7250=Beclomethasone Sample solution
Dipropionate=B-Monos.pdf) RS = peak height ratio of beclomethasone
dipropionate to the internal standard from the
Standard solution
CS = concentration of USP Beclomethasone
Dipropionate RS in the Standard solution
(mg/mL)
C28H37ClO7 521.04 IMPURITIES

Pregna-1,4-diene-3,20-dione, 9-chloro-11-hydroxy-16- Inorganic Impurities
methyl-17,21-bis(1-oxopropoxy)-, (11,16)-; RESIDUE ON IGNITION 281: NMT 0.1%
9-Chloro-11,17,21-trihydroxy-16-methylpregna-1,4-
diene-3,20-dione 17,21-dipropionate [5534-09-8]. SPECIFIC TESTS
Monohydrate 539.07 SPECIFIC ROTATION 781S: +88 to +94
Sample solution: 10 mg/mL, in dioxane
DEFINITION LOSS ON DRYING 731: Dry it at 105 for 3 h: the anhydrous
Beclomethasone Dipropionate is anhydrous or contains one form loses NMT 0.5% of its weight; the monohydrate form
molecule of water of hydration. It contains NLT 97.0% and loses between 2.8% and 3.8% of its weight.
NMT 103.0% of C28H37ClO7, calculated on the dried basis.
IDENTIFICATION PACKAGING AND STORAGE: Preserve in well-closed containers.
INFRARED ABSORPTION 197M
USP Beclomethasone Dipropionate RS
ASSAY USP Testosterone Propionate RS
PROCEDURE
Mobile phase: Acetonitrile and water (3:2)
Internal standard solution: 1.2 mg/mL of USP Belladonna Leaf
Testosterone Propionate RS in methanol
Standard stock solution: 1.4 mg/mL of USP (Comment on this Monograph)id=m7410=Belladonna Leaf=B-
Beclomethasone Dipropionate RS in methanol Monos.pdf)
Standard solution: Transfer 4.0 mL of Standard stock
solution to a suitable vial, and add 4.0 mL of Internal DEFINITION
standard solution to obtain a solution having a known Belladonna Leaf consists of the dried leaf and flowering or
concentration of about 0.7 mg/mL of USP Beclomethasone fruiting top of Atropa belladonna Linne or of its variety
Dipropionate RS and 0.6 mg/mL of USP Testosterone acuminata Royle ex Lindley (Fam. Solanaceae). Belladonna Leaf
Dipropionate RS. yields NLT 0.35% of the alkaloids of belladonna leaf.
Sample stock solution: 1.4 mg/mL of Beclomethasone
Dipropionate in methanol
USP 32 Official Monographs / Belladonna 19
ASSAY vigorously, allow the layers to separate, and discard the

PROCEDURE chloroform layer.
Phosphate buffer: 34.8 g of dibasic potassium phosphate [NOTEIf emulsions are formed, a mixed solvent consisting
in 900 mL of water of chloroform-isopropyl alcohol (10:3) may be substituted
Adjust to a pH of 9.5 by the addition of 3 N hydrochloric for chloroform throughout the extraction procedure.]
acid or sodium hydroxide, with mixing. Add another 15 mL of chloroform, and extract again,
Diluent: Dilute sulfuric acid (1 in 350) discarding the chloroform phase. Add 15 mL of Phosphate
Internal standard solution: 0.8 mg/mL of USP buffer and sufficient 1 N sodium hydroxide to yield a final
Homatropine Hydrobromide RS in Diluent pH between 9.0 and 9.5. Add 15 mL of chloroform, shake
[NOTEPrepare fresh on the day of use.] vigorously, and allow the layers to separate. Filter the
Standard stock solution A: 1.0 mg/mL of USP organic phase through 10 g of anhydrous sodium sulfate
Scopolamine Hydrobromide RS in Diluent (see Suitability for alkaloid assays under Reagents, Indicators,
Standard stock solution B: Dissolve 20 mg of USP Atropine and SolutionsSodium Sulfate, Anhydrous), previously
Sulfate RS in 25 mL of Diluent in a 50-mL volumetric flask, washed with chloroform and supported in a funnel with a
add 2.0 mL of Standard stock solution A, and mix. Add small pledget of glass wool, into a suitable container.
Diluent to volume. Extract again with two 15-mL portions of chloroform, again
[NOTEPrepare fresh on the day of use.] collecting the clarified organic phase. Wash the sodium
Standard solutions: Pipet into three separate 60-mL sulfate and the tip of the funnel with 5 mL of chloroform.
separators 1.0-, 2.0-, and 3.0-mL portions, respectively, of Evaporate the combined organic phases under reduced
Standard stock solution A, and add 9.0, 8.0, and 7.0 mL, pressure, at a temperature below 45, add 1 mL of
respectively, of Diluent. Add 1.0 mL of Internal standard chloroform, and mix to dissolve the alkaloids, taking care
solution, then add 15 mL of chloroform, shake vigorously, to wet the sides of the container.
allow the layers to separate, and discard the chloroform Extraction blank: Place 10 mL of Diluent in a 60-mL
layer. separator. Proceed as directed under Sample solution,
[NOTEIf emulsions are formed, a mixed solvent consisting beginning with then add 15 mL of chloroform. The blank
of chloroform-isopropyl alcohol (10:3) may be substituted chromatogram contains no significant interferences at the
for chloroform throughout the procedure.] locus of atropine, scopolamine, or homatropine.
Add another 15 mL of chloroform, and extract again, Chromatographic system
discarding the chloroform phase. Add 15 mL of Phosphate (See Chromatography 621, System Suitability.)
buffer and sufficient 1 N sodium hydroxide to yield a final Mode: GC
pH between 9.0 and 9.5. Add 15 mL of chloroform, shake Detector: Flame ionization
vigorously, and allow the layers to separate. Filter the Column: 1.2-m 4-mm; glass column packed with 3% G3
organic phase through 10 g of anhydrous sodium sulfate on S1AB
previously washed with chloroform and supported in a [NOTEThe column may be cured and conditioned as
funnel with a small pledget of glass wool, into a suitable specified under Chromatography 621, Gas
container. Extract again with two 15-mL portions of Chromatography.]
chloroform, again collecting the clarified organic phase. Temperature
Wash the sodium sulfate and the tip of the funnel with 5 Column: 215
mL of chloroform. Evaporate the combined organic phases Injector port: 240
under reduced pressure, at a temperature below 45, add 1 Detector: 240
mL of chloroform, and mix to dissolve the alkaloids, taking Carrier gas: Dry helium
care to wet the sides of the container. Flow rate: 65 mL/min
Sample solution: Moisten 10 g, previously reduced to a Injection size: 5 L
moderately coarse powder, with a mixture of 8 mL of System suitability
ammonium hydroxide, 10 mL of alcohol, and 20 mL of Sample: Sample solution (610 injections)
ether, and extract the alkaloids by either Method I or Method Suitability requirements
II below. If necessary, reduce the volume of the extract to Resolution: NLT 3.0 between aH and aA (R)
100 mL by evaporation on a steam bath. Tailing factor: NMT 2.0 (the sum of the distances from
Method I: Place the moistened drug in a continuous- peak center to the leading edge and to the tailing edge
extraction thimble, and allow maceration to proceed divided by twice the distance from peak center to the
overnight, then extract with ether for 3 h, or longer if leading edge), measured at 5% of the peak height of aA
necessary to effect complete extraction. Relative standard deviation: The analytical system is
Method II: Place the moistened drug in a small percolator, suitable for conducting this Assay if the relative standard
and allow maceration to proceed overnight. Percolate deviation for the ratio, RA, is NMT 2.0%, calculated as
slowly with a mixture of 3 volumes of ether and 1 volume follows:
of chloroform. Continue the percolation until the residue
from 3 to 4 mL of percolate last passed, when dissolved in (standard deviation/mean ratio) 100
dilute sulfuric acid (1 in 70) and treated with mercuric
iodide TS, shows NMT a faint turbidity. Analysis
Transfer the extract from Method I or Method II to a Samples: Standard solutions and Sample solution
separator with the aid of ether. Extract with five 15-mL Measure the areas, aA, aH, and aS, of the atropine,
portions of dilute sulfuric acid (1 in 70), filtering each homatropine, and scopolamine peaks, respectively, in
portion drawn off into a 100-mL volumetric flask. Wash the each chromatogram, and calculate the ratios AA and AS:
filter with dilute sulfuric acid (1 in 70), and collect the
washings in the flask. Add dilute sulfuric acid (1 in 70) to Result = aA/aH and aS/aH
volume, and mix. Dilute 20.0 mL of the resulting solution
with the same dilute acid to 100.0 mL. Pipet 10 mL of this Plot the standard curves of the values of RA and RS against
solution into a 60-mL separator. Add 1.0 mL of Internal the amounts, in mg, of atropine and scopolamine in the
standard solution, then add 15 mL of chloroform, shake solutions. (The ratio of the molecular weight of atropine
20 Belladonna / Official Monographs USP 32
to that of anhydrous atropine sulfate is 0.8551, and the having three germinal furrows and rows of pits between
ratio of the molecular weight of scopolamine to that of the ridges on the exine.
anhydrous scopolamine hydrobromide is 0.7894.) Inject a Fruit: The epicarp exhibits polygonal epidermal cells with a
portion of the Sample solution into the chromatograph, striated cuticle and stomata. The mesocarp consists of large
obtain the chromatogram area ratios, measure the peak pulp cells some of which contain rosette aggregate crystals
areas, and calculate the area ratios, as with the Standard of calcium oxalate.
solutions. Record from the standard curves the quantities, Seed: The seed is characterized by an epidermis of large,
in mg, of atropine and scopolamine in the weight of the wavy-walled cells with prominent ridges over the anticlinal
specimen taken. Add the quantity, in mg, of atropine and walls.
scopolamine, and multiply by 50 to obtain the weight, in Powdered belladonna leaf: Light olive-brown to moderate
mg, of alkaloids in the portion of Belladonna Leaf taken. olive-green in color. The following are among the elements
Acceptance criteria: NLT 0.35% of the alkaloids of of identification: the separate microcrystals, the dark gray
belladonna leaf crystal cells, the cuticular striping of the epidermal cells, the
vessels with ellipsoidal bordered pits, the fibers of the stem,
IMPURITIES and occasional hairs and pollen grains. Rosette aggregates of
ARTICLES OF BOTANICAL ORIGIN, Acid-Insoluble Ash 561: calcium oxalate and fragments of the seed occur when the
NMT 3.0% drug contains belladonna fruits. Examine Belladonna Leaf for
BELLADONNA STEMS: The proportion of belladonna stems hairs having a papillose cuticle and for raphides of calcium
over 10 mm in diameter does not exceed 3.0%. oxalate: their presence indicates adulteration.
SPECIFIC TESTS ADDITIONAL REQUIREMENTS
BOTANIC CHARACTERISTICS PACKAGING AND STORAGE: Preserve in well-closed containers
Belladonna leaf: Usually partly matted together, crumpled and avoid long exposure to direct sunlight. Preserve
or broken leaves, together with some smaller stems and a powdered Belladonna Leaf in light-resistant containers.
number of flowers and fruits USP REFERENCE STANDARDS 11
The leaves are thin and brittle, mostly light green to USP Atropine Sulfate RS
moderate olive-green. The lamina is mostly from 5 to 25 USP Homatropine Hydrobromide RS
cm in length and from 4 to 12 cm in width and possesses USP Scopolamine Hydrobromide RS
an ovate-lanceolate to broadly ovate outline, an acute to
acuminate apex, an entire margin, an acute to somewhat
decurrent base and slightly hairy surface, the hairs being
more abundant along the veins; when broken transversely, Belladonna Extract
it shows numerous light-colored dots (crystal cells) visible (Comment on this Monograph)id=m7330=Belladonna
with a lens. The petiole is slender and usually up to 4 cm Extract=B-Monos.pdf)
in length. The flowers possess a campanulate corolla with
five small, reflexed lobes, purplish to yellowish purple, DEFINITION
becoming faded to brown or dusky yellow or yellow; a Belladonna Extract contains, in each 100 g, NLT 1.15 g and
green, five-lobed calyx; five epipetalous stamens; and a NMT 1.35 g of the alkaloids of belladonna leaf.
superior, bilocular ovary with numerous ovules. The fruit is Pilular Belladonna Extract
subglobular, dark yellow to yellowish brown to dusky red Prepare the extract by percolating 1000 g of Belladonna Leaf,
or black, up to 12 mm in width, and sometimes subtended using a mixture of 3 volumes of alcohol and 1 volume of
by the persistent calyx and containing numerous flattened, water as the menstruum. Macerate the drug for 16 h, and
somewhat reniform seeds, the latter up to 2 mm in width. then percolate it at a moderate rate. Evaporate the percolate
The stems are more or less flattened and hollow and finely under reduced pressure and at a temperature not exceeding
hairy when young. 60 to a pilular consistency, and adjust the remaining extract,
Histology after assaying, by dilution with liquid glucose so that the
Leaf: The epidermis of the lamina possesses wavy finished Extract will contain, in each 100 g, 1.25 g of the
anticlinal walls and a distinctly striated cuticle. Stomata are alkaloids of belladonna leaf.
more numerous in the lower epidermis and are surrounded Powdered Belladonna Extract
by three or four neighboring cells, one of which is smaller Prepare the extract by percolating 1000 g of Belladonna Leaf,
than the others. The nonglandular hairs are uniseriate and using alcohol as the menstruum. Macerate the drug for 16 h,
up to six-celled. Short club-shaped glandular hairs with a and then percolate it slowly. Evaporate the percolate under
one-celled stalk and multicellular head and long glandular reduced pressure and at a temperature not exceeding 60 to a
hairs with a uniseriate stalk and unicellular head occur on soft extract, add 50 g of dry starch, and continue the
both epidermises. The mesophyll consists of a single layer evaporation, at the same temperature, until the product is dry.
of palisade parenchyma beneath which occurs spongy Powder the residue. The extract may be deprived of its fat by
parenchyma, the latter with scattered cells filled with treating either the soft extract first obtained, or the dry and
microcrystals. The midrib contains an arc of bicollateral powdered extract, as directed under Pharmaceutical Dosage
bundles, collenchyma beneath upper epidermis, and Forms 1151, Extracts. Assay the powdered residue, and add
scattered parenchyma cells with microcrystals. sufficient starch, previously dried at 100, to obtain a finished
Stem: The stem shows an epidermis with striated cuticle Extract containing 1.25 g of the alkaloids of belladonna leaf in
and few hairs; a distinct endodermis; small strands of long, each 100 g. Mix the powders, and pass the Extract through a
thin-walled, slightly lignified pericyclic fibers; and a circle of fine sieve.
bicollateral bundles. The parenchyma of the cortex and
pith is interspersed with crystal cells. ASSAY
Flower: The calyx possesses numerous glandular hairs with PROCEDURE
uniseriate stalks and one- to three-celled glandular heads. Phosphate buffer: 34.8 g of dibasic potassium phosphate
The corolla shows a papillose inner epidermis and an outer in 900 mL of water
epidermis with glandular hairs similar to those of the calyx. Adjust to a pH of 9.5 by the addition of 3 N hydrochloric
The pollen grains, when mounted in chloral hydrate acid or sodium hydroxide, with mixing.
solution, are subspherical, 40 m in diameter, tricolpate,
Diluent: Dilute sulfuric acid (1 in 350) with 5 mL of chloroform. Evaporate the combined organic
Internal standard solution: 0.8 mg/mL of USP phases under reduced pressure, at a temperature below 45,
Homatropine Hydrobromide RS in Diluent add 1 mL of chloroform, and mix to dissolve the alkaloids,
[NOTEPrepare fresh on the day of use.] taking care to wet the sides of the container.
Standard stock solution A: 1.0 mg/mL of USP Chromatographic system
Scopolamine Hydrobromide RS in Diluent (See Chromatography 621, System Suitability.)
Standard stock solution B: Dissolve 20 mg of USP Atropine Mode: GC
Sulfate RS in 25 mL of Diluent in a 50-mL volumetric flask, Detector: Flame ionization
and add 2.0 mL of Standard stock solution A. Add Diluent to Column: 1.2-m 4-mm; glass column packed with 3% G3
volume. on S1AB
[NOTEPrepare fresh on the day of use.] [NOTEThe column may be cured and conditioned as
Standard solutions: Pipet into three separate 60-mL specified under Chromatography 621, Gas
separators 1.0-, 2.0-, and 3.0-mL portions, respectively, of Chromatography.]
Standard solution A, and add 9.0, 8.0, and 7.0 mL, Temperature
respectively, of Diluent. Add 1.0 mL of Internal standard Column: 215
solution, then add 15 mL of chloroform, shake vigorously, Injector port: 240
allow the layers to separate, and discard the chloroform Detector: 240
layer. (If emulsions are formed, a mixed solvent consisting of Carrier gas: Dry helium
chloroform-isopropyl alcohol (10:3) may be substituted for Flow rate: 65 mL/min
chloroform throughout the procedure). Add another 15 mL Injection size: 5 L
of chloroform, and extract again, discarding the chloroform System suitability
phase. Add 15 mL of Phosphate buffer and sufficient 1 N Sample: Standard solutions, six to ten injections
sodium hydroxide to yield a final pH between 9.0 and 9.5. Suitability requirements
Add 15 mL of chloroform, shake vigorously, and allow the Resolution: NLT 3.0 between aH and aA (R)
layers to separate. Filter the organic phase through 10 g of Tailing factor: NMT 2.0 (the sum of the distances from
anhydrous sodium sulfate previously washed with peak center to the leading edge and to the tailing edge
chloroform and supported in a funnel with a small pledget divided by twice the distance from peak center to the
of glass wool, into a suitable container. Extract again with leading edge), measured at 5% of the peak height of aA
two 15-mL portions of chloroform, again collecting the Relative standard deviation: The analytical system is
clarified organic phase. Wash the sodium sulfate and the tip suitable for conducting this assay if the relative standard
of the funnel with 5 mL of chloroform. Evaporate the deviation for the ratio, RA, is NMT 2.0% calculated as
combined organic phases under reduced pressure, at a follows: (standard deviation/mean ratio) 100
temperature below 45, add 1 mL of chloroform, and mix Analysis
to dissolve the alkaloids, taking care to wet the sides of the Samples: Standard solutions and Sample solution
container. Measure the areas, aA, aH, and aS, of the atropine,
Extraction blank: Place 10 mL of Diluent in a 60-mL homatropine, and scopolamine peaks, respectively, in
separator. Proceed as directed under Sample solution, each chromatogram, and calculate the ratios AA and AS:
beginning with then add 15 mL of chloroform. The blank
chromatogram contains no significant interferences at the Result = aA/aH and aS/aH
locus of atropine, scopolamine, or homatropine.
Sample solution: Transfer 0.5 g of Extract to a 125-mL Plot the curves of the Standard solutions of the values of RA
conical flask, and add 40 mL of Diluent. Heat to a and RS versus the amounts, in mg, of atropine and
temperature not above 45, and stir to hasten solution. Filter scopolamine in the solutions. (The ratio of the molecular
the solution through filter paper into a 100-mL volumetric weight of atropine to that of anhydrous atropine sulfate is
flask. Wash the flask and the filter with two 20-mL portions 0.8551, and the ratio of the molecular weight of
of warmed Diluent, and collect the washings in the 100-mL scopolamine to that of anhydrous scopolamine
volumetric flask. Add Diluent to volume. Pipet 10 mL of this hydrobromide is 0.7894.) Inject a portion of the Sample
solution into a 60-mL separator. To the separator, add 1.0 solution into the chromatograph, obtain the
mL of Internal standard solution, then add 15 mL of chromatogram area ratios, measure the peak areas, and
chloroform, shake vigorously, allow the layers to separate, calculate the area ratios, as with the Standard solutions.
and discard the chloroform layer. (If emulsions are formed, a Record from the curves of the Standard solutions the
mixed solvent consisting of chloroform and isopropyl alcohol quantities, in mg, of atropine and scopolamine in the
(10:3) may be substituted for chloroform throughout the volume of specimen taken. Add the quantity, in mg, of
extraction procedure.) Add another 15 mL of chloroform, atropine and scopolamine, and multiply by 10 to obtain
and extract again, discarding the chloroform phase. Add 15 the weight, in mg, of alkaloids in the portion of Extract
mL of Phosphate buffer and sufficient 1 N sodium hydroxide taken.
to yield a final pH between 9.0 and 9.5. Add 15 mL of Acceptance criteria: 1.15 g1.35 g of the alkaloids of
chloroform, shake vigorously, and allow the layers to belladonna leaf/100 g
separate. Filter the organic phase through 10 g of anhydrous
sodium sulfate (see Suitability for alkaloid assays under ADDITIONAL REQUIREMENTS
Reagents, Indicators, and SolutionsSodium Sulfate, PACKAGING AND STORAGE: Preserve in tight containers, at a
Anhydrous), previously washed with chloroform and temperature not exceeding 30.
supported in a funnel with a small pledget of glass wool, USP REFERENCE STANDARDS 11
into a suitable container. Extract again with two 15-mL USP Atropine Sulfate RS
portions of chloroform, again collecting the clarified organic USP Homatropine Hydrobromide RS
phase. Wash the sodium sulfate and the tip of the funnel USP Scopolamine Hydrobromide RS
Belladonna Extract Tablets mL of chloroform, and mix to dissolve the alkaloids, taking
(Comment on this Monograph)id=m7360=Belladonna Extract care to wet the sides of the container.
Tablets=B-Monos.pdf) Extraction blank: Place 10 mL of Diluent in a 60-mL
separator. Proceed as directed under Sample solution,
DEFINITION beginning with then add 15 mL of chloroform. The blank
Belladonna Extract Tablets contain NLT 90.0% and NMT chromatogram contains no significant interferences at the
110.0% of the labeled amount of the alkaloids of belladonna locus of atropine, scopolamine, or homatropine.
leaf. Sample solution: Transfer an equivalent to 600 g of
atropine and scopolamine, from weighed and finely
IDENTIFICATION powdered Tablets (NLT 20), to a 60-mL separator, add 10.0
[NOTEThe Sample to be used in Identification tests A and B is mL of Diluent, and sonicate to dissolve as much as possible
prepared as follows.] of the specimen Add 1.0 mL of Internal standard solution,
Sample: Macerate a quantity of powdered Tablets, equivalent then add 15 mL of chloroform, shake vigorously, allow the
to 5 mg of the alkaloids of belladonna extract, with 20 mL of layers to separate, and discard the chloroform layer.
water, and transfer to a separator. Render the solution alkaline [NOTEIf emulsions are formed, a mixed solvent consisting
with 6 N ammonium hydroxide, and extract the alkaloids with of chloroform and isopropyl alcohol (10:3) may be
50 mL of chloroform. Filter the chloroform layer, divide it into substituted for chloroform throughout the extraction
two equal portions, and evaporate to dryness. procedure.]
A. PROCEDURE Add another 15 mL of chloroform, and extract again,
Analysis: To one portion of the Sample add 2 drops of nitric discarding the chloroform phase. Add 15 mL of Phosphate
acid, evaporate on a steam bath to dryness, and add a few Buffer and sufficient 1 N sodium hydroxide to yield a final
drops of alcoholic potassium hydroxide TS. pH between 9.0 and 9.5. Add 15 mL of chloroform, shake
Acceptance criteria: A violet color is produced. vigorously, and allow the layers to separate. Filter the
B. PROCEDURE organic phase through 10 g of anhydrous sodium sulfate
Analysis: Dissolve the other portion of the Sample in 1 mL (see Suitability for alkaloid assays under Reagents, Indicators,
of dilute hydrochloric acid (1 in 120), and add gold chloride and SolutionsSodium Sulfate, Anhydrous), previously
TS, dropwise with shaking, until a definite precipitate washed with chloroform and supported in a funnel with a
separates. Slowly heat until the precipitate dissolves, and small pledget of glass wool, into a suitable container.
allow the solution to cool. Extract again with two 15-mL portions of chloroform, again
Acceptance criteria: A lusterless precipitate is produced. collecting the clarified organic phase. Wash the sodium
sulfate and the tip of the funnel with 5 mL of chloroform.
ASSAY Evaporate the combined organic phases under reduced
PROCEDURE pressure, at a temperature below 45, add 1 mL of
Phosphate buffer: 34.8 g of dibasic potassium phosphate chloroform, and mix to dissolve the alkaloids, taking care
in 900 mL of water to wet the sides of the container.
Adjust to a pH of 9.5 by the addition of 3 N hydrochloric Chromatographic system
acid or sodium hydroxide, with mixing (See Chromatography 621, System Suitability.)
Diluent: Dilute sulfuric acid (1 in 350) Mode: GC
Internal standard solution: 0.8 mg/mL of USP Detector: Flame ionization
Homatropine Hydrobromide RS in Diluent Column: 1.2-m 4-mm; glass column packed with 3% G3
[NOTEPrepare fresh on the day of use.] on S1AB
Standard stock solution A: 1.0 mg/mL of USP [NOTEThe column may be cured and conditioned as
Scopolamine Hydrobromide RS in Diluent specified under Chromatography 621, Gas
Standard stock solution B: Dissolve 20 mg of USP Atropine Chromatography.]
Sulfate RS in 25 mL of Diluent in a 50-mL volumetric flask, Temperature
and add 2.0 mL of Standard stock solution A. Add Diluent to Column: 215
volume. Injector port: 240
[NOTEPrepare fresh on the day of use.] Detector: 240
Standard solutions: Pipet into three separate 60-mL Carrier gas: Dry helium
separators 1.0-, 2.0-, and 3.0-mL portions, respectively, of Flow rate: 65 mL/min
Standard stock solution A, and add 9.0, 8.0, and 7.0 mL, Injection size: 5 L
respectively, of Diluent. Add 1.0 mL of Internal standard System suitability
solution, then add 15 mL of chloroform, shake vigorously, Sample: Standard solutions for six to ten injections
allow the layers to separate, and discard the chloroform Suitability requirements
layer. Resolution: NLT 3.0 between aH and aA (R)
[NOTEIf emulsions are formed, a mixed solvent consisting Tailing factor: NMT 2.0 (the sum of the distances from
of chloroform-isopropyl alcohol (10:3) may be substituted peak center to the leading edge and to the tailing edge
for chloroform throughout the procedure.] divided by twice the distance from peak center to the
Add another 15 mL of chloroform, and extract again, leading edge), measured at 5% of the peak height of aA
discarding the chloroform phase. Add 15 mL of Phosphate Relative standard deviation: The analytical system is
buffer and sufficient 1 N sodium hydroxide to yield a final suitable for conducting this Assay if the relative standard
pH between 9.0 and 9.5. Add 15 mL of chloroform, shake deviation for the ratio, RA, is NMT 2.0%, calculated by the
vigorously, and allow the layers to separate. Filter the formula: RA = (standard deviation/mean ratio) 100
organic phase through 10 g of anhydrous sodium sulfate Analysis
previously washed with chloroform and supported in a Samples: Standard solutions and Sample solution
funnel with a small pledget of glass wool, into a suitable Measure the areas, aA, aH, and aS, of the atropine,
container. Extract again with two 15-mL portions of homatropine, and scopolamine peaks, respectively, in
chloroform, again collecting the clarified organic phase. each chromatogram, and calculate the ratios AA and AS:
Wash the sodium sulfate and the tip of the funnel with 5
mL of chloroform. Evaporate the combined organic phases Result = aA/aH and aS/aH
under reduced pressure, at a temperature below 45, add 1
Plot the curves of the Standard solutions of the values of RA allow the layers to separate, and discard the chloroform
and RS versus the amounts, in mg, of atropine and layer.
scopolamine in the solutions. (The ratio of the molecular [NOTEIf emulsions are formed, a mixed solvent consisting
weight of atropine to that of anhydrous atropine sulfate is of chloroform-isopropyl alcohol (10:3) may be substituted
0.8551, and the ratio of the molecular weight of for chloroform throughout the procedure.]
scopolamine to that of anhydrous scopolamine Add another 15 mL of chloroform and extract again,
hydrobromide is 0.7894.) Inject a portion of the Sample discarding the chloroform phase. Add 15 mL of Phosphate
solution into the chromatograph, obtain the buffer and sufficient 1 N sodium hydroxide to yield a final
chromatogram area ratios, measure the peak areas, and pH of 9.09.5. Add 15 mL of chloroform, shake vigorously,
calculate the area ratios, as with the Standard solutions. and allow the layers to separate. Filter the organic phase
Record from the curves of the Standard solutions the through 10 g of anhydrous sodium sulfate previously
quantities, in mg, of atropine and scopolamine in the washed with chloroform and supported in a funnel with a
weight of specimen taken. small pledget of glass wool, into a suitable container.
Acceptance criteria: 90.0%110.0% Extract again with two 15-mL portions of chloroform, again
collecting the clarified organic phase. Wash the sodium
PERFORMANCE TESTS sulfate and the tip of the funnel with 5 mL of chloroform.
DISINTEGRATION 701: 30 min Evaporate the combined organic phases under reduced
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements pressure, at a temperature below 45, add 1 mL of
chloroform, and mix to dissolve the alkaloids, taking care
ADDITIONAL REQUIREMENTS to wet the sides of the container.
PACKAGING AND STORAGE: Preserve in tight, light-resistant Extraction blank: Place 10 mL of Diluent in a 60-mL
containers. separator. Proceed as directed under Sample solution,
USP REFERENCE STANDARDS 11 beginning with then add 15 mL of chloroform. The blank
USP Atropine Sulfate RS chromatogram contains no significant interferences at the
USP Homatropine Hydrobromide RS locus of atropine, scopolamine, or homatropine.
USP Scopolamine Hydrobromide RS Sample solution: Moisten 10 g, previously reduced to a
moderately coarse powder with a mixture of 8 mL of
ammonium hydroxide, 10 mL of alcohol, and 20 mL of
Belladonna Tincture ether, and extract the alkaloids by either Method I or Method
II below. If necessary, reduce the volume of the extract to
(Comment on this Monograph)id=m7440=Belladonna 100 mL by evaporation on a steam bath.
Tincture=B-Monos.pdf) Method I: Place the moistened drug in a continuous-
DEFINITION extraction thimble, and allow maceration to proceed
Belladonna Tincture yields, from each 100 mL, NLT 27 mg and overnight, then extract with ether for 3 h, or longer if
NMT 33 mg of the alkaloids of belladonna leaf. necessary to effect complete extraction.
Method II: Place the moistened drug in a small percolator,
and allow maceration to proceed overnight. Percolate
Belladonna Leaf, in moderately coarse powder 100 g slowly with a mixture of 3 volumes of ether and 1 volume
To make 1000 mL of chloroform. Continue the percolation until the residue
from 3 to 4 mL of percolate last passed, when dissolved in
Prepare a tincture by Process P as modified for assayed Tinctures dilute sulfuric acid (1 in 70) and treated with mercuric
(see Pharmaceutical Dosage Forms 1151), using a mixture of 3 iodide TS, shows not more than a faint turbidity.
volumes of alcohol and 1 volume of water as the menstruum. Transfer the extract to a separator with the aid of ether.
Finally adjust the Tincture to contain, in each 100 mL, 30 mg Extract with five 15-mL portions of dilute sulfuric acid (1
of the alkaloids of belladonna leaf. in 70), filtering each portion drawn off into a 100-mL
volumetric flask. Wash the filter with dilute sulfuric acid (1
ASSAY in 70), and collect the washings in the flask. Add dilute
PROCEDURE sulfuric acid (1 in 70) to volume, and mix. Dilute 20.0 mL
Phosphate buffer: 34.8 g of dibasic potassium phosphate of the resulting solution with the same dilute acid to
in 900 mL of water 100.0 mL. Pipet 2 mL of Tincture into a 60-mL separator
Adjust to a pH of 9.5 by the addition of 3 N hydrochloric containing 10 mL of Diluent. Add 1.0 mL of Internal
acid or sodium hydroxide, with mixing. standard solution, then add 15 mL of chloroform, shake
Diluent: Dilute sulfuric acid (1 in 350) vigorously, allow the layers to separate, and discard the
Internal standard solution: 0.8 mg/mL of USP chloroform layer.
Homatropine Hydrobromide RS in Diluent [NOTEIf emulsions are formed, a mixed solvent consisting
[NOTEPrepare fresh on the day of use.] of chloroform-isopropyl alcohol (10:3) may be substituted
Standard stock solution A: 1.0 mg/mL of USP for chloroform throughout the extraction procedure.]
Scopolamine Hydrobromide RS in Diluent Add another 15 mL of chloroform, and extract again,
Standard stock solution B: Dissolve 20 mg of USP Atropine discarding the chloroform phase. Add 15 mL of Phosphate
Sulfate RS in 25 mL of Diluent in a 50-mL volumetric flask. buffer and sufficient 1 N sodium hydroxide to yield a final
Add 2.0 mL of Standard stock solution A. Add Diluent to pH of 9.09.5. Add 15 mL of chloroform, shake
volume. vigorously, and allow the layers to separate. Filter the
[NOTEPrepare fresh on the day of use.] organic phase through 10 g of anhydrous sodium sulfate
Standard solutions: Pipet into three separate 60-mL (see Suitability for alkaloid assays under Reagents,
separators 1.0-, 2.0-, and 3.0-mL portions, respectively, of Indicators, and SolutionsSodium Sulfate, Anhydrous)
Standard stock solution A, and add 9.0, 8.0, and 7.0 mL, previously washed with chloroform and supported in a
respectively, of Diluent. Add 1.0 mL of Internal standard funnel with a small pledget of glass wool, into a suitable
solution, then add 15 mL of chloroform, shake vigorously,
container. Extract again with two 15-mL portions of USP REFERENCE STANDARDS 11
chloroform, again collecting the clarified organic phase. USP Atropine Sulfate RS
Wash the sodium sulfate and the tip of the funnel with 5 USP Homatropine Hydrobromide RS
mL of chloroform. Evaporate the combined organic phases USP Scopolamine Hydrobromide RS
under reduced pressure, at a temperature below 45, add
1 mL of chloroform, and mix to dissolve the alkaloids,
taking care to wet the sides of the container.
Chromatographic system Benazepril Hydrochloride
(See Chromatography 621, System Suitability.) (Comment on this Monograph)id=m7490=Benazepril
Mode: GC Hydrochloride=B-Monos.pdf)
Column: 1.2-m 4-mm; glass column packed with 3% G3
on S1AB
[NOTEThe column may be cured and conditioned as
specified under Chromatography 621, Gas
Chromatography.]
Temperature
Column: 215
Injector port: 240 C24H28N2O5 HCl 460.95
Detector: 240 1H-1-Benzazepine-1-acetic acid, 3-[[1-(ethoxycarbonyl)-3-
Carrier gas: Dry helium phenylpropyl]amino]-2,3,4,5-tetrahydro-2-oxo-,
Flow rate: 65 mL/min monohydrochloride, [S-(R*,R*)]-;
Injection size: 5 L (3S)-3-[[(1S)-1-Carboxy-3-phenylpropyl]amino]-2,3,4,5-
System suitability tetrahydro-2-oxo-1H-1-benzazepine-1-acetic acid, 3-ethyl
Sample: Sample solution for 610 injections ester, monohydrochloride [86541-74-4].
Resolution: NLT 3.0 between aH and aA (R) DEFINITION
Tailing factor NMT 2.0, (the sum of the distances from Benazepril Hydrochloride contains NLT 98.0% and NMT
peak center to the leading edge and to the tailing edge 102.0% of C24H28N2O5 HCl, calculated on the dried basis.
divided by twice the distance from peak center to the IDENTIFICATION
leading edge) measured at 5% of the peak height of aA A. INFRARED ABSORPTION 197M
Relative standard deviation: The analytical system is B. The retention time of the Sample solution corresponds to
suitable for conducting this Assay if the relative standard that of the Standard solution, as obtained in the Assay.
deviation for the ratio, RA, is NMT 2.0%, calculated by the C. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the
formula: requirements
(standard deviation/mean ratio) 100 ASSAY
PROCEDURE
Analysis Solution A: 0.81 g of tetrabutylammonium bromide in 360
Samples: Standard solutions and Sample solution mL of water containing 0.2 mL of glacial acetic acid
Measure the areas, aA, aH, and aS, of the atropine, Mobile phase: Methanol and Solution A (16:9)
homatropine, and scopolamine peaks, respectively, in System suitability solution: 0.4 mg/mL each of USP
each chromatogram, and calculate the ratios AA and AS: Benazepril Hydrochloride RS and USP Benazepril Related
Compound B RS in Mobile phase
Result = aA/aH and aS/aH Standard solution: 0.2 mg/mL of USP Benazepril
Plot the Standard curves of the values of RA and RS against Hydrochloride RS in Mobile phase
the amounts, in mg, of atropine and scopolamine in the Sample solution: Transfer about 10.0 mL of the Sample
solutions. (The ratio of the molecular weight of atropine solution (from either Procedure 1 or Procedure 2), prepared as
to that of anhydrous atropine sulfate is 0.8551, and the directed in the tests for Impurities, Organic Impurities to a 50-
ratio of the molecular weight of scopolamine to that of mL volumetric flask, and dilute with Mobile phase to volume.
anhydrous scopolamine hydrobromide is 0.7894.) Inject a Chromatographic system
portion of the Sample solution into the chromatograph, (See Chromatography 621, System Suitability.)
obtain the chromatogram area ratios, measure the peak Mode: LC
areas, and calculate the area ratios, as with the Standard Detector: UV 240 nm
solutions. Record from the Standard curve the quantities, Column: 3.9-mm 30-cm; packing L1
in mg, of atropine and scopolamine in the specimen. Add Guard column: 4.6-mm 3-cm; packing L1
the quantity, in mg, of atropine and scopolamine, and Flow rate: 1 mL/min
multiply by 50 to obtain the weight, in mg, of alkaloids/ Injection size: 25 L
100 mL. System suitability
Acceptance criteria: 2733 mg of the alkaloids of Sample: System suitability solution
belladonna leaf/100 g Suitability requirements
Resolution: NLT 1.7 between benazepril hydrochloride
OTHER COMPONENTS and benazepril related compound B
ALCOHOL DETERMINATION, Method II 611: 65.0%70.0% of Relative standard deviation: NMT 2.0% for both
C2H5OH, determined by the gas-liquid chromatographic benazepril hydrochloride and benazepril related
procedure, acetone being used as the internal standard compound B
containers, and avoid exposure to direct sunlight and to
excessive heat.
USP 32 Official Monographs / Benazepril 25
Analysis rU = peak response of benazepril related compound

Samples: Sample solution and Standard solution A from the Sample solution
Calculate the percentage of C24H28N2O5 HCl in the portion rS = peak response of benazepril related compound
of Benazepril Hydrochloride taken: A from the Standard solution
CS = concentration of USP Benazepril Related
Result = (rU/rS) (CS/CU) 100 Compound A RS in the Standard solution
(mg/mL)
rU = peak response from the Sample solution CU = nominal concentration of Benazepril
rS = peak response from the Standard solution Hydrochloride in the Sample solution (mg/mL)
CS = concentration of USP Benazepril Hydrochloride Acceptance criteria
RS in the Standard solution (mg/mL) Individual impurities: NMT 0.1%
CU = nominal concentration of benazepril in the PROCEDURE 2: TEST FOR BENAZEPRIL RELATED COMPOUNDS B, C,
Sample solution (mg/mL) D, E, F, and G
Acceptance criteria: 98.0%102.0% Solution A, Mobile phase, System suitability solution,
Chromatographic system and System suitability:
IMPURITIES Proceed as directed in the Assay.
Inorganic Impurities Standard solution: 1 g/mL of USP Benazepril
RESIDUE ON IGNITION 281: Ignite at 600. NMT 0.1% Hydrochloride RS, 10 g/mL each of related compound
residue is found. USP Benazepril Related Compound B RS, USP Benazepril
HEAVY METALS, Method II 231: NMT 10 ppm Related Compound C RS, USP Benazepril Related
Organic Impurities Compound D RS, USP Benazepril Related Compound E RS,
PROCEDURE 1: TEST FOR BENAZEPRIL RELATED COMPOUND A USP Benazepril Related Compound F RS, and USP
Buffer solution: 9.66 mg/mL of monobasic potassium Benazepril Related Compound G RS in Mobile phase
phosphate and 2.68 mg/mL of dibasic sodium phosphate, Sample solution: 1.0 mg/mL of Benazepril Hydrochloride
heptahydrate in Mobile phase
Mobile phase: Methanol and Buffer solution (1:4) Analysis
System suitability solution: 1.0 mg/mL of USP Benazepril Samples: Sample solution and Standard solution
Hydrochloride RS and 0.005 mg/mL of USP Benazepril Calculate the percentage of benazepril related compounds
Related Compound A RS in Mobile phase in the portion of Benazepril Hydrochloride taken:
Standard stock solution: 0.05 mg/mL of USP Benazepril
Related Compound A RS in Mobile phase Result = (rU/rS) (CS/CU) 100
Standard solution A: 5 g/mL of USP Benazepril Related
Compound A in Mobile phase, from the Standard stock rU = peak response for the relevant benazepril related
solution compound from the Sample solution
Standard solution B: 1 g/mL of USP Benazepril Related rS = peak response for the relevant benazepril related
Compound A in Mobile phase, from the Standard stock compound from the Standard solution
solution CS = concentration of relevant USP Reference
Sample solution: 1.0 mg/mL of Benazepril Hydrochloride Standard in the Standard solution (mg/mL)
in Mobile phase CU = nominal concentration of Benazepril
Chromatographic system Hydrochloride in the Sample solution (mg/mL)
(See Chromatography 621, System Suitability.) Acceptance criteria
Mode: LC Individual impurities: See Impurity Table 1
Detector: UV 240 nm [NOTEIn addition to not exceeding the limits for
Column: 4.0-mm 10-cm; packing L41 benazepril related compounds in Impurity Table 1, NMT
Temperature: 30 0.1% of any other single impurity is found.]
Flow rate: 0.9 mL/min For calculating any other single unspecified impurity:
Injection size: 50 L CS = concentration of the USP Benazepril
System suitability Hydrochloride RS in the Standard solution
Samples: System suitability solution, Standard solution A, Total impurities: NMT 2.0% of total impurities
and Standard solution B (excluding benazepril related compound A from Procedure
[NOTEThe relative retention time of benazepril related 1) is found.
compound A1 is about 2.3.]
Impurity Table 1
Resolution: NLT 2.0 between benazepril hydrochloride
and benazepril related compound A Relative Acceptance
Signal-to-noise ratio: NLT 10:1, Standard solution B Retention Criteria,
Relative standard deviation: NMT 10.0%, Standard Name Time NMT (%)
solution A Benazepril Related Compound E1 0.4 0.2
Analysis
Samples: Sample solution and Standard solution Benazepril Related Compound F2 0.5 0.2
Calculate the percentage of benazepril related compound Benazepril Related Compound C3 0.6 0.3
A in the portion of Benazepril Hydrochloride taken: Benazepril Related Compound B4 1.5 0.5
Result = (rU/rS) (CS/CU) 100 Benazepril Related Compound D5 1.7 0.2
1((3R)-3-[[(1R)-1-(ethoxycarbonyl)-3-phenylpropyl]amino]-2,3,4,5-
tetrahydro-2-oxo-1H-1-benzazepine-1-acetic acid, monohydrochloride.
26 Benazepril / Official Monographs USP 32
Impurity Table 1 (continued) B. The retention time of the Sample solution corresponds to
Relative Acceptance
Retention Criteria, ASSAY
Name Time NMT (%) PROCEDURE
Benazepril Related Compound G6 2.0 0.2 Solution A: 0.81 g of tetrabutylammonium bromide in 360
13-Amino-2,3,4,5-tetrahydro-2-oxo-1H-1-(3S)-benzazepine-1-acetic acid. mL of water containing 0.2 mL of glacial acetic acid
2t-Butyl-3-amino-2,3,4,5-tetrahydro-2-oxo-1H-1-(3S)-benzazepine-1-acetic Mobile phase: Methanol and Solution A (16:9)
acid. System suitability solution: 0.4 mg/mL each of USP
33-(1-Carboxy-3-phenyl-(1S)-propyl)amino-2,3,4,5-tetrahydro-2-oxo-1H-1- Benazepril Hydrochloride RS and USP Benazepril Related
(3S)-benzazepine)-1-acetic acid. Compound B RS in Mobile phase
4Mixture of diastereoisomers (3-(1-ethoxycarbonyl-3-phenyl-(1R)- Standard solution: 0.2 mg/mL of USP Benazepril
propyl)amino-2,3,4,5-tetrahydro-2-oxo-1H-1-(3S)-benzazepine)-1-acetic Hydrochloride RS in Mobile phase
acid and (3-(1-ethoxycarbonyl-3-phenyl-(1S)-propyl)amino-2,3,4,5- Sample solution: Finely powder NLT 20 Tablets, and
tetrahydro-2-oxo-1H-1-(3R)-benzazepine)-1-acetic acid. transfer a portion of the powder, equivalent to 50 mg of
53-(1-Ethoxycarbonyl-3-cyclohexyl-(1S)-propyl)amino-2,3,4,5- benazepril hydrochloride, to a 250-mL volumetric flask. Add
tetrahydro-2-oxo-1H-1-(3S)-benzazepine)-1-acetic acid about 150 mL of Mobile phase, and shake by mechanical
monohydrochloride. means for 30 min. Dilute with Mobile phase to volume, mix,
63-(1-Ethoxycarbonyl-3-phenyl-(1S)-propyl)amino-2,3,4,5-tetrahydro-2- and centrifuge. Pass an aliquot of the supernatant through a
oxo-1H-1-(3S)-benzazepine)-1-acetic acid ethyl ester. suitable filter, discarding the first 6 mL of the filtrate.
SPECIFIC TESTS Mode: LC
LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT Detector: UV 240 nm
1.5% of its weight. Column: 3.9-mm 30-cm; packing L1
ABSORBANCE OF SOLUTION: The absorbance of a solution (1 in Guard column: 4.6-mm 3-cm; packing L1
100) of it in methanol, determined in a 1-cm cell at 420 nm, Flow rate: 1 mL/min
is NMT 0.015, methanol being used as the blank. Injection size: 25 L
ABSORPTIVITY System suitability
Sample solution: 0.025 mg/mL of Benazepril Hydrochloride Sample: System suitability solution
in methanol Suitability requirements
Analysis: Proceed as directed under Spectrophotometry and Resolution: NLT 1.7 between benazepril hydrochloride
Light-Scattering 851, and measure the absorbance at 238 and benazepril related compound B
nm. Relative standard deviation: NMT 2.0% for each from
Acceptance criteria: 21.023.2 benazepril hydrochloride and benazepril related
compound B
PACKAGING AND STORAGE: Preserve in well-closed containers, Samples: Standard solution and Sample solution
and store at a temperature below 30, preferably between Calculate the percentage of C24H28N2O5 HCl in the portion
15 and 30. of Tablets taken:
USP Benazepril Hydrochloride RS Result = (rU/rS) (CS/CU) 100
USP Benazepril Related Compound A RS
USP Benazepril Related Compound B RS rU = peak response from the Sample solution
USP Benazepril Related Compound C RS rS = peak response from the Standard solution
USP Benazepril Related Compound D RS CS = concentration of USP Benazepril Hydrochloride
USP Benazepril Related Compound E RS RS in the Standard solution (mg/mL)
USP Benazepril Related Compound F RS CU = nominal concentration of benazepril
USP Benazepril Related Compound G RS hydrochloride in the Sample solution (mg/mL)
PERFORMANCE TESTS
Benazepril Hydrochloride Tablets DISSOLUTION 711
(Comment on this Monograph)id=m7495=Benazepril Medium: Water; 500 mL
Hydrochloride Tablets=B-Monos.pdf) Apparatus 2: 50 rpm
Time: 30 min
DEFINITION Determine the amount of C24H28N2O5 HCl dissolved by
Benazepril Hydrochloride Tablets contain NLT 90.0% and NMT employing the following method.
110.0% of the labeled amount of benazepril hydrochloride Solution A, Mobile phase, System suitability solution,
(C24H28N2O5 HCl). Chromatographic system, and System suitability:
Proceed as directed in the Assay.
IDENTIFICATION Analysis: Inject 60 L, or an amount of a filtered portion of
A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 the solution under test, equivalent to 1.2 g of benazepril,
Sample solution: Equivalent to 1.0 mg/mL of benazepril into the chromatograph. The amount of benazepril injected
hydrochloride from powdered Tablets (NLT 20) in methanol should not exceed 1.5 g. Record the chromatogram, and
[NOTEShake by mechanical means for 15 min. Dilute with measure the responses for the major peaks. Determine the
methanol to volume, mix, and centrifuge. Pass an aliquot percentage of C24H28N2O5 HCl dissolved in comparison with
of the supernatant through a suitable filter, discarding the a Standard solution having a known concentration of USP
first 6 mL of the filtrate.] Benazepril Hydrochloride RS in the same Medium and
Application volume: 20 L similarly chromatographed.
Developing solvent system: Ethyl acetate, methanol, and
ammonium hydroxide (80:20:15)
USP 32 Official Monographs / Bendroflumethiazide 27
Tolerances: NLT 80% (Q) of the labeled amount of Acceptance criteria

C24H28N2O5 HCl Individual impurities: NMT 3.0% of benazepril related
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements compound C is found; NMT 1.0% of any other individual
Analysis for content uniformity impurity is found
Solution A, Mobile phase, System suitability solution, Total impurities: NMT 2.0% (excluding benazepril related
Standard solution, Chromatographic system, and compound C)
System suitability: Proceed as directed in the Assay.
Sample solution: Transfer 1 Tablet to a suitable volumetric ADDITIONAL REQUIREMENTS
flask, add a volume of Mobile phase equivalent to 50% of PACKAGING AND STORAGE: Preserve in well-closed containers.
the volume of the flask, sonicate for 5 min, and then shake USP REFERENCE STANDARDS 11
by mechanical means for NLT 10 min. Dilute quantitatively, USP Benazepril Hydrochloride RS
and stepwise if necessary, with Mobile phase to obtain a USP Benazepril Related Compound B RS
final concentration of nominally 0.2 mg/mL, mix, and pass USP Benazepril Related Compound C RS
a portion of the solution through a suitable filter,
discarding the first 6 mL of the filtrate.
Analysis Bendroflumethiazide
Calculate the percentage of C24H28N2O5 HCl in the Tablet: (Comment on this
Monograph)id=m7520=Bendroflumethiazide=B-Monos.pdf)
CS = concentration of USP Benazepril Hydrochloride
RS in the Standard solution (mg/mL)
CU = nominal concentration of benazepril
C15H14F3N3O4S2 421.42
IMPURITIES 2H-1,2,4-Benzothiadiazine-7-sulfonamide, 3,4-dihydro- 3-
Organic Impurities (phenylmethyl)-6-(trifluoromethyl)-, 1,1-dioxide, ()-;
PROCEDURE ()-3-Benzyl-3,4-dihydro-6-(trifluoromethyl)-2H-1,2,4-
Solution A, Mobile phase, System suitability solution, and benzothiadiazine-7-sulfonamide 1,1-dioxide [73-48-3].
System suitability: Proceed as directed in the Assay.
Standard solution: 0.006 mg/mL of USP Benazepril DEFINITION
Related Compound C RS in Mobile phase Bendroflumethiazide contains NLT 98.0% and NMT 102.0% of
Sample solution: Use the Sample solution as directed in C15H14F3N3O4S2, calculated on the anhydrous basis.
the Assay.
(See Chromatography 621, System Suitability.) A. INFRARED ABSORPTION 197K: Previously dried over silica
Proceed as directed under Assay, except use following gel for 4 h
Injection size. B. ULTRAVIOLET ABSORPTION 197U
Injection size: 80 L Analytical wavelength: 271 nm
Analysis Solution: 10 g/mL in methanol
Samples: Standard solution and Sample solution Acceptance criteria: Absorptivities, calculated on the
Record the chromatograms, and measure the responses of anhydrous basis, do not differ by more than 4.0%.
the peaks for benazepril related compound C. C. PROCEDURE
Calculate the percentage of benazepril related compound Sample solution: Mix 5 mL of dilute hydrochloric acid (1 in
C in the portion of Tablets taken: 2) with 20 mg of Bendroflumethiazide, boil gently for 1
min, and cool in an ice bath.
Result = (rU/rS) (CS/CU) 100 Analysis: To the Sample solution, add in succession, 0.5 mL
of sodium nitrite solution (1 in 1000), 0.5 mL of ammonium
rU = peak response from the Sample solution sulfamate solution (1 in 200), and 0.5 mL of N-(1-
rS = peak response from the Standard solution naphthyl)ethylenediamine dihydrochloride solution (1 in
CS = concentration of USP Benazepril Related 1000).
Compound C RS in the Standard solution Acceptance criteria: A deep red color is produced.
(mg/mL)
CU = nominal concentration of benazepril ASSAY
hydrochloride in the Sample solution (mg/mL) PROCEDURE
Calculate the percentage of each impurity (other than Sample: 190 mg of Bendroflumethiazide
benazepril related compound C) in the portion of Analysis: Dissolve the Sample in 80 mL of pyridine in a 250-
Tablets: mL tall-form beaker in a well-ventilated hood. Add 3 drops
of a saturated solution of azo-violet in methanol, cover the
Result = (ru/rT) 100 beaker, and gently bubble nitrogen through the solution for
5 min, being careful to avoid any contact between the
ru = peak response for each impurity from the solution and the cover. Raise the nitrogen delivery tube
Sample solution above the solution surface and, maintaining a gentle
rT = sum of the responses of all the peaks (including flushing with nitrogen and stirring with a magnetic or
benazepril related compound C) from the mechanical stirring device, add 0.1 N sodium methoxide VS
Sample solution from a 10-mL buret inserted through an opening in the
28 Bendroflumethiazide / Official Monographs USP 32
cover. Titrate to a blue endpoint, approaching the endpoint USP 2,4-Disulfamyl-5-trifluoromethylaniline RS

at a rate of 1 or 2 drops/s. Perform a blank determination,
and make any necessary correction (see Titrimetry 541).
Each mL of 0.1 N sodium methoxide is equivalent to 21.07
mg of C15H14F3N3O4S2. Bendroflumethiazide Tablets
Acceptance criteria: 98.0%102.0% (Comment on this Monograph)id=m7550=Bendroflumethiazide
Tablets=B-Monos.pdf)
IMPURITIES
RESIDUE ON IGNITION 281: NMT 0.2% Bendroflumethiazide Tablets contain NLT 90.0% and NMT
SELENIUM 291: The absorbance from the Sample solution 110.0% of the labeled amount of C15H14F3N3O4S2.
prepared with 100 mg of Bendroflumethiazide and 100 mg
of magnesium oxide, is NMT one-half that from the IDENTIFICATION
Standard solution (NMT 30 ppm). The retention time of the major peak of the Sample solution
HEAVY METALS, Method II 231: NMT 20 ppm corresponds to that of the Standard solution, as obtained in
Organic Impurities the Assay.
LIMIT OF 2,4-DISULFAMYL-5-TRIFLUOROMETHYLANILINE
[NOTEUse low-actinic glassware for the Sample solution and ASSAY
the Standard solution.] PROCEDURE
Mobile phase: Dissolve 5.62 g of sodium chloride and [NOTEUse low-actinic glassware for the Sample solution and
1.97 g of anhydrous sodium sulfate in 1000 mL of water in the Standard solution.]
a 2-L volumetric flask. Add 4.0 mL of glacial acetic acid and Mobile phase: Dissolve 5.62 g of sodium chloride and 1.97
800 mL of methanol, and dilute with water to volume. g of anhydrous sodium sulfate in 1000 mL of water in a 2-L
Standard solution: 0.75 g/mL of USP 2,4-Disulfamyl-5- volumetric flask. Add 4.0 mL of glacial acetic acid and 800
trifluoromethylaniline RS in methanol mL of methanol, and dilute with water to volume.
Sample solution: 50 g/mL of Bendroflumethiazide in Standard solution: 50 g/mL of USP Bendroflumethiazide
methanol RS in methanol
Chromatographic system Sample solution: Weigh and finely powder NLT 20 Tablets.
(See Chromatography 621, System Suitability.) Transfer a portion of the powder, nominally equivalent to
Mode: LC about 5 mg of bendroflumethiazide, to a 100-mL volumetric
Detector: UV 270 nm flask, add about 70 mL of methanol, and sonicate for 15
Column: 4.6-mm 30-cm; packing L11 min, with occasional shaking. Dilute with methanol to
Temperature: 35 5 volume, mix, and centrifuge a portion of the solution for 15
Flow rate: 1.5 mL/min min.
Injection size: 20 L Chromatographic system
System suitability (See Chromatography 621, System Suitability.)
Sample: Standard solution Mode: LC
Suitability requirements Detector: UV 270 nm
Resolution: NLT 1.4 between the methanol and 2,4- Column: 4.6-mm 30-cm; packing L11
disulfamyl-5-trifluoromethylaniline Temperature: 35 5
Relative standard deviation: NMT 3.0% for five Flow rate: 1.5 mL/min
replicate injections Injection size: 20 L
Analysis System suitability
Samples: Sample solution and Standard solution Sample: Standard solution
Calculate the percentage of 2,4-disulfamyl-5- Suitability requirements
trifluoromethylaniline in the portion of Tailing factor: NMT 2.0
Bendroflumethiazide taken: Relative standard deviation: NMT 3.0% for five replicate
injections
Result = (rU/rS) (CS/CU) 100 Analysis
rU = peak response from the Sample solution Calculate the percentage of C15H14F3N3O4S2 in the portion
rS = peak response from the Standard solution of Tablets taken:
CS = concentration of USP 2,4-Disulfamyl-5-
trifluoromethylaniline RS in the Standard Result = (rU/rS) (CS/CU) 100
solution (g/mL)
CU = concentration of bendroflumethiazide in the rU = peak response from the Sample solution
Sample solution (g/mL) rS = peak response from the Standard solution
Acceptance criteria: NMT 1.5% CS = concentration of USP Bendroflumethiazide RS in
the Standard solution (g/mL)
SPECIFIC TESTS CU = nominal concentration of bendroflumethiazide in
WATER DETERMINATION, Method I 921: NMT 0.5% the Sample solution (mg/mL)
USP Bendroflumethiazide RS
USP 32 Official Monographs / Benoxinate 29

PERFORMANCE TESTS IMPURITIES
DISSOLUTION 711 Inorganic Impurities
[NOTEProtect solutions from light throughout this test.] RESIDUE ON IGNITION 281: NMT 0.2%
Medium: 0.01 N hydrochloric acid; 900 mL Organic Impurities
Apparatus 2: 50 rpm ORDINARY IMPURITIES 466
Time: 45 min Sample solution: Methanol
Detector: UV 271 nm Standard solution: Methanol
Sample solutions: Sample per Dissolution 711 Eluant: A mixture of chloroform, cyclohexane, and
Standard solution: USP Bendroflumethiazide RS in Medium diethylamine (5:4:1)
Analysis: Determine the amount of C15H14F3N3O4S2 Visualization: 12
dissolved by employing UV absorption on filtered portions of
the Sample solution, suitably diluted with water, if necessary, SPECIFIC TESTS
in comparison with a Standard solution having a known PH 791: 5.06.0, in a solution (1 in 100)
concentration of USP Bendroflumethiazide RS. LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT
Tolerances: NLT 75% (Q) of the labeled amount of 1.0% of its weight.
C15H14F3N3O4S2
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS
PACKAGING AND STORAGE: Preserve in tight containers. USP Benoxinate Hydrochloride RS
USP Bendroflumethiazide RS
Benoxinate Hydrochloride Ophthalmic
Solution
Benoxinate Hydrochloride (Comment on this Monograph)id=m7630=Benoxinate
(Comment on this Monograph)id=m7600=Benoxinate Hydrochloride Ophthalmic Solution=B-Monos.pdf)
Hydrochloride=B-Monos.pdf)
DEFINITION
Benoxinate Hydrochloride Ophthalmic Solution is a sterile
solution of Benoxinate Hydrochloride in water. It contains NLT
95.0% and NMT 105.0% of the labeled amount of C17H28N2O3
HCl.
IDENTIFICATION
IDENTIFICATIONORGANIC NITROGENOUS BASES 181
C17H28N2O3 HCl 344.88 Sample solution: Equivalent to 50 mg of benoxinate
Benzoic acid, 4-amino-3-butoxy-, 2-(diethylamino)ethyl ester, hydrochloride from a volume of Solution in 25 mL 0.01 N
monohydrochloride; hydrochloric acid
2-(Diethylamino)ethyl 4-amino-3-butoxybenzoate Analysis: Proceed as directed under IdentificationOrganic
monohydrochloride [5987-82-6]. Nitrogenous Bases 181, beginning with Transfer the liquid
to a separator.
DEFINITION Acceptance criteria: The solution meets the requirements
Benoxinate Hydrochloride contains NLT 98.5% and NMT of the test.
101.5% of C17H28N2O3 HCl, calculated on the dried basis.
ASSAY
IDENTIFICATION PROCEDURE
A. INFRARED ABSORPTION 197K: Previously dried Standard solution: 400 g/mL of USP Benoxinate
B. ULTRAVIOLET ABSORPTION 197U Hydrochloride RS in 0.1 N hydrochloric acid
Sample solution: 15 g/mL in water Sample solution: Transfer the equivalent of 20 mg of
C. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the benoxinate hydrochloride from a volume of Ophthalmic
requirements of the tests Solution, to a separator containing 15 mL of water, add 1
Sample solution: 10 mg/mL mL of ammonium hydroxide, and extract with five 20-mL
portions of ether. Wash the combined ether extracts with 10
ASSAY mL of water, extract the water washing with 10 mL of ether,
PROCEDURE and add this ether extract to the main extract. Extract the
Sample solution: Dissolve 250 mg of Benoxinate ether solution with three 5-mL portions of 0.1 N
Hydrochloride in a mixture of 20 mL of glacial acetic acid hydrochloric acid, collect the acid extracts in a 50-mL
and 20 mL of acetic anhydride. volumetric flask, and dilute with 0.1 N hydrochloric acid to
Analysis: Titrate with 0.1 N perchloric acid VS, determining volume.
the endpoint potentiometrically. Perform a blank Blank: 0.1 N hydrochloric acid
determination, and make any necessary correction (see Spectrometric conditions
Titrimetry 541). Each mL of 0.1 N perchloric acid is (See Spectrophotometry and Light-Scattering 851.)
equivalent to 34.49 mg of C17H28N2O3 HCl.
30 Benoxinate / Official Monographs USP 32
Mode: UV mixture for 10 min. Cool, add 0.2 g of sodium nitrite to 1

Analytical wavelength: 308 nm mL of the clear liquid, and add this mixture to 20 mg of
Cell: 1 cm naphthol dipotassium disulfonate or naphthol disodium
Analysis disulfonate in 1 mL of ammonium hydroxide.
Samples: Blank, Sample solution, and Standard solution Acceptance criteria: The solution turns orange-red, and a
Transfer 5.0 mL each of the Standard solution, Sample brown precipitate may be formed.
solution, and Blank, to separate 200-mL volumetric flasks.
Dilute the contents of each flask with water to volume. ASSAY
Concomitantly determine the absorbances of the PROCEDURE
solutions, using the Blank to set the instrument. Sample: 0.3 g of Benzethonium Chloride
Calculate the percentage of C17H28N2O3 HCl in each mL of Analysis: Dissolve the Sample in 75 mL of water contained
the Ophthalmic Solution: in a glass-stoppered, 250-mL flask. Add 0.4 mL of
bromophenol blue solution (1 in 2000), 10 mL of
Result = (AU/AS) (CS/CU) 100 chloroform, and 1 mL of 1 N sodium hydroxide. Titrate with
0.02 M sodium tetraphenylboron VS until the blue color
AU = absorbance from the Sample solution disappears from the chloroform layer. Add the last portions
AS = absorbance from the Standard solution of the sodium tetraphenylboron solution dropwise, agitating
CS = concentration of USP Benoxinate Hydrochloride vigorously after each addition. Each mL of 0.02 M sodium
RS in the Standard solution (g/mL) tetraphenylboron is equivalent to 8.962 mg of C27H42ClNO2.
CU = nominal concentration of benoxinate Acceptance criteria: 97.0%103.0%
SPECIFIC TESTS RESIDUE ON IGNITION 281: NMT 0.1%
STERILITY TESTS 71: Meets the requirements Organic Impurities
PH 791: 3.06.0 LIMIT OF AMMONIUM COMPOUNDS: To 5 mL of a solution (1 in
50), add 3 mL of 1 N sodium hydroxide, and heat to
ADDITIONAL REQUIREMENTS boiling: the odor of ammonia is not perceptible.
USP REFERENCE STANDARDS 11 SPECIFIC TESTS
USP Benoxinate Hydrochloride RS MELTING RANGE OR TEMPERATURE 741: 158163, the
specimen having been dried previously
LOSS ON DRYING 731: Dry at 105 for 4 h: it loses NMT
5.0% of its weight.
Benzethonium Chloride
(Comment on this Monograph)id=m7960=Benzethonium ADDITIONAL REQUIREMENTS
Chloride=B-Monos.pdf) PACKAGING AND STORAGE: Preserve in tight, light-resistant
containers.
Benzethonium Chloride Concentrate

(Comment on this Monograph)id=m7965=Benzethonium
Chloride Concentrate=B-Monos.pdf)
C27H42ClNO2 448.08 DEFINITION

Benzenemethanaminium, N,N-dimethyl-N-[2-[2-[4-(1,1,3,3- Benzethonium Chloride Concentrate contains NLT 94.0% and
tetramethylbutyl)phenoxy]ethoxy]ethyl]-, chloride; NMT 106.0% of the labeled amount of benzethonium chloride
Benzyldimethyl[2-[2-[p-(1,1,3,3- (C27H42ClNO2).
tetramethylbutyl)phenoxy]ethoxy]ethyl]ammonium chloride
[121-54-0]. IDENTIFICATION
A. PROCEDURE
DEFINITION Sample solution: Evaporate a volume of Concentrate,
Benzethonium Chloride contains NLT 97.0% and NMT 103.0% equivalent to 200 mg of benzethonium chloride, on a steam
of C27H42ClNO2, calculated on the dried basis. bath.
Analysis: To the residue, add 2 mL of alcohol, 0.5 mL of 2
IDENTIFICATION N nitric acid, and 1 mL of silver nitrate TS.
A. PROCEDURE Acceptance criteria: A white precipitate, which is insoluble
Sample solution: 10 mg/mL in 2 N nitric acid but soluble in 6 N ammonium hydroxide,
Analysis: To 1 mL of the Sample solution, add 2 mL of is formed.
alcohol, 0.5 mL of 2 N nitric acid, and 1 mL of silver nitrate B. PROCEDURE: Evaporate a volume of Concentrate,
TS. equivalent to 200 mg of benzethonium chloride, on a steam
Acceptance criteria: A white precipitate, which is insoluble bath.
in 2 N nitric acid but soluble in 6 N ammonium hydroxide, Acceptance criteria: The residue so obtained forms
is formed. precipitates with 2 N nitric acid and with mercuric chloride
B. A solution (1 in 100) forms precipitates with 2 N nitric TS, both of which dissolve upon the addition of alcohol.
acid and with mercuric chloride TS, both of which dissolve C. PROCEDURE
upon the addition of alcohol. Sample solution: Evaporate a volume of Concentrate,
C. PROCEDURE equivalent to 200 mg of benzethonium chloride, on a steam
Sample solution: 0.1 g in 1 mL of sulfuric acid bath.
Analysis: Add 0.1 g of potassium nitrate, and heat on a Analysis: To the residue, add 0.1 g of potassium nitrate,
steam bath for 3 min. Cautiously dilute the solution with and heat on a steam bath for 3 min. Cautiously dilute the
water to 10 mL, add 0.5 g of granulated zinc, and warm the solution with water to 10 mL, add 0.5 g of granulated zinc,
USP 32 Official Monographs / Benzethonium 31
and warm the mixture for 10 min. Cool, add 0.2 g of Acceptance criteria: The residue so obtained, forms
sodium nitrite to 1 mL of the clear liquid, and add this precipitates with 2 N nitric acid and with mercuric chloride
mixture to 20 mg of naphthol dipotassium disulfonate or TS, both of which dissolve upon the addition of alcohol.
naphthol disodium disulfonate in 1 mL of ammonium C. PROCEDURE
hydroxide. Sample solution: Evaporate a volume of Topical Solution,
Acceptance criteria: The solution turns orange-red, and a equivalent to 200 mg of benzethonium chloride, on a steam
brown precipitate may be formed. bath.
Analysis: To the residue, add 0.1 g of potassium nitrate,
ASSAY and heat on a steam bath for 3 min. Cautiously dilute the
PROCEDURE solution with water to 10 mL, add 0.5 g of granulated zinc,
Sample solution: Equivalent to 200 mg of benzethonium and warm the mixture for 10 min. Cool, add 0.2 g of
chloride from a volume of Concentrate, in a glass-stoppered sodium nitrite to 1 mL of the clear liquid, and add this
flask mixture to 20 mg of naphthol dipotassium disulfonate or
Analysis: Add 0.4 mL of bromophenol blue solution (1 in naphthol disodium disulfonate in 1 mL of ammonium
2000), 10 mL of chloroform, and 1 mL of 1 N sodium hydroxide.
hydroxide. Acceptance criteria: The solution turns orange-red, and a
Titrate with 0.02 M sodium tetraphenylboron VS until the brown precipitate may be formed.
blue color disappears from the chloroform layer. Add the
last portions of the sodium tetraphenylboron solution ASSAY
dropwise, agitating vigorously after each addition. Each mL PROCEDURE
of 0.02 M sodium tetraphenylboron is equivalent to 8.962 Sample solution: Equivalent to 200 mg of benzethonium
mg of C27H42ClNO2. chloride from a volume of Topical Solution, in a glass-
Acceptance criteria: 94.0%106.0% stoppered flask.
Analysis: Add 0.4 mL of bromophenol blue solution (1 in
IMPURITIES 2000), 10 mL of chloroform, and 1 mL of 1 N sodium
Inorganic Impurities hydroxide.
LIMIT OF NITRITES: To 1 drop of Concentrate on a spot plate, Titrate with 0.02 M sodium tetraphenylboron VS until the
add 1 drop each of glacial acetic acid, sulfanilic acid in blue color disappears from the chloroform layer. Add the
acetic acid solution (1 in 100), and 1-naphthylamine-acetic last portions of the sodium tetraphenylboron solution
acid solution (prepared by boiling 30 mg of 1- dropwise, agitating vigorously after each addition. Each mL
naphthylamine in 70 mL of water, decanting the colorless of 0.02 M sodium tetraphenylboron is equivalent to 8.962
solution from the blue-violet residue, and mixing with 30 mg of C27H42ClNO2.
mL of glacial acetic acid): no red color develops in the Acceptance criteria: 95.0%105.0%
resulting solution within 10 min.
IMPURITIES
OXIDIZING SUBSTANCES: To 5 mL of Concentrate, add 0.5 mL LIMIT OF NITRITES: To 1 drop of Topical Solution on a spot
of potassium iodide TS and a few drops of 3 N hydrochloric plate, add 1 drop each of glacial acetic acid, sulfanilic acid
acid: the solution does not acquire a yellow color. in acetic acid (1 in 100), and 1-naphthylamine-acetic acid
solution (prepared by boiling 30 mg of 1-naphthylamine in
ADDITIONAL REQUIREMENTS 70 mL of water, decanting the colorless solution from the
PACKAGING AND STORAGE: Preserve in tight, light-resistant blue-violet residue, and mixing with 30 mL of glacial acetic
containers. Store at room temperature. acid): no red color develops in the resulting solution within
LABELING: The label states that this article is not intended for 10 min.
direct administration to humans or animals.
SPECIFIC TESTS
OXIDIZING SUBSTANCES: To 5 mL, add 0.5 mL of potassium
Benzethonium Chloride Topical iodide TS and a few drops of 3 N hydrochloric acid: the
solution does not acquire a yellow color.
Solution
(Comment on this Monograph)id=m7990=Benzethonium ADDITIONAL REQUIREMENTS
Chloride Topical Solution=B-Monos.pdf) PACKAGING AND STORAGE: Preserve in tight, light-resistant
containers.
DEFINITION
Benzethonium Chloride Topical Solution contains NLT 95.0%
and NMT 105.0% of the labeled amount of benzethonium
chloride (C27H42ClNO2). Benzethonium Chloride Tincture
(Comment on this Monograph)id=m8000=Benzethonium
IDENTIFICATION Chloride Tincture=B-Monos.pdf)
A. PROCEDURE
Sample solution: Evaporate a volume of Topical Solution, DEFINITION
equivalent to 200 mg of benzethonium chloride, on a steam Benzethonium Chloride Tincture contains, in each 100 mL, NLT
bath. 190 mg and NMT 210 mg of C27H42ClNO2.
Analysis: To the residue, add 2 mL of alcohol, 0.5 mL of 2 Benzethonium Chloride Tincture is prepared as follows.
N nitric acid, and 1 mL of silver nitrate TS.
Acceptance criteria: A white precipitate, which is insoluble Benzethonium Chloride 2g
in 2 N nitric acid but soluble in 6 N ammonium hydroxide,
is formed. Alcohol 685 mL
B. Evaporate a volume of Topical Solution, equivalent to
200 mg of benzethonium chloride, on a steam bath.
32 Benzethonium / Official Monographs USP 32
Acetone 100 mL Analysis: From the respective chromatograms obtained as

Purified Water a sufficient quantity described previously, calculate the ratios of peak areas for
alcohol to internal standard and for acetone to internal
To make 1000 mL standard.
Calculate the percentage of alcohol and of acetone in the
Dissolve the Benzethonium Chloride in a mixture of the Alcohol portion of Tincture taken:
and the Acetone. Add sufficient Purified Water to make 1000
mL. Result = [A(Y Z) + B(Z X)]/(Y X)
[NOTEBenzethonium Chloride Tincture may be colored by the
addition of any suitable color or combination of colors certified A = percentage of alcohol, or of acetone, in the
by the FDA for use in drugs.] lower standards
B = percentage of alcohol, or of acetone, in the
IDENTIFICATION higher standards
A. PROCEDURE X = ratio of the alcohol peak areas, or the acetone
Sample: 50 mL peak areas, to the internal standard peak areas
Analysis: To the residue obtained by evaporating the for the lower standard
Sample on a steam bath, add 2 mL of alcohol, 0.5 mL of 2 Y = ratio of the alcohol peak areas, or the acetone
N nitric acid, and 1 mL of silver nitrate TS. peak areas, to the internal standard peak areas
Acceptance criteria: A white precipitate, which is insoluble for the higher standard
in 2 N nitric acid but soluble in 6 N ammonium hydroxide, Z = ratio of the alcohol peak areas, or the acetone
is formed. peak areas, to the internal standard peak areas
B. Evaporate 50 mL on a steam bath. for the Sample solution
Acceptance criteria: The residue obtained forms Acceptance criteria
precipitates with 2 N nitric acid and with mercuric chloride C2H5OH: 62.0%68.0%
TS, both of which dissolve upon the addition of alcohol. C3H6O: 9.0%11.0%
PROCEDURE SPECIFIC GRAVITY 841: 0.8680.876
Sample solution: Place 50.0 mL of Tincture in a 150-mL
beaker, and add, with continuous stirring, 10 mL of sodium ADDITIONAL REQUIREMENTS
tetraphenylboron solution (1 in 40). Cover, and allow to PACKAGING AND STORAGE: Preserve in tight, light-resistant
stand for 16 h. Decant the supernatant into a tared sintered- containers.
glass crucible, applying vacuum filtration. Suspend the
precipitate in 20 mL of water.
Analysis: Transfer the precipitate to the crucible, washing
well with water. Dry the precipitate and the crucible at 105 Benzocaine
for 1 h, cool, and weigh. The weight of the precipitate so (Comment on this Monograph)id=m8050=Benzocaine=B-
obtained, multiplied by 0.6122, represents its equivalent of Monos.pdf)
C27H42ClNO2.
Acceptance criteria: 190210 mg
OTHER COMPONENTS
ALCOHOL AND ACETONE CONTENT
Standard solution A: Dehydrated alcohol, methanol as the
internal standard, and water (11:5:84)
Standard solution B: Dehydrated alcohol, methanol as the
internal standard, and water (14:5:81) C9H11NO2 165.19
Standard solution C: Acetone, methanol as the internal Benzoic acid, 4-amino-, ethyl ester;
standard, and water (1.7:5:93.3) Ethyl p-aminobenzoate [94-09-7].
Standard solution D: Acetone, methanol as the internal
standard, and water (2.2:5:92.8) DEFINITION
Sample solution: Benzethonium Chloride Tincture, Benzocaine, dried over phosphorus pentoxide for 3 h, contains
methanol as the internal standard, and water (4:1:15) NLT 98.0% and NMT 102.0% of C9H11NO2.
(See Chromatography 621, System Suitability.) IDENTIFICATION
Mode: GC A. INFRARED ABSORPTION 197K: Previously dried over
Detector: Flame ionization phosphorus pentoxide for 3 h
Column: 120-cm 4-mm column packed with a suitable B. ULTRAVIOLET ABSORPTION 197U
type of support, such as 80- to 100-mesh S3 Wavelength: 278 nm
Carrier gas: Dry helium Solution: 5 g/mL in chloroform
Temperatures Acceptance criteria: Absorptivities, calculated on the dried
Column: 120 basis, do not differ by more than 3.0%.
Injection port: 240 C. PROCEDURE
Detector block: 240 Sample solution: 20 mg in 10 mL of water with the aid of
Flow rate: 90 mL/min a few drops of 3 N hydrochloric acid
Injection size: 0.8 L Analysis: Add 5 drops of a solution of sodium nitrite (1 in
Analysis 10), followed by 2 mL of a solution of 100 mg of 2-
Samples: Standard solutions A, B, C, and D and Sample naphthol in 5 mL of 1 N sodium hydroxide.
solution
USP 32 Official Monographs / Benzocaine 33
Acceptance criteria: An orange-red precipitate is formed. ADDITIONAL REQUIREMENTS

PROCEDURE USP Benzocaine RS
Solution A: Acetic acid, triethylamine, and water (20:1:980)
The pH should be between 2.953.0 (adjust as needed).
Mobile phase: Methanol and Solution A (2:3)
Standard solution: 0.024 mg/mL of USP Benzocaine RS in Benzocaine Topical Aerosol
Mobile phase (Comment on this Monograph)id=m8060=Benzocaine Topical
Sample solution: 0.024 mg/mL of Benzocaine in Mobile Aerosol=B-Monos.pdf)
phase
(See Chromatography 621, System Suitability.) Benzocaine Topical Aerosol is a solution of Benzocaine in a
Mode: LC pressurized container. It contains NLT 90.0% and NMT
Detector: UV 285 nm 110.0% of the labeled amount of C9H11NO2.
System suitability Sample solution: Spray a portion of Topical Aerosol,
Sample: Standard solution equivalent to 5 mg of benzocaine, into a beaker, and heat
Suitability requirements on a steam bath for a few min to expel residual propellant.
Tailing factor: NMT 2.0 Add 20 mL of 0.5 N hydrochloric acid, warm gently to
Relative standard deviation: NMT 2.0% disperse, cool, and filter.
Analysis: Analysis: To 10 mL of the filtrate, add 5 drops of sodium
Samples: Standard solution and Sample solution nitrite solution (1 in 10) and 2 drops of methyl red TS, and
Calculate the percentage of C9H11NO2 in the portion taken: neutralize with 1 N sodium hydroxide. Add 2 mL of a
solution of 100 mg of 2-naphthol in 5 mL of 1 N sodium
Result = (rU/rS) (CS/CU) 100 hydroxide.
Acceptance criteria: An orange-red precipitate is formed.
CS = concentration of USP Benzocaine RS in the PROCEDURE
Standard solution (mg/mL) Sample solution: Spray a portion of the Topical Aerosol
CU = concentration of the Sample solution (mg/mL) into a beaker, and heat on a steam bath for a few minutes
Acceptance criteria: 98.0%102.0% to expel residual propellant. Cool, and transfer a portion of
the solution, equivalent to 200 mg of benzocaine, to a 250-
IMPURITIES mL beaker. Add 50 mL of water and 5 mL of hydrochloric
Inorganic Impurities acid, and stir. Cool the solution in an ice bath to 10.
RESIDUE ON IGNITION 281: NMT 0.1% Analysis: Titrate slowly with 0.1 M sodium nitrite VS, using
CHLORIDE: To a solution of 200 mg in 5 mL of alcohol, a calomelplatinum electrode system. Perform a blank
previously acidified with a few drops of diluted nitric acid, determination, and make any necessary correction (see
add a few drops of silver nitrate TS: no turbidity is produced Titrimetry 541). Each mL of 0.1 M sodium nitrite is
immediately. equivalent to 16.52 mg of C9H11NO2.
HEAVY METALS, Method II 231: NMT 10 ppm Acceptance criteria: 90.0%110.0%
Organic Impurities
ORDINARY IMPURITIES 466 SPECIFIC TESTS
Sample solution: Dehydrated alcohol OTHER REQUIREMENTS: It meets the requirements for Aerosols,
Standard solution: Dehydrated alcohol Nasal Sprays, Metered-Dose Inhalers, and Dry Powder Inhalers
Application volume: 10 L 601, Pressure Test, Minimum Fill, and Leakage Test.
Eluant: Chloroform containing about 0.75% of dehydrated
alcohol as a preservative, in a nonequilibrated chamber ADDITIONAL REQUIREMENTS
Visualization: 1 PACKAGING AND STORAGE: Preserve in pressurized containers,
Limit: The total of any ordinary impurities observed does and avoid exposure to excessive heat.
not exceed 1%.
READILY CARBONIZABLE SUBSTANCES TEST 271: Dissolve 500
mg in 5 mL of sulfuric acid TS: the solution has no more Benzocaine Cream
color than Matching Fluid A.
(Comment on this Monograph)id=m8070=Benzocaine
SPECIFIC TESTS Cream=B-Monos.pdf)
MELTING RANGE OR TEMPERATURE, Class I 741: 8892, but
the range between beginning and end of melting does not DEFINITION
exceed 2 Benzocaine Cream contains NLT 90.0% and NMT 110.0% of
REACTION: Dissolve 1.0 g in 10 mL of neutralized alcohol: a the labeled amount of C9H11NO2 in a suitable cream base.
clear solution results. Dilute this solution with 10 mL of IDENTIFICATION
water, and add 2 drops of phenolphthalein TS and 1 drop of PROCEDURE
0.10 N sodium hydroxide: a red color is produced. Sample solution: Place an amount equivalent to 50 mg of
LOSS ON DRYING 731: Dry over phosphorus pentoxide for 3 benzocaine from Cream in a small beaker, add 20 mL of 0.5
h: it loses NMT 1.0% of its weight.
34 Benzocaine / Official Monographs USP 32
N hydrochloric acid, warm gently to disperse the Cream, filtrate. Transfer 1.0 mL of the clear solution to a second
cool, and filter. 100-mL volumetric flask, and dilute with Diluent to volume.
Analysis: To 10 mL of the filtrate add 5 drops of a solution Chromatographic system
of sodium nitrite (1 in 10) and 2 drops of methyl red TS, (See Chromatography 621, System Suitability.)
and neutralize with 1 N sodium hydroxide. Add 2 mL of a Mode: LC
20 mg/mL solution of 2-naphthol in 1 N sodium hydroxide. Detector: UV 294 nm
Acceptance criteria: An orange-red precipitate is formed. Column: 3.9-mm 30-cm; packing L1
ASSAY Injection size: 50 L
PROCEDURE System suitability
Sample solution: An amount of Cream, nominally Sample: Standard solution
equivalent to 200 mg of benzocaine in a tared 250-mL Suitability requirements
beaker Relative standard deviation: NMT 2.0%
Analysis: Add 50 mL of water and 5 mL of hydrochloric Analysis
acid, and stir by mechanical means, with gentle warming, Samples: Sample solution and Standard solution
until a solution is effected. Cool the solution in an ice bath Calculate the percentage of C9H11NO2 in the portion of Gel
to about 10. Titrate slowly with 0.1 M sodium nitrite VS, taken:
using a calomel-platinum electrode system. Each mL of 0.1
M sodium nitrite is equivalent to 16.52 mg of C9H11NO2. Result = (rU/rS) (CS/CU) 100
PERFORMANCE TESTS rS = peak area from the Standard solution
MINIMUM FILL 755: Meets the requirements CS = concentration of USP Benzocaine RS in the
SPECIFIC TESTS CU = nominal concentration of the Sample solution
MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED (mg/mL)
MICROORGANISMS 62: It meets the requirements of the Acceptance criteria: 90.0%110.0%
aeruginosa. PERFORMANCE TESTS
PACKAGING AND STORAGE: Preserve in tight containers, SPECIFIC TESTS
protected from light, and avoid prolonged exposure to MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED
temperatures exceeding 30. MICROORGANISMS 62: It meets the requirements of the
aeruginosa.
Benzocaine Gel ADDITIONAL REQUIREMENTS
(Comment on this Monograph)id=m8073=Benzocaine Gel=B- PACKAGING AND STORAGE: Preserve in well-closed containers.
Monos.pdf) USP REFERENCE STANDARDS 11
USP Benzocaine RS
DEFINITION
Benzocaine Gel contains NLT 90.0% and NMT 110.0% of the
labeled amount of C9H11NO2.
Benzocaine Lozenges
IDENTIFICATION (Comment on this Monograph)id=m8077=Benzocaine
A. PROCEDURE Lozenges=B-Monos.pdf)
Sample solution: Place an amount equivalent to 5 mg of
benzocaine from Gel in a beaker, add 20 mL of 0.5 N DEFINITION
hydrochloric acid, warm gently to disperse the Gel, cool, Benzocaine Lozenges contain NLT 85.0% and NMT 120.0% of
and filter, if necessary, to obtain a clear solution. the labeled amount of C9H11NO2.
Analysis: To 10 mL of the clear solution, add 5 drops of a
solution of sodium nitrite (1 in 10) and 2 drops of methyl IDENTIFICATION
red TS, and neutralize with 1 N sodium hydroxide. Add 2 A. PROCEDURE
mL of a solution of 100 mg of 2-naphthol in 5 mL of 1 N Sample solution: Place an amount equivalent to 20 mg of
sodium hydroxide. benzocaine from powdered Lozenges, in 10 mL of water
Acceptance criteria: An orange-red precipitate is formed. with the aid of a few drops of 3 N hydrochloric acid. Filter,
B. The retention time of the Sample solution corresponds to if necessary, to obtain a clear solution.
that of the Standard solution, as obtained in the Assay. Analysis: Add 5 drops of a 100 mg/mL sodium nitrite
solution, followed by 2 mL of a 20 mg/mL solution of 2-
ASSAY naphthol in 1 N sodium hydroxide.
PROCEDURE Acceptance criteria: An orange-red precipitate is formed.
Diluent: Methanol and water (1:1) B. The retention time of the Sample solution corresponds to
Mobile phase: Methanol, glacial acetic acid, and water that of the Standard solution, as obtained in the Assay.
(14:1:10)
Standard solution: 0.02 mg/mL of USP Benzocaine RS in ASSAY
Diluent PROCEDURE
Sample solution: Place an amount equivalent to 200 mg of Mobile phase: Acetonitrile, 1.0 M monobasic potassium
benzocaine from Gel, in a 100-mL volumetric flask, add 80 phosphate solution previously adjusted with phosphoric acid
mL of Diluent, and shake by mechanical means for 15 min. to a pH of 3.0, and water (5:1:14)
Dilute with Diluent to volume, and filter, if necessary, to Standard solution A: 0.01 mg/mL of USP Benzocaine RS in
obtain a clear solution, discarding the first 5 mL of the 0.1 N hydrochloric acid
Standard solution B: 0.01 mg/mL of USP Benzocaine RS in IDENTIFICATION

a mixture of acetonitrile and water (1:1) OINTMENTS HAVING WATER-SOLUBLE BASES
Sample stock solution A: Add the equivalent of 40 mg of Sample solution: Transfer a portion of Ointment, equivalent
benzocaine from powdered Lozenges (NLT 20 Lozenges), to to 5 mg of benzocaine, to a small beaker. Add 20 mL of 0.5
a 200-mL volumetric flask, add 150 mL of 0.1 N N hydrochloric acid, warm gently to disperse the Ointment,
hydrochloric acid, and stir for NLT 2 h. Dilute with 0.1 N cool, and filter, if necessary.
hydrochloric acid to volume. Analysis: To 10 mL of the filtrate, add 5 drops of a solution
Sample solution A: Equivalent to 0.02 mg/mL of of sodium nitrite (1 in 10) and 2 drops of methyl red TS,
benzocaine in 0.1 N hydrochloric acid from Sample stock and neutralize with 1 N sodium hydroxide. Add 2 mL of a
solution A 20 mg/mL solution of 2-naphthol in sodium hydroxide TS.
Sample stock solution B: Add the equivalent of 40 mg of Acceptance criteria: An orange-red precipitate is formed.
benzocaine from powdered Lozenges (NLT 20 Lozenges), to OINTMENTS HAVING WATER-INSOLUBLE BASES
a 200-mL volumetric flask, add 150 mL of a mixture of Sample solution: Transfer a portion of Ointment, equivalent
acetonitrile and water (1:1), and stir for NLT 30 min. Dilute to 2.5 mg of benzocaine, to a small separator, and dissolve
with the mixture of acetonitrile and water (1:1) to volume. in 20 mL of ether. Extract with 10 mL of 0.5 N hydrochloric
Sample solution B: Equivalent to 0.02 mg/mL of acid, and filter the aqueous phase through paper into a
benzocaine in a mixture of acetonitrile and water (1:1), from small beaker.
Sample stock solution B Analysis: Add 5 drops of a solution of sodium nitrite (1 in
Chromatographic system 10) and 2 drops of methyl red TS, and neutralize with 1 N
(See Chromatography 621, System Suitability.) sodium hydroxide. Add 2 mL of a 20 mg/mL solution of 2-
Mode: LC naphthol in 1 N sodium hydroxide.
Detector: UV 280 nm Acceptance criteria: An orange-red precipitate is formed.
Injection size: 20 L OINTMENTS HAVING WATER-SOLUBLE BASES
System suitability Sample solution: Ointment, nominally equivalent to 200
Samples: Standard solution A and Standard solution B mg of benzocaine, in a tared 250-mL beaker
Suitability requirements Analysis: Add 50 mL of water and 5 mL of hydrochloric
Tailing factor: NMT 1.5 acid, and stir by mechanical means until a solution is
Relative standard deviation: NMT 2.0% effected. Cool the solution in an ice bath to about 10.
Analysis Titrate slowly with 0.1 M sodium nitrite VS, using a
Samples: Sample solution A, Sample solution B, Standard calomelplatinum electrode system. Perform a blank
solution A, and Standard solution B determination, and make any necessary correction (see
Calculate the percentage of total C9H11NO2 taken: Titrimetry 541). Each mL of 0.1 M sodium nitrite is
equivalent to 16.52 mg of C9H11NO2.
OINTMENTS HAVING WATER-INSOLUBLE BASES
rU = peak response from the Sample solution A Sample solution: Ointment, nominally equivalent to 200
rS = peak response from the Standard solution A mg of benzocaine
CS = concentration of USP Benzocaine RS in Standard Analysis: Transfer to a 125-mL separator. Suspend the
solution A (mg/mL) Ointment in 50 mL of ether by shaking, and extract with
CU = nominal concentration of Sample solution A three successive 25-mL portions of 1 N hydrochloric acid,
(mg/mL) filtering each portion into a 250-mL beaker. Cool the
Calculate the percentage of free C9H11NO2 taken: solution in an ice bath to about 10. Titrate slowly with 0.1
M sodium nitrite VS, using a calomelplatinum electrode
Result = (rU/rS) (CS/CU) 100 system. Perform a blank determination, and make any
necessary correction (see Titrimetry 541). Each mL of 0.1 M
rU = peak response from Sample solution B sodium nitrite is equivalent to 16.52 mg of C9H11NO2.
rS = peak response from Standard solution B Acceptance criteria: 90.0%110.0%
CS = concentration of USP Benzocaine RS in Standard
solution B (mg/mL) PERFORMANCE TESTS
CU = nominal concentration of Sample solution B MINIMUM FILL 755: Meets the requirements
(mg/mL)
MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED
ADDITIONAL REQUIREMENTS MICROORGANISMS 62: It meets the requirements of the
PACKAGING AND STORAGE: Preserve in well-closed containers. tests for absence of Staphylococcus aureus and Pseudomonas
USP REFERENCE STANDARDS 11 aeruginosa.
USP Benzocaine RS
PACKAGING AND STORAGE: Preserve in tight containers,
protected from light, and avoid prolonged exposure to
Benzocaine Ointment temperatures exceeding 30.
(Comment on this Monograph)id=m8080=Benzocaine
Ointment=B-Monos.pdf)
DEFINITION
Benzocaine Ointment contains NLT 90.0% and NMT 110.0% of
the labeled amount of C9H11NO2 in a suitable ointment base.
Benzocaine Otic Solution Analysis: Add 50 mL of water and 5 mL of hydrochloric

(Comment on this Monograph)id=m8090=Benzocaine Otic acid, and stir. Cool the solution in an ice bath to about 10.
Solution=B-Monos.pdf) Titrate slowly with 0.1 M sodium nitrite VS using a calomel-
platinum electrode system. Perform a blank determination,
DEFINITION and make any necessary correction (See Titrimetry 541).
Benzocaine Otic Solution contains NLT 90.0% and NMT Each mL of 0.1 M sodium nitrite is equivalent to 16.52 mg
110.0% of the labeled amount of C9H11NO2. of C9H11NO2.
IDENTIFICATION
PROCEDURE SPECIFIC TESTS
Sample solution: Place an amount equivalent to 2.5 mg of MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED
benzocaine from a volume of Otic Solution in 10 mL of MICROORGANISMS 62: It meets the requirements of the
water, and dissolve with the aid of a few drops of 3 N tests for absence of Staphylococcus aureus and Pseudomonas
hydrochloric acid. aeruginosa.
Analysis: Add 5 drops of a sodium nitrite solution (1 in 10),
then add 2 mL of a 20 mg/mL solution of 2-naphthol in 1 N ADDITIONAL REQUIREMENTS
sodium hydroxide. PACKAGING AND STORAGE: Preserve in tight containers,
Acceptance criteria: An orange-red precipitate is formed. protected from light, and avoid prolonged exposure to
temperatures exceeding 30.
ASSAY
PROCEDURE
Sample: A volume of Otic Solution nominally equivalent to
400 mg of benzocaine Benzocaine, Butamben, and Tetracaine
Analysis: Add 150 mL of cold 1.6 N hydrochloric acid, and Hydrochloride Topical Aerosol
stir until a fine dispersion is obtained. Cool the solution in (Comment on this Monograph)id=m8120=Benzocaine,
an ice bath to 10. Titrate slowly with 0.1 M sodium nitrite Butamben, and Tetracaine Hydrochloride Topical Aerosol=B-
VS, using a calomelplatinum electrode system. Perform a Monos.pdf)
blank determination, and make any necessary correction
(See Titrimetry 541). Each mL of 0.1 M sodium nitrite is DEFINITION
equivalent to 16.52 mg of C9H11NO2. Benzocaine, Butamben, and Tetracaine Hydrochloride Topical
Acceptance criteria: 90.0%110.0% Aerosol is Benzocaine, Butamben, and Tetracaine
Hydrochloride Topical Solution packaged in a pressurized
SPECIFIC TESTS container with a suitable inert propellant. It contains NLT
MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED 90.0% and NMT 110.0% of the labeled amounts of
MICROORGANISMS 62: It meets the requirements of the benzocaine (C9H11NO2), butamben (C11H15NO2), and tetracaine
tests for absence of Salmonella species and Escherichia coli hydrochloride (C15H24N2O2 HCl).
and for absence of Staphylococcus aureus and Pseudomonas
aeruginosa; the total aerobic microbial count is less than 100 IDENTIFICATION
cfu/mL. The retention times of the Sample solution correspond to
those of the Standard solution, as obtained in the Assay.
PACKAGING AND STORAGE: Preserve in tight, light-resistant ASSAY
containers. PROCEDURE
Diluent: Methanol and water (3:2)
Mobile phase: Methanol, 0.25 M sodium 1-
Benzocaine Topical Solution heptanesulfonate, and water (25:1:25)
Standard solution: Transfer 140 mg of USP Benzocaine RS
(Comment on this Monograph)id=m8100=Benzocaine Topical to a 100-mL volumetric flask. Swirl with the aid of 25 mL of
Solution=B-Monos.pdf) methanol, and transfer 140 J mg of USP Butamben RS to
the same volumetric flask with the aid of 25 mL of water, J
DEFINITION being the ratio of the labeled amount, as a percentage, of
Benzocaine Topical Solution is a solution of Benzocaine in a butamben to the labeled amount, as a percentage, of
suitable solvent. It contains NLT 90.0% and NMT 110.0% of benzocaine in the Topical Aerosol. Transfer 140 J mg of USP
the labeled amount of C9H11NO2. It contains a suitable Tetracaine Hydrochloride RS into the same volumetric flask
antimicrobial agent. with the aid of 25 mL of water, J being the ratio of the
IDENTIFICATION labeled amount, as a percentage, of tetracaine hydrochloride
PROCEDURE to the labeled amount, as a percentage, of benzocaine in
Sample solution: Transfer an amount of Topical solution, the Topical Aerosol. Sonicate for 1 min, and dilute with
equivalent to 5 mg of benzocaine, to a small beaker and Diluent to volume.
add 20 mL of 0.5 N hydrochloric acid, warm gently to Sample stock solution: Fit the valve of the Topical Aerosol
disperse, cool, and filter. container with a cannula, and expel the contents of the
Analysis: To 10 mL of the filtrate, add 5 drops of sodium container into a 100-mL, glass-stoppered, round-bottom
nitrite solution (1 in 10) and 2 drops of methyl red TS, and flask. Attach the flask to a rotary evaporator, and evaporate
neutralize with 1 N sodium hydroxide. Add 2 mL of a 20 under a vacuum of 600 mm of mercury for 2.5 h. Transfer a
mg/mL solution of 2-naphthol in 1 N sodium hydroxide. portion of the material in the round-bottom flask, equivalent
Acceptance criteria: An orange-red precipitate is formed. to 1400 mg of benzocaine, to a 100-mL volumetric flask.
Add 75 mL of methanol, and mix. Sonicate for 1 min, dilute
ASSAY with methanol to volume, and mix.
PROCEDURE
Sample solution: Topical Solution equivalent to 200 mg of
benzocaine in a 250-mL separator
Sample solution: 1.4 mg/mL from the Sample stock solution ASSAY
in Diluent PROCEDURE
[NOTEPrepare by adding a quantity of Sample stock Diluent: Methanol and water (3:2)
solution corresponding to 10% of the total volume to a Mobile phase: Methanol, 0.25 M sodium 1-
quantity of Diluent corresponding to 75% of the total heptanesulfonate, and water (25:1:25)
volume. Shake, and dilute with Diluent to volume.] Standard solution: Transfer 140 mg of USP Benzocaine RS,
Chromatographic system in a 100-mL volumetric flask with the aid of 25 mL of
(See Chromatography 621, System Suitability.) methanol, and swirl. Transfer 140 J mg of USP Butamben RS
Mode: LC to the same volumetric flask with the aid of 25 mL of water,
Detector: UV 313 nm J being the ratio of the labeled amount, in percentage of
Column: 3.9-mm 30-cm; packing L1 butamben to the labeled amount, in percentage of
Flow rate: 2 mL/min benzocaine in the topical solution. Transfer 140 J mg of USP
Injection size: 10 L Tetracaine Hydrochloride RS, into the same volumetric flask
System suitability with the aid of 25 mL of water, J being the ratio of the
Sample: Standard solution labeled amount, in percentage of tetracaine hydrochloride
[NOTEThe relative retention times for benzocaine and to the labeled amount, in percentage of benzocaine in the
butamben are 0.3 and 0.8, respectively.] topical solution. Sonicate for 1 min, and dilute with Diluent
Suitability requirements to volume.
Resolution: NLT 2 between the benzocaine peak and the Sample stock solution: Nominally equivalent to 14 mg/mL
butamben peak and between the butamben peak and the of benzocaine from Gel in methanol
tetracaine peak [NOTESonicate for about 1 min.]
Relative standard deviation: NMT 2.0% for each of the Sample solution: Nominally 1.4 mg/mL from the Sample
three analyte peaks stock solution diluted with Diluent
Samples: Standard solution and Sample solution (See Chromatography 621, System Suitability.)
Calculate the individual percentages of C9H11NO2, Mode: LC
C11H15NO2, and C15H24N2O2 HCl in the portion of the Detector: UV 313 nm
residue of Topical Aerosol taken from the round-bottom Column: 3.9-mm 30-cm; packing L1
flask: Flow rate: 2 mL/min
Result = (RU/RS) (CS/CU) 100 System suitability
RU = peak response from the corresponding analyte of [NOTEThe relative retention times for benzocaine,
the Sample solution butamben, and tetracaine are about 0.3, 0.8, and 1.0,
RS = peak response from the corresponding analyte of respectively.]
the Standard solution Suitability requirements
CS = concentration of the appropriate USP Reference Resolution: NLT 2 between the benzocaine peak and the
Standard in the Standard solution (mg/mL) butamben peak, and between the butamben peak and
CU = nominal concentration of the individual the tetracaine peak
quantities in the Sample solution (mg/mL) Relative standard deviation: NMT 2.0% for each of the
Acceptance criteria: 90.0%110.0% three analyte peaks
Analysis
SPECIFIC TESTS Samples: Sample solution and Standard solution
OTHER REQUIREMENTS: It meets the requirements for Aerosols, Calculate the individual percentages of C9H11NO2,
Nasal Sprays, Metered-Dose Inhalers, and Dry Powder Inhalers C11H15NO2, and C15H24N2O2 HCl in the portion of Gel
601, Pressure Test, Minimum Fill, and Leakage Test. taken:
PACKAGING AND STORAGE: Preserve in pressurized containers, Result = (rU/rS) (CS/CU) 100
and avoid exposure to excessive heat. rU = peak area in the corresponding analyte from the
USP REFERENCE STANDARDS 11 Sample solution
USP Benzocaine RS rS = peak area in the corresponding analyte from the
USP Butamben RS Standard solution
USP Tetracaine Hydrochloride RS CS = concentration of the appropriate Reference
CU = nominal concentration of the analyte in the
Benzocaine, Butamben, and Tetracaine Sample solution (mg/mL)
Hydrochloride Gel Acceptance criteria: 90.0%110.0%
(Comment on this Monograph)id=m8104=Benzocaine, PERFORMANCE TESTS
Butamben, and Tetracaine Hydrochloride Gel=B-Monos.pdf) MINIMUM FILL 755: Meets the requirements
Benzocaine, Butamben, and Tetracaine Hydrochloride Gel is PACKAGING AND STORAGE: Preserve in tight containers, and
Benzocaine, Butamben, and Tetracaine Hydrochloride in a avoid freezing.
suitable gel base. It contains NLT 90.0% and NMT 110.0% of USP REFERENCE STANDARDS 11
the labeled amounts of benzocaine (C9H11NO2), butamben USP Benzocaine RS
(C11H15NO2), and tetracaine hydrochloride (C15H24N2O2 HCl). USP Butamben RS
USP Tetracaine Hydrochloride RS
IDENTIFICATION
The retention times of the Sample solution correspond to
those of the Standard solution, as obtained in the Assay.
Benzocaine, Butamben, and Tetracaine Analysis

Hydrochloride Ointment Samples: Sample solution and Standard solution
Calculate the individual percentages of C9H11NO2,
(Comment on this Monograph)id=m8106=Benzocaine, C11H15NO2, and C15H24N2O2 HCl in the portion of
Butamben, and Tetracaine Hydrochloride Ointment=B- Ointment taken:
Monos.pdf)
DEFINITION Result = (rU/rS) (CS/CU) 100
Benzocaine, Butamben, and Tetracaine Hydrochloride Ointment rU = peak response from the corresponding analyte
is Benzocaine, Butamben, and Tetracaine Hydrochloride in a from the Sample solution
suitable ointment base. It contains NLT 90.0% and NMT rS = peak response from the corresponding analyte
110.0% of the labeled amounts of benzocaine (C9H11NO2), from the Standard solution
butamben (C11H15NO2), and tetracaine hydrochloride CS = concentration of the appropriate Reference
(C15H24N2O2 HCl). Standard in the Standard solution (mg/mL)
IDENTIFICATION CU = nominal concentration of the analyte in the
The retention times of the Sample solution correspond to Sample solution (mg/mL)
those of the Standard solution, as obtained in the Assay. Acceptance criteria: 90.0%110.0%

PROCEDURE MINIMUM FILL 755: Meets the requirements
Diluent: Methanol and water (3:2) ADDITIONAL REQUIREMENTS
Mobile phase: Methanol, 0.25 M sodium 1- PACKAGING AND STORAGE: Preserve in tight containers, and
heptanesulfonate, and water (25:1:25) avoid freezing.
Standard solution: Transfer 140 mg of USP Benzocaine RS, USP REFERENCE STANDARDS 11
in a 100-mL volumetric flask with the aid of 25 mL of USP Benzocaine RS
methanol, and swirl. Transfer 140 J mg of USP Butamben RS USP Butamben RS
to the same volumetric flask with the aid of 25 mL of water, USP Tetracaine Hydrochloride RS
J being the ratio of the labeled amount, as a percentage, of
butamben to the labeled amount, as a percentage, of
benzocaine in the Ointment. Transfer 140 J mg of USP
Tetracaine Hydrochloride RS, into the same volumetric flask Benzocaine, Butamben, and Tetracaine
with the aid of 25 mL of water, J being the ratio of the
labeled amount, as a percentage, of tetracaine hydrochloride
Hydrochloride Topical Solution
to the labeled amount, as a percentage, of benzocaine in (Comment on this Monograph)id=m8108=Benzocaine,
the Ointment. Sonicate for about 1 min, and dilute with Butamben, and Tetracaine Hydrochloride Topical Solution=B-
Diluent to volume. Monos.pdf)
Sample stock solution: Equivalent to 14 mg/mL of DEFINITION
benzocaine from Ointment in methanol Benzocaine, Butamben, and Tetracaine Hydrochloride Topical
Prepare by adding to a quantity of methanol corresponding Solution contains NLT 90.0% and NMT 110.0% of the labeled
to 75% of the total volume. Sonicate for about 1 min, and amounts of benzocaine (C9H11NO2), butamben (C11H15NO2),
dilute to volume. and tetracaine hydrochloride (C15H24N2O2 HCl).
Sample solution: 1.4 mg/mL from the Sample stock solution
in Diluent IDENTIFICATION
Prepare by adding a quantity of the Sample stock solution The retention times of the Sample solution correspond to
corresponding to 10% of the total volume to a quantity of those of the Standard solution, as obtained in the Assay.
Diluent corresponding to 75% of the total volume. Shake,
and dilute to volume with Diluent. ASSAY
(See Chromatography 621, System Suitability.) Diluent: Methanol and water (3:2)
Mode: LC Mobile phase: Methanol, 0.25 M sodium 1-
Detector: UV 313 nm heptanesulfonate, and water (25:1:25)
Column: 3.9-mm 30-cm; packing L1 Standard solution: Transfer 140 mg of USP Benzocaine RS,
Flow rate: 2 mL/min in a 100-mL volumetric flask with the aid of 25 mL of
Injection size: 10 L methanol, and swirl. Transfer 140 J mg of USP Butamben RS
System suitability to the same volumetric flask with the aid of 25 mL of water,
Sample: Standard solution J being the ratio of the labeled amount, in percentage of
[NOTEThe relative retention times for benzocaine, butamben to the labeled amount, in percentage of
butamben, and tetracaine are 0.3, 0.8, and 1.0, benzocaine in the Topical Solution. Transfer 140 J mg of USP
respectively.] Tetracaine Hydrochloride RS, into the same volumetric flask
Suitability requirements with the aid of 25 mL of water, J being the ratio of the
Resolution: NLT 2 between the benzocaine peak and the labeled amount, in percentage of tetracaine hydrochloride
butamben peak and between the butamben peak and the to the labeled amount, in percentage of benzocaine in the
tetracaine peak Topical Solution. Sonicate for 1 min, and dilute with Diluent
Relative standard deviation: NMT 2.0% for each of the to volume.
three analyte peaks
Sample stock solution: Equivalent to 14 mg/mL of IDENTIFICATION

benzocaine from a volume of Topical Solution in methanol A. PROCEDURE
Prepare by adding to a quantity of methanol corresponding Sample solution: Transfer an amount of Topical Aerosol,
to 75% of the total volume. Sonicate for 1 min, and dilute equivalent to 5 mg of benzocaine, to a beaker, add 20 mL
to volume. of 0.5 N hydrochloric acid, warm gently to disperse the
Sample solution: 1.4 mg/mL from the Sample stock solution solution, cool, and filter, if necessary, to obtain a clear
in Diluent solution.
Prepare by adding a quantity of the Sample stock solution Analysis: To 10 mL of the clear Sample solution, add 5
corresponding to 10% of the total volume to a quantity of drops of a solution of sodium nitrite (1 in 10), and 2 drops
Diluent corresponding to 75% of the total volume, shake, of methyl red TS, and neutralize with 1 N sodium hydroxide.
and dilute to volume with Diluent. Add 2 mL of 20 mg 2-naphthol/mL 1 N sodium hydroxide.
Chromatographic system Acceptance criteria: An orange-red precipitate is formed.
(See Chromatography 621, System Suitability.) B. The retention time of the major peak for benzocaine of
Mode: LC the Sample solution corresponds to that of the Standard
Detector: UV 313 nm solution, as obtained in the Assay.
Column: 3.9-mm 30-cm; packing L1 C. The retention time of the major peak for menthol of the
Flow rate: 2 mL/min Sample solution corresponds to that of the Standard solution,
Injection size: 10 L as obtained in the Assay.
System suitability
[NOTEThe relative retention times for benzocaine, BENZOCAINE
butamben, and tetracaine are 0.3, 0.8, and 1.0, Diluent: Methanol and water (1:1)
respectively.] Mobile phase: Methanol, glacial acetic acid, and water
Suitability requirements (14:1:10)
Resolution: NLT 2 between the benzocaine peak and the Standard solution: 0.02 mg/mL of USP Benzocaine RS in
butamben peak and between the butamben peak and the Diluent
tetracaine peak Sample stock solution: Spray the Topical Aerosol into a
Relative standard deviation: NMT 2.0% for each of the flask, and dilute a volume of this solution with Diluent to
three analyte peaks prepare a nominal 2 mg/mL solution.
Analysis Sample solution: Equivalent to 0.02 mg/mL from the
Samples: Sample solution and Standard solution Sample stock solution, diluted with Diluent
Calculate the individual percentages of C9H11NO2, Chromatographic system
C11H15NO2, and C15H24N2O2 HCl in the portion of Topical (See Chromatography 621, System Suitability.)
Solution taken: Mode: LC
Detector: UV 294 nm
Result = (rU/rS) (CS/CU) 100 Column: 3.9-mm 30-cm; packing L1
rU = peak response from the corresponding analyte Injection size: 50 L
from the Sample solution System suitability
rS = peak response from the corresponding analyte Sample: Standard solution
from the Standard solution Suitability requirements
CS = concentration of the appropriate Reference Relative standard deviation: NMT 2.0%
Standard in the Standard solution (mg/mL) Analysis
CU = nominal concentration of the analyte in the Samples: Standard solution and Sample solution
Sample solution (mg/mL) Calculate the percentage of C9H11NO2 in the portion of
Acceptance criteria: 90.0%110.0% Topical Aerosol taken:
SPECIFIC TESTS Result = (rU/rS) (CS/CU) 100
ADDITIONAL REQUIREMENTS rS = peak response of the Standard solution
PACKAGING AND STORAGE: Preserve in tight containers, and CS = concentration of USP Benzocaine RS in the
avoid freezing. Standard solution (mg/mL)
USP REFERENCE STANDARDS 11 CU = nominal concentration in the Sample solution
USP Benzocaine RS (mg/mL)
USP Butamben RS Acceptance criteria: 90.0%110.0%
USP Tetracaine Hydrochloride RS MENTHOL
Internal standard solution: 1 mg/mL of decanol in n-
hexane
Benzocaine and Menthol Topical Standard stock solution: 1 mg/mL of USP Menthol RS in
n-hexane
Aerosol Standard solution: Transfer 5.0 mL of Standard stock
(Comment on this Monograph)id=m8102=Benzocaine and solution and 5.0 mL of Internal standard solution to a 100-mL
Menthol Topical Aerosol=B-Monos.pdf) volumetric flask, and dilute with ether to volume.
Sample stock solution: Spray the Topical Aerosol into a
DEFINITION flask. Transfer an amount nominally equivalent to 50 mg of
Benzocaine and Menthol Topical Aerosol is a solution of menthol to a separator, and extract with three 15-mL
Benzocaine and Menthol with suitable propellants in a portions of ether, collecting the ether extracts in a 50-mL
pressurized container. It contains NLT 90.0% and NMT volumetric flask. Dilute with ether to volume.
110.0% of the labeled amounts of benzocaine (C9H11NO2) and
menthol (C10H20O).
Sample solution: Transfer 5.0 mL of the Sample stock Benzoic Acid

solution to a 100-mL volumetric flask, add 5.0 mL of n- (Comment on this Monograph)id=m8130=Benzoic Acid=B-
hexane and 5.0 mL of Internal standard solution, and dilute Monos.pdf)
with ether to volume.
Mode: GC
Column: 2-mm 1.8-m column packed with 10% phase
G16 on support S1AB
Carrier gas: Dry helium
Temperature C7H6O2 122.12
Column: 170 Benzoic acid;
Injection port: 260 Benzoic acid [65-85-0].
Detector: 240
Flow rate: 50 mL/min DEFINITION
Injection size: 2 L Benzoic Acid contains NLT 99.5% and NMT 100.5% of C7H6O2,
System suitability calculated on the anhydrous basis.
[NOTEThe relative retention times for menthol and IDENTIFICATION
decanol are 0.7 and 1.0, respectively.] PROCEDURE
Suitability requirements Sample solution: Prepare a saturated solution of benzoic
Resolution: NLT 2.5 between the menthol and decanol acid in water, and filter twice.
peaks Analysis 1: To one portion of the filtrate, add ferric chloride
Relative standard deviation: NMT 2.0% of the ratio of TS.
the peak response of menthol to that of decanol Acceptance criteria 1: A salmon-colored precipitate is
Analysis formed.
Samples: Standard solution and Sample solution Analysis 2: To a separate 10-mL portion of the filtrate, add
Calculate the percentage C10H20O in each mL of Topical 1 mL of 7 N sulfuric acid and cool the mixture.
Aerosol taken: Acceptance criteria 2: A white precipitate forms in 10 min;
this precipitate is soluble in ether.
ASSAY
RU = peak response ratio of menthol to decanol from PROCEDURE
the Sample solution Sample: 500 mg of benzoic acid
RS = peak response ratio of menthol to decanol from Analysis: Dissolve in 25 mL of diluted alcohol that
the Standard solution previously has been neutralized with 0.1 N sodium
CS = concentration of USP Menthol RS in the hydroxide, add phenolphthalein TS, and titrate with 0.1 N
Standard solution (mg/mL) sodium hydroxide VS to a pink color. Each mL of 0.1 N
CU = nominal concentration of menthol in the Sample sodium hydroxide is equivalent to 12.21 mg of C7H6O2.
solution (mg/mL) Acceptance criteria: 99.5%100.5%
PERFORMANCE TESTS Inorganic Impurities
MINIMUM FILL 755: Meets the requirements RESIDUE ON IGNITION 281: NMT 0.05%
HEAVY METALS 231: NMT 10 ppm
SPECIFIC TESTS Sample solution: Dissolve 2.0 g in 25 mL of acetone, and
MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED add 2 mL of water.
MICROORGANISMS 62: It meets the requirements for Analysis: Add 1.2 mL of thioacetamide-glycerin base TS
absence of Staphylococcus aureus and Pseudomonas and 2 mL of pH 3.5 Acetate Buffer, and allow to stand for 5
aeruginosa. min.
OTHER REQUIREMENTS: It meets the requirements under Acceptance criteria: Any color produced is not darker
Aerosols, Nasal Sprays, Metered-Dose Inhalers, and Dry Powder than that of a control made with 25 mL of acetone, 2.0 mL
Inhalers 601, Pressure Test and Leakage Test. of Standard Lead Solution, and treated in the same manner.
ADDITIONAL REQUIREMENTS SPECIFIC TESTS
PACKAGING AND STORAGE: Preserve in tight, pressurized CONGEALING TEMPERATURE 651: 121123
containers, and avoid exposure to excessive heat. WATER DETERMINATION, Method I 921: NMT 0.7%, a 1 in
USP REFERENCE STANDARDS 11 2 solution of methanol in pyridine being used as the solvent
USP Benzocaine RS
USP Menthol RS
USP 32 Official Monographs / Benzoic 41
READILY CARBONIZABLE SUBSTANCES TEST 271: 500 mg in 5 Similarly, mix 4 g of chromatographic siliceous earth with 3
mL of sulfuric acid TS: the solution has no more color than mL of Ferric chloride-urea reagent, and pack uniformly over
Matching Fluid Q. the first layer. Cover the column with a pad of glass wool.
READILY OXIDIZABLE SUBSTANCES: Add 1.5 mL of sulfuric acid Column B: Insert a small pledget of glass wool above the
to 100 mL of water, heat to boiling, and add 0.1 N stem constriction of a second 20- 2.5-cm chromatographic
potassium permanganate, dropwise, until the pink color tube. Mix 4 g of chromatographic siliceous earth with 2 mL
persists for 30 s. Dissolve 1.00 g of benzoic acid in the hot of sodium bicarbonate solution (1 in 12), prepared just prior
solution, and titrate with 0.1 N potassium permanganate VS to use, to a uniform, fluffy mixture, and pack evenly over
to a pink color that persists for 15 s: NMT 0.50 mL of 0.10 the glass wool. Cover the column with a pad of glass wool.
N potassium permanganate is consumed. Diluent: Mixture of chloroform and glacial acetic acid
(97:3)
ADDITIONAL REQUIREMENTS Standard solution A: 20 g/mL of USP Salicylic Acid RS in
PACKAGING AND STORAGE: Preserve in well-closed containers. Diluent
Standard solution B: 40 g/mL of USP Benzoic acid RS in
Diluent
Benzoic and Salicylic Acids Ointment Sample solution: Transfer an amount of the Ointment
equivalent to 100 mg of benzoic acid and 50 mg of salicylic
(Comment on this Monograph)id=m8160=Benzoic and Salicylic acid to a 250-mL volumetric flask, and dissolve in 150 mL of
Acids Ointment=B-Monos.pdf) chloroform by warming on a steam bath. Cool, dilute with
chloroform to volume to obtain a solution having a nominal
DEFINITION concentration of 200 g/mL of salicylic acid and 400 g/mL
Benzoic and Salicylic Acids Ointment is Benzoic Acid and of benzoic acid.
Salicylic Acid, present in a ratio of 2 to 1, in a suitable Analysis
ointment base. It contains NLT 90.0% and NMT 110.0% of Samples: Standard solution A, Standard solution B, and
the labeled amounts of benzoic acid (C7H6O2) and salicylic Sample solution
acid (C7H6O3). Mount Column A directly over Column B, then pipet 10 mL
IDENTIFICATION of Sample solution onto Column A, and allow it to pass into
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 the column. Wash the columns with two 40-mL portions
Diluent: Mixture of chloroform and methanol (1:1) of chloroform, allowing the first portion to recede to the
Standard solution A: 2.4 mg/mL of USP Benzoic Acid RS in top of each column before adding the second portion.
Diluent Discard the eluates, and separate the columns.
Standard solution B: 1.2 mg/mL of USP Salicylic Acid RS in Salicylic acid content: Elute Column A with 95 mL of
Diluent Diluent collecting the eluate in a 100-mL volumetric flask.
Sample solution: Equivalent to 60 mg of benzoic acid and Dilute the contents of the flask with Diluent to volume, and
30 mg of salicylic acid from Ointment, in 25 mL of Diluent mix. Concomitantly determine the absorbances of the
Chromatographic system eluate and Standard solution A in 1-cm cells at the
(See Chromatography 621, Thin-Layer Chromatography.) wavelength of maximum absorbance at 311 nm, with a
Mode: TLC suitable spectrophotometer, using the Diluent as the blank.
Adsorbent: 0.25-mm layer of chromatographic silica gel Calculate the percentage of C7H6O3 in the portion of
mixture Ointment:
Application volume: 5 L of each solution at separate
points 2.5 cm from the bottom edge of a 20- 20-cm Result = (AU/AS) (CS/CU) F 100
thin-layer chromatographic plate AU = absorbance of the diluted eluate from Column A
Developing solvent system: Chloroform, acetone, AS = absorbance of Standard solution A
isopropyl alcohol, methanol, and ammonium hydroxide CS = concentration of USP Salicylic Acid RS in
(30:30:15:15:10) Standard solution A (g/mL)
Analysis CU = nominal concentration of the salicylic acid in the
Samples: Standard solution A, Standard solution B, and Sample solution (g/mL)
Sample solution F = sample dilution factor, 10
Develop the chromatogram in the Developing solvent system Acceptance criteria: 90.0%110.0%
until the solvent front has moved three-fourths of the Benzoic acid content: Elute Column B with 95 mL of a
length of the plate. Remove the plate from the solution (3 in 100) of glacial acetic acid in chloroform,
chromatographic chamber, mark the solvent front, and collecting the eluate in a 100-mL volumetric flask. Dilute
allow the solvent to evaporate. View the chromatogram the contents of the flask with the same solvent to volume,
under short-wavelength (254 nm) UV radiation. and mix. Concomitantly determine the absorbances of
Acceptance criteria: The two major fluorescent spots from eluate and Standard solution B in 1-cm cells at the
the Sample solution correspond in color and in RF value to wavelength of maximum absorbance at 275 nm, with a
those from the respective Standard solution A and B. suitable spectrophotometer, using the Diluent as the blank.
ASSAY Calculate the percentage of C7H6O2 in the portion of
PROCEDURE Ointment taken:
Ferric chloride-urea reagent: On the day of use, dissolve,
without heating, 18 g of urea in a mixture of 2.5 mL of Result = (AU/AS) (CS/CU) F 100
ferric chloride solution (6 in 10) and 12.5 mL of 0.05 N AU = absorbance of the diluted eluate from Column B
hydrochloric acid. AS = absorbance of Standard solution B
Column A: Insert a small pledget of glass wool above the CS = concentration of USP Benzoic Acid RS in
stem constriction of a 20- 2.5-cm chromatographic tube. Standard solution B (g/mL)
Mix 1 g of chromatographic siliceous earth with 0.5 mL of CU = nominal concentration of benzoic acid in the
dilute phosphoric acid (3 in 10) to form a uniform, fluffy Sample solution (g/mL)
mixture, transfer to the chromatographic tube, and pack F = sample dilution factor,10
evenly over the glass wool, exerting gentle pressure.
42 Benzoic / Official Monographs USP 32
Acceptance criteria: 90.0%110.0% the Benzoin taken for the Assay, and subtract it from the
original weight of the Benzoin taken. The difference
PERFORMANCE TESTS between this result and the weight of the residue in the
MINIMUM FILL 755: Meets the requirements extraction thimble represents the alcohol-soluble extractive.
Acceptance criteria: The alcohol-soluble extractive is NLT
ADDITIONAL REQUIREMENTS 75.0% in Sumatra Benzoin and NLT 90.0% in Siam Benzoin.
and avoid exposure to temperatures exceeding 30. OTHER COMPONENTS
LABELING: Label Ointment to indicate the concentrations of CONTENT OF BENZOIC ACID
Benzoic Acid and Salicylic Acid and to indicate whether the Sample solution: 1 g of powdered Benzoin with 15 mL of
ointment base is water-soluble or water-insoluble. warm carbon disulfide
USP REFERENCE STANDARDS 11 Filter through a small pledget of cotton, wash the cotton
USP Benzoic Acid RS with an additional 5 mL of carbon disulfide, and allow the
USP Salicylic Acid RS filtrate to evaporate spontaneously.
Acceptance criteria: Compared to the weight of Benzoin
taken, the weight of the residue is NLT 6.0% in Sumatra
Benzoin Benzoin and NLT 12.0% in Siam Benzoin. This residue meets
the requirements for Identification TestsGeneral 191,
(Comment on this Monograph)id=m8190=Benzoin=B- Benzoate.
Monos.pdf)
SPECIFIC TESTS
DEFINITION ARTICLES OF BOTANICAL ORIGIN, Acid-Insoluble Ash 561:
Benzoin is the balsamic resin obtained from Styrax benzoin NMT 1.0% in Sumatra Benzoin; NMT 0.5% in Siam Benzoin
Dryander or Styrax paralleloneurus Perkins, known in commerce ARTICLES OF BOTANICAL ORIGIN, Foreign Organic Matter 561:
as Sumatra Benzoin, or from Styrax tonkinensis (Pierre) Craib ex NMT 1.0% in Siam Benzoin
Hartwich, or other species of the Section Anthostyrax of the
genus Styrax, known in commerce as Siam Benzoin (Fam. ADDITIONAL REQUIREMENTS
Styraceae). PACKAGING AND STORAGE: Preserve in well-closed containers.
Sumatra Benzoin yields NLT 75.0% of alcohol-soluble extractive, LABELING: Label it to indicate whether it is Sumatra Benzoin
and Siam Benzoin yields NLT 90.0% of alcohol-soluble or Siam Benzoin.
extractive.
Botanic characteristics
Sumatra Benzoin: Blocks or lumps of varying size, made up
of tears, compacted together, with a reddish brown, reddish Compound Benzoin Tincture
gray, or grayish brown resinous mass; the tears are (Comment on this Monograph)id=m8220=Compound Benzoin
externally yellowish or rusty brown, milky white on fresh Tincture=B-Monos.pdf)
fracture; hard and brittle at ordinary temperatures, but
softened by heat and becoming gritty on chewing. DEFINITION
Siam Benzoin: Pebble-like tears of variable size and shape, Prepare Compound Benzoin Tincture as follows.
compressed, yellowish brown to rusty brown externally,
milky white on fracture, separate or very slightly Benzoin, in moderately coarse powder 100 g
agglutinated; hard and brittle at ordinary temperatures, but Aloe, in moderately coarse powder 20 g
softened by heat and becoming plastic on chewing.
Storax 80 g
IDENTIFICATION Tolu Balsam 40 g
A. A solution in alcohol becomes milky upon the addition of
water, and the mixture is acid to litmus paper. To make 1000 mL
B. Heat a few fragments in a test tube. Sumatra Benzoin
evolves a sublimate consisting of plates and small, rod-like Prepare a Tincture by Process M (see Pharmaceutical Dosage
crystals of cinnamic acid and its esters that strongly polarize Forms 1151), using alcohol as the menstruum.
light. Siam Benzoin evolves a sublimate directly above the OTHER COMPONENTS
melted mass, consisting of numerous long, rod-shaped Alcohol Determination, Method II 611: 74.0%80.0% of
crystals of benzoic acid that do not strongly polarize light. C2H5OH, the dilution to approximately 2% alcohol being
C. Heat 500 mg in a test tube with 10 mL of potassium made with methanol instead of with water
permanganate TS: only the Sumatra variety develops a faint
odor of benzaldehyde. SPECIFIC TESTS
SPECIFIC GRAVITY 841: 0.8700.885
ASSAY LIMIT OF NONVOLATILE RESIDUE: Evaporate 3 mL of Tincture
PROCEDURE in a suitable tared dish on a steam bath, and dry the residue
Sample: Place 2 g of Benzoin in a tared extraction thimble, at 100 for 2 h.
and insert the thimble in a continuous-extraction apparatus. Acceptance criteria: The weight of the residue is NLT 525
Place 100 mg of sodium hydroxide in the receiving flask of mg and NMT 675 mg.
the apparatus, and extract the Benzoin with alcohol for 5 h,
or until completely extracted. Dry the extraction thimble ADDITIONAL REQUIREMENTS
containing the insoluble residue at 105 for 2 h. PACKAGING AND STORAGE: Preserve in tight, light-resistant
Analysis: On a separate portion of Benzoin, determine the containers, and avoid exposure to direct sunlight and to
water content as directed for Water Determination 921, excessive heat.
Method II. Calculate the weight of water in the quantity of
USP 32 Official Monographs / Benzonatate 43
LABELING: Label it to indicate that it is flammable. Benzonatate Capsules

(Comment on this Monograph)id=m8260=Benzonatate
Capsules=B-Monos.pdf)
Benzonatate DEFINITION
(Comment on this Monograph)id=m8250=Benzonatate=B- Benzonatate Capsules contain NLT 90.0% and NMT 110.0% of
Monos.pdf) the labeled amount of benzonatate (C30H53NO11 av.).
IDENTIFICATION
A. INFRARED ABSORPTION 197F
[NOTEIf a difference is observed, or if excipients are
present, use the equivalent of 100 mg of benzonatate from
Capsules. Mix with 25 mL of 0.01 N hydrochloric acid, and
proceed as directed under IdentificationOrganic
Nitrogenous Bases 181, beginning with Transfer the liquid
C30H53NO11 (av.) 603.00 (av.) to a separator.]
Benzoic acid, 4-(butylamino)-, 2,5,8,11,14,17,20,23,26- B. ULTRAVIOLET ABSORPTION 197U
nonaoxaoctacos-28-yl ester; Solution: 15 g/mL from Capsules
2,5,8,11,14,17,20,23,26-Nonaoxaoctacosan-28-yl p-
(butylamino)benzoate [104-31-4]. ASSAY
PROCEDURE
DEFINITION Standard solution: 0.5 mg/mL of USP Benzonatate RS
Benzonatate contains NLT 95.0% and NMT 105.0% of Sample stock solution: Mix a number of capsules,
C30H53NO11. equivalent to 500 mg of benzonatate, with 40 mL of
chloroform in a suitable high speed blender, and dilute with
IDENTIFICATION chloroform to 100 mL.
A. INFRARED ABSORPTION 197F Sample solution: 10.0 mL of Sample stock solution, in a
B. ULTRAVIOLET ABSORPTION 197U 100-mL volumetric flask, evaporate the chloroform on a
Solution: 15 g/mL steam bath with the aid of a current of air. Dissolve the
ASSAY residue in water and dilute to volume.
PROCEDURE Blank: Water
Sample: 5 g of Benzonatate Spectrometric conditions
Analysis: Reflux with 25.0 mL of 0.5 N sodium hydroxide Mode: UV-Vis
VS for 1 h. Cool, add 25 mL of water and 10 drops of Analytical wavelength: 500 nm
bromothymol blue TS, and titrate the excess alkali with 0.5 Cell: 1 cm
N hydrochloric acid VS. Perform a blank determination (see Analysis
Titrimetry 541, Residual Titrations). Each mL of 0.5 N Samples: Standard solution, Sample solution, and Blank
sodium hydroxide is equivalent to 301.5 mg of C30H53NO11. Transfer 4.0 mL each of the Standard solution, the Sample
Acceptance criteria: 95.0%105.0% solution, and Blank, to separate test tubes. To each tube
add in succession 1.0 mL of 1 M hydroxylamine
IMPURITIES hydrochloride and 1.0 mL of 3.5 N sodium hydroxide,
Inorganic Impurities mixing after each addition. Allow to stand for 10 min,
RESIDUE ON IGNITION 281: NMT 0.1% accurately timed, then add 1.0 mL of 3.5 N hydrochloric
CHLORIDE AND SULFATE, Chloride 221: Mix 20 mL of a acid, mix, add 1.0 mL of an 80 mg/mL ferric chloride
solution (1 in 10) with 20 mL of water and 1 mL of nitric solution, and mix. Allow to stand for 30 min, accurately
acid, shake for 1 h, and allow to stand for 1 h. Pass through timed. Gently swirl the tubes for 1 min to remove any gas
a filter having a porosity of 0.2 m, and to the filtrate add 1 bubbles present, then concomitantly determine the
mL of silver nitrate TS. Dilute with water to 50 mL, mix, and absorbances of the solution
allow to stand protected from light for 10 min. Calculate the percentage of C30H53NO11 av in the number
Acceptance criteria: the turbidity does not exceed that of Capsules:
produced by 0.10 mL of 0.020 N hydrochloric acid (35
ppm). Result = (AU/AS) (CS/CU) 100
CHLORIDE AND SULFATE, Sulfate 221: Mix 5 mL of a solution
(1 in 20) with 5 mL of water and 1 mL of 3 N hydrochloric AU = absorbance of the Sample solution
acid, shake for 1 h, and allow to stand for 1 h. Pass through AS = absorbance of the Standard solution
a filter having a porosity of 0.2 m, and to the filtrate add 1 CS = concentration of USP Benzonatate RS in the
mL of barium chloride TS. Mix, and allow to stand for 10 Standard solution (g/mL)
min. CU = nominal concentration of benzonatate in the
Acceptance criteria: the turbidity does not exceed that Sample solution (g/mL)
produced by 0.10 mL of 0.020 N sulfuric acid (400 ppm). Acceptance criteria: 90.0%110.0%
HEAVY METALS, Method II 231: NMT 10 ppm PERFORMANCE TESTS
SPECIFIC TESTS DISSOLUTION 711
REFRACTIVE INDEX 831: 1.5091.511 at 20 Medium: Water; 900 mL
WATER DETERMINATION, Method I 921: NMT 0.3% Apparatus 2: 50 rpm
Time: 30 min
ADDITIONAL REQUIREMENTS Determine the amount of benzonatate dissolved by
PACKAGING AND STORAGE: Preserve in tight, light-resistant employing the following method.
containers. Mobile phase: Acetonitrile and 0.04 M monobasic
USP REFERENCE STANDARDS 11 potassium phosphate (3:1)
USP Benzonatate RS
44 Benzonatate / Official Monographs USP 32
Standard solution: 0.1 mg/mL of USP Benzonatate RS Sample solution: 10 mg/mL of benzoyl peroxide in
[NOTESonicate as necessary.] methanol
Sample solutions: Sample per Dissolution 711. Dilute with Mode: TLC
Medium to a concentration that is similar to the Standard Adsorbent: 0.25-mm layer of chromatographic silica gel
solution. Pass a portion of the solution under test through a mixture
0.45-m filter. Application volume: 5 L
Chromatographic system Developing solvent system: Toluene, dichloromethane,
(See Chromatography 621, System Suitability.) and glacial acetic acid (50:2:1)
Mode: LC Analysis
Detector: UV 310 nm Samples: Standard solution and Sample solution
Column: 3.9-mm 30-cm; packing L1 Analysis: Place the plate in a developing chamber
Flow rate: 1.5 mL/min containing, and equilibrated with the Developing solvent
Injection size: 15 L system. Develop the chromatogram until the solvent front
System suitability has moved three-fourths of the length of the plate. Remove
Sample: Standard solution the plate, and allow the solvent to evaporate. Observe the
Suitability requirements plate under short-wavelength UV light.
Relative standard deviation: NMT 2.0% Aceptance criteria: The RF value of the principal spot of
Analysis the Sample solution corresponds to that of the Standard
Samples: Standard solution and Sample solution solution.
Calculate the percentage of benzonatate dissolved: B. The Sample solution in the Organic Impurities test exhibits
a major peak for benzoyl peroxide, the retention time of
Result = (rU/rS) (CS/CU) 100 which corresponds to that exhibited by the Standard
solution.
CS = concentration of USP Benzonatate RS in the PROCEDURE
Standard solution (mg/mL) Sample: 300 mg of previously mixed Hydrous Benzoyl
CU = nominal concentration of benzonatate in the Peroxide in a conical flask fitted with a ground-glass stopper,
Sample solution (mg/mL) and weigh again to obtain the weight of the Sample.
Tolerances: NLT 80% (Q) of the labeled amount of Analysis: Add 30 mL of glacial acetic acid, previously
benzonatate sparged with carbon dioxide for not less than 2 min just
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements before use, and swirl the flask gently to effect solution. Add
5 mL of potassium iodide solution (1 in 2), and mix. Allow
ADDITIONAL REQUIREMENTS the solution to stand for 1 min. Titrate the liberated iodine
PACKAGING AND STORAGE: Preserve in tight, light-resistant with 0.1 N sodium thiosulfate VS. As the endpoint is
containers. approached, add 1 drop of starch iodide paste TS, or
USP REFERENCE STANDARDS 11 equivalent, and continue the titration to the discharge of the
USP Benzonatate RS blue color. Perform a blank determination, and make any
necessary correction (see Titrimetry 541). Each mL of 0.1 N
sodium thiosulfate is equivalent to 12.11 mg of C14H10O4.
Hydrous Benzoyl Peroxide Acceptance criteria: 65.0%82.0%
(Comment on this Monograph)id=m8310=Hydrous Benzoyl IMPURITIES
Peroxide=B-Monos.pdf) Organic Impurities
PROCEDURE
Solution A: Prepare a mixture of acetonitrile and glacial
acetic acid (1000:1).
Solution B: Prepare a mixture of water and glacial acetic
acid (1000:1).
Mobile phase: Use variable mixtures of Solution A and
Solution B as directed for Chromatographic system.
Sample solution: 0.32 mg/mL of benzoyl peroxide in
C14H10O4 (anhydrous) 242.23 acetonitrile
Peroxide, dibenzoyl; Standard solution: Dissolve a quantity of Hydrous Benzoyl
Benzoyl peroxide [94-36-0]. Peroxide, previously subjected to the Assay, in acetonitrile
to obtain a solution containing 0.32 mg/mL.
DEFINITION System suitability solution: 100 g/mL of benzoic acid
Hydrous Benzoyl Peroxide contains NLT 65.0% and NMT and 60 g/mL of methylparaben in acetonitrile
82.0% of C14H10O4. It contains 26% of water for the purpose Chromatographic system
of reducing flammability and shock sensitivity. (See Chromatography 621, System Suitability.)
[CAUTIONHydrous Benzoyl Peroxide may explode at Mode: LC
temperatures higher than 60 or cause fires in the presence of Detector: UV 235 nm
reducing substances. Store it in the original container, treated Column: 4.6-mm 25-cm; packing L1
to reduce static charges.] Flow rate: 1.2 mL/min
A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 The chromatograph is programmed as follows.
Standard solution: 10 mg/mL of Hydrous Benzoyl
Peroxide, previously subjected to the Assay, in methanol
USP 32 Official Monographs / Benzoyl 45
Time Solution A Solution B Sample stock solution: Equivalent to 40 mg of benzoyl

(min) (%) (%) peroxide from Gel. In a 50-mL volumetric flask, add 40 mL
0 18 82 of acetonitrile, and shake until the material is thoroughly
dispersed. Sonicate the mixture for 5 min, dilute with
20 60 40 acetonitrile to volume, mix, and filter.
30 60 40 Sample solution: 10 mL of Sample stock solution and 5 mL
of Internal standard solution. Dilute with acetonitrile to 25
System suitability mL.
Sample: System suitability solution Chromatographic system
Suitability requirements (See Chromatography 621, System Suitability.)
Resolution: NLT 2.0 between benzoic acid and Mode: LC
methylparaben Detector: UV 254 nm
Tailing factors: NMT 2.0 for the benzoic acid and Column: 3.9-mm 30-cm; packing L1
methylparaben peaks Flow rate: 1 mL/min
Analysis Injection size: 10 L
Samples: Sample solution and Standard solution System suitability
Calculate the area, as a percentage, of each peak in the Sample: Standard solution (three replicate injections)
chromatogram of the Sample solution: [NOTEThe lowest and highest peak response ratios (RS)
agree within 2.0%. The retention times for ethyl benzoate
Result = (rU/rT) 100 and benzoyl peroxide are 7 and 14 min, respectively.]
rU = peak response for any individual peak other Resolution: NLT 2.0 between ethyl benzoate and benzoyl
than the principal peak in the Sample solution peroxide
rT = sum of the peak responses of all the individual Tailing factors: NMT 2.0 for the ethyl benzoate and
peaks including the principal peak in the benzoyl peroxide peaks
Sample solution Analysis
Acceptance criteria: The area of any individual peak other Samples: Sample solution and Standard solution
than the principal peak is NMT 1.5% of the total area. The Calculate the percentage of C14H10O4 in the portion of Gel
sum of the areas of all peaks other than the principal peak taken:
is NMT 2.0% of the total area.
PACKAGING AND STORAGE: Store in the original container, at RU = peak response ratios of benzoyl peroxide to ethyl
room temperature. [NOTEDo not transfer Hydrous Benzoyl benzoate from the Sample solution
Peroxide to metal or glass containers fitted with friction tops. RS = peak response ratios of benzoyl peroxide to ethyl
Do not return unused material to its original container, but benzoate from the Standard solution
destroy it by treatment with sodium hydroxide solution (1 in CS = concentration of benzoyl peroxide in the
10) until addition of a crystal of potassium iodide results in Standard solution (mg/mL)
no release of free iodine.] CU = nominal concentration of benzoyl peroxide in
the Sample solution (mg/mL)
Benzoyl Peroxide Gel IMPURITIES
(Comment on this Monograph)id=m8320=Benzoyl Peroxide Organic Impurities
Gel=B-Monos.pdf) PROCEDURE
Solution A: Acetonitrile and glacial acetic acid (1000:1)
DEFINITION Solution B: Glacial acetic acid and water (1:1000)
Benzoyl Peroxide Gel is benzoyl peroxide in a suitable gel base. Mobile phase: See the gradient table below.
It contains NLT 90.0% and NMT 125.0% of the labeled
amount of benzoyl peroxide (C14H10O4).
IDENTIFICATION (min) (%) (%)
The retention time of the major peak for benzoyl peroxide of 0 18 82
the Sample solution corresponds to that of the Standard 20 60 40
solution, as obtained in the Assay.
30 60 40
ASSAY
PROCEDURE System suitability solution: 100 g/mL of benzoic acid
Mobile phase: Acetonitrile in water (5 in 10) and 60 g/mL of methylparaben in acetonitrile
Internal standard solution: 3.6 mg/mL of ethyl benzoate Standard solution A: 500 g/mL of benzoic acid in
in acetonitrile acetonitrile
Standard stock solution: A suitable quantity of benzoyl Standard solution B: 20 g/mL of ethyl benzoate in
peroxide, recently subjected to the Assay under Hydrous acetonitrile
Benzoyl Peroxide, in a weighed conical flask fitted with a Standard solution C: 20 g/mL of benzaldehyde in
glass stopper. Weigh again to obtain the weight of the acetonitrile
specimen, and quantitatively dissolve in acetonitrile to Standard solution D: Equivalent to 40 g/mL of
obtain a solution containing 0.8 mg/mL. anhydrous benzoyl peroxide, previously subjected to the
Standard solution: 10 mL of Standard stock solution and 5 Assay under Hydrous Benzoyl Peroxide, in acetonitrile
mL of Internal standard solution Sample solution: Equivalent to 100 mg of benzoyl
Dilute with acetonitrile to 25 mL. peroxide from Gel. To a 50-mL volumetric flask, add 25 mL
[NOTEThis Standard solution contains 0.32 mg/mL of of acetonitrile, shake vigorously to disperse the specimen,
benzoyl peroxide.]
46 Benzoyl / Official Monographs USP 32
sonicate for 5 min, dilute with acetonitrile to volume, and Acceptance criteria: The RF value of the principal spot from
filter. the solution under test corresponds to that from the
Chromatographic system Standard solution.
(See Chromatography 621, System Suitability.) B. The retention time of the major peak for benzoyl
Mode: LC peroxide of the Sample solution corresponds to that of the
Detector: UV 235 nm Standard solution, as obtained in the Assay.
System suitability Mobile phase: Acetonitrile in water (5 in 10)
Sample: System suitability solution Internal standard solution: 3.6 mg/mL of ethyl benzoate
Suitability requirements in acetonitrile
Resolution: NLT 2.0 between benzoic acid and Standard stock solution: A suitable quantity of benzoyl
methylparaben peroxide, recently subjected to the Assay under Hydrous
Tailing factors: NMT 2.0 for the benzoic acid and Benzoyl Peroxide, in a weighed conical flask fitted with a
methylparaben peaks glass stopper. Weigh again to obtain the weight of the
Analysis specimen, and quantitatively dissolve in acetonitrile to
Samples: Sample solution and Standard solution obtain a solution containing 0.8 mg/mL.
Acceptance criteria: The responses of any peaks from the Standard solution: 10 mL of Standard stock solution and 5
Sample solution corresponding to benzoic acid, ethyl mL of Internal standard solution. Dilute with acetonitrile to
benzoate, and benzaldehyde are NMT those of the main 25 mL.
peaks from Standard solution A (25%), Standard solution B [NOTEThis Standard solution contains 0.32 mg/mL of
(1%), and Standard solution C (1%), respectively. The benzoyl peroxide.]
response of any other impurity peak from the Sample Sample stock solution: Equivalent to 40 mg of benzoyl
solution, other than the main benzoyl peroxide peak, any peroxide from Lotion. In a 50-mL volumetric flask, add 40
benzoic acid, ethyl benzoate, benzaldehyde, mL of acetonitrile. Shake vigorously to disperse the
methylparaben, or propylparaben peak, and any solvent specimen, and sonicate until the material is thoroughly
peak, is NMT that from Standard solution D (2%); and the dispersed. Sonicate the mixture for 5 min, dilute with
sum of the responses of all the impurity peaks, other than acetonitrile to volume, mix, and filter.
those of benzoic acid, ethyl benzoate, and benzaldehyde, is Sample solution: 10 mL of Sample stock solution and 5 mL
NMT that from Standard solution D (2%). of Internal standard solution. Dilute with acetonitrile to 25
mL.
SPECIFIC TESTS Chromatographic system
PH 791: 2.86.6 (See Chromatography 621, System Suitability.)
Mode: LC
ADDITIONAL REQUIREMENTS Detector: UV 254 nm
PACKAGING AND STORAGE: Preserve in tight containers. Column: 3.9-mm 30-cm; packing L1
Flow rate: 1 mL/min
Benzoyl Peroxide Lotion System suitability
Sample: Standard solution (three replicate injections)
(Comment on this Monograph)id=m8330=Benzoyl Peroxide [NOTEThe lowest and highest peak response ratios (RS)
Lotion=B-Monos.pdf) agree within 2.0%. The retention times for ethyl benzoate
and benzoyl peroxide are 7 and 14 min, respectively.]
Benzoyl Peroxide Lotion is benzoyl peroxide in a suitable lotion Resolution: NLT 2.0 between ethyl benzoate and benzoyl
base. It contains NLT 90.0% and NMT 110.0% of the labeled peroxide
amount of C14H10O4. Tailing factors: NMT 2.0 for the ethyl benzoate and
IDENTIFICATION benzoyl peroxide peaks
A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Analysis
Adsorbent: 0.25-mm layer of chromatographic silica gel Samples: Sample solution and Standard solution
mixture Calculate the percentage of C14H10O4 in the portion of
Standard solution: 10 mg/mL of Hydrous Benzoyl Lotion taken:
Peroxide, previously subjected to the Assay, in methanol
Sample solution: Equivalent to 10 mg/mL of benzoyl Result = (RU/RS) (CS/CU) 100
peroxide from Lotion in acetone RU = peak response ratios of benzoyl peroxide to ethyl
Application volume: 5 L benzoate from the Sample solution
Developing solvent system: Toluene, dichloromethane, RS = peak response ratios of benzoyl peroxide to ethyl
and glacial acetic acid (50:2:1) benzoate from the Standard solution
Analysis CS = concentration of benzoyl peroxide in the
Sample: Sample solution and Standard solution Standard solution (mg/mL)
Place the plate in a developing chamber containing, and CU = nominal concentration of benzoyl peroxide in
equilibrated, with Developing solvent system. Develop the the Sample solution (mg/mL)
chromatogram until the solvent front has moved three-
fourths of the length of the plate. Remove the plate, and
allow the solvent to evaporate. Observe the plate under
short-wavelength UV light.
USP 32 Official Monographs / Benztropine 47

PH 791: 2.86.6
IMPURITIES
Organic Impurities ADDITIONAL REQUIREMENTS
PROCEDURE PACKAGING AND STORAGE: Preserve in tight containers.
Solution A: Acetonitrile and glacial acetic acid (1000:1)
Solution B: Glacial acetic acid and water (1:1000)
Benztropine Mesylate
Time Solution A Solution B (Comment on this Monograph)id=m8500=Benztropine
(min) (%) (%) Mesylate=B-Monos.pdf)
0 18 82
20 60 40
30 60 40
System suitability solution: 100 g/mL of benzoic acid

and 60 g/mL of methylparaben in acetonitrile
Standard solution A: 500 g/mL of benzoic acid in C21H25NO CH4O3S 403.54
acetonitrile 8-Azabicyclo[3.2.1]octane, 3-(diphenylmethoxy)-, endo-,
Standard solution B: 20 g/mL of ethyl benzoate in methanesulfonate;
acetonitrile 3-(Diphenylmethoxy)-1H,5H-tropane methanesulfonate
Standard solution C: 20 g/mL of benzaldehyde in [132-17-2].
acetonitrile
Standard solution D: Equivalent to 40 g/mL of DEFINITION
anhydrous benzoyl peroxide, previously subjected to the Benztropine Mesylate contains NLT 98.0% and NMT 100.5% of
Assay under Hydrous Benzoyl Peroxide, in acetonitrile C21H25NO CH4O3S, calculated on the dried basis.
Sample solution: Equivalent to 100 mg of benzoyl
peroxide from Lotion. In a 50-mL volumetric flask, add 25 IDENTIFICATION
mL of acetonitrile, shake vigorously to disperse the INFRARED ABSORPTION 197K
specimen, sonicate for 5 min, dilute with acetonitrile to
volume, mix, and filter. ASSAY
(See Chromatography 621, System Suitability.) Sample: 60 mg of Benztropine Mesylate
Mode: LC Analysis: Dissolve in 25 mL of water, add 5 mL of sodium
Detector: UV 235 nm carbonate TS, and extract with four 10-mL portions of
Column: 4.6-mm 25-cm; packing L1 chloroform. Wash the combined chloroform extracts with
Flow rate: 1.2 mL/min about 10 mL of water, and extract the wash solution with 5
Injection size: 10 L mL of chloroform. Filter the combined chloroform extracts
System suitability through a tightly packed pledget of cotton, and wash the
Sample: System suitability solution cotton with about 5 mL of chloroform. Add methyl red TS,
Suitability requirements and titrate the chloroform solution with 0.01 N perchloric
Resolution: NLT 2.0 between benzoic acid and acid in dioxane VS. Perform a blank determination, and
methylparaben make any necessary correction (see Titrimetry 541). Each
Tailing factors: NMT 2.0 for the benzoic acid and mL of 0.01 N perchloric acid is equivalent to 4.035 mg of
methylparaben peaks C21H25NO CH4O3S.
Acceptance criteria: The responses of any peaks from the IMPURITIES
Sample solution corresponding to benzoic acid, ethyl Inorganic Impurities
benzoate, and benzaldehyde are NMT those of the main RESIDUE ON IGNITION 281: NMT 0.1%
peaks from Standard solution A (25%), Standard solution B
(1%), and Standard solution C (1%), respectively. The SPECIFIC TESTS
response of any other impurity peak from the Sample MELTING RANGE OR TEMPERATURE 741: 141148
solution, other than the main benzoyl peroxide peak, any LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT
benzoic acid, ethyl benzoate, benzaldehyde, 5.0% of its weight.
methylparaben, or propylparaben peak, and any solvent ADDITIONAL REQUIREMENTS
peak, is NMT that from Standard solution D (2%); and the PACKAGING AND STORAGE: Preserve in tight containers.
sum of the responses of all the impurity peaks, other than USP REFERENCE STANDARDS 11
those of benzoic acid, ethyl benzoate, and benzaldehyde, is USP Benztropine Mesylate RS
NMT that from Standard solution D (2%).
48 Benztropine / Official Monographs USP 32
Benztropine Mesylate Injection Injection size: 25 L

(Comment on this Monograph)id=m8530=Benztropine System suitability
Mesylate Injection=B-Monos.pdf) Sample: Standard solution
DEFINITION Relative standard deviation: NMT 2.0%
Benztropine Mesylate Injection is a sterile solution of Analysis
Benztropine Mesylate in Water for Injection. It contains NLT Samples: Sample solution and Standard solution
90.0% and NMT 110.0% of the labeled amount of C21H25NO Calculate the percentage of C21H25NO CH4O3S in each mL
CH4O3S. of the Injection:
IDENTIFICATION Result = (rU/rS) (CS/CU) 100

THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
Adsorbent: 0.25-mm layer of chromatographic silica gel rU = peak response from the Sample solution
Standard stock solution: 10 mg of USP Benztropine rS = peak response from the Standard solution
Mesylate RS in 50 mL in a separator (0.2 mg/mL) CS = concentration of USP Benztropine Mesylate RS in
Standard solution: To the separator containing the the Standard solution (mg/mL)
Standard stock solution, add 2 mL of 1 N sodium hydroxide. CU = nominal concentration of benztropine mesylate
Extract with three 10-mL portions of chloroform, collecting in the Sample solution (mg/mL)
the chloroform extracts to a 50-mL beaker. Evaporate the Acceptance criteria: 90.0%110.0%
chloroform extracts with the aid of gentle heat and a
current of air to dryness, and dissolve the residue in 1 mL of SPECIFIC TESTS
chloroform. BACTERIAL ENDOTOXINS TEST 85: NMT 55.6 USP Endotoxin
Sample stock solution: Equivalent to 10 mg of Benztropine Units/mg of benztropine mesylate
Mesylate from a volume of Injection to 50 mL in a separator PH 791: 5.08.0
(0.2 mg/mL) OTHER REQUIREMENTS: Meets the requirements under
Sample solution: Repeat the procedure for the Standard Injections 1
solution, using the Sample stock solution. ADDITIONAL REQUIREMENTS
Application volume: 1 L PACKAGING AND STORAGE: Preserve in single-dose or in
Developing solvent system: Chloroform, methanol, and a multiple-dose containers, preferably of Type I glass.
1-in-4 solution of ammonium hydroxide (40:10:1) USP REFERENCE STANDARDS 11
Analysis USP Benztropine Mesylate RS
Samples: Sample solution and Standard solution USP Endotoxin RS
Allow the applications to dry, and develop the
chromatogram in the Developing solvent system, until the
solvent front has moved about three-fourths of the length
of the plate. Remove the plate from the developing Benztropine Mesylate Tablets
chamber, mark the solvent front, and allow the solvent to (Comment on this Monograph)id=m8540=Benztropine
evaporate. Locate the spots on the plate by lightly spraying Mesylate Tablets=B-Monos.pdf)
with potassium iodoplatinate TS.
Acceptance criteria: The RF value of the principal spot from DEFINITION
the Sample solution corresponds to that from the Standard Benztropine Mesylate Tablets contain NLT 90.0% and NMT
solution. 110.0% of the labeled amount of C21H25NO CH4O3S.
ASSAY IDENTIFICATION
PROCEDURE THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
Solution A: 0.005 M Octylamine phosphate buffer Adsorbent: 0.25-mm layer of chromatographic silica gel
Transfer 0.83 mL of octylamine to a 1-L volumetric flask, Standard stock solution: 10 mg of USP Benztropine
dilute to volume, adjust with phosphoric acid to a pH of Mesylate RS in 50 mL in a separator (0.2 mg/mL)
3.0, and pass the solution through a membrane filter. Standard solution: To the separator containing the
Mobile phase: Acetonitrile and Solution A (13:7) Standard stock solution, add 2 mL of 1 N sodium hydroxide.
Standard solution: 1 mg/mL of USP Benztropine Mesylate Extract with three 10-mL portions of chloroform, collecting
RS the chloroform extracts to a 50-mL beaker. Evaporate the
Sample solution: Equivalent to 1 mg/mL of benztropine chloroform extracts with the aid of gentle heat and a
mesylate from the volume of Injection current of air to dryness, and dissolve the residue in 1 mL of
Chromatographic system chloroform.
(See Chromatography 621, System Suitability.) Sample stock solution: Add equivalent to 10 mg of
Mode: LC Benztropine Mesylate from a portion of finely powdered
Detector: UV 259 nm Tablets in 50 mL, shake by mechanical means for 30 min,
Column: 4.6-mm 25-cm; packing L7 and filter into a separator (0.2 mg/mL).
Flow rate: 1 mL/min Sample solution: Repeat the procedure for the Standard
[NOTEAdjust the flow rate to obtain a retention time of 7 solution, using the Sample stock solution.
min for benztropine mesylate.]
USP 32 Official Monographs / Benzyl 49
Application volume: 1 L Solution A: Proceed as directed in the Assay.

Developing solvent system: Chloroform, methanol, and a Mobile phase: Acetonitrile and Solution A (13:7)
1-in-4 solution of ammonium hydroxide (40:10:1) Sample solution: Sample per Dissolution 711. Use a
Analysis filtered portion of the solution under test from the
Samples: Sample solution and Standard solution dissolution vessel.
Allow the applications to dry, and develop the Standard solution: USP Benztropine Mesylate RS in
chromatogram in the Developing solvent system, until the Medium. Dilute to obtain a solution having a known
solvent front has moved about three-fourths of the length concentration similar to that of the Sample solution.
of the plate. Remove the plate from the developing Chromatographic system
chamber, mark the solvent front, and allow the solvent to (See Chromatography 621, System Suitability.)
evaporate. Locate the spots on the plate by lightly spraying Mode: LC
with potassium iodoplatinate TS. Detector: UV 220 nm
Acceptance criteria: The RF value of the principal spot from Column: 4.6-mm 25-cm; packing L7
the Sample solution corresponds to that from the Standard Flow rate: 2 mL/min
solution. Injection size: 300 L
Analysis
ASSAY Samples: Sample solution and Standard solution
PROCEDURE Calculate the percentage of C21H25NO CH4O3S dissolved:
Solution A: Transfer 0.83 mL of octylamine to a 1-L
volumetric flask, dilute to volume, adjust with phosphoric Result = (rU/rS) (CS/CU) 100
acid to a pH of 3.0, and pass the solution through a
membrane filter. rU = peak response from the Sample solution
Diluent: Isopropyl alcohol, phosphoric acid, and water rS = peak response from the Standard solution
(40:0.1:60) CS = concentration of USP Benztropine Mesylate RS in
Mobile phase: Acetonitrile and Solution A (9:11) the Standard solution (mg/mL)
Standard solution: 0.04 mg/mL of USP Benztropine CU = nominal concentration of benztropine mesylate
Mesylate RS in Diluent in the Sample solution (mg/mL)
Sample solution: Equivalent to 0.04 mg/mL of benztropine Acceptance criteria: NLT 80% (Q) of the labeled amount
mesylate from powdered Tablets (NLT 20 Tablets) in Diluent of C21H25NO CH4O3S
[NOTEAdd powdered tablets to a portion of Diluent UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
corresponding to 60% of the final volume. Mix by
mechanical means for NLT 60 min, and dilute with Diluent ADDITIONAL REQUIREMENTS
to volume. Centrifuge a portion of this mixture, and filter PACKAGING AND STORAGE: Preserve in well-closed containers.
the supernatant layer.] USP REFERENCE STANDARDS 11
Chromatographic system USP Benztropine Mesylate RS
Mode: LC
Detector: UV 259 nm Benzyl Benzoate
Flow rate: 0.7 mL/min (Comment on this Monograph)id=m8600=Benzyl Benzoate=B-
Injection size: 50 L Monos.pdf)
System suitability
Analysis
Calculate the percentage of C21H25NO CH4O3S in the
portion of Tablets: C14H12O2 212.24
Benzoic acid, phenylmethyl ester;
Result = (rU/rS) (CS/CU) 100 Benzyl benzoate [120-51-4].
rU = peak response from the Sample solution DEFINITION

rS = peak response from the Standard solution Benzyl Benzoate contains NLT 99.0% and NMT 100.5% of
CS = concentration of USP Benztropine Mesylate RS in C14H12O2.
the Standard solution (mg/mL) IDENTIFICATION
CU = nominal concentration of benztropine mesylate
in the Sample solution (mg/mL) INFRARED ABSORPTION 197F
Acceptance criteria: 90.0%110.0% ASSAY
DISSOLUTION 711 Sample: 2 g of Benzyl Benzoate
Medium: 0.1 N hydrochloric acid; 900 mL Analysis: Transfer to a conical flask fitted with a reflux
Apparatus 2: 50 rpm condenser, add 50.0 mL of 0.5 N alcoholic potassium
Time: 30 min hydroxide VS, and boil gently for 1 h. Cool, add
Determine the amount of C21H25NO CH4O3S dissolved by phenolphthalein TS, and titrate with 0.5 N hydrochloric acid
using the following method. VS. Perform a blank determination (see Titrimetry 541).
50 Benzyl / Official Monographs USP 32
Each mL of 0.5 N alcoholic potassium hydroxide is Acceptance criteria: 26.0%30.0% (w/w)

equivalent to 106.1 mg of C14H12O2.
PH 791: 8.59.2
SPECIFIC TESTS
SPECIFIC GRAVITY 841: 1.1161.120 ADDITIONAL REQUIREMENTS
CONGEALING TEMPERATURE 651: NLT 18.0 PACKAGING AND STORAGE: Preserve in tight containers.
Congelation may be brought about by the addition of a
fragment of previously congealed Benzyl Benzoate when
the temperature has reached the expected congealing Benzylpenicilloyl Polylysine
temperature.
REFRACTIVE INDEX 831: 1.5681.570 at 20 Concentrate
ALDEHYDE: Transfer 10 g to a 125-mL conical flask (Comment on this Monograph)id=m8650=Benzylpenicilloyl
containing 50 mL of alcohol and 5 mL of hydroxylamine Polylysine Concentrate=B-Monos.pdf)
hydrochloride solution (3.5 in 100), mix, and allow to stand
for 10 min. Add 1 mL of bromophenol blue TS, and titrate DEFINITION
with 0.1 N sodium hydroxide VS to a light green endpoint. Benzylpenicilloyl Polylysine Concentrate has a molar
Perform a blank determination, and match the color of the concentration of benzylpenicilloyl moeity (C16H19N2O5S) of NLT
endpoint with that of the titrated Sample solution. The net 0.0125 M and NMT 0.020 M. It contains one or more suitable
volume of 0.1 N sodium hydroxide consumed does not buffers.
exceed 0.50 mL (0.05% as benzaldehyde).
ACIDITY: Add 2 drops of phenolphthalein TS to 25 mL of ASSAY
alcohol, and add 0.020 N sodium hydroxide until a pink PROCEDURE
color is produced. Add 5.0 g of Benzyl Benzoate, and titrate Solution A: 9 g of sodium chloride and 1.38 g of
with 0.020 N sodium hydroxide: NMT 1.5 mL of 0.020 N monobasic sodium phosphate in 900 mL. Adjust with 5 N
sodium hydroxide is required to restore the pink color. sodium hydroxide or phosphoric acid to a pH of 7.6. Dilute
to 1000 mL.
ADDITIONAL REQUIREMENTS Solution B: 0.07 mg/mL of mercuric chloride
PACKAGING AND STORAGE: Preserve in tight, well-filled, light- Sample solution: Concentrate and Solution A (1.0 in 500)
resistant containers, and avoid exposure to excessive heat. Blank: Solution A
USP Benzyl Benzoate RS Sample: Blank and Sample solution
Transfer 3.0 mL of Sample solution to a spectrophotometric
cell. Using a suitable spectrophotometer and using Blank,
determine the initial absorbance at the wavelength of
Benzyl Benzoate Lotion maximum absorbance at 282 nm. Add 0.02 mL of
(Comment on this Monograph)id=m8630=Benzyl Benzoate Solution B to the Sample solution in the
Lotion=B-Monos.pdf) spectrophotometric cell and determine the absorbance at
the same wavelength after 1 and 3 min. Repeat the
DEFINITION addition of 0.02-mL portions of Solution B until a
Benzyl Benzoate Lotion contains NLT 26.0% and NMT 30.0% maximum absorbance reading is obtained.
(w/w) of C14H12O2. Calculate the molar concentration of benzylpenicilloyl
moiety in the Concentrate:
Benzyl Benzoate 250 mL
Result = 500{[AM(3 + 0.02N)/3] AI}/MAB
Triethanolamine 5g
Oleic Acid 20 g AM = highest absorbance observed
N = number of 0.02-mL portions of Solution B added
Purified Water 750 mL
to the Sample solution to obtain the maximum
To make about 1000 mL absorbance
AI = initial absorbance
Mix the Triethanolamine with the Oleic Acid, and add the MA = molar absorptivity of the penamaldate formed
Benzyl Benzoate. Transfer the mixture to a suitable container by the reaction of benzylpenicilloyl with
of 2000-mL capacity, add 250 mL of Purified Water, and shake mercuric chloride at pH 7.6, 22,325
the mixture thoroughly. Finally add the remaining Purified B = length of the cell (cm)
Water, and again shake thoroughly. Acceptance criteria: 0.01250.020 M
ASSAY OTHER COMPONENTS
PROCEDURE BENZYLPENICILLOYL SUBSTITUTION
Sample: 5 g of Lotion in a conical flask Solution A: 19.69 g of sodium citrate dihydrate, 0.1 mL of
Analysis: Add 25 mL of alcohol and 2 drops of pentachlorophenol, and 5 mL of 2,2-thiodiethanol in 900
phenolphthalein TS. Cool the solution to 15, and titrate mL of 0.2 N hydrochloric acid. Adjust with hydrochloric acid
quickly with 0.1 N sodium hydroxide to a slight pink color. to a pH of 2.2, and dilute with water to 1000 mL.
Add 50.0 mL of 0.5 N alcoholic potassium hydroxide VS, Ninhydrin reagent: 18 g of ninhydrin and 0.7 g of
connect the flask to a reflux condenser, and boil gently for 1 hydrindantin in 675 mL of dimethyl sulfoxide. Add 225 mL
h. Cool, promptly add phenolphthalein TS and titrate with of 4 M lithium acetate solution previously adjusted with
0.5 N hydrochloric acid VS. Perform a blank determination glacial acetic acid to a pH of 5.2.
(see Titrimetry 541). Each mL of 0.5 N alcoholic potassium
hydroxide is equivalent to 106.1 mg of C14H12O2.
USP 32 Official Monographs / Benzylpenicilloyl 51
Standard solution: 91 g/mL (5 104 M) of USP L-Lysine Acceptance criteria: NMT 0.00020 M
Hydrochloride RS in Solution A Calculate the molar concentration of penamaldate taken:
Sample solution: 1.0 mL of Concentrate in a 10-mL
volumetric flask, and dilute with water to volume. Transfer Result = 50A282/MA2B
1.0 mL of this solution to an ampul, add 1.5 mL of 6 N
hydrochloric acid, and seal the ampul under nitrogen. Heat A282 = absorbance at 282 nm
the ampul at 110 for 22 h. Transfer the contents of the MA2 = molar absorptivity of the penamaldate moiety at
ampul to a round-bottom, 50-mL flask, and dry by vacuum pH 7.6, 22,325
rotary evaporation. Dissolve the residue three times, using 5- B = length of the cell (cm)
mL portions, evaporating to dryness after each dissolution. Acceptance criteria: NMT 0.00060 M
Dissolve the residue in 10 mL of Solution A.
Chromatographic system SPECIFIC TESTS
(See Chromatography 621, System Suitability.) PH 791: 6.58.5, using the undiluted Concentrate
Detector: 570 nm PACKAGING AND STORAGE: Preserve in tight containers.
Column: 1.75-mm 50-cm; packing of 8-m 8% cross- LABELING: The label states that this article is not intended for
linked sulfonated divinylbenzene polystyrene cation- direct administration to humans or animals.
exchange resin [NOTEThe column effluent is mixed USP REFERENCE STANDARDS 11
continuously with flowing Ninhydrin reagent, and the USP L-Lysine Hydrochloride RS
flowing mixture is heated at 130 for 1.5 min in a reaction
coil. The absorbance of the reaction mixture is measured
continuously.]
Injection size: 20 L Benzylpenicilloyl Polylysine Injection
System suitability (Comment on this Monograph)id=m8654=Benzylpenicilloyl
Sample: Standard solution Polylysine Injection=B-Monos.pdf)
Column efficiency: NLT 1800 theoretical plates DEFINITION
Relative standard deviation: NMT 4.0% Benzylpenicilloyl Polylysine Injection has a molar concentration
Analysis of benzylpenicilloyl moiety (C16N2H19O5S) of NLT 5.4 105 M
Sample: Standard solution and Sample solution and NMT 7.0 105 M. It contains one or more suitable
[NOTEThe retention time for L-lysine is 57 min.] buffers.
Calculate the molar concentration of lysine in the
Concentrate: ASSAY
PROCEDURE
Result = (rU/rS) (C/1000 * Mr) F Solution A: 9 g of sodium chloride and 1.38 g of
monobasic sodium phosphate in 900 mL. Adjust with 5 N
rU = peak response of the Sample solution sodium hydroxide or phosphoric acid to a pH of 7.6. Dilute
rS = peak response of the Standard solution to 1000 mL.
C = concentration of USP L-Lysine Hydrochloride RS Solution B: 0.07 mg/mL of mercuric chloride
in the Standard solution (g/mL) Sample solution: Combine the contents of a sufficient
Mr = molecular weight of anhydrous lysine number of containers to obtain NLT 3 mL of Injection.
hydrochloride, 182.65 Transfer 3.0 mL of Injection to a 10-mL volumetric flask, and
F = dilution factor of the Sample solution (100) dilute with Solution A to volume.
Calculate the percentage of benzylpenicilloyl substitution: Blank: Solution A
Analysis
Result = (B/L) 100 Samples: Blank and Sample solution
Transfer 3.0 mL of Sample solution to a spectrophotometric
B = molar concentration of benzylpenicilloyl moiety cell. Using a suitable spectrophotometer and using the
in the Concentrate, as determined in the Assay Blank, determine the initial absorbance at the wavelength
L = molar concentration of lysine in the Concentrate of maximum absorbance at 282 nm. Add 0.02 mL of
Acceptance criteria: 50%70% Solution B to the Sample solution in the spectrophotometric
IMPURITIES cell, and determine the absorbance at the same
Organic Impurities wavelength after 1 and 3 min. Repeat the addition of 0.02-
PROCEDURE: LIMIT OF PENICILLENATE AND PENAMALDATE mL portions of Solution B until a maximum absorbance
Sample solution: 1 mL of Concentrate in a 50-mL reading is obtained.
volumetric flask. Dilute with Solution A, prepared as Calculate the molar concentration of benzylpenicilloyl
directed in the Assay, to volume. Using a suitable moiety in the Injection:
spectrophotometer and using Solution A as a Blank, Result = (10/3){[AM(3 + 0.02N)/3] AI}/MAB
determine the absorbances at the wavelengths of
maximum absorption at 322 nm and 282 nm. AM = highest absorbance observed
Calculate the molar concentration of penicillenate taken: N = number of 0.02-mL portions of Solution B added
to the Sample solution to obtain the maximum
Result = 50A322/MA1B absorbance
A322 = absorbance at 322 nm AI = initial absorbance
MA1 = molar absorptivity of the penicillenate moiety at MA = molar absorptivity of the penamaldate formed by
pH 7.6, 26,600 the reaction of benzylpenicilloyl with mercuric
B = length of the cell (cm) chloride at pH 7.6: 22,325
52 Benzylpenicilloyl / Official Monographs USP 32
B = length of the cell (cm) Acceptance criteria: 96.0%101.0%

Acceptance criteria: NLT 5.4 105 M and NMT 7.0 105
M IMPURITIES
BACTERIAL ENDOTOXINS TEST 85: NMT 5833.0 USP [NOTE2 g of specimen being used]
Endotoxin Units/mL HEAVY METALS, Method II 231: NMT 10 ppm
STERILITY TESTS 71: Meets the requirements when tested as
directed under Test for Sterility of the Product to Be Examined, SPECIFIC TESTS
Membrane Filtration MELTING RANGE OR TEMPERATURE 741: 176182, with
PH 791: 6.58.5 decomposition
LOSS ON DRYING 731: Dry it in a vacuum over phosphorus
ADDITIONAL REQUIREMENTS pentoxide at 40 for 4 h: It loses NMT 0.2% of its weight.
PACKAGING AND STORAGE: Preserve in single-dose or in
multiple-dose containers, preferably of Type I glass, in a ADDITIONAL REQUIREMENTS
refrigerator. PACKAGING AND STORAGE: Preserve in tight, light-resistant
USP REFERENCE STANDARDS 11 containers
USP Endotoxin RS
Beta Carotene Capsules

Beta Carotene (Comment on this Monograph)id=m8735=Beta Carotene
(Comment on this Monograph)id=m8730=Beta Carotene=B- Capsules=B-Monos.pdf)
Monos.pdf)
DEFINITION
Beta Carotene Capsules contain NLT 90.0% and NMT 125.0%
of the labeled amount of C40H56.
IDENTIFICATION
Grind a portion of the Capsule contents, equivalent to 10 mg
of beta carotene, transfer to a centrifuge tube, add 5 mL of
chloroform, shake for 1 min, and centrifuge for 3 min. Filter
C40H56 536.87 the supernatant layer, collecting 2 mL of the filtrate in a 25-
,-Carotene; mL conical flask. Add 5 mL of antimony trichloride TS to the
all-trans--Carotene; filtrate: A transient purple or blue color forms.
(all-E)-1,1-(3,7,12,16-Tetramethyl-1,3,5,7,9,11,13,15,17- ASSAY
octadecanonaene-1,18-diyl)bis[2,6,6-trimethylcyclohexene] PROCEDURE
[7235-40-7]. [NOTEPerform this analysis in subdued light, using low-
DEFINITION actinic glassware.]
Beta Carotene contains NLT 96.0% and NMT 101.0% of C40H56. Cyclohexane: Spectrophotometric grade, or material that
has been purified by being passed through a column of
IDENTIFICATION activated silica gel and distilled.
A. Determine the absorbances of the Assay Sample solution Iodine solution: 0.01 mg/mL of iodine in cyclohexane.
at 455 nm and at 483 nm: The ratio of the absorbance at [NOTEPrepare this solution fresh daily.]
455 nm to that at 483 nm is 1.141.18. Sample solution 1: Combine the contents of NLT 20
B. Determine the absorbance of the Assay Sample solution Capsules, and grind, using a freezer mill, to a fine powder of
at 455 nm and that of the Assay Sample stock solution at 340 uniform color. Transfer a quantity of the finely ground
nm: The ratio of the absorbance at 455 nm to that at 340 Capsule contents, equivalent to 75 mg of beta carotene, to
nm is NLT 1.5. a 1000-mL volumetric flask, add 500 mL of water, and heat
at 60 for 15 min. Cool to ambient temperature, and dilute
ASSAY with water to volume.
PROCEDURE Sample solution 2: Transfer 5.0 mL of Sample solution 1 to
[NOTEPerform this procedure in subdued light, using low- a glass-stoppered, 50-mL centrifuge tube. Add 3 g of
actinic glassware.] sodium sulfate decahydrate, 2 mL of 1 N hydrochloric acid,
Sample stock solution: Dissolve 50 mg of Beta Carotene in and 20.0 mL of chloroform. Shake by mechanical means for
10 mL of acid-free chloroform in a 100-mL volumetric flask. 10 min, centrifuge for 5 min, and remove the aqueous layer
Dilute with cyclohexane to volume. Pipet 5 mL into a 100- without disturbing the chloroform layer. Add 2 g of
mL volumetric flask, and dilute with cyclohexane. anhydrous sodium sulfate to the chloroform layer, shake
Sample solution: Sample stock solution with cyclohexane (1 vigorously, and allow to settle.
in 10) Sample solution 3: Transfer 5.0 mL of Sample solution 2 to
Spectrometric conditions a 50-mL volumetric flask, and add 30 mL of cyclohexane.
Analytical wavelength: 455 nm Add 0.05 mL of Iodine solution, dilute with cyclohexane to
Blank: Cyclohexane volume, and allow to stand for 3 h. Transfer 20 mL of this
Sample: Sample solution solution to a centrifuge tube, and centrifuge for 2 min.
Analysis: Determine the absorbance of Sample solution Standard solution 1: 17 mg of beta carotene, previously
using the Blank. Calculate the percentage of C40H56 in the subjected to the Assay and previously dried in vacuum over
portion of Beta Carotene taken: phosphorus pentoxide at 40 for 4 h, and transfer to a
1000-mL volumetric flask. Add 10 mL of water, heat at 60
Result = 400 (AU/250) 100 for 15 min, and cool to room temperature. Add 3 g of
sodium sulfate decahydrate and 2 mL of 1 N hydrochloric
AU = absorbance of the Sample solution acid, and shake by mechanical means for 10 min. Add 200
250 = absorptivity of pure beta carotene
USP 32 Official Monographs / Betahistine 53
mL of chloroform, and shake for 10 min. Add 750 mL of Standard solution: 0.38 mg/mL of USP Betahistine
chloroform, shake, and dilute with chloroform to volume, Hydrochloride RS in Mobile phase
disregarding the aqueous layer. Sample solution: 0.38 mg/mL of Betahistine Hydrochloride
Standard solution 2: Shake Standard solution 1 vigorously, in Mobile phase
and allow the layers to separate completely. Transfer 20 mL Chromatographic system
of the chloroform layer to a centrifuge tube, add 2 g of (See Chromatography 621, System Suitability.)
anhydrous sodium sulfate, shake vigorously, and allow to Mode: LC
settle. Transfer 5.0 mL to a 50 mL volumetric flask, add 30 Detector: UV 254 nm
mL of cyclohexane. Add 0.05 mL of Iodine solution, dilute Column: 3.0-mm 15-cm; packing L1
with cyclohexane to volume, and allow to stand for 3 h. Temperature: 40
Transfer 20 mL of Standard solution 2 to a centrifuge tube, Flow rate: 0.5 mL/min
and centrifuge for 2 min. Injection size: 10 L
Spectrometric conditions System suitability
Analytical wavelength: 452 nm Sample: Standard solution
Blank: Cyclohexane Suitability requirements
Samples: Sample solution 3 and Standard solution 2 Tailing factor: NMT 2.0
Measure the absorbances using the Blank. Relative standard deviation: NMT 2.0%
Calculate the percentage, of C40H56 in the portion of Analysis
Capsule contents taken: Samples: Sample solution and Standard solution
Calculate the percentage of C8H12N2 2HCl in the portion
Result = (AU/AS) (CS/CU) 100 of Betahistine Hydrochloride taken:
AU = absorbance of Sample solution 3 Result = (rU/rS) (CS/CU) 100
AS = absorbance of Standard solution 2
CS = concentration of beta carotene in the Standard rU = peak response from the Sample solution
solution (g/mL) rS = peak response from the Standard solution
CU = nominal concentration of beta carotene in CS = concentration of USP Betahistine Hydrochloride
Sample solution 3 (g/mL) RS in the Standard solution (mg/mL)
Acceptance criteria: 90.0%125.0% CU = concentration of Betahistine Hydrochloride taken
to prepare the Sample solution (mg/mL)
PERFORMANCE TESTS Acceptance criteria: 99.0%101.0%
IMPURITIES
ADDITIONAL REQUIREMENTS Inorganic Impurities
PACKAGING AND STORAGE: Preserve in tight, light-resistant RESIDUE ON IGNITION 281: NMT 0.1%
containers. Organic Impurities
PROCEDURE
Mobile phase and Chromatographic system: Proceed as
Betahistine Hydrochloride directed in the Assay.
Sample solution: 0.38 mg/mL of Betahistine
(Comment on this Monograph)id=m8750=Betahistine Hydrochloride in Mobile phase
Hydrochloride=B-Monos.pdf) Analysis
of Betahistine Hydrochloride taken:
Result = (ru/rT) F 100
ru = peak response for each impurity
rT = sum of the responses of all of the peaks,
C8H12N2 2HCl 209.12 adjusted for the relative response factor
2-Pyridineethanamine, N-methyl-, dihydrochloride; F = response factor of the respective impurity (see
2-[2-(Methylamino)ethyl]pyridine; Impurity Table) and 1.0 for all other peaks
dihydrochloride [5579-84-0]. Acceptance criteria
Individual impurities: See Impurity Table.
DEFINITION Total impurities: NMT 0.5%
Betahistine Hydrochloride contains NLT 99.0% and NMT
101.0% of C8H12N2 2HCl, calculated on the dried basis.
Impurity Table
IDENTIFICATION Relative Relative Acceptance
A. INFRARED ABSORPTION 197K Retention Response Criteria,
B. The retention time of the major peak of the Sample Name Time Factor NMT (%)
obtained in the Assay. 2-(2-Hydroxyethyl) 0.3 0.5 0.2
pyridine
ASSAY 2-Vinylpyridine 0.4 0.4 0.2
PROCEDURE
Solution A: 0.69 mg/mL of ammonium acetate in water,
adjust with glacial acetic acid to a pH of 4.7
Mobile phase: Acetonitrile and Solution A (7:13), containing
2.88 mg/mL of sodium lauryl sulfate
54 Betahistine / Official Monographs USP 32
Impurity Table (continued) Acceptance criteria: 98.0%100.5%

Relative Relative Acceptance IMPURITIES
Retention Response Criteria, Inorganic Impurities
Name Time Factor NMT (%) RESIDUE ON IGNITION 281: NMT 0.1%
N-Methyl-N,N-bis(2- 2.4 1.4 0.2 HEAVY METALS 231: NMT 10 ppm
pyridin-2-yl-ethyl)
amine SPECIFIC TESTS
Any other individual 0.1
impurities
SPECIFIC TESTS USP REFERENCE STANDARDS 11
PH 791: 2.03.0, in a solution (1 in 10) USP Betaine Hydrochloride RS
LOSS ON DRYING 731: Dry it between 100 and 105 to
constant weight: it loses NMT 1.0% of its weight.
Betamethasone
USP Betahistine Hydrochloride RS (Comment on this Monograph)id=m8760=Betamethasone=B-
Monos.pdf)
Betaine Hydrochloride
(Comment on this Monograph)id=m8755=Betaine
Hydrochloride=B-Monos.pdf)
C22H29FO5 392.46
Pregna-1,4-diene-3,20-dione, 9-fluoro-11,17,21-trihydroxy-16-
methyl-, (11,16)-;
9-Fluoro-11,17,21-trihydroxy-16-methylpregna-1,4-
diene-3,20-dione [378-44-9].
C5H11NO2 HCl 153.61 DEFINITION
Methanaminium, 1-carboxy-N, N, N-trimethyl-, chloride; Betamethasone contains NLT 97.0% and NMT 103.0% of
Betaine hydrochloride; C22H29FO5, calculated on the dried basis.
(Carboxymethyl)trimethylammonium
chloride [590-46-5]. IDENTIFICATION
DEFINITION B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
Betaine Hydrochloride contains NLT 98.0% and NMT 100.5% Sample solution: 0.5 mg/mL Betamethasone in dehydrated
of C5H11NO2 HCl, calculated on the anhydrous basis. alcohol
IDENTIFICATION Developing solvent system: Chloroform and diethylamine
A. INFRARED ABSORPTION 197K (2:1)
B. IDENTIFICATION TESTSGENERAL, Chloride 191: A 50 Analysis: Proceed as directed in the chapter, except to
mg/mL solution meets the requirements. locate the spots by lightly spraying with dilute sulfuric acid
(1 in 2) and heating on a hot plate or under a lamp until
ASSAY spots appear.
PROCEDURE
Sample solution: Transfer 400 mg of Betaine Hydrochloride ASSAY
to a conical flask, add 50 mL of glacial acetic acid, and heat PROCEDURE
gently with swirling until solution is complete. Add 25 mL of Mobile phase: Acetonitrile and water (37:63)
mercuric acetate TS, cool, and add 2 drops of crystal violet Internal standard stock solution: 0.25 mg/mL
TS. propylparaben in alcohol
Analysis: Titrate with 0.1 N perchloric acid VS to a green Standard stock solution: 0.2 mg/mL USP Betamethasone
endpoint. Perform a blank determination, and make any RS in alcohol
necessary correction (see Titrimetry 541). Each mL of 0.1 N Standard solution: Internal standard stock solution and
perchloric acid is equivalent to 15.36 mg of C5H11NO2 HCl. Standard stock solution (1:1)
USP 32 Official Monographs / Betamethasone 55
Sample stock solution: 0.2 mg/mL Betamethasone in IDENTIFICATION

alcohol THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
Sample solution: Internal standard stock solution and Sample solution: 1 mg/mL, prepared by concentrating 10
Sample stock solution (1:1) mL of the Sample solution from the Assay on a steam bath to
Chromatographic system 1 mL
(See Chromatography 621, System Suitability.) Standard solution: 1 mg/mL of USP Betamethasone RS in
Mode: LC dehydrated alcohol
Detector: UV 240 nm Developing solvent system: Chloroform and diethylamine
Column: 4.6-mm 25-cm; packing L1 (2:1)
Flow rate: 1 mL/min Spray reagent: Methanol, sulfuric acid, and nitric acid
Injection size: 10 L (10:10:1)
System suitability Analysis
Sample: Standard solution Samples: Sample solution and Standard solution
[NOTEThe relative retention times for betamethasone and Proceed as directed in the chapter, except to spray the
propylparaben are 1.0 and 1.4, respectively.] plate with the Spray reagent, and heat at 105 for 10 min.
Resolution: NLT 3.0 between betamethasone and ASSAY
propylparaben PROCEDURE
Relative standard deviation: NMT 2.0% Mobile phase: Acetonitrile and water (37:63)
Analysis Internal standard solution: 0.25 mg/mL of propylparaben
Samples: Standard solution and Sample solution in alcohol
Calculate the percentage of C22H29FO5 taken: Standard stock solution: 0.2 mg/mL USP Betamethasone
RS in alcohol
Result = (RU/RS) (CS/CU) 100 Standard solution: Internal Standard solution and Standard
stock solution (1:1)
RU = peak height ratio of the betamethasone peak to Sample solution: To a portion of Cream equivalent to 2 mg
the internal standard peak in the Sample of betamethasone, add 10.0 mL of Internal standard solution
solution and 10.0 mL of alcohol. Mix by rotation for 20 min.
RS = peak height ratio of the betamethasone peak to Centrifuge at 2500 rpm for 10 min. Transfer a portion of the
the internal standard peak in the Standard supernatant to a suitable vial.
solution Chromatographic system
CS = concentration of USP Betamethasone RS in the (See Chromatography 621, System Suitability.)
Standard solution (mg/mL) Mode: LC
CU = concentration of Betamethasone in the Sample Detector: UV 240 nm
solution (mg/mL) Column: 4.6-mm 25-cm; packing L1
Acceptance criteria: 97.0%103.0% Flow rate: 1 mL/min
IMPURITIES System suitability
Inorganic Impurities Sample: Standard solution
RESIDUE ON IGNITION 281: NMT 0.2%, a platinum crucible [NOTEThe relative retention times for betamethasone and
being used propylparaben are 1.0 and 1.4, respectively.]
Organic Impurities Suitability requirements
PROCEDURE: ORDINARY IMPURITIES 466 Resolution: NLT 3.0 between betamethasone and
Sample solution: Methanol propylparaben
Standard solution: Methanol Relative standard deviation: NMT 2.0%
Eluant: Toluene, acetone, methyl ethyl ketone, and formic Samples: Sample solution and Standard solution
acid (55:20:20:5), in a nonequilibrated chamber Calculate the label claim percentage of C22H29FO5 in the
Visualization: 5 portion of Cream taken:
SPECIFIC TESTS Result = (RU/RS) (CS/CU) 100
OPTICAL ROTATION, Specific Rotation 781S: +118 to +126,
calculated on the dried basis RU = peak height ratio of the betamethasone peak to
Sample solution: 5 mg/mL, in methanol the internal standard peak from the Sample
LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT solution
1.0% of its weight. RS = peak height ratio of the betamethasone peak to
the internal standard peak from the Standard
ADDITIONAL REQUIREMENTS solution
PACKAGING AND STORAGE: Preserve in tight containers. Store CS = concentration of USP Betamethasone RS in the
between 2 and 30. Standard solution (mg/mL)
USP REFERENCE STANDARDS 11 CU = nominal concentration of betamethasone in the
USP Betamethasone RS Sample solution (mg/mL)
Betamethasone Cream SPECIFIC TESTS

(Comment on this Monograph)id=m8770=Betamethasone MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED
Cream=B-Monos.pdf) MICROORGANISMS 62: It meets the requirements of the
tests for the absence of Staphylococcus aureus and
DEFINITION Pseudomonas aeruginosa.
Betamethasone Cream contains NLT 90.0% and NMT 115.0%
of the labeled amount of C22H29FO5 in a suitable cream base.
56 Betamethasone / Official Monographs USP 32
MINIMUM FILL 755: Meets the requirements rotate for 20 min. Centrifuge to clarify. Transfer 10-mL
portions of the supernatant to separate, stoppered tubes.
ADDITIONAL REQUIREMENTS To each tube add 1 mL of blue tetrazolium TS, followed
PACKAGING AND STORAGE: Preserve in collapsible tubes or in by 1 mL of Reagent, and mix. Heat in a 35 water bath for
tight containers. 1 h. Remove from the water bath, add 1 mL of glacial
USP REFERENCE STANDARDS 11 acetic acid to each tube, and mix. Cool to room
USP Betamethasone RS temperature. Concomitantly determine the absorbances of
the solutions in 1-cm cells at the wavelength of maximum
absorbance at 525 nm, with a suitable
Betamethasone Oral Solution spectrophotometer.
Calculate the percentage of C22H29FO5 in each mL of the
(Comment on this Monograph)id=m8780=Betamethasone Oral Oral Solution taken:
Solution=B-Monos.pdf)
Result = (AU AB)/(AS AB) (CS/CU) 100
DEFINITION
Betamethasone Oral Solution contains NLT 90.0 % and NMT AU =
absorbance of the Sample solution
115.0 % of the labeled amount of betamethasone (C22H29FO5). AB =
absorbance of the blank
AS =
absorbance of the Standard solution
IDENTIFICATION CS =
concentration of USP Betamethasone RS in the
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution (mg/mL)
Sample solution: Evaporate 1 mL of the Sample solution, CU = nominal concentration of betamethasone in the
prepared as directed in the Assay, on a steam bath just to Sample solution (mg/mL)
dryness, and dissolve the residue in 0.5 mL of alcohol. Acceptance criteria: 90.0%115.0%
Developing solvent system: Chloroform and diethylamine
(2:1) ADDITIONAL REQUIREMENTS
Analysis PACKAGING AND STORAGE: Store between 2 and 25,
Samples: Sample solution and Standard solution excursions permitted up to 30, protected from light.
Proceed as directed in the chapter. Locate the spots by Preserve in a tight container. Protect from freezing.
lightly spraying with dilute sulfuric acid (1 in 2) and USP REFERENCE STANDARDS 11
heating on a hot plate or under a lamp until spots appear. USP Betamethasone RS
ASSAY
PROCEDURE
Adsorbent: 0.25-mm layer of chromatographic silica gel Betamethasone Tablets
Standard solution: 0.6 mg/mL of USP Betamethasone RS in (Comment on this Monograph)id=m8790=Betamethasone
a mixture of chloroform and methanol (1:1) Tablets=B-Monos.pdf)
Sample solution: Equivalent to 1.2 mg of Betamethasone
from Oral Solution in a 50-mL centrifuge tube. Rinse the DEFINITION
pipet with 15 mL of 0.1 N hydrochloric acid, then with 20 Betamethasone Tablets contain NLT 90.0% and NMT 110.0%
mL of ethyl acetate, and add the rinsings to the centrifuge of the labeled amount of betamethasone (C22H29FO5).
tube. Rotate for 10 min, or shake manually for 1 min.
[NOTEDo not use a mechanical shaker.] IDENTIFICATION
Centrifuge to separate the phases. Transfer the upper phase THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
(ethyl acetate) to a small, pear-shaped flask. Extract the Sample solution: Evaporate 50 mL of the Sample solution,
aqueous phase twice more with 20-mL portions of ethyl prepared as directed in the Assay, on a steam bath just to
acetate, and add the extracts to the pear-shaped flask. dryness, and dissolve the residue in 1 mL of chloroform.
Evaporate the combined extracts on a steam bath under a Developing solvent system: Chloroform and diethylamine
gentle stream of nitrogen to dryness. Allow to cool to (2:1)
room temperature. Dissolve the residue in 0.5 mL of Application volume: 10 L
chloroform and methanol (1:1), using a vortex mixer. Analysis
Transfer the solution to a 2-mL volumetric flask, with small Sample: Sample solution
portions of a mixture of chloroform and methanol (1:1), Proceed as directed in the chapter, except to locate the
and dilute with the mixture of chloroform and methanol spots by lightly spraying with dilute sulfuric acid (1 in 2)
(1:1) to volume. and heating on a hot plate or under a lamp until spots
Application volume: 200 L appear.
Developing solvent system: Chloroform, methanol, and
ammonium hydroxide (175:20:1) ASSAY
Reagent: Tetramethylammonium hydroxide TS in alcohol (1 PROCEDURE
in 5) Mobile phase: Acetonitrile and water (1:2)
Analysis Internal standard solution: 0.125 mg/mL of
Samples: Sample solution and Standard solution beclomethasone in methanol
Proceed as directed for Chromatography 621, Thin-Layer Standard stock solution: 0.1 mg/mL of USP
Chromatography. Allow the spots to dry, and develop the Betamethasone RS in methanol
chromatogram, using the Developing solvent system, until Standard solution: 0.05 mg/mL from a mixture of Standard
the solvent front has moved 15 cm. Remove the plates stock solution and Internal standard solution (1:1)
from the developing chamber, and allow them to dry for Sample solution: Equivalent to 0.5 mg of betamethasone
15 min. Mark the betamethasone bands, using short- from powdered Tablets (NLT 20 Tablets), into a 125-mL
wavelength UV light, to include similar zones of silica gel separator. Add 25 mL of water, and shake by mechanical
for the Sample solution, the Standard solution, and a zone means for 15 min. Add 5.0 mL of Internal standard solution.
containing no betamethasone for the blank. Scrape off Extract with four 25-mL portions of chloroform. Filter the
these zones, and transfer them to separate 50-mL chloroform extracts through 4 g of chloroform-washed
centrifuge tubes. Add 15 mL of alcohol to each, and anhydrous sodium sulfate, collecting the extracts in a 150-
mL beaker. Evaporate the extracts on a steam bath with the Tolerances: NLT 75% (Q) of the labeled amount of
aid of a stream of nitrogen to dryness, taking care to avoid C22H29FO5
overheating. Dissolve the residue in 2 mL of methanol, and UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
transfer to a 10-mL volumetric flask. Rinse the beaker with Analysis for content uniformity
small portions of methanol, transferring the rinses to the Standard solution: 12 g/mL instead of 10 g/mL of USP
same flask. Dilute with methanol to volume. Betamethasone RS, as directed under Assay for Steroids
Chromatographic system 351
(See Chromatography 621, System Suitability.) Sample solution: Weigh and finely powder 1 Tablet.
Mode: LC Transfer to a 125-mL separator, add 20 mL of water, and
Detector: UV 254 nm shake. Extract the betamethasone completely, using three
Column: 4-mm 30-cm; packing L1 15-mL portions of chloroform and filtering each extract
Flow rate: 1.2 mL/min through chloroform-washed cotton into a 50-mL
Injection size: 10 L volumetric flask. Dilute with chloroform to volume, and
System suitability mix. Transfer 20.0 mL of this solution to a glass-stoppered,
Sample: Standard solution 50-mL conical flask, evaporate the chloroform on a steam
[NOTEThe relative retention times for beclomethasone bath just to dryness, cool, and dissolve the residue in 20.0
and betamethasone are 1.4 and 1.0, respectively.] mL of alcohol.
Suitability requirements Analysis: Proceed as directed under Assay for Steroids
Resolution: NLT 1.7 between the analyte and internal 351, except to keep the flasks in a constant-temperature
standard peaks bath at 45 1 for 90 min, then add 1.0 mL of glacial
Relative standard deviation: NMT 2.0% acetic acid, and cool.
Analysis Calculate the percentage of C22H29FO5 in the Tablet:
Calculate the percentage of C22H29FO5 in the portion of (AU/AS) (CS/CU) 100
Tablets taken:
Result = (RU/RS) (CS/CU) 100 AS = absorbance of the Standard solution
CS = concentration of USP Betamethasone RS in the
RU = peak height ratio of the Sample solution Standard solution (g/mL)
RS = peak height ratio of the Standard solution CU = concentration of the Sample solution, based the
CS = concentration of USP Betamethasone RS in the extent of dilution (g/mL)
CU = nominal concentration of betamethasone in the ADDITIONAL REQUIREMENTS
Sample solution (mg/mL) PACKAGING AND STORAGE: Preserve in tight containers. Store
Acceptance criteria: 90.0%110.0% between 2 and 25, excursions permitted between 15 and
30. [NOTEProtect the 21-tablet pack from excessive
PERFORMANCE TESTS moisture.]
DISSOLUTION, Procedure for a Pooled Sample 711 USP REFERENCE STANDARDS 11
Medium: 900 mL water USP Betamethasone RS
Add 1.0 mL of Internal standard solution to each vessel.
Apparatus 2: 50 rpm
Time: 45 min
Mobile phase: Methanol and water (3:2) Betamethasone Acetate
Internal standard solution: 0.5 mg/mL of testosterone in (Comment on this Monograph)id=m8820=Betamethasone
methanol Acetate=B-Monos.pdf)
Standard stock solution: 0.5 mg/mL of USP C24H31FO6 434.50
Betamethasone RS in methanol Pregna-1,4-diene-3,20-dione, 9-fluoro-11,17-dihydroxy-16-
Standard solution: 1 mL of Standard stock solution, and 1 methyl-21-(acetyloxy)-, (11,16)-;
mL of Internal standard solution. Dilute to 900 mL. 9-Fluoro-11,17,21-trihydroxy-16-methylpregna-1,4-
Sample solution: Sample per Dissolution 711. Dilute with diene-3,20-dione 21-acetate [987-24-6].
Medium to a concentration that is similar to that of the
Standard solution. DEFINITION
Chromatographic system Betamethasone Acetate contains NLT 97.0% and NMT 103.0%
(See Chromatography 621, System Suitability.) of C24H31FO6, calculated on the anhydrous basis.
Mode: LC
Flow rate: 2 mL/min B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
Injection size: 200 L Sample solution: 0.5 mg/mL in dehydrated alcohol
System suitability Developing solvent system: Chloroform and diethylamine
Sample: Standard solution (2:1)
[NOTEThe relative retention times for betamethasone and Analysis: Proceed as directed in the chapter. Locate the
testosterone are 0.5 and 1.0, respectively.] spots on the plate by lightly spraying with 10% sulfuric acid
Suitability requirements in alcohol and heating on a hot plate or under a lamp until
Resolution: NLT 1.5 between betamethasone and the spots appear.
testosterone
Analysis PROCEDURE
Samples: Standard solution and Sample solution Mobile phase: Acetonitrile, glacial acetic acid, and water
Calculate the quantity of C22H29FO5 dissolved in comparison (700:1.5:800)
with the Standard solution, similarly chromatographed.
Internal standard solution: 0.7 mg/mL of progesterone in Betamethasone Benzoate

Mobile phase (Comment on this Monograph)id=m8870=Betamethasone
Standard stock solution: 0.5 mg/mL of USP Benzoate=B-Monos.pdf)
Betamethasone Acetate RS in Mobile phase
Standard solution: 10.0 mL Internal standard solution and C29H33FO6 496.58
10.0 mL Standard stock solution. Dilute to 50 mL with Mobile Pregna-1,4-diene-3,20-dione, 17-(benzoyloxy)-9-fluoro-11,21-
phase. The concentration of USP Betamethasone Acetate RS dihydroxy-16-methyl-, (11,16)-;
is 0.1 mg/mL. 9-Fluoro-11,17,21-trihydroxy-16-methylpregna-1,4-
Sample stock solution: 0.5 mg/mL of Betamethasone diene-3,20-dione 17-benzoate [22298-29-9].
Acetate in Mobile phase
Sample solution: 10.0 mL Internal standard solution and DEFINITION
10.0 mL Sample stock solution. Dilute to 50 mL with Mobile Betamethasone Benzoate contains NLT 98.0% and NMT
phase. 102.0% of C29H33FO6, calculated on the dried basis.
Mode: LC INFRARED ABSORPTION 197M
Column: 4-mm 30-cm; packing L1 PROCEDURE
Flow rate: 1 mL/min Mobile phase: Acetonitrile and water (3:2)
Injection size: 25 L Internal standard solution: 0.6 mg/mL of betamethasone
System suitability dipropionate in methanol
Sample: Standard solution Standard stock solution: 0.6 mg/mL of USP
[NOTEThe relative retention times for progesterone and Betamethasone Benzoate RS in methanol
betamethasone acetate are 3 and 1.0, respectively.] Standard solution: 0.2 mg/mL from Standard stock solution
Suitability requirements and Internal standard solution (1:2)
Resolution: NLT 2 between the analyte and internal Sample stock solution: 0.6 mg/mL of Betamethasone
standard peaks Benzoate in methanol
Relative standard deviation: NMT 2.0% Sample solution: 0.2 mg/mL from Sample stock solution
Analysis and Internal standard solution (1:2)
Samples: Standard solution and Sample solution Chromatographic system
Calculate the percentage of C24H31FO6 in the portion taken: (See Chromatography 621, System Suitability.)
Result = (RU/RS) (CS/CU) 100 Mode: LC
Detector: UV 254 nm
RU = peak response ratio of the Sample solution Column: 4-mm 30-cm; 5-m packing L1
RS = peak response ratio of the Standard solution Flow rate: 1 mL/min
CS = concentration of USP Betamethasone Acetate RS Injection size: 15 L
in the Standard solution (mg/mL) System suitability
CU = concentration of Betamethasone in the Sample Sample: Standard solution (three replicate injections)
solution (mg/mL) [NOTEThe retention times for betamethasone benzoate
Acceptance criteria: 97.0%103.0% and betamethasone dipropionate are 5 min and 7 min,
respectively.]
IMPURITIES Suitability requirements
Inorganic Impurities Resolution: NLT 3, between betamethasone benzoate
RESIDUE ON IGNITION 281: NMT 0.2%, a platinum crucible and the internal standard
being used Relative standard deviation: NMT 2.0%
Organic Impurities Analysis
PROCEDURE: ORDINARY IMPURITIES 466 Samples: Sample solution and Standard solution
Sample solution: Methanol Calculate the percentage of C29H33FO6 in the portion of
Standard solution: Methanol Betamethasone Benzoate taken:
Eluant: A mixture of toluene and isopropyl alcohol Result = (RU/RS) (CS/CU) 100
(90:10), in a nonequilibrated chamber
Visualization: 5 RU = peak response ratios of the betamethasone
benzoate peak to the internal standard peak
SPECIFIC TESTS from the Sample solution
OPTICAL ROTATION, Specific Rotation 781S: +120 to +128 RS = peak response ratios of the betamethasone
Sample solution: 10 mg/mL, in dioxane benzoate peak to the internal standard peak
WATER DETERMINATION, Method I 921: NMT 4.0% from the Standard solution
CS = concentration of USP Betamethasone Benzoate
PACKAGING AND STORAGE: Preserve in tight containers. Store CU = concentration of Betamethasone Benzoate in the
between 2 and 30. Sample solution (mg/mL)
USP Betamethasone Acetate RS
Acceptance criteria: 98.0%102.0% Sample solution: Transfer the equivalent of 0.5 mg of

betamethasone benzoate from the Gel to a 125-mL
IMPURITIES separatory funnel, add 20 mL of water and 2 mL of
Organic Impurities saturated sodium acetate solution, shake to disperse the Gel,
PROCEDURE: RELATED STEROIDS and add 2 mL of Internal standard solution. Extract this
Sample solution: 20.0 mg/mL in methanol solution with one 50-mL portion of chloroform followed by
Standard solution A: 5 mg/mL of USP Betamethasone three 40-mL portions of chloroform. Discard the aqueous
Benzoate RS in methanol layer. Wash the chloroform extract with 10 mL of water,
Standard solution B: 100 g/mL from Standard solution A allow to stand for 10 min, then pass through chloroform-
in methanol wetted glass fiber filter paper and anhydrous sodium sulfate
Chromatographic system into a suitable container. Evaporate to dryness under a
(See Chromatography 621, Thin-Layer Chromatography.) vacuum at 30. Dissolve the residue in 10 mL of methanol.
Mode: TLC Chromatographic system
Adsorbent: 0.25-mm layer of chromatographic silica gel (See Chromatography 621, System Suitability.)
mixture Mode: LC
Application volume: 10 L Detector: UV 236 nm
Developing solvent system: Toluene, acetone, and Column: 3.9-mm 30-cm; packing L1
methanol (75:25:4) Temperature: 30
Analysis Flow rate: 1.5 mL/min
Samples: Sample solution and Standard solutions Injection size: 10 L
Examine the plate under short-wavelength UV light. System suitability
Acceptance criteria: The principal spot from the Sample Sample: Standard solution
solution corresponds in RF value to that of Standard solution [NOTEThe relative retention times for methyltestosterone
A; and the Sample solution shows NMT 3 additional spots, and betamethasone benzoate are about 1.0 and 1.33,
the intensity and size of which do not exceed those of the respectively.]
spot from Standard solution B. Suitability requirements
Resolution: NLT 3.0 between methyltestosterone and
SPECIFIC TESTS betamethasone benzoate
OPTICAL ROTATION, Specific Rotation 781S: +60 to +66 Relative standard deviation: NMT 1.0%
Sample solution: 40 mg/mL, in dioxane Analysis
LOSS ON DRYING 731: Dry 200 mg at 105 for 3 h: it loses Samples: Sample solution and Standard solution
NMT 0.5% of its weight. Calculate the percentage label claim of C29H33FO6 in the
ADDITIONAL REQUIREMENTS portion of Gel taken:
between 2 and 30. Result = (RU/RS) (CS/CU) 100
USP REFERENCE STANDARDS 11 RU = peak response ratio of betamethasone benzoate
USP Betamethasone Benzoate RS to methyltestosterone from the Sample solution
RS = peak response ratio of betamethasone benzoate
to methyltestosterone from the Standard
Betamethasone Benzoate Gel solution
CS = concentration of USP Betamethasone Benzoate
(Comment on this Monograph)id=m8890=Betamethasone RS in the Standard solution (g/mL)
Benzoate Gel=B-Monos.pdf) CU = nominal concentration of betamethasone
DEFINITION benzoate in the Sample solution (mg/mL)
Betamethasone Benzoate Gel contains an amount of Acceptance criteria: 90.0%110.0%
betamethasone benzoate (C29H33FO6) equivalent to NLT 90.0% SPECIFIC TESTS
and NMT 110.0% of the labeled amount of betamethasone MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED
benzoate (C29H33FO6). MICROORGANISMS 62: It meets the requirements of the
IDENTIFICATION tests for absence of Staphylococcus aureus and Pseudomonas
The retention time of the major peak of the Sample solution aeruginosa.
corresponds to that of the Standard solution, both relative to MINIMUM FILL 755: Meets the requirements
the internal standard, as obtained in the Assay. ADDITIONAL REQUIREMENTS
ASSAY PACKAGING AND STORAGE: Preserve in tight containers. Store
PROCEDURE at 25, excursions permitted between 15 and 30. Protect
Mobile phase: Methanol, acetonitrile, and water (23:9:18) from freezing.
Internal standard solution: 250 g/mL of USP USP REFERENCE STANDARDS 11
Methyltestosterone RS in methanol USP Betamethasone Benzoate RS
Standard stock solution: 0.5 mg/mL of USP USP Methyltestosterone RS
Betamethasone Benzoate RS in methanol
Standard solution: 5.0 mL of Standard stock solution, 10
mL of Internal standard solution, diluted with methanol to 50
mL
Betamethasone Dipropionate the peak obtained from the Internal standard in the
(Comment on this Monograph)id=m8960=Betamethasone Standard solution is 0.6 full-scale.]
Dipropionate=B-Monos.pdf) Detector: UV 254 nm or 240 nm
Temperature: Room temperature
Column pressure: Capable up to 3500 psi
Injection size: 5 L25 L
System suitability
Sample: Standard solution (three successive injections)
Relative standard deviation: NMT 2.0% between the
C28H37FO7 504.60 lowest and highest peak area ratios
Pregna-1,4-diene-3,20-dione, 9-fluoro-11-hydroxy-16- Analysis
methyl-17,21-bis(1-oxopropoxy)-, (11,16); Sample: Standard solution and Sample solution
9-Fluoro-11,17,21-trihydroxy-16- Determine the ratio of the peak heights, at equivalent
methylpregna-1,4-diene-3,20-dione 17,21-dipropionate retention times, obtained with the Sample solution and the
[5593-20-4]. Standard solution.
Calculate the percentage of C28H37FO7 in the portion of
DEFINITION Betamethasone Dipropionate taken:
Betamethasone Dipropionate contains NLT 97.0% and NMT
103.0% of C28H37FO7, calculated on the dried basis. Result = (RU/RS) (CS/CU) 100
IDENTIFICATION RU = peak height ratios of betamethasone
A. INFRARED ABSORPTION 197M dipropionate to the internal standard of the
B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution
Sample solution: 1 mg/mL, in chloroform RS = peak height ratios of betamethasone
Developing solvent system: Chloroform and acetone (7:1) dipropionate to the internal standard of the
Standard solution
ASSAY CS = concentration of USP Betamethasone
PROCEDURE Dipropionate RS in the Standard solution
Mobile phase: Acetonitrile solution (1 in 2), degassed by (mg/mL)
ultrasonic vibration for 5 to 10 min, such that the retention CU = nominal concentration of Betamethasone
time of betamethasone dipropionate is approximately 14 Dipropionate in the Sample solution (mg/mL)
min and that of beclomethasone dipropionate is Acceptance criteria: 97.0%103.0%
approximately 18 min. [NOTEDo not leave the Mobile
phase in the column overnight, but flush the system after IMPURITIES
use with water for 15 min, followed by methanol for 15 Inorganic Impurities
min.] RESIDUE ON IGNITION 281: NMT 0.2%, a platinum crucible
Internal standard solution: 0.9 mg/mL of USP being used
Beclomethasone Dipropionate RS in a solution of acetic acid Organic Impurities
in methanol (1 in 1000) PROCEDURE
Standard stock solution: 0.6 mg/mL of USP Mobile phase: Acetonitrile and water (13:7)
Betamethasone Dipropionate RS in a solution of acetic acid System suitability solution: 0.05 mg/mL of each USP
in methanol (1 in 1000) Betamethasone Dipropionate RS and USP Betamethasone
Standard solution: Standard stock solution and Internal Valerate RS in Mobile phase
standard solution (1:1) Sample solution: 0.3 mg/mL of Betamethasone
[NOTEThe concentrations for the Standard solution are 0.3 Dipropionate in Mobile phase, and shake until dissolved
mg/mL of betamethasone dipropionate and 0.45 mg/mL Chromatographic system
of beclomethasone dipropionate.] (See Chromatography 621, System Suitability.)
Sample stock solution: 0.6 mg/mL of Betamethasone Mode: LC
Dipropionate in a solution of acetic acid in methanol (1 in Detector: UV 254 nm
1000) Column: 4.6-mm 15-cm; packing L1
Sample solution: Sample stock solution and Internal Flow rate: 1 mL/min
standard solution (1:1) Injection size: 10 L
Chromatographic system System suitability
(See Chromatography 621, System Suitability.) Sample: System suitability solution
[NOTESeparately inject equal volumes (5 L25 L) of the Resolution: NLT 4.0 between betamethasone valerate
Sample solution and Standard solution by means of a and betamethasone dipropionate
suitable microsyringe or sampling valve, adjusting the
specimen size and other operating parameters such that
Column efficiency: NLT 8000 theoretical plates Diluent: Isopropyl alcohol containing acetic acid (1 in
Analysis 1000)
Sample: Sample solution Internal standard solution: 0.90 mg/mL of USP
Calculate the percentage of each impurity in the portion Beclomethasone Dipropionate RS in Diluent
of Betamethasone Dipropionate taken: Standard stock solution: 0.642 mg/mL of USP
Betamethasone Dipropionate RS in Diluent
Result = (ru/rT) 100 Standard solution: 10.0 mL Standard stock solution and
10.0 mL Internal standard solution, dilute with Diluent to 100
ru = peak response for each impurity mL
rT = sum of the responses for all the peaks [NOTEThe concentrations for the Standard solution are
Acceptance criteria 0.09 mg/mL of beclomethasone dipropionate and 0.0642
Individual impurities: NMT 1.0% of any individual mg/mL of betamethasone dipropionate.]
impurity is found Sample solution: Discharge the entire contents of the
Total impurities: NMT 2.0% container of Topical Aerosol into a 100-mL volumetric flask.
Allow the solution to warm to room temperature slowly to
SPECIFIC TESTS prevent it from boiling out of the flask, then evaporate the
OPTICAL ROTATION, Specific Rotation 781S: +63 to +70 propellant by swirling the flask in a water bath at 25 until
Sample solution: 10 mg/mL, in dioxane the solution stops bubbling. Add 10.0 mL of Internal
LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT standard solution, and dilute with Diluent to volume. Pass the
1.0% of its weight. solution through a 0.45-m filter.
ADDITIONAL REQUIREMENTS Chromatographic system
PACKAGING AND STORAGE: Preserve in tight containers. Store (See Chromatography 621, System Suitability.)
at 25, excursions permitted between 15 and 30. Mode: LC
USP REFERENCE STANDARDS 11 [NOTESeparately inject equal volumes (525 L) of the
USP Beclomethasone Dipropionate RS Standard solution and Sample solution by means of a
USP Betamethasone Dipropionate RS suitable microsyringe or sampling valve, adjusting the
USP Betamethasone Valerate RS specimen size and other operating parameters such that
the peak obtained from the internal standard in the
Standard solution is 0.6 full-scale.]
Detector: UV 254 nm or 240 nm
Betamethasone Dipropionate Topical Column: 4-mm 30-cm; packing L1
Aerosol Temperature: Room temperature
Column pressure: Capable up to 3500 psi
(Comment on this Monograph)id=m8970=Betamethasone Injection size: 525 L
Dipropionate Topical Aerosol=B-Monos.pdf) System suitability
DEFINITION Sample: Standard solution [NOTEthree successive
Betamethasone Dipropionate Topical Aerosol is a solution, in injections]
suitable propellants in a pressurized container, of Suitability requirements
betamethasone dipropionate (C28H37FO7) equivalent to NLT Relative standard deviation: NMT 2.0% between the
90.0% and NMT 110.0% of the labeled amount of lowest and highest peak area ratios
betamethasone (C22H29FO5). Analysis
IDENTIFICATION Calculate the percentage of C22H29FO5 equivalent to the
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 quantity of C28H37FO7 in the container of the Topical
Standard solution: 3.2 mg/mL of USP Betamethasone Aerosol:
Dipropionate RS in methanol
Sample solution: Place the container in a dry icemethanol Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100
bath for 5 min. Open the can by means of a tube-cutter,
and allow the propellant to evaporate under a gentle stream RU = peak height ratios of the betamethasone
of nitrogen for 1 h. Transfer 3 mL of the residue to a 50-mL dipropionate to beclomethasone dipropionate
centrifuge tube. Add 10 mL of a mixture of methanol and peaks from the Sample solution
water (4:1), and shake vigorously. Centrifuge to clarify. RS = peak height ratios of the betamethasone
Application volume: 25 L dipropionate to beclomethasone dipropionate
Developing solvent system: Toluene and ethyl acetate peaks from the Standard solution
(1:1) CS = concentration of USP Betamethasone
Analysis Dipropionate RS in the Standard solution
Samples: Standard solution and Sample solution (mg/mL)
Proceed as directed in the chapter. Spray the plate with a CU = nominal concentration of the Sample solution
mixture of sulfuric acid, methanol, and nitric acid (mg/mL)
(10:10:1), and heat at 105 for 15 min. Mr1 = molecular weight of betamethasone, 392.46
Mr2 = molecular weight of betamethasone
ASSAY dipropionate, 504.60
PROCEDURE Acceptance criteria: 90.0%110.0%
Mobile phase: Acetonitrile solution (1 in 2), degassed by
ultrasonic vibration for 510 min, such that the retention SPECIFIC TESTS
time of betamethasone dipropionate is approximately 14 OTHER REQUIREMENTS: It meets the requirements for Aerosols,
min and that of beclomethasone dipropionate is Nasal Sprays, Metered-Dose Inhalers, and Dry Powder Inhalers
approximately 18 min. [NOTEDo not leave the Mobile 601, Pressure Test, Minimum Fill, and Leakage Test.
phase in the column overnight, but flush the system after
use with water for 15 min, followed by methanol for 15
min.]
ADDITIONAL REQUIREMENTS for 5 min. Transfer a portion of the supernatant to a suitable

PACKAGING AND STORAGE: Preserve in tight, pressurized vial.
containers, and avoid exposure to excessive heat. Store at Chromatographic system
25, excursions permitted between 15 and 30. (See Chromatography 621, System Suitability.)
USP Beclomethasone Dipropionate RS [NOTESeparately inject equal volumes (5 L25 L) of the
USP Betamethasone Dipropionate RS Sample solution and Standard solution by means of a
suitable microsyringe or sampling valve, adjusting the
specimen size and other operating parameters such that
the peak obtained from the Internal standard solution in
Betamethasone Dipropionate Cream the Standard solution is 0.6 full-scale.]
(Comment on this Monograph)id=m8980=Betamethasone Detector: UV 254 nm or 240 nm
Dipropionate Cream=B-Monos.pdf) Column: 4-mm 30-cm; packing L1
Temperature: Room temperature
DEFINITION Column pressure: Capable up to 3500 psi
Betamethasone Dipropionate Cream contains an amount of Injection size: 5 L25 L
betamethasone dipropionate (C28H37FO7) equivalent to NLT System suitability
90.0% and NMT 110.0% of the labeled amount of Sample: Standard solution (three successive injections)
betamethasone (C22H29FO5), in a suitable cream base. Suitability requirements
IDENTIFICATION lowest and highest peak area ratios
A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Analysis
Standard solution: 150 g/mL of USP Betamethasone Samples: Sample solution and Standard solution
Dipropionate RS in chloroform Calculate the percentage of C22H29FO5 in the portion of
Sample solution: 1.5 g of Cream to a glass-stoppered, 50- Cream taken:
mL centrifuge tube. Add 15 mL of a methanolhydrochloric
acid solution prepared by mixing 1 volume of dilute Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100
hydrochloric acid (1 in 120) with 4 volumes of methanol.
Shake to obtain a homogeneous mixture. Add 30 mL of RU = peak height ratio of the betamethasone
solvent hexane, mix for 10 min, and centrifuge. Using a dipropionate peak and the internal standard
suitable syringe, transfer the lower aqueous phase to a peak obtained from the Sample solution
second centrifuge tube, add 20 mL of water. Extract this RS = peak height ratio of the betamethasone
aqueous mixture with chloroform by shaking, centrifuging, dipropionate peak and the internal standard
and removing the lower, chloroform phase with a syringe. peak obtained from the Standard solution
Evaporate the chloroform on a steam bath with the aid of a CS = concentration of USP Betamethasone
stream of nitrogen to dryness, cool, and dissolve the residue Dipropionate RS in the Standard solution
in chloroform to obtain a solution containing 150 g/mL of (mg/mL)
betamethasone dipropionate. CU = nominal concentration of betamethasone
Application volume: 40 L dipropionate in the Sample solution (mg/mL)
Developing solvent system: Chloroform and acetone (7:1) Mr1 = molecular weight of betamethasone, 392.46
Analysis Mr2 = molecular weight of betamethasone
Samples: Sample solution and Standard solution dipropionate, 504.60
Proceed as directed in the chapter. Acceptance criteria: 90.0%110.0%
PROCEDURE MINIMUM FILL 755: Meets the requirements
Mobile phase: Acetonitrile solution (1 in 2), degassed by
ultrasonic vibration for 5 to 10 min, such that the retention ADDITIONAL REQUIREMENTS
time of betamethasone dipropionate is approximately 14 PACKAGING AND STORAGE: Preserve in collapsible tubes or
min and that of beclomethasone dipropionate is tight containers. Store at 25; excursions permitted between
approximately 18 min. [NOTEDo not leave the Mobile 15 and 30. Protect from freezing.
phase in the column overnight, but flush the system after USP REFERENCE STANDARDS 11
use with water for 15 min, followed by methanol for 15 USP Beclomethasone Dipropionate RS
min.] USP Betamethasone Dipropionate RS
Diluent: Acetic acid in methanol (1 in 1000)
Internal standard solution: 0.45 mg/mL of USP
Beclomethasone Dipropionate RS in Diluent
Standard stock solution: 0.2 mg/mL of USP Betamethasone Dipropionate Lotion
Betamethasone Dipropionate RS in Diluent (Comment on this Monograph)id=m8990=Betamethasone
Standard solution: Standard stock solution and Internal Dipropionate Lotion=B-Monos.pdf)
standard solution (2:1)
[NOTEThe concentrations for the Standard solution are DEFINITION
0.133 mg/mL of betamethasone dipropionate and 0.15 Betamethasone Dipropionate Lotion contains an amount of
mg/mL of beclomethasone dipropionate.] betamethasone dipropionate (C28H37FO7) equivalent to NLT
Sample solution: Equivalent to 2 mg of betamethasone 90.0% and NMT 110.0% of the labeled amount of
dipropionate from Cream, into a capped 50-mL centrifuge betamethasone (C22H29FO5), in a suitable lotion base.
tube. Add 5.0 mL of Internal standard solution and 10.0 mL
of Diluent. Heat in a water bath at 60, shaking IDENTIFICATION
intermittently, until the specimen melts. Remove from the THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
bath, and shake vigorously until the specimen has Standard solution: 150 g/mL of USP Betamethasone
resolidified. Repeat the heating and shaking. Freeze in an Dipropionate RS in chloroform
icemethanol bath for 15 min, and centrifuge at 2500 rpm
Sample solution: Equivalent to 0.6 mg of betamethasone RU = peak height ratios of the betamethasone
dipropionate from Lotion. In a 50-mL vial, add 10 mL of 0.1 dipropionate peak to the internal standard
N hydrochloric acid, then add 4 mL of chloroform. Disperse peak from the Sample solution
on a vortex mixer for about 1 min, then shake vigorously for RS = peak height ratios of the betamethasone
10 min, and centrifuge at 2000 rpm for about 5 min. dipropionate peak to the internal standard
Transfer the chloroform layer to a suitable vial. peak from the Standard solution
Application volume: 40 L CS = concentration of USP Betamethasone
Developing solvent system: Chloroform and acetone (7:1) Dipropionate RS in the Standard solution
Analysis (mg/mL)
Samples: Sample solution and Standard solution CU = nominal concentration of betamethasone
Proceed as directed in the chapter. dipropionate in the Sample solution (mg/mL)
Mr1 = molecular weight of betamethasone, 392.46
ASSAY Mr2 = molecular weight of betamethasone
PROCEDURE dipropionate, 504.60
Mobile phase: Acetonitrile solution (about 1 in 2), Acceptance criteria: 90.0%110.0%
degassed by ultrasonic vibration for 5 to 10 min, such that
the retention time of betamethasone dipropionate is SPECIFIC TESTS
approximately 14 min and that of beclomethasone MINIMUM FILL 755: Meets the requirements
dipropionate is approximately 18 min
[NOTEDo not leave the Mobile phase in the column ADDITIONAL REQUIREMENTS
overnight, but flush the system after use with water for 15 PACKAGING AND STORAGE: Preserve in tight containers. Store
min, followed by methanol for 15 min.] at 25; excursions permitted between 15 and 30. Protect
Internal standard solution: 0.9 mg/mL of USP from light and freezing.
Beclomethasone Dipropionate RS in chloroform USP REFERENCE STANDARDS 11
Standard stock solution: 0.6 mg/mL of USP USP Beclomethasone Dipropionate RS
Betamethasone Dipropionate RS in chloroform USP Betamethasone Dipropionate RS
Standard solution: Standard stock solution and Internal
[NOTEThe concentrations for the Standard solution are 0.3 Betamethasone Dipropionate Ointment
mg/mL of betamethasone dipropionate and 0.45 mg/mL
of beclomethasone dipropionate.] (Comment on this Monograph)id=m9020=Betamethasone
To 10.0 mL of 0.1 N hydrochloric acid in a capped 5-mL Dipropionate Ointment=B-Monos.pdf)
centrifuge tube add 4.0 mL of the solution, and treat this
solution as follows. Cap, and shake vigorously for about 2 DEFINITION
min, or disperse on a vortex mixer for about 1 min. Betamethasone Dipropionate Ointment contains an amount of
Centrifuge at 2500 rpm for about 3 min. Transfer the betamethasone dipropionate (C28H37FO7) equivalent to NLT
chloroform phase to a suitable vial. Evaporate the 90.0% and NMT 110.0% of the labeled amount of
chloroform under a stream of nitrogen at a slightly betamethasone (C22H29FO5), in a suitable ointment base.
elevated temperature to dryness. Cool the vial to room IDENTIFICATION
temperature, add 4.0 mL of methanol, and swirl to dissolve A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
the residue. Standard solution: 150 g/mL of USP Betamethasone
Sample solution: Equivalent to 1.2 mg of betamethasone Dipropionate RS in chloroform
dipropionate from Lotion. In a capped 50-mL centrifuge Sample solution: 1.5 g of Ointment to a glass-stoppered,
tube, add 10.0 mL of 0.1 N hydrochloric acid, shake to 50-mL centrifuge tube. Add 15 mL of
disperse, then add 2.0 mL of Internal standard solution and methanolhydrochloric acid solution prepared by mixing 1
2.0 mL of chloroform. Cap, and shake vigorously for about volume of dilute hydrochloric acid (1 in 120) with 4
2 min, or disperse on a vortex mixer for about 1 min. volumes of methanol. Shake to obtain a homogeneous
Centrifuge at 2500 rpm for about 3 min. Transfer the mixture. Add 30 mL of solvent hexane, mix for 10 min, and
chloroform phase to a suitable vial. Evaporate the centrifuge. Using a suitable syringe, transfer the lower
chloroform under a stream of nitrogen at a slightly elevated aqueous phase to a second centrifuge tube, add 20 mL of
temperature to dryness. Cool the vial to room temperature, water, and mix. Extract this aqueous mixture with
add 4.0 mL of methanol, and swirl to dissolve the residue. chloroform by shaking, centrifuging, and removing the
Chromatographic system lower, chloroform phase with a syringe. Evaporate the
(See Chromatography 621, System Suitability.) chloroform on a steam bath with the aid of a stream of
Mode: LC nitrogen to dryness, cool, and dissolve the residue in
Detector: UV 254 or 240 nm chloroform to obtain a solution containing 150 g/mL of
Column: 4-mm 30-cm; packing L1 betamethasone dipropionate.
Temperature: Room temperature Application volume: 40 L
Column pressure: Capable up to 3500 psi Developing solvent system: Chloroform and acetone (7:1)
System suitability Samples: Sample solution and Standard solution
Samples: Standard solution [NOTEthree successive Proceed as directed in the chapter.
injections]
Suitability requirements ASSAY
Relative standard deviation: NMT 2.0% between the PROCEDURE
lowest and highest peak area ratios Mobile phase: Acetonitrile solution (1 in 2), degassed by
Analysis ultrasonic vibration for 510 min, such that the retention
Samples: Sample solution and Standard solution time of betamethasone dipropionate is approximately 14
Calculate the percentage of C22H29FO5 in the portion of min and that of beclomethasone dipropionate is
Lotion taken: approximately 18 min. [NOTEDo not leave the Mobile
phase in the column overnight, but flush the system after
Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100
use with water for 15 min, followed by methanol for 15 USP Betamethasone Dipropionate RS
min.]
Diluent: Acetic acid in alcohol (1 in 1000)
Internal standard solution: 0.45 mg/mL of USP
Beclomethasone Dipropionate RS in Diluent Betamethasone Sodium Phosphate
Standard stock solution: 0.2 mg/mL of USP (Comment on this Monograph)id=m9070=Betamethasone
Betamethasone Dipropionate RS in Diluent Sodium Phosphate=B-Monos.pdf)
Standard solution: Standard stock solution and Internal C22H28FNa2O8P 516.40
standard solution (2:1) Pregna-1,4-diene-3,20-dione, 9-fluoro-11,17-dihydroxy-16-
[NOTEThe concentrations for the Standard solution are methyl-21-(phosphonooxy)-, disodium salt, (11,16)-;
0.133 mg/mL of betamethasone dipropionate and 0.15 9-Fluoro-11,17,21-trihydroxy-16-methylpregna-1,4-
mg/mL of beclomethasone dipropionate.] diene-3,20-dione 21-(disodium phosphate) [151-73-5].
Sample solution: Equivalent to 2 mg of betamethasone
dipropionate from Ointment, in a capped 50-mL centrifuge DEFINITION
tube. Add 5.0 mL of Internal standard solution and 10.0 mL Betamethasone Sodium Phosphate contains NLT 97.0% and
of Diluent. Heat in a water bath at 70, shaking NMT 103.0% of C22H28FNa2O8P, calculated on the anhydrous
intermittently until the sample melts. Remove from the bath, basis.
and shake vigorously until the ointment has solidified.
Repeat the heating and shaking operation. Freeze in an IDENTIFICATION
icemethanol bath for 15 min, and centrifuge at 2500 rpm A. INFRARED ABSORPTION 197M
for 5 min. Transfer a portion of the supernatant to a suitable B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
vial. Standard solution: 1 mg/mL of USP Betamethasone
Chromatographic system Sodium Phosphate RS in methanol
(See Chromatography 621, System Suitability.) Sample solution: 1 mg/mL
Mode: LC Developing solvent system: 500 mL of butyl alcohol and
[NOTEUse a suitable microsyringe or sampling valve, and 200 mL of dilute hydrochloric acid (1 in 12)
adjust the specimen size and other operating parameters Place in a separatory funnel, and mix. Use the organic layer
such that the peak obtained from the internal standard in as the developing solvent.
the Standard solution is 0.6 full-scale.] Spray reagent: A mixture of sulfuric acid, methanol, and
Detector: UV 254 nm or 240 nm nitric acid (10:10:1)
Column: 4-mm 30-cm; packing L1 Analysis
Temperature: Room temperature Samples: Sample solution and Standard solution
Column pressure: Capable up to 3500 psi Proceed as directed in Thin-Layer Chromatographic
Injection size: 525 L Identification Test 201, except to spray the plate with
System suitability Spray reagent, and heat at 105 for 10 min.
Sample: Standard solution [NOTEthree successive C. IDENTIFICATION TESTSGENERAL, Sodium 191 AND
injections] Phosphate 191: Ignite it at 800 (see Residue on Ignition
Suitability requirements 281): the residue meets the requirements.
lowest and highest peak area ratios ASSAY
Analysis PROCEDURE
Samples: Sample solution and Standard solution Mobile phase: Methanol and 0.07 M anhydrous monobasic
Calculate the percentage of C22H29FO5 in the portion of potassium phosphate (3:2)
Ointment: Standard solution: 0.17 mg/mL of USP Betamethasone
Sodium Phosphate RS in a mixture of methanol and water
Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 (3:2)
Sample solution: 0.17 mg/mL of Betamethasone Sodium
RU = peak height ratio of the betamethasone Phosphate in a mixture of methanol and water (3:2)
dipropionate peak and the internal standard Chromatographic system
peak from the Sample solution (See Chromatography 621, System Suitability.)
RS = peak height ratio of the betamethasone Mode: LC
dipropionate peak and the internal standard Detector: UV 254 nm
peak from the Standard solution Column: 3.9-mm 30-cm; packing L1
CS = concentration of USP Betamethasone Flow rate: 1.5 mL/min
Dipropionate RS in the Standard solution Injection size: 20 L
(mg/mL) System suitability
CU = nominal concentration of the Sample solution Sample: Standard solution
(mg/mL) Suitability requirements
Mr1 = molecular weight of betamethasone, 392.46 Tailing factor: NMT 2.0
Mr2 = molecular weight of betamethasone Relative standard deviation: NMT 3.0%
dipropionate, 504.60 Analysis
Acceptance criteria: 90.0%110.0% Samples: Sample solution and Standard solution
Calculate the percentage of C22H28FNa2O8P in the portion
PERFORMANCE TESTS of Betamethasone Sodium Phosphate taken:
PACKAGING AND STORAGE: Preserve in collapsible tubes or rU = peak response from the Sample solution
tight containers. Store at 25, excursions permitted between rS = peak response from the Standard solution
15 and 30. Protect from freezing.
USP Beclomethasone Dipropionate RS
CS = concentration of USP Betamethasone Sodium A = absorbance of the Sample solution

Phosphate RS in the Standard solution Acceptance criteria: 250 g (1.0%)
(mg/mL)
CU = concentration of Betamethasone Sodium SPECIFIC TESTS
Phosphate in the Sample solution (mg/mL) SPECIFIC ROTATION 781S: +99 to +105
Acceptance criteria: 97.0%103.0% Sample solution: 10 mg/mL
IMPURITIES
Inorganic Impurities ADDITIONAL REQUIREMENTS
LIMIT OF PHOSPHATE IONS PACKAGING AND STORAGE: Preserve in tight containers.
Standard phosphate solution and Phosphate reagent A: USP REFERENCE STANDARDS 11
Prepare as directed under Phosphate in Reagents (see USP Betamethasone Sodium Phosphate RS
Reagents, Indicators, and SolutionsGeneral Tests for
Reagents).
Phosphate reagent B: 350 mg of p-methylaminophenol Betamethasone Sodium Phosphate
sulfate in 50 mL of water. Add 20 g of sodium
metabisulfite, mix to dissolve, and dilute with water to 100 Injection
mL. (Comment on this Monograph)id=m9080=Betamethasone
Sample solution: 50 mg of Betamethasone Sodium Sodium Phosphate Injection=B-Monos.pdf)
Phosphate in a mixture of 10 mL of water and 5 mL of 2 N
sulfuric acid contained in a 25-mL volumetric flask, by DEFINITION
warming if necessary. Add 1 mL each of Phosphate reagent Betamethasone Sodium Phosphate Injection is a sterile solution
A and Phosphate reagent B, dilute with water to 25.0 mL, of Betamethasone Sodium Phosphate in Water for Injection. It
mix, and allow to stand at room temperature for 30 min. contains an amount of betamethasone sodium phosphate
Standard solution: 5.0 mL of Standard phosphate solution (C22H28FNa2O8P) equivalent to NLT 90.0% and NMT 110.0%
in a mixture of 10 mL of water and 5 mL of 2 N sulfuric of the labeled amount of betamethasone (C22H29FO5).
acid contained in a 25-mL volumetric flask, by warming if
necessary. Add 1 mL each of Phosphate reagent A and IDENTIFICATION
Phosphate reagent B, dilute with water to 25.0 mL, mix, THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
and allow to stand at room temperature for 30 min. Adsorbent: Chromatographic silica gel mixture
Blank: Water Standard solution: 2 mg/mL of USP Betamethasone
Spectrometric conditions Sodium Phosphate RS in methanol
Mode: Spectrophotometry Sample solution: 2 mg/mL of betamethasone sodium
Analytical wavelength: 730 nm phosphate from Injection in methanol
Cell: 1 cm Application volume: 10 L
Analysis Analysis
Samples: Sample solution and Standard solution Samples: Sample solution and Standard solution
Acceptance criteria: The absorbance of the Sample Develop the chromatogram in an equilibrated chamber
solution is NMT that of the Standard solution. The limit is containing n-butyl alcohol previously shaken with 1 N
1.0% of phosphate (PO4). hydrochloric acid, until the solvent front has moved three-
Organic Impurities fourths of the length of the plate. Remove the plate from
PROCEDURE: LIMIT OF FREE BETAMETHASONE the developing chamber, air-dry, then spray with a
Sample solution: 25.0 mg of Betamethasone Sodium mixture of sulfuric acid, methanol, and nitric acid
Phosphate in water to make 25.0 mL. Transfer 5.0 mL of (10:10:1). Heat the plate at 105 for 10 min.
the solution to a separator, and extract with three 25-mL Acceptance criteria: The RF value of the principal spot of
portions of chloroform. Filter each extract through a the Sample solution corresponds to that of the Standard
chloroform-saturated cotton pledget, combining the solution.
filtrates in a conical flask. Evaporate the chloroform on a ASSAY
steam bath with the aid of a current of air to dryness, and PROCEDURE
dissolve the residue in methanol to make 25.0 mL. Mobile phase: Methanol and 0.05 M monobasic potassium
Blank solution: 5.0 mL of water transferred to a separator phosphate (1:1)
Proceed as directed under Sample solution. The methanolic Internal standard solution: 1 mg/mL of butylparaben in
solution so obtained is the Blank solution. methanol
Spectrometric conditions Standard stock solution: 4 mg/mL of USP Betamethasone
Mode: Spectrophotometry Sodium Phosphate RS
Analytical wavelength: 239 nm Standard solution: 3.0 mL of Standard stock solution and
Blank: Blank solution 5.0 mL of Internal standard solution. Dilute to 25 mL.
Cell: 1 cm Sample solution: Equivalent to 9 mg of betamethasone
Analysis from a volume of Injection, in a 25-mL volumetric flask. Add
Sample: Sample solution 5.0 mL of the Internal standard solution, and dilute to
Calculate the quantity, in mg, of free betamethasone in volume.
the portion of Betamethasone Sodium Phosphate taken: Chromatographic system
Result = A 3.125
Detector: UV 254 nm A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
Column: 3.9-mm 30-cm; packing L1 Absorbent: 0.25-mm layer of chromatographic silica gel
Flow rate: 2 mL/min Standard solution: 2 mg/mL of USP Betamethasone
Injection size: 20 L Sodium Phosphate RS in methanol and water solution (1:1)
System suitability Sample solution: Dilute 2 mL with 2 mL of methanol.
Sample: Standard solution Application volume: 10 L
[NOTEThe relative retention times for betamethasone Developing solvent system: Place 500 mL of butyl alcohol
sodium phosphate and butylparaben are 1.0 and 2.4, and 200 mL of dilute hydrochloric acid (1 in 12) in a
respectively.] separatory funnel, and mix. Use the organic layer as the
Suitability requirements developing solvent.
Resolution: NLT 5 between the analyte and internal Spray reagent: A mixture of sulfuric acid, methanol, and
standard peaks nitric acid (10:10:1)
Relative standard deviation: NMT 2.0% Analysis
Analysis Samples: Sample solution and Standard solution
Samples: Standard solution and Sample solution Proceed as directed in the chapter except to spray the
Calculate the percentage of C22H29FO5 in each mL of plate with Spray reagent, and heat at 105 for 10 min.
Injection taken: B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
Absorbent: 0.25-mm layer of chromatographic silica gel
Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 Standard solution: 1.5 mg/mL of USP Betamethasone
Acetate RS in methanol and water solution (1:1)
RU = peak response ratio of the Sample solution Sample solution: Dilute 2 mL with 2 mL of methanol.
RS = peak response ratio of the Standard solution Application volume: 10 L
CS = concentration of USP Betamethasone Sodium Developing solvent system: Chloroform and diethylamine
Phosphate RS in the Standard solution (mg/mL) (2:1)
CU = nominal concentration of betamethasone in the Analysis
Sample solution (mg/mL) Samples: Sample solution and Standard solution
Mr1 = molecular weight of betamethasone, 392.47 Proceed as directed in the chapter, except to locate the
Mr2 = molecular weight of betamethasone sodium spots by lightly spraying with dilute sulfuric acid (1 in 2),
phosphate, 516.41 and heating on a hot plate or under a lamp until spots
Acceptance criteria: 90.0%110.0% appear.
SPECIFIC TESTS ASSAY
BACTERIAL ENDOTOXINS TEST 85: NMT 29.2 USP Endotoxin PROCEDURE
Units/mg of betamethasone Mobile phase: Methanol and 0.075 M monobasic
PH 791: 8.09.0 potassium phosphate (7:5)
PARTICULATE MATTER IN INJECTIONS 788: Meets the Internal standard solution: 1 mg/mL of methyltestosterone
requirements for small-volume injections in methanol
OTHER REQUIREMENTS: It meets the requirements under Standard stock solution A: 2.52 mg/mL of USP
Injections 1. Betamethasone Sodium Phosphate RS in Mobile phase
Standard stock solution B: 1.8 mg/mL of USP
ADDITIONAL REQUIREMENTS Betamethasone Acetate RS in methanol
PACKAGING AND STORAGE: Preserve in single-dose or in Standard solution: 5 mL Standard stock solution A, 5 mL
multiple-dose containers, preferably of Type I glass. Standard stock solution B, 10 mL Internal Standard solution.
USP REFERENCE STANDARDS 11 Dilute to 100 mL with Mobile phase.
USP Betamethasone Sodium Phosphate RS [NOTEThe Standard solution has a known concentration of
USP Endotoxin RS 126 g/mL USP Betamethasone Sodium Phosphate RS and
90 g/mL USP Betamethasone Acetate RS.]
Sample solution: Equivalent to 9 mg of betamethasone
Betamethasone Sodium Phosphate and acetate. Using a To contain pipet, transfer a volume of the
Betamethasone Acetate Injectable well-mixed Injectable Suspension to a 100-mL volumetric
flask. Rinse the pipet with 10 mL of Mobile phase, collecting
Suspension the rinse in the volumetric flask. Add 10 mL of Internal
(Comment on this Monograph)id=m9125=Betamethasone standard solution, and dilute with Mobile phase to volume.
Sodium Phosphate and Betamethasone Acetate Injectable Chromatographic system
Suspension=B-Monos.pdf) (See Chromatography 621, System Suitability.)
Mode: LC
DEFINITION Detector: UV 254 nm
Betamethasone Sodium Phosphate and Betamethasone Acetate Column: 3.9-mm 30-cm; packing L1
Injectable Suspension is a sterile solution of Betamethasone Flow rate: 1.2 mL/min
Sodium Phosphate in solution and Betamethasone Acetate in Injection size: 20 L
suspension in Water for Injection. It contains an amount of System suitability
betamethasone sodium phosphate (C22H28FNa2O8P) equivalent Sample: Standard solution
to NLT 90.0% and NMT 115.0% of the labeled amount of [NOTEThe relative retention times for betamethasone
betamethasone (C22H29FO5), and NLT 90.0% and NMT phosphate, methyltestosterone, and betamethasone
115.0% of the labeled amount of betamethasone acetate acetate are 0.5, 1.7, and 1.0, respectively.]
(C24H31FO6).
Suitability requirements USP Betamethasone Sodium Phosphate RS

Resolution: NLT 5.0 between the betamethasone USP Endotoxin RS
phosphate and betamethasone acetate peaks; NLT 3.0
between the betamethasone acetate and internal standard
peaks
Relative standard deviation: NMT 2.0% Betamethasone Valerate
Analysis (Comment on this Monograph)id=m9180=Betamethasone
Samples: Sample solution and Standard solution Valerate=B-Monos.pdf)
Calculate the percentage of C24H31FO6 in each mL of
Injectable Suspension taken:
RU = peak response ratio for betamethasone acetate
and methyltestosterone from the Sample
solution
RS = peak response ratio for betamethasone acetate C27H37FO6 476.59
and methyltestosterone from the Standard Pregna-1,4-diene-3,20-dione, 9-fluoro-11,21-dihydroxy-16-
solution methyl-17-[(1-oxopentyl)oxy]-, (11,16)-;
CS = concentration of USP Betamethasone Acetate RS 9-Fluoro-11,17,21-trihydroxy-16-methylpregna-1,4-
in the Standard solution (g/mL) diene-3,20-dione 17-valerate [2152-44-5].
(mg/mL) DEFINITION
Calculate the percentage of C22H29FO5 equivalent to the Betamethasone Valerate contains NLT 97.0% and NMT 103.0%
quantity of C22H28FNa2O8P in each mL of Injectable of C27H37FO6, calculated on the dried basis.
Suspension taken:
IDENTIFICATION
Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 A. INFRARED ABSORPTION 197M
B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
RU = peak response ratio for betamethasone Sample solution: 1 mg/mL, in alcohol
phosphate and methyltestosterone from the Developing solvent system: Toluene and ethyl acetate
Sample solution (1:1)
RS = peak response ratio for betamethasone Analysis
phosphate and methyltestosterone from the Samples: Sample solution and Standard solution
Standard solution Proceed as directed in the chapter. Spray the plate with a
CS = concentration of USP Betamethasone Sodium mixture of sulfuric acid, methanol, and nitric acid
Phosphate RS in the Standard solution (10:10:1), and heat at 105 for 15 min.
(mg/mL)
CU = nominal concentration of the Sample solution ASSAY
(mg/mL) PROCEDURE
Mr1 = molecular weight of betamethasone, 392.46 Mobile phase: Acetonitrile and water (3:2)
Mr2 = molecular weight of betamethasone sodium Diluent: Glacial acetic acid in methanol (1 in 1000)
phosphate, 516.41 Internal standard solution: 0.4 mg/mL of USP
Acceptance criteria: 90.0%115.0% Beclomethasone Dipropionate RS in Diluent
SPECIFIC TESTS Betamethasone Valerate RS in Diluent
BACTERIAL ENDOTOXINS TEST 85: NMT 29.2 USP Endotoxin Standard solution: Standard stock solution and Internal
Units/mg of betamethasone standard solution (1:2)
PH 791: 6.87.2 [NOTEThis solution has a concentration of 0.2 mg/mL of
OTHER REQUIREMENTS: It meets the requirements under USP Betamethasone Valerate RS.]
Injections 1. Sample stock solution: 0.6 mg/mL of Betamethasone
Valerate in Diluent
ADDITIONAL REQUIREMENTS Sample solution: Sample stock solution and Internal
PACKAGING AND STORAGE: Preserve in multiple-dose standard solution (1:2)
containers, preferably of Type I glass. Chromatographic system
USP Betamethasone Acetate RS

Detector: UV 254 nm PACKAGING AND STORAGE: Preserve in tight containers.
Column: 4-mm 30-cm; packing L1 USP REFERENCE STANDARDS 11
Flow rate: 1.2 mL/min USP Beclomethasone Dipropionate RS
Injection size: 10 L USP Betamethasone Valerate RS
System suitability
[NOTEThe relative retention times for betamethasone
valerate and beclomethasone dipropionate are 1.0 and Betamethasone Valerate Cream
1.7, respectively.] (Comment on this Monograph)id=m9200=Betamethasone
Suitability requirements Valerate Cream=B-Monos.pdf)
Resolution: NLT 4.5 between betamethasone valerate
and beclomethasone dipropionate DEFINITION
Relative standard deviation: NMT 2.0% Betamethasone Valerate Cream contains an amount of
Analysis betamethasone valerate (C27H37FO6) equivalent to NLT 90.0%
Samples: Standard solution and Sample solution and NMT 110.0% of the labeled amount of betamethasone
Calculate the percentage of C27H37FO6 in the portion of (C22H29FO5), in a suitable cream base.
Betamethasone Valerate taken:
IDENTIFICATION
Result = (RU/RS) (CS/CU) 100 THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
RU = peak response ratio of the Sample solution betamethasone from Cream to a separator, add 20 mL of
RS = peak response ratio of the Standard solution water and 2 mL of dilute hydrochloric acid (1 in 120), and
CS = concentration of USP Betamethasone Valerate RS mix. Extract with four 50-mL portions of chloroform, and
in the Standard solution (mg/mL) combine the extracts. Filter through a cotton pledget,
CU = concentration of the Sample solution (mg/mL) previously layered over with anhydrous sodium sulfate.
Acceptance criteria: 97.0%103.0% Evaporate the filtrates on a steam bath under a stream of
dry nitrogen to dryness. Dissolve the residue in alcohol to
IMPURITIES obtain a solution containing about 1 mg/mL.
Inorganic Impurities Developing solvent system: Toluene and ethyl acetate
RESIDUE ON IGNITION 281: NMT 0.2%, using a platinum (1:1)
crucible Application volume: 10 L
Organic Impurities Analysis
PROCEDURE Samples: Sample solution and Standard solution
Mobile phase: Acetonitrile, glacial acetic acid, and water Proceed as directed in the chapter. Spray the plate with a
(550:1:450) mixture of sulfuric acid, methanol, and nitric acid
Sample solution: 0.4 mg/mL of Betamethasone Valerate in (10:10:1), and heat at 105 for 15 min.
Mobile phase. Shake until dissolved.
(See Chromatography 621, System Suitability.) PROCEDURE
Mode: LC Mobile phase: Acetonitrile and water (3:2)
Detector: UV 254 nm Diluent: Glacial acetic acid in methanol (1 in 1000)
Column: 4.6-mm 15-cm; packing L1 Internal standard solution: 0.4 mg/mL of USP
Flow rate: 1 mL/min Beclomethasone Dipropionate RS in Diluent
Injection size: 10 L Standard stock solution: 0.6 mg/mL of USP
System suitability Betamethasone Valerate RS in Diluent
Sample: Sample solution Standard solution: Standard stock solution and Internal
Suitability requirements standard solution (1:2)
Resolution: NLT 1.5 between betamethasone valerate [NOTEThis solution has a concentration of 0.2 mg/mL of
and any impurity USP Betamethasone Valerate RS.]
Column efficiency: NLT 9000 theoretical plates Sample solution: Equivalent to 2.5 mg of betamethasone
Analysis from Cream, in a 50-mL centrifuge tube. Add 10.0 mL of
Sample: Sample solution the Internal standard solution and 5.0 mL of Diluent. Insert
Calculate the percentage of each impurity in the portion the stopper into the tube, and place in a water bath held at
of Betamethasone Valerate taken: 60 until the specimen melts. Remove from the bath, and
shake vigorously until the specimen resolidifies. Repeat the
Result = (rU/rT) 100 heating and shaking two more times. Place the tube in an
icemethanol bath for 20 min, then centrifuge to separate
rU = peak response for each impurity the phases. Decant the clear supernatant into a suitable
rT = sum of all the peak responses stoppered flask, and allow to warm to room temperature.
Acceptance criteria Chromatographic system
Individual impurities: NMT 1.0% (See Chromatography 621, System Suitability.)
SPECIFIC TESTS
OPTICAL ROTATION, Specific Rotation 781S: +75 to +82
Sample solution: 10 mg/mL, in dioxane
0.5% of its weight.
Mode: LC Analysis
Detector: UV 254 nm Samples: Sample solution and Standard solution
Column: 4-mm 30-cm; packing L1 Allow the spots to dry, and develop the chromatogram in a
Flow rate: 1.2 mL/min solvent system, until the solvent front has moved three-
Injection size: 10 L fourths of the length of the plate. Remove the plate from
System suitability the developing chamber, mark the solvent front, and allow
Sample: Standard solution the solvent to evaporate. View the spots under UV light.
[NOTEThe relative retention times for beclomethasone Acceptance criteria: The RF value of the principal spot from
dipropionate and betamethasone valerate are 1.7 and 1.0, the Sample solution corresponds to that from the Standard
respectively.] solution.
Resolution: NLT 4.5 between betamethasone valerate ASSAY
and beclomethasone dipropionate PROCEDURE
Relative standard deviation: NMT 2.0% Mobile phase: Acetonitrile and water (3:2)
Analysis Diluent: Glacial acetic acid in methanol (1 in 1000)
Samples: Sample solution and Standard solution Internal standard solution: 2 mg/mL of beclomethasone
Calculate the percentage of C22H29FO5 in the portion of dipropionate in chloroform
Cream taken: Standard stock solution: 1.6 mg/mL of USP
Betamethasone Valerate RS in chloroform
Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 Standard solution: Pipet 2 mL of Standard stock solution
into a 50-mL centrifuge tube, add 10 mL of 0.1 N
RU = peak response ratio from the Sample solution hydrochloric acid, then add 2.0 mL of Internal standard
RS = peak response ratio from the Standard solution solution. Insert the stopper into the tube, shake vigorously
CS = concentration of USP Betamethasone Valerate RS for 2 min, and centrifuge to separate the phases. Using a
in the Standard solution (mg/mL) syringe, transfer the lower chloroform phase to a small
CU = nominal concentration of the Sample solution stoppered vial. Evaporate the chloroform on a steam bath,
(mg/mL) at low heat, with the aid of a stream of nitrogen. Add 4.0
Mr1 = molecular weight of betamethasone, 392.46 mL of Diluent, and swirl to dissolve the residue.
Mr2 = molecular weight of betamethasone valerate, Sample solution: Equivalent to 2.5 mg of betamethasone
476.59 from Lotion in a stoppered, 50-mL centrifuge tube. Add
Acceptance criteria: 90.0%110.0% 10.0 mL of 0.1 N hydrochloric acid, insert the stopper, and
shake to disperse the specimen. Add 2.0 mL of chloroform
SPECIFIC TESTS and 2.0 mL of Internal standard solution. Insert the stopper
MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED into the tube, shake vigorously for 2 min, and centrifuge to
MICROORGANISMS 62: It meets the requirements of the separate the phases. Using a syringe, transfer the lower
tests for absence of Staphylococcus aureus and Pseudomonas chloroform phase to a small stoppered vial. Evaporate the
aeruginosa. chloroform on a steam bath, at low heat, with the aid of a
MINIMUM FILL 755: Meets the requirements stream of nitrogen. Add 4.0 mL of Diluent, and swirl to
dissolve the residue.
PACKAGING AND STORAGE: Preserve in collapsible tubes or in (See Chromatography 621, System Suitability.)
tight containers. Mode: LC
USP Beclomethasone Dipropionate RS Column: 4-mm 30-cm; packing L1
USP Betamethasone Valerate RS Flow rate: 1.2 mL/min
System suitability
Betamethasone Valerate Lotion Sample: Standard solution
(Comment on this Monograph)id=m9210=Betamethasone valerate and beclomethasone dipropionate are 1.0 and
Valerate Lotion=B-Monos.pdf) 1.7, respectively.]
Betamethasone Valerate Lotion contains an amount of Resolution: NLT 4.5 between betamethasone valerate
Betamethasone Valerate (C27H37FO6) equivalent to NLT 95.0% and beclomethasone dipropionate
and NMT 115.0% of the labeled amount of betamethasone Relative standard deviation: NMT 2.0%
(C22H29FO5). Analysis
IDENTIFICATION Calculate the percentage of C22H29FO5 in the portion of
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Lotion taken:
mixture Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100
Standard solution: 0.6 mg/mL of USP Betamethasone
Valerate RS in a mixture of methanol and chloroform (2:1) RU = peak response ratio from the Sample solution
Sample solution: Equivalent to 0.5 mg/mL of RS = peak response ratio from the Standard solution
betamethasone from Lotion, in a mixture of methanol and CS = concentration of USP Betamethasone Valerate RS
chloroform (2:1) in the Standard solution (mg/mL)
Application volume: 20 L CU = nominal concentration of the Sample solution
Developing solvent system: Chloroform and ethyl acetate (mg/mL)
(1:1) Mr1 = molecular weight of betamethasone, 392.46
Mr2 = molecular weight of betamethasone valerate, the phases. Decant the clear supernatant into a suitable
476.59 stoppered flask, and allow to warm to room temperature.
Acceptance criteria: 95.0%115.0% Chromatographic system
SPECIFIC TESTS Mode: LC
MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED Detector: UV 254 nm
MICROORGANISMS 62: Meets the requirements of the tests Column: 4-mm 30-cm; packing L1
for absence of Staphylococcus aureus and Pseudomonas Flow rate: 1.2 mL/min
aeruginosa Injection size: 10 L
MINIMUM FILL 755: Meets the requirements System suitability
PH 791: 4.06.0 Sample: Standard solution
ADDITIONAL REQUIREMENTS valerate and beclomethasone dipropionate are 1.0 and
PACKAGING AND STORAGE: Preserve in tight, light-resistant 1.7, respectively.]
containers, and store at controlled room temperature. Suitability requirements
USP REFERENCE STANDARDS 11 Resolution: NLT 4.5 between betamethasone valerate
USP Betamethasone Valerate RS and beclomethasone dipropionate
Analysis
Betamethasone Valerate Ointment Sample: Sample solution and Standard solution
Calculate the percentage of C22H29FO5 in the portion of
(Comment on this Monograph)id=m9220=Betamethasone Ointment taken:
Valerate Ointment=B-Monos.pdf)
DEFINITION Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100
Betamethasone Valerate Ointment contains an amount of RU = peak response ratio from the Sample solution
betamethasone valerate (C27H37FO6) equivalent to NLT 90.0% RS = peak response ratio from the Standard solution
and NMT 110.0% of the labeled amount of betamethasone CS = concentration of USP Betamethasone Valerate RS
(C22H29FO5), in a suitable ointment base. in the Standard solution (mg/mL)
IDENTIFICATION CU = nominal concentration of betamethasone in the
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution (mg/mL)
Sample solution: Transfer the equivalent to 2 mg of Mr1 = molecular weight of betamethasone, 392.46
betamethasone from Ointment to a separator. Add 20 mL of Mr2 = molecular weight of betamethasone valerate,
water and 2 mL of dilute hydrochloric acid (1 in 120). 476.59
Extract with four 50-mL portions of chloroform, and Acceptance criteria: 90.0%110.0%
combine the extracts. Filter through a cotton pledget, SPECIFIC TESTS
previously layered over with anhydrous sodium sulfate. MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED
Evaporate the filtrates on a steam bath under a stream of MICROORGANISMS 62: Meets the requirements of the tests
dry nitrogen to dryness. Dissolve the residue in alcohol to for absence of Staphylococcus aureus and Pseudomonas
obtain a solution containing 1 mg/mL. aeruginosa
Developing solvent system: Toluene and ethyl acetate MINIMUM FILL 755: Meets the requirements
(1:1)
Application volume: 10 L ADDITIONAL REQUIREMENTS
Analysis PACKAGING AND STORAGE: Preserve in collapsible tubes or in
Samples: Sample solution and Standard solution tight containers, and avoid exposure to excessive heat.
Proceed as directed in the chapter. Spray the plate with a USP REFERENCE STANDARDS 11
mixture of sulfuric acid, methanol, and nitric acid USP Betamethasone Valerate RS
(10:10:1), and heat at 105 for 15 min.
ASSAY
PROCEDURE Betaxolol Hydrochloride
Mobile phase: Acetonitrile and water (3:2) (Comment on this Monograph)id=m9230=Betaxolol
Diluent: Glacial acetic acid in methanol (1 in 1000) Hydrochloride=B-Monos.pdf)
Internal standard solution: 0.4 mg/mL of beclomethasone
dipropionate in Diluent
Betamethasone Valerate RS in Diluent
Standard solution: Standard stock solution and Internal
[NOTEThis solution has a known concentration of 0.2
mg/mL of USP Betamethasone Valerate RS.]
Sample solution: Equivalent to 2.5 mg of betamethasone C18H29NO3 HCl 343.89
from Ointment, in a 50-mL centrifuge tube. Add 10.0 mL of 2-Propanol, 1-[4-[2-(cyclopropylmethoxy)ethyl]phenoxy]-3-(1-
the Internal standard solution and 5.0 mL of a solution of methylethyl)amino-, hydrochloride, ()-;
glacial acetic acid in alcohol (1 in 1000). Insert the stopper ()-1-[p-[2-(Cyclopropylmethoxy) ethyl]phenoxy]-3-
into the tube, and place in a water bath held at 70 until (isopropylamino)-2-propanol hydrochloride. [63659-18-7].
the specimen melts. Remove from the bath, and shake
vigorously until the specimen resolidifies. Repeat the heating DEFINITION
and shaking two more times. Place the tube in an Betaxolol Hydrochloride contains NLT 98.5% and NMT 101.5%
icemethanol bath for 20 min, then centrifuge to separate of C18H29NO3 HCl, calculated on the dried basis.
USP 32 Official Monographs / Betaxolol 71
IDENTIFICATION Betaxolol Ophthalmic Solution

A. INFRARED ABSORPTION 197K (Comment on this Monograph)id=m9235=Betaxolol
B. IDENTIFICATION TESTSGENERAL, Chloride 191: It meets Ophthalmic Solution=B-Monos.pdf)
the requirements of the test, when tested as directed for
alkaloidal hydrochlorides. DEFINITION
Betaxolol Ophthalmic Solution is a sterile, aqueous, isotonic
ASSAY solution of Betaxolol Hydrochloride. It contains a suitable
PROCEDURE antimicrobial preservative. It contains the equivalent of NLT
Sample: 300 mg of Betaxolol Hydrochloride 90.0% and NMT 110.0% of the labeled amount of betaxolol
Analysis: Dissolve in 50 mL of glacial acetic acid. Add 7 mL (C18H29NO3).
of mercuric acetate TS and titrate with 0.1 N perchloric acid
VS. Perform a blank determination, and make any necessary IDENTIFICATION
correction (see Titrimetry 541). Each mL of 0.1 N perchloric THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
acid is equivalent to 34.39 mg of C18H29NO3 HCl. Standard solution: 2.75 mg/mL of USP Betaxolol
Acceptance criteria: 98.5%101.5% Hydrochloride RS in water
Sample solution: Equivalent to 2.5 mg/mL of betaxolol
IMPURITIES from a portion of Ophthalmic Solution diluted with water
Inorganic Impurities Mode: TLC
RESIDUE ON IGNITION 281: NMT 0.1% Adsorbent: 0.25-mm layer of silica gel
HEAVY METALS, Method II 231: NMT 20 ppm Application volume: 5 L
Organic Impurities Developing solvent system: Chloroform, isopropyl alcohol,
PROCEDURE and ammonium hydroxide (70:30:2)
Solution A: 0.025 M monobasic potassium phosphate Spray reagent: A solution (1 in 1000) of ninhydrin in
containing 0.1% (w/v) of tetrabutylammonium bromide. isopropyl alcohol
Adjust with 0.025 M phosphoric acid to a pH of 3.0. Analysis
Mobile phase: Acetonitrile and Solution A (3:17) Samples: Standard solution and Sample solution
System suitability solution: 2 mg/mL of USP Betaxolol Allow the spots to dry, and develop the chromatogram in a
Hydrochloride RS and 1 mg/mL of alprenolol hydrochloride chromatographic chamber, using the Developing solvent
in Mobile phase system until the solvent front has moved three-fourths of
Sample solution: 2.0 mg/mL of Betaxolol Hydrochloride in the length of the plate. Spray the plate with Spray reagent
Mobile phase and heat the plate at 105 for 10 min. Locate the spots on
Chromatographic system the plate.
(See Chromatography 621, System Suitability.) Acceptance criteria: The RF value of the principal spot of
Mode: LC the Sample solution corresponds to that of the Standard
Detector: UV 273 nm solution.
System suitability Solution A: 7.1 g/L of anhydrous dibasic sodium phosphate
Sample: System suitability solution in water, adjust with phosphoric acid to a pH of 3.0
[NOTEThe relative retention times for alprenolol and Mobile phase: Acetonitrile and Solution A (1:1)
betaxolol are 0.9 and 1.0, respectively.] Standard solution: 0.11 mg/mL of USP Betaxolol
Suitability requirements Hydrochloride RS in Solution A
Resolution: NLT 1.0 between the two peaks Sample solution: Nominally 0.1 mg/mL. Transfer a volume
Tailing factor: NMT 2.0 for the two peaks of Ophthalmic Solution, equivalent to about 10 mg of
Relative standard deviation: NMT 2.0% betaxolol, to a 100-mL volumetric flask. Dilute with Solution
Analysis A to volume.
Samples: Sample solution Chromatographic system
[NOTEAllow the elution to continue for five times the (See Chromatography 621, System Suitability.)
elution time of the betaxolol peak before making the Mode: LC
next injection.] Detector: UV 280 nm
Calculate the percentage of each impurity in the portion Column: 4-mm 25-cm; packing L1
of Betaxolol Hydrochloride taken: Flow rate: 1.1 mL/min
Result = (rU/rT) 100 System suitability
rU = area of each individual peak, other than the Suitability requirements
main betaxolol peak of the Sample solution Capacity factor, k: For the main betaxolol peak,
rT = sum of the areas of all the peaks of the Sample between 1 and 3
solution Column efficiency: NLT 750 theoretical plates
Acceptance criteria: Sum of all impurities, NMT 1.0% Tailing factor: 0.82.0
SPECIFIC TESTS Relative standard deviation: NMT 2.0%
MELTING RANGE OR TEMPERATURE, Class I 741: 113117 Analysis
PH 791: 4.56.5, in a solution (1 in 50) Samples: Standard solution and Sample solution
LOSS ON DRYING 731: Dry in vacuum at 65 for 2 h: it Calculate the percentage of C18H29NO3 in each mL of the
loses NMT 1.0% of its weight. Ophthalmic Solution taken:
ADDITIONAL REQUIREMENTS Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100

USP Betaxolol Hydrochloride RS rS = peak response from the Standard solution
72 Betaxolol / Official Monographs USP 32
CS = concentration of USP Betaxolol Hydrochloride RS rU = peak response from the Sample solution
in the Standard solution (mg/mL) rS = peak response from the Standard solution
CU = nominal concentration of the Sample solution CS = concentration of USP Betaxolol Hydrochloride RS
(mg/mL) in the Standard solution (mg/mL)
Mr1 = molecular weight of betaxolol, 307.43 CU = nominal concentration of betaxolol
Mr2 = molecular weight of betaxolol hydrochloride, hydrochloride in the Sample solution (mg/mL)
343.89 Acceptance criteria: 90.0%110.0%
PERFORMANCE TESTS
SPECIFIC TESTS DISSOLUTION 711
STERILITY TESTS 71: It meets the requirements when tested Medium: 0.01 N hydrochloric acid; 500 mL
as directed for Test for Sterility of the Product to be Examined, Apparatus 2: 50 rpm
Membrane Filtration. Time: 30 min
PH 791: 4.08.0 Detector: UV 274 nm
Sample solutions: Sample per Dissolution 711. Dilute with
ADDITIONAL REQUIREMENTS Medium to a concentration that is similar to that of the
PACKAGING AND STORAGE: Preserve in tight containers. Standard solution.
USP REFERENCE STANDARDS 11 Standard solution: USP Betaxolol Hydrochloride RS in
USP Betaxolol Hydrochloride RS Medium
[NOTEA 5-cm pathlength cell may be used for lower
dosage levels.]
Betaxolol Tablets Tolerances: NLT 80% (Q) of the labeled amount of
C18H29NO3 HCl
(Comment on this Monograph)id=m9238=Betaxolol Tablets=B- UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Monos.pdf) Analysis for content uniformity
Standard solution: 0.1 mg/mL of USP Betaxolol
DEFINITION Hydrochloride RS in 0.1 N hydrochloric acid
Betaxolol Tablets contain an amount of Betaxolol Hydrochloride Sample solution: Place 1 Tablet in a volumetric flask of
equivalent to NLT 90.0% and NMT 110.0% of the labeled appropriate size to obtain a concentration of 0.1 mg/mL
amount of betaxolol hydrochloride (C18H29NO3 HCl). based on the labeled claim. Add an amount of 0.1 N
IDENTIFICATION hydrochloric acid equal to 70% of the volume of the flask,
The retention time of the major peak of the Sample solution shake by mechanical means until dissolved, dilute with 0.1
corresponds to that of the Standard solution, as obtained in N hydrochloric acid to volume, and mix. Filter the mixture,
the Assay. discarding the first 20 mL of the filtrate.
Blank: 0.1 N hydrochloric acid
ASSAY Mode: Spectrophotometry
PROCEDURE Analytical wavelength: 274 nm
Diluent: Acetonitrile and water (1:1) Cell: 1 cm
Mobile phase: Acetonitrile, methanol, and 0.025 M pH 6.0 Analysis
ammonium phosphate buffer (7:6:7) Samples: Blank, Standard solution, and Sample solution
Mix, and degas under vacuum while stirring. Calculate the percentage of C18H29NO3 HCl in the Tablet
Standard solution: 2 mg/mL of USP Betaxolol taken:
Hydrochloride RS in Diluent
Sample solution: Dissolve NLT 20 Tablets in an (AU/AS) (CS/CU) 100
appropriate, accurately measured volume of Diluent so that
the final concentration is nominally 2 mg/mL of betaxolol AU = absorbance of the Sample solution
hydrochloride. Sonicate until the Tablets are disintegrated. AS = absorbance of the Standard solution
Cool to room temperature, dilute with Diluent to volume, CS = concentration of USP Betaxolol Hydrochloride
and filter. Use the clear filtrate. RS in the Standard solution (mg/mL)
Chromatographic system CU = nominal concentration of betaxolol
(See Chromatography 621, System Suitability.) hydrochloride in the Sample solution (mg/mL)
Column: 4.6-mm 15-cm; packing L1 LABELING: Label the Tablets to state both the content of the
Flow rate: 1.5 mL/min betaxolol active moiety and the content of betaxolol
Injection size: 10 L hydrochloride used in formulating them.
System suitability USP REFERENCE STANDARDS 11
Sample: Standard solution USP Betaxolol Hydrochloride RS
Analysis
Calculate the percentage of C18H29NO3 HCl in each Tablet:
USP 32 Official Monographs / Bethanechol 73
Bethanechol Chloride Suitability requirements

(Comment on this Monograph)id=m9350=Bethanechol Resolution: NLT 0.8 between 2-hydroxypropyltrimethyl
Chloride=B-Monos.pdf) ammonium chloride and bethanechol, System suitability
solution
Tailing factor: NMT 3.5, Standard solution
solution
Analysis
Calculate the percentage of C7H17ClN2O2 in the portion of
Bethanechol Chloride taken:
C7H17ClN2O2 196.67
1-Propanaminium, 2-(aminocarbonyl)oxy-N,N,N-trimethyl-, Result = (rU/rS) (CS/CU) 100
chloride, ()-;
()-(2-Hydroxypropyl)trimethylammonium chloride carbamate rU = peak response from the Sample solution
[590-63-6]. rS = peak response from the Standard solution
CS = concentration of USP Bethanechol Chloride RS in
DEFINITION the Standard solution (mg/mL)
Bethanechol Chloride contains NLT 98.0% and NMT 101.5% of CU = concentration of Bethanechol Chloride in the
C7H17ClN2O2, calculated on the dried basis. Sample solution (mg/mL)
IDENTIFICATION
A. INFRARED ABSORPTION 197M OTHER COMPONENTS
B. PROCEDURE CONTENT OF CHLORIDE
Sample solution: 50 mg in 2 mL Sample: 400 mg, previously dried
Analysis: To the Sample solution, add 0.1 mL of cobaltous Analysis: Dissolve in 30 mL of water, and add 40.0 mL of
chloride solution (1 in 100), then add 0.1 mL of potassium 0.1 N silver nitrate VS, then add 3 mL of nitric acid and 5
ferrocyanide TS. mL of nitrobenzene. Shake for a few min, add 2 mL of ferric
Acceptance criteria: An emerald-green color is produced, ammonium sulfate TS, and titrate the excess silver nitrate
and almost entirely fades in 510 min (as distinguished from with 0.1 N ammonium thiocyanate VS. Each mL of 0.1 N
choline chloride, which gives the same reaction but the silver nitrate is equivalent to 3.545 mg of Cl.
color does not fade). Acceptance criteria: 17.7%18.3%
C. PROCEDURE
Sample solution: 10 mg/mL IMPURITIES
Analysis: To 1 mL of a solution, add 0.1 mL of iodine TS. Inorganic Impurities
Acceptance criteria: A brown precipitate is formed, and it RESIDUE ON IGNITION 281: NMT 0.1%
rapidly changes to a dark olive-green color. HEAVY METALS, Method I 231: NMT 30 ppm
Sample solution: 667 mg in 10 mL, add 2 mL of 1 N
D. IDENTIFICATION TESTSGENERAL, Chloride 191 acetic acid, and dilute to 25 mL
PROCEDURE PROCEDURE
Solution A: Transfer 29 mg of edetic acid to a 1000-mL Solution A: 0.48 mg/mL of methanesulfonic acid
volumetric flask, and dissolve in 500 mL. Add 300 L of Mobile phase, System suitability solution, and Sample
nitric acid to the volumetric flask, and dilute to volume. Pass solution: Proceed as directed in the Assay.
through a 0.45-m nylon membrane filter. Standard solution: 1 g/mL of USP Bethanechol Chloride
Mobile phase: Acetonitrile and Solution A (1:19) RS in Mobile phase
System suitability solution: 25 mg of Bethanechol Chloride Chromatographic system
in 10 mL of 0.1 N sodium hydroxide in a 250-mL volumetric (See Chromatography 621, System Suitability.)
flask. Allow to stand for 15 min. Add 10 mL of 0.1 N Mode, Detector, Column, Temperature, and Flow rate:
hydrochloric acid. Dissolve in and dilute with Mobile phase Proceed as directed in the Assay.
to volume. Injection size: 50 L
Standard solution: 0.1 mg/mL of USP Bethanechol System suitability: Proceed as directed in the Assay.
Chloride RS in Mobile phase Suitability requirements
Sample solution: 0.1 mg/mL of Bethanechol Chloride in Resolution: Proceed as directed in the Assay.
Mobile phase Relative standard deviation: NMT 10.0% for
Chromatographic system bethanechol chloride in the Standard solution
(See Chromatography 621, System Suitability.) Analysis
Mode: LC Samples: Sample solution and Standard solution
Detector: Conductivity Calculate the percentage of each impurity in the portion
Column: 3.9- 150-mm; packing L55 of Bethanechol Chloride:
Temperature Result = (rU/rS) (CS/CU) F 100
Detector: 35
Column: 30 rU = peak response for any impurity in the Sample
Flow rate: 1 mL/min solution
Injection size: 25 L rS = peak response of USP Bethanechol Chloride RS
System suitability in the Standard solution
Samples: System suitability solution and Standard solution CS = concentration of USP Bethanechol Chloride RS
[NOTEThe relative retention times for 2- in the Standard solution (mg/mL)
hydroxypropyltrimethyl ammonium chloride and
bethanechol are 0.9 and 1.0, respectively.]
74 Bethanechol / Official Monographs USP 32
CU = concentration of Bethanechol Chloride taken to Mode: LC

prepare the Sample solution (mg/mL) Detector: Conductivity
F = relative response factor, 0.79 for 2- Column: 4-mm 25-cm; packing L53
hydroxypropyltrimethyl ammonium and 1.0 Flow rate: 1 mL/min
for any other impurity Injection size: 50 L
Individual impurities: NMT 1.0% of 2- Sample: System suitability solution
hydroxypropyltrimethyl ammonium is found; NMT 0.1% [NOTEThe relative retention times for sodium,
of any other impurity is found. magnesium, calcium, 2-hydroxypropyltrimethyl
Total impurities: NMT 1.5% ammonium chloride, and bethanechol are 1.0, 1.4, 1.6,
2.0, and 2.8, respectively. ]
PH 791: 5.56.5, in a solution (1 in 100) Resolution: NLT 2.0, between the calcium ion and 2-
LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT hydroxypropyltrimethyl ammonium chloride
1.0% of its weight. Column efficiency: NLT 350 theoretical plates,
determined from the bethanechol peak
ADDITIONAL REQUIREMENTS Tailing factor: NMT 4.5
PACKAGING AND STORAGE: Preserve in tight containers. Relative standard deviation: NMT 2.0%
USP Bethanechol Chloride RS Samples: Sample solution and Standard solution
Calculate the percentage of C7H17ClN2O2 in each mL of the
Injection taken:
Bethanechol Chloride Injection Result = (rU/rS) (CS/CU) 100
(Comment on this Monograph)id=m9400=Bethanechol
Chloride Injection=B-Monos.pdf) rU = peak response from the Sample solution
DEFINITION CS = concentration of USP Bethanechol Chloride RS in
Bethanechol Chloride Injection is a sterile solution of the Standard solution (mg/mL)
Bethanechol Chloride in Water for Injection. It contains NLT CU = nominal concentration of the Sample solution
95.0% and NMT 105.0% of the labeled amount of (mg/mL)
C7H17ClN2O2. Acceptance criteria: 95.0%105.0%
IDENTIFICATION IMPURITIES
A. PROCEDURE Organic Impurities
Sample solution: 25 mg/mL of bethanechol chloride from PROCEDURE: LIMIT OF 2-HYDROXYPROPYLTRIMETHYL AMMONIUM
the Injection CHLORIDE
Analysis: To the Sample solution, add 0.1 mL of cobaltous Diluent, Mobile phase, System suitability solution, and
chloride solution (1 in 100), then add 0.1 mL of potassium Chromatographic system: Prepare as directed in the
ferrocyanide TS. Assay.
Acceptance criteria : An emerald-green color is produced, 2-Hydroxypropyltrimethyl ammonium chloride solution:
and almost entirely fades in 5 to 10 min (distinction from Transfer 50 mg of bethanechol chloride to a 50-mL
choline chloride, which gives the same reaction but the volumetric flask. Add 40 mL of 0.1 N sodium hydroxide,
color does not fade). and sonicate until fully dissolved. Dilute with 0.1 N sodium
B. PROCEDURE hydroxide to volume, and allow to stand for 5 days to
Sample solution : 10 mg/mL from the Injection allow adequate time for conversion from bethanechol to 2-
Analysis: To 1 mL of Sample solution add 0.1 mL of iodine hydroxypropyltrimethyl ammonium chloride.
TS. Chromatograph as directed in Analysis to verify the
Acceptance criteria: A brown precipitate is formed, and it presence and location of the peak for 2-
rapidly changes to a dark olive-green color. hydroxypropyltrimethyl ammonium chloride.
C. IDENTIFICATION TESTSGENERAL, Chloride 191 Standard solution: Use the Standard solution, prepared as
ASSAY Sample solution: Use the Sample solution, prepared as
PROCEDURE directed in the Assay.
Diluent: 0.1 mg/mL of calcium chloride and 0.1 mg/mL of System suitability: Proceed as directed in the Assay.
magnesium chloride Analysis
Mobile phase: 20 mM methanesulfonic acid Samples: 2-Hydroxypropyltrimethyl ammonium chloride
System suitability solution: To 25 mg of USP Bethanechol solution, Sample solution, and Standard solution
Chloride RS add 15 mL of water, 2 mL of Diluent, and 0.5 Calculate the percentage of 2-hydroxypropyltrimethyl
mL of 0.1 N of sodium hydroxide. Dilute with water to 25 ammonium chloride in each mL of Injection taken:
mL.
Standard solution: 1 mg/mL of USP Bethanechol Chloride Result = (rU/rS) (CS/CU) 100
RS
Sample solution: 1 mg/mL of Bethanechol Chloride from a rU = peak response for 2-hydroxypropyltrimethyl
volume of Injection ammonium chloride from the Sample solution
USP 32 Official Monographs / Bethanechol 75
rS = peak response for bethanechol from the Suitability requirements

Standard solution Relative standard deviation: NMT 3.1%
CS = concentration of USP Bethanechol Chloride RS Analysis
in the Standard solution (mg/mL) Samples: Standard solution and Sample solution
CU = concentration of bethanechol chloride in the Calculate the percentage of C7H17ClN2O2 in the volume of
Sample solution (mg/mL) Oral Solution:
SPECIFIC TESTS
BACTERIAL ENDOTOXINS TEST 85: NMT 25.0 USP Endotoxin rU = peak response from the Sample solution
Units/mg of bethanechol chloride rS = peak response from the Standard solution
PH 791: 5.57.5 CS = concentration of USP Bethanechol Chloride RS in
OTHER REQUIREMENTS: It meets the requirements under the Standard solution (g/mL)
Injections 1. CU = nominal concentration of bethanechol chloride
in the Sample solution (g/mL)
preferably of Type I glass. SPECIFIC TESTS
USP REFERENCE STANDARDS 11 PH 791: 3.94.9
USP Bethanechol Chloride RS
USP Endotoxin RS ADDITIONAL REQUIREMENTS
containers. Store at room temperature or in a cold place.
LABELING: Label it to state the beyond-use date.
Bethanechol Chloride Oral Solution BEYOND-USE DATE: 60 days after the day on which it was
(Comment on this Monograph)id=m1598=Bethanechol compounded.
Chloride Oral Solution=B-Monos.pdf) USP REFERENCE STANDARDS 11
USP Bethanechol Chloride RS
DEFINITION
Bethanechol Chloride Oral Solution contains NLT 90.0% and
NMT 110.0% of the labeled amount of bethanechol chloride
(C7H17ClN2O2). Bethanechol Chloride Oral Suspension
Prepare Bethanechol Chloride Oral Solution 5 mg/mL as follows (Comment on this Monograph)id=m1381=Bethanechol
(See Pharmaceutical CompoundingNonsterile Preparations Chloride Oral Suspension=B-Monos.pdf)
795).
DEFINITION
Bethanechol Chloride Oral Suspension contains NLT 90.0% and
Bethanechol Chloride 500 mg
NMT 110.0% of the labeled amount of bethanechol chloride
Vehicle for Oral Solution (regular or sugar- A sufficient quantity (C7H17ClN2O2).
free), NF Prepare Bethanechol Chloride Oral Suspension 5 mg/mL as
To make 100 mL follows (See Pharmaceutical CompoundingNonsterile
Preparations 795).
Add Bethanechol Chloride powder and about 20 mL of Vehicle
to a mortar, and mix. Add the Vehicle in small portions almost Bethanechol Chloride 500 mg
to volume, and mix thoroughly after each addition. Transfer Vehicle: a mixture of Vehicle for Oral Solution,
the contents of the mortar, stepwise and quantitatively, to a (regular or sugar-free), NF and Vehicle for Oral A sufficient
calibrated bottle. Add enough Vehicle to bring to final volume, Suspension, NF (1:1) quantity
and mix well.
To make 100 mL
ASSAY
PROCEDURE If using Bethanechol Chloride Tablets, add to a suitable mortar
Mobile phase: Acetonitrile and water (33:67) and comminute to a fine powder, or add the Bethanechol
Standard solution: 500 g/mL of USP Bethanechol Chloride powder to the mortar. Add about 20 mL of the
Chloride RS in Mobile phase Vehicle, and mix to a uniform paste. Add the Vehicle in small
Sample solution: Agitate the container of Oral Solution for portions almost to volume, and mix thoroughly after each
30 min on a rotating mixer, remove a 10-mL sample, and addition. Transfer the contents of the mortar, stepwise and
store in a clear glass vial at 70 until analyzed. At the time quantitatively, to a calibrated bottle. Add sufficient Vehicle to
of analysis, remove the sample from the freezer, allow it to final volume, and mix well.
reach room temperature, and mix with a vortex mixer for
30 s. Pipet 2.0 mL of the sample into a 20-mL volumetric ASSAY
flask, and dilute with Mobile phase to volume. PROCEDURE
Chromatographic system Mobile phase: Acetonitrile and water (33:67)
(See Chromatography 621, System Suitability.) Standard solution: 500 g/mL of USP Bethanechol
Mode: LC Chloride RS in Mobile phase
Detector: UV 200 nm Sample solution: Agitate the container of Oral Suspension
Column: 4.6-mm 25-cm; 5-m packing L1 for 30 min on a rotating mixer, remove a 10-mL sample,
Flow rate: 0.7 mL/min and store in a clear glass vial at 70 until analyzed. At the
Injection size: 20 L time of analysis, remove the sample from the freezer, allow
System suitability it to reach room temperature, and mix with a vortex mixer
Sample: Standard solution [NOTERetention time: about 3 for 30 s. Pipet 5.0 mL of the sample into a 50-mL
min] volumetric flask, and dilute with Mobile phase to volume.
76 Bethanechol / Official Monographs USP 32
Chromatographic system 0.1 N hydrochloric acid. Dissolve in and dilute with Mobile
(See Chromatography 621, System Suitability.) phase to volume.
Mode: LC Standard solution: 0.1 mg/mL of USP Bethanechol
Detector: UV 200 nm Chloride RS in Mobile phase
Column: 4.6-mm 25-cm; 5-m packing L1 Sample solution: Equivalent to 0.1 mg/mL of bethanechol
Flow rate: 0.7 mL/min chloride from powdered Tablets (NLT 20 Tablets) in Mobile
Injection size: 20 L phase
System suitability [NOTEIn a suitable volumetric flask, transfer a portion of
Sample: Standard solution [NOTERetention time: about 3 the powder equivalent to 1 Tablet. Add an amount of
min] Mobile phase, 60% to 70% of the total volume of the
Suitability requirements flask. Sonicate for 20 min. Shake by mechanical means for
Relative standard deviation: NMT 3.1% 15 min. Dilute with Mobile phase to volume, and mix.
Analysis Allow to stand for 10 min, and pass the solution through
Samples: Standard solution and Sample solution a 1-m glass filter, discarding the first 3 mL of the filtrate.]
Calculate the percentage of C7H17ClN2O2 in the volume of Chromatographic system
Oral Suspension: (See Chromatography 621, System Suitability.)
Mode: LC
Result = (rU/rS) (CS/CU) 100 Detector: Conductivity
Column: 3.9- 150-mm; packing L55
rU = peak response from the Sample solution Temperature
rS = peak response from the Standard solution Detector: 35
CS = concentration of USP Bethanechol Chloride RS in Column: 30
the Standard solution (g/mL) Flow rate: 1 mL/min
CU = nominal concentration of bethanechol chloride Injection size: 50 L
in the Sample solution (g/mL) System suitability
Acceptance criteria: 90.0%110.0% Samples: System suitability solution and Standard solution
[NOTEThe relative retention times for 2-
SPECIFIC TESTS hydroxypropyltrimethyl ammonium chloride and
PH 791: 3.94.9 bethanechol are 0.9 and 1.0, respectively.]
PACKAGING AND STORAGE: Preserve in tight, light-resistant Resolution: NLT 0.8 between 2-hydroxypropyltrimethyl
containers. Store at room temperature, or in a cold place. ammonium chloride and bethanechol, System suitabillity
LABELING: Label it to state that it is to be well shaken, and solution
to state the beyond-use date. Tailing factor: NMT 3.5, Standard solution
Beyond-Use Date: 60 days after the day on which it was Relative standard deviation: NMT 3.0%, Standard
compounded. solution
USP Bethanechol Chloride RS Samples: Sample solution and Standard solution
Calculate the percentage of C7H17ClN2O2 in the portion of
Tablets taken:
Bethanechol Chloride Tablets Result = (rU/rS) (CS/CU) 100

(Comment on this Monograph)id=m9450=Bethanechol rU = peak response from the Sample solution
Chloride Tablets=B-Monos.pdf) rS = peak response from the Standard solution
DEFINITION CS = concentration of USP Bethanechol Chloride RS in
Bethanechol Chloride Tablets contain NLT 90.0% and NMT the Standard solution (mg/mL)
110.0% of the labeled amount of bethanechol chloride CU = concentration of the Sample solution (mg/mL)
(C7H17ClN2O2). Acceptance criteria: 90.0%110.0%
IDENTIFICATION PERFORMANCE TESTS

INFRARED ABSORPTION 197M DISSOLUTION 711
Sample solution: Equivalent to 100 mg of bethanechol Medium: 0.1 N hydrochloric acid; 900 mL
chloride from pulverized Tablets. Add 15 mL of ether, and Apparatus 2: 50 rpm
allow to digest for 15 min. Decant the ether, again extract Time: 30 min
the residue with 10 mL of ether, and discard the ether Determine the amount of C7H17ClN2O2 dissolved using the
extracts. To the residue add 30 mL of alcohol, shake for 10 following method.
min, and allow to stand for 1 h with frequent agitation. Solution A, Mobile phase, and Chromatographic system:
Filter with suction, and evaporate the filtrate on a steam Proceed as directed in the Assay.
bath to dryness: the bethanechol chloride so obtained is Sample solution: Sample per Dissolution 711. Dilute with
recrystallized from alcohol and dried at 105 for 2 h. Medium to a concentration that is similar to that of the
Standard solution.
ASSAY Standard solution: USP Bethanechol Chloride RS in Medium
PROCEDURE System suitability: Proceed as directed in the Assay.
Solution A: Transfer 29 mg of edetic acid to a 1000-mL Analysis
volumetric flask, and dissolve in 500 mL. Samples: Sample solution and Standard solution
Add 300 L of nitric acid to the volumetric flask, and dilute Calculate the quantity of C7H17ClN2O2 dissolved based on
to volume. Pass through a 0.45-m nylon membrane filter. the peak responses obtained from the Sample solution and
Mobile phase: Acetonitrile and Solution A (1:19) the Standard solution.
System suitability solution: Transfer 25 mg of bethanechol Tolerances: NLT 80% (Q) of the labeled amount of
chloride in 10 mL of 0.1 N sodium hydroxide to a 250-mL C7H17ClN2O2
volumetric flask. Allow to stand for 15 min. Add 10 mL of
USP 32 Official Monographs / Bicalutamide 77
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Total impurities: NMT 1.5%
IMPURITIES ADDITIONAL REQUIREMENTS
Organic Impurities PACKAGING AND STORAGE: Preserve in tight containers.
Solution A: 0.48 mg/mL of methanesulfonic acid USP Bethanechol Chloride RS
System suitability solution: Transfer 25 mg of
bethanechol chloride in 10 mL of 0.1 N sodium hydroxide
to a 250 mL volumetric flask. Allow to stand for 15 min. Bicalutamide
Add 10 mL of 0.1 N hydrochloric acid. Dissolve in and (Comment on this Monograph)id=m2641=Bicalutamide=B-
dilute with Mobile phase to volume. Monos.pdf)
Standard solution: 1 g/mL of USP Bethanechol Chloride
RS in Mobile phase
Sample solution: Equivalent to 0.1 mg/mL of bethanechol
chloride from powdered Tablets (NLT 20 Tablets) in Mobile
phase
[NOTEIn a suitable volumetric flask, transfer a portion of
the powder equivalent to 1 Tablet. Add an amount of
Mobile phase, 60% to 70% of the total volume of the
flask. Sonicate for 20 min. Shake by mechanical means C18H14F4N2O4S 430.37
for 15 min. Dilute with Mobile phase to volume, and mix. Propanamide, N-[4-cyano-3-(trifluoromethyl)phenyl]-3-[(4-
Allow to stand for 10 min, and pass the solution through fluorophenyl)sulfonyl]-2-hydroxy-2-methyl-,(+)-
a 1-m glass filter, discarding the first 3 mL of the (+)-4-Cyano-,,-trifluoro-3-[(p-fluorophenyl)
filtrate.] sulfonyl]-2-methyl-m-lactotoluidide [90357-06-5].
Mode: LC Bicalutamide contains NLT 98.0% and NMT 102.0% of
Detector: Conductivity C18H14F4N2O4S, calculated on the anhydrous and solvent-free
Column: 3.9- 150-mm; packing L55 basis.
Temperature IDENTIFICATION
Detector: 35 A. INFRARED ABSORPTION 197M
Column: 30 B. The retention time of the major peak from the Sample
Flow rate: 1 mL/min solution corresponds to that of the Standard solution, as
Injection size: 50 L obtained in the Assay.
System suitability
Samples: System suitability solution and Standard solution ASSAY
[NOTEThe relative retention times for 2- PROCEDURE
hydroxypropyltrimethyl ammonium chloride and Mobile phase: Methanol, tetrahydrofuran, and water
bethanechol are 0.9 and 1.0, respectively.] (6:3:11)
Suitability requirements Standard solution: 0.05 mg/mL of USP Bicalutamide RS in
Resolution: NLT 0.8 between 2-hydroxypropyltrimethyl a minimum amount of tetrahydrofuran, and diluted with
ammonium chloride and bethanechol, System suitability Mobile phase
solution Sample solution: 0.05 mg/mL of Bicalutamide in a
Relative standard deviation: NMT 10.0% for minimum amount of tetrahydrofuran, and diluted with
bethanechol chloride, Standard solution Mobile phase
Samples: Sample solution and Standard solution (See Chromatography 621, System Suitability.)
Calculate the percentage of each impurity in the portion Mode: LC
of Tablets taken: Detector: UV 270 nm
Result = (ru/rS) (CS/CU) F 100 Temperature: 3540
ru = peak response for any impurity in the Sample Injection size: 10 L
solution System suitability
rS = peak response of USP Bethanechol Chloride RS Sample: Standard solution
in the Standard solution Suitability requirements
CS = concentration of USP Bethanechol Chloride RS Relative standard deviation: NMT 2.0%
in the Standard solution (mg/mL) Analysis
CU = concentration of bethanechol chloride in the Samples: Sample solution and Standard solution
portion taken for the Sample solution as Calculate the percentage of C18H14F4N2O4S in the portion of
determined in the Assay (mg/mL) Bicalutamide taken:
F = relative response factor equal to 0.79 for 2-
hydroxypropyltrimethyl ammonium and 1.0 Result = (rU/rS) (CS/CU) 100
for any other impurity
Acceptance criteria rU = peak response from the Sample solution
Individual impurities: NMT 1.0% of 2- rS = peak response from the Standard solution
hydroxypropyltrimethyl ammonium chloride is found;
NMT 0.2% of any other impurity is found
78 Bicalutamide / Official Monographs USP 32
CS = concentration of USP Bicalutamide RS in the Impurity Table (continued)

Standard solution (mg/mL) Relative Acceptance
CU = concentration of the Sample solution (mg/mL) Retention Criteria,
Acceptance criteria: 98.0%102.0% Name Time NMT (%)
IMPURITIES Individual unspecified impurity 0.1
Inorganic Impurities Total unspecified impurities 0.3
1(RS)-4-cyano-3-phenylsulfonyl-2-hydroxy-2-methyl-3-(trifluoromethyl)-
Organic Impurities propionanilide.
2(RS)-4-cyano-3-(2-fluorophenylsulfonyl)-2-hydroxy-2-methyl-3-
PROCEDURE
Mobile phase: Proceed as directed in the Assay. (trifluoromethyl)-propionanilide.
3(RS)-4-cyano-3-(4-fluorophenylsulfonyl)-2-methyl-3-
System suitability solution: 0.05 mg/mL of USP
Bicalutamide RS and 0.02 mg/mL of USP Bicalutamide (trifluoromethyl)propionanilide.
Related Compound B RS in a minimum amount of SPECIFIC TESTS
tetrahydrofuran, and dilute with Mobile phase WATER, Method I 921: NMT 0.2%
Standard solution: 0.02 mg/mL of USP Bicalutamide RS in
a minimum amount of tetrahydrofuran, and dilute with ADDITIONAL REQUIREMENTS
Mobile phase PACKAGING AND STORAGE: Preserve in tight containers, and
Sample solution: 4.0 mg/mL of Bicalutamide in a store at room temperature.
minimum amount of tetrahydrofuran, and dilute with USP REFERENCE STANDARDS 11
Mobile phase USP Bicalutamide RS
Chromatographic system USP Bicalutamide Related Compound B RS
Mode: LC
Detector: UV 270 nm
Temperature: 3540 Add the following:
System suitability
Bicalutamide Tablets
Sample: System suitability solution (Comment on this Monograph)id=m9465=Bicalutamide
Suitability requirements Tablets=B-Monos.pdf)
Resolution: NLT 2.0 between bicalutamide and
bicalutamide related compound B DEFINITION
Tailing factor: NMT 1.3 for bicalutamide Bicalutamide Tablets contain NLT 90.0% and NMT 110.0% of
Relative standard deviation: NMT 4.0% for the labeled amount of bicalutamide (C18H14F4N2O4S).
bicalutamide
Samples: Standard solution and Sample solution The retention time of the major peak of the Sample solution
Calculate the percentage of each impurity in the portion corresponds to that of the Standard solution, as obtained in
of Bicalutamide taken: the Assay.
PROCEDURE
rU = peak response for each impurity from the Mobile phase: Tetrahydrofuran, acetonitrile, and water
Sample solution (20:15:65).
rS = peak response for bicalutamide from the System suitability stock solution: 0.8 mg/mL of USP
Standard solution Bicalutamide RS and 0.4 mg/mL of USP Bicalutamide
CS = concentration of USP Bicalutamide RS in the Related Compound B RS in tetrahydrofuran
Standard solution (mg/mL) System suitability solution: 0.04 mg/mL of USP
CU = concentration of the Sample solution (mg/mL) Bicalutamide RS and 0.02 mg/mL of USP Bicalutamide
Acceptance criteria Related Compound B RS, from System suitability stock
Individual impurities: See Impurity Table. solution, in Mobile phase
Total impurities: NMT 0.5% Standard stock solution: 0.8 mg/mL of USP Bicalutamide
RS in tetrahydrofuran
Impurity Table Standard solution: 0.04 mg/mL of USP Bicalutamide RS,
from Standard stock solution, in Mobile phase
Relative Acceptance Sample stock solution: Equivalent to 50 mg of
Retention Criteria, bicalutamide, from powdered Tablets, in 50 mL of
Name Time NMT (%) tetrahydrofuran. Sonicate for NLT 10 min to complete
Des-fluoro analog1 0.74 0.2 dissolution, and allow to cool. Dilute with tetrahydrofuran to
2-Fluoro isomer2 0.78 0.2
100.0 mL, and pass through a suitable filter having a
porosity of 0.45 m.
Des hydroxy analog3 1.19 0.2 Sample solution: 0.04 mg/mL of bicalutamide, from
1(RS)-4-cyano-3-phenylsulfonyl-2-hydroxy-2-methyl-3-(trifluoromethyl)-
Sample stock solution, in Mobile phase
propionanilide.
2(RS)-4-cyano-3-(2-fluorophenylsulfonyl)-2-hydroxy-2-methyl-3-
(trifluoromethyl)-propionanilide.
3(RS)-4-cyano-3-(4-fluorophenylsulfonyl)-2-methyl-3-
(trifluoromethyl)propionanilide.
USP 32 Official Monographs / Bicalutamide 79
Mode: LC porosity of 0.45 m, and dilute 10.0 mL with Diluent to

Detector: UV 270 nm 100.0 mL.
Column: 5-mm 12.5-cm; 3-m packing L1 Analysis
Temperature: 50 Samples: Standard solution, Sample solution, and Blank
Flow rate: 1.5 mL/min Calculate the percentage of C18H14F4N2O4S in each Tablet:
System suitability Result = (AU/AS) (CS/CU) 100
Suitability requirements AU = absorbance of the Sample solution
Resolution: NLT 1.9 between bicalutamide and AS = absorbance of the Standard solution
bicalutamide related compound B CS = concentration of the Standard solution (mg/mL)
Tailing factor: Less than 1.3 for bicalutamide CU = nominal concentration of the Sample solution
Relative standard deviation: NMT 2.0% for bicalutamide (mg/mL)
Analysis
Samples: Standard solution and Sample solution IMPURITIES
Calculate the percentage of C18H14F4N2O4S in the portion of Organic Impurities
Tablets taken: PROCEDURE: LIMIT OF 4-AMINO-2-(TRIFLUOROMETHYL)BENZONITRILE
Mobile phase and System suitability solution: Proceed as
Result = (rU/rS) (CS/CU) 100 directed in the Assay.
Standard stock solution: 0.2 mg/mL of USP Bicalutamide
rU = peak area from the Sample solution RS in tetrahydrofuran
rS = peak area from the Standard solution Standard solution: 0.02 mg/mL of USP Bicalutamide RS,
CS = concentration of the Standard solution (mg/mL) from Standard stock solution, in Mobile phase
CU = concentration of the Sample solution (mg/mL) Sample solution: Transfer an equivalent to 50 mg of
bicalutamide, from powdered Tablets, to a 25-mL
Acceptance criteria: 90.0%110.0% volumetric flask. Add 2.0 mL of tetrahydrofuran, and allow
PERFORMANCE TESTS to stand for 5 min. Add 20 mL of Mobile phase, sonicate
DISSOLUTION 711 for NLT 10 min, and allow to cool. Dilute with Mobile
Medium: 1.0% (w/v) sodium lauryl sulfate in water; 1000 phase to volume, and pass through a suitable filter having
mL a porosity of 0.2 m.
Apparatus 2: 50 rpm Chromatographic system
Time: 45 min (See Chromatography 621, System Suitability.)
Detector: UV 270 nm Mode: LC
Standard solution: 0.05 mg/mL of USP Bicalutamide RS in Detector: UV 220 nm
Medium [NOTEDissolve the RS in a volume of Column: 5-mm 12.5-cm; 3-m packing L1
tetrahydrofuran equal to 1% of the final solution volume Temperature: 50
before dilution with Medium.] Flow rate: 1.5 mL/min
Sample solution: Sample per Dissolution 711. Pass Injection size: 10 L
through a suitable filter having a porosity of 0.45 m. System suitability
Blank: Medium Sample: System suitability solution
Analysis [NOTEThe relative retention times of 4-amino-2-
Samples: Standard solution and Sample solution (trifluoromethyl)benzonitrile and bicalutamide related
Determine the amount of C18H14F4N2O4S dissolved: compound B are about 0.4 and about 1.1, respectively.]
Result = (AU/AS) (CS V)/L 100 Resolution: NLT 1.9 between bicalutamide and
bicalutamide related compound B
AU = absorbance of the Sample solution Tailing factor: Less than 1.3 for bicalutamide
AS = absorbance of the Standard solution Relative standard deviation: NMT 2.0% for
CS = concentration of the Standard solution (mg/mL) bicalutamide
V = volume of Medium (1000 mL) Analysis
L = label claim (mg) Sample: Sample solution and Standard solution
Tolerances: NLT 80% (Q) of the labeled amount of Calculate the percentage of 4-amino-2-
C18H14F4N2O4S (trifluoromethyl)benzonitrile in the portion of Tablets
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements taken:
Analysis for content uniformity
Spectrometric conditions Result = (rU/rS) (CS/CU) (1/RRF) x 100
Analytical wavelength: UV 270 nm
Blank: Diluent rU = peak response of 4-amino-2-
Diluent: 10 mg/mL of sodium lauryl sulfate in water (trifluoromethyl)benzonitrile from the Sample
Standard solution: 0.05 mg/mL of USP Bicalutamide RS in solution
Medium [NOTEDissolve the RS in a minimum volume of rS = peak response of bicalutamide from the
tetrahydrofuran before dilution with Diluent.] Standard solution
Sample solution: Transfer 1 Tablet to a 100-mL volumetric CS = concentration of the Standard solution (mg/mL)
flask, add about 10 mL of water, and sonicate for CU = concentration of the Sample solution (mg/mL)
approximately 30 min. Add about 80 mL of RRF = relative response factor of 4-amino-2-
tetrahydrofuran, and sonicate for 30 min to complete (trifluoromethyl)benzonitrile (1.4)
dissolution of the bicalutamide. Allow to cool to room Acceptance criteria: NMT 0.1% of 4-amino-2-
temperature, and dilute with tetrahydrofuran to volume. (trifluoromethyl)benzonitrile
Pass a portion of the solution through a filter having a
80 Bicalutamide / Official Monographs USP 32
SPECIFIC TESTS TOTAL VIABLE SPORE COUNT

WATER DETERMINATION, Method I 921: NMT 5.0% Analysis: Proceed as directed for Total Viable Spore Count.
Acceptance criteria: The requirements of the test are met if
ADDITIONAL REQUIREMENTS the log average number of viable spores per carrier is NLT
PACKAGING AND STORAGE: Preserve in tight containers, and 0.3 log of the labeled spore count per carrier and does not
store at controlled room temperature. exceed the log labeled spore count per carrier by 0.48.
USP Bicalutamide RS SPECIFIC TESTS
USP Bicalutamide Related Compound B RSUSP32 PURITY
Presence of contamination by other microorganisms: By
examination of the spores on a suitable plate culture
medium, there is no evidence of contamination with other
Biological Indicator for Dry-Heat microorganisms.
Sterilization, Paper Carrier ADDITIONAL REQUIREMENTS
(Comment on this Monograph)id=m9497=Biological Indicator PACKAGING AND STORAGE: Preserve in the original package
for Dry-Heat Sterilization, Paper Carrier=B-Monos.pdf) under the conditions recommended on the label, and
DEFINITION protect from light, toxic substances, excessive heat, and
Biological Indicator for Dry-Heat Sterilization, Paper Carrier, is a moisture. The packaging and container materials do not
defined solution of viable spores made from a culture derived adversely affect the performance of the article used as
from a specified strain of Bacillus subtilis subspecies niger, on a directed in the labeling.
suitable grade of paper carrier, individually packaged in a EXPIRATION DATE: The expiration date is determined on the
container readily penetrable by dry heat, and characterized for basis of stability studies and is NLT 18 months from the date
predictable resistance to dry-heat sterilization. The packaged of manufacture. The date of manufacture is the date on
Biological Indicator for Dry-Heat Sterilization, Paper Carrier, which the first determination of the total viable spore count
has a particular labeled spore count per carrier of NLT 104 and was made.
NMT 109 spores. When labeled for and subjected to dry-heat LABELING: Label it to state that it is a Biological Indicator for
sterilization conditions at a particular temperature, it has a Dry-Heat Sterilization, Paper Carrier; to indicate its D value
survival time and kill time appropriate to the labeled spore and the method used to determine such D value, i.e., by
count and to the decimal reduction value (D value, in min) of spore count or fraction negative procedure after graded
the solution, specified by: exposures to the sterilization conditions; the survival time
Survival time (in min) = NLT (labeled D value) (log labeled and kill time under the specified sterilization conditions
spore count per carrier 2); and stated on the label; its particular total viable spore count,
Kill time (in min) = NMT (labeled D value) (log labeled spore with a statement that such count has been determined after
count per carrier + 4) preliminary heat treatment; and its recommended storage
conditions. State in the labeling the size of the paper carrier,
IDENTIFICATION the strain and ATCC number from which the spores were
The biological indicator organism complies substantially with derived, and instructions for spore recovery and for safe
the morphological, cultural, and biochemical characteristics disposal of the indicator. Indicate in the labeling that the
of the strain of Bacillus subtilis, ATCC No. 9372, designated stated D value is reproducible only under the exact
subspecies niger, detailed for that biological indicator conditions under which it was determined, that the user
organism under Biological Indicator for Ethylene Oxide would not necessarily obtain the same result, and that the
Sterilization, Paper Carrier. user should determine the suitability of the biological
indicator for the particular use.
PERFORMANCE TESTS DISPOSAL: Before destruction or discard, sterilize it by steam
[NOTESee Biological IndicatorsResistance Performance Tests at 121 for NLT 30 min, or by NLT an equivalent method
55] recommended by the manufacturer. This includes a test strip
D VALUE used in any test procedure for the strips themselves.
Analysis: Proceed as directed for D Value Determination.
Acceptance criteria: The requirements of the test are met if
the determined D value is within 20% of the labeled D value
for the selected sterilizing temperature, and if the Biological Indicator for Ethylene Oxide
confidence limits of the estimate are within 10% of the Sterilization, Paper Carrier
determined D value. (Comment on this Monograph)id=m9500=Biological Indicator
SURVIVAL TIME and KILL TIME for Ethylene Oxide Sterilization, Paper Carrier=B-Monos.pdf)
Analysis: Proceed as directed for Survival Time and Kill Time.
Acceptance criteria: The requirements of the test are met if DEFINITION
all of the specimens subjected to dry-heat sterilization for Biological Indicator for Ethylene Oxide Sterilization, Paper
the survival time show evidence of growth, while none of Carrier, is a defined solution of viable spores made from a
the specimens subjected to dry-heat sterilization for the kill culture derived from a specified strain of Bacillus subtilis
time show growth. If for either the survival time test or the subspecies niger on a suitable grade of paper carrier,
kill time test NMT one specimen out of both groups fails the individually packaged in a suitable container readily penetrable
survival requirement or the kill requirement (whichever is by ethylene oxide sterilizing gas mixture, and characterized for
applicable), continue the corresponding test with four predictable resistance to sterilization with such gas mixture.
additional groups, each consisting of 20 specimens, The packaged Biological Indicator for Ethylene Oxide
according to the procedure described. If all of the additional Sterilization, Paper Carrier, has a particular labeled spore count
specimens subjected to dry-heat sterilization either meet the per carrier of NLT 104 and NMT 109 spores. Where labeled for
survival requirement for the survival time test, or meet the and subjected to particular ethylene oxide sterilization
kill requirement for the kill time test, whichever is applicable, conditions of a stated gaseous mixture, temperature, and
the requirements of the test are met. relative humidity, it has a survival time and kill time
USP 32 Official Monographs / Biological 81
appropriate to the labeled spore count and to the decimal SPECIFIC TESTS
reduction value (D value, in min) of the solution, specified by: PURITY (Presence of contamination by other
Survival time (in min) = NLT (labeled D value) (log labeled microorganisms): By examination of the spores on a
spore count per carrier 2), and suitable plate culture medium, there is no evidence of
Kill time (in min) = NMT (labeled D value) (log labeled spore contamination with other microorganisms.
count per carrier + 4).
IDENTIFICATION PACKAGING AND STORAGE: Preserve in the original package
The biological indicator organism complies substantially with under the conditions recommended on the label, and
the morphological, cultural, and biochemical characteristics protect it from light, toxic substances, excessive heat, and
of the strain of Bacillus subtilis, ATCC No. 9372, designated moisture. The packaging and container material shall be
subspecies niger: under microscopic examination, it consists such that it does not adversely affect the performance of the
of Gram-positive rods of width 0.7 to 0.8 m, and length 2 article used as directed in the labeling.
to 3 m; the endospores are oval and central, and the cells EXPIRATION DATE: The expiration date is determined on the
are not swollen; when incubated aerobically in appropriate basis of stability studies and is NLT 18 months from the date
media at 3035, growth occurs within 24 h, and similar of manufacture, the date of manufacture being the date on
inoculated media incubated concomitantly at 5560 show which the first determination of the total viable spore count
no evidence of growth in the same period; agar colonies was made.
have a dull appearance and may be cream or brown- LABELING: Label it to state that it is a Biological Indicator for
colored; when incubated in nutrient broth, it develops a Ethylene Oxide Sterilization, Paper Carrier; to indicate its D
pellicle and shows little or no turbidity; when examined value, the method used to determine such D value, i.e., by
under conventional biochemical tests for microbial spore count or fraction negative procedure after graded
characterization, develops a black pigment with tyrosine, it exposures to the sterilization conditions; the Survival time
liquefies gelatin, utilizes citrate but not propionate or and Kill time under specified sterilization conditions stated on
hippurate, reduces nitrate, and hydrolyzes both starch and the label; its particular Total viable spore count, with a
glucose with no gas production; it shows a positive catalase statement that such count has been determined after
reaction and gives a positive result with the Voges-Proskauer preliminary heat treatment; and its recommended storage
test. conditions. State in the labeling the size of the paper carrier,
the strain, and ATCC number from which the spores were
PERFORMANCE TESTS derived, and instructions for spore recovery and for safe
[NOTESee Biological IndicatorsResistance Performance Tests disposal of the indicator. Indicate in the labeling that the
55.] stated D value is reproducible only under the exact
D VALUE conditions under which it was determined, that the user
Analysis: Proceed as directed in the relevant procedure for would not necessarily obtain the same result, and that the
D value. user should determine the suitability of the biological
Acceptance criteria: The requirements of the test are met indicator for the particular use.
if the determined D value is within 20% of the labeled D DISPOSAL: Prior to destruction or discard, sterilize it by
value for the selected sterilizing temperature, and if the steam at 121 for NLT 30 min, or by NLT an equivalent
confidence limits of the estimate are within 10% of the method recommended by the manufacturer. This includes a
determined D value. strip used in test procedures for strips themselves.
SURVIVAL TIME and KILL TIME
Analysis: Proceed as directed for Survival Time and Kill
Time, using the procedure for Biological Indicator for
Ethylene Oxide Sterilization, Paper Carrier. Biological Indicators for Moist Heat,
Acceptance criteria: The requirements of the test are met Dry Heat, and Gaseous Modes of
if all of the specimens subjected to the ethylene oxide
sterilization conditions for the survival time show evidence
Sterilization, Liquid Spore Suspensions
of growth, while none of the specimens subjected to the (Comment on this Monograph)id=m825=Biological Indicators
ethylene oxide sterilization conditions for the kill time for Moist Heat, Dry Heat, and Gaseous Modes of Sterilization,
shows evidence of growth. If for either the survival time Liquid Spore Suspensions=B-Monos.pdf)
test or the kill time test, NMT 1 specimen out of both DEFINITION
groups fails the survival requirement or the kill requirement Liquid spore suspensions may be used to prepare biological
(whichever is applicable), continue the corresponding test indicators for moist heat, dry heat, and gaseous modes of
with 4 additional groups, each consisting of 20 specimens, sterilization. On the basis of the intended sterilization use, the
according to the procedure described. If all the additional suspension is prepared inoculated from a culture of viable
specimens subjected to ethylene oxide sterilization meet spores derived from one of several sterilization resistant
either the survival requirement for the Survival time test or microorganisms. Cultures used for liquid spore suspensions
the kill requirement for the Kill time test, whichever is include, among others, the following: Clostridium sporogenes,
applicable, the requirements of the test are met. Geobacillus stearothermophilus (formerly B. stearothermophilus),
TOTAL VIABLE SPORE COUNT Bacillus atrophaeus (formerly B. subtilis), Bacillus subtilis, or
Analysis: Proceed as directed for Total Viable Spore Count, Bacillus coagulans. Each tube or container containing the spore
using the procedure for Biological Indicator for Ethylene suspension is individually packaged for use. The packaged
Oxide Sterilization, Paper Carrier. biological indicator spore suspension has a particular labeled
Acceptance criteria: The requirements of the test are met spore count of NLT 103, and NMT 109, spores/mL of
if the log average number of viable spores per carrier is suspension. The suspending medium or vehicle is identified
NLT 0.3 log of the labeled spore count per carrier and does according to chemical composition. It has a survival time and
not exceed the log labeled spore count per carrier by 0.48.
82 Biological / Official Monographs USP 32
kill time appropriate to the labeled spore count, and to the toxic substances, and excessive heat. The materials of
decimal reduction value (the D value, in min), specified by the composition of the tube or container must not adversely
following: affect the performance of the spore suspension.
Survival time (in min) = NLT (labeled D value) (log of labeled EXPIRATION DATE: The expiration date is determined on the
spore count per mL from 1:100 dilution of original suspension basis of stability studies. The date of manufacture is the date
2); and on which the first determination of the total viable count
Kill time (in min) = NMT (labeled D value) (log of labeled was made.
spore count per mL from 1:100 dilution of original suspension SHIPMENT: Spore suspensions must be shipped following EPA
+ 4). requirements for the shipment of biological and/or
etiological agents.
IDENTIFICATION DISPOSAL: Spore suspensions that a user or manufacturer
Identification for the biological indicator is of lesser wishes to dispose of are first sterilized by moist heat by a
importance than the more relevant concerns of population process that achieves temperatures of approximately 121
and resistance to the sterilization processes. The for NLT 30 min. Alternative sterilization methods yielding
manufacturer should identify the species used. equivalent or greater levels of lethality may be used.
LABELING: Label the spore suspension tube or container or
PERFORMANCE TESTS package insert to state that it is a biological indicator spore
[NOTESee Biological IndicatorsResistance Performance Tests suspension for use in label specified applications for moist-
55, Total Viable Spore Count] heat, dry-heat, and/or gaseous sterilization. State the
D VALUE biological indicator D value obtained under defined
Analysis: If the biological indicators are being used in exposures to stated sterilization conditions using the Survival
moist-heat or dry-heat sterilization, proceed as directed in Curve Method of D value analysis. State the Survival time
the relevant procedure in D Value Determination. and kill time for the biological indicator suspension under
Acceptance criteria: The requirements of the test are met if specified conditions on the label. The total viable spore
the determined D value is within 20% of the labeled D value count per mL of the suspension following heat shock
for the selected sterilizing conditions, and if the confidence treatment must also appear on the label. State in the
limits of the estimate are within 10% of the determined D labeling the strain and ATCC number of the microorganisms
value. The D value determination method used should be used in the spore suspension and instructions for spore
that identified by the biological indicator manufacturer. recovery and for safe disposal of the suspension. Indicate in
SURVIVAL TIME and KILL TIME the labeling that the stated D value is reproducible only
Analysis: Follow the procedure under Survival Time and Kill under the exact conditions determined by the manufacturer
Time in D Value Determination. The test is conducted using and that the user would not necessarily obtain the same
1:100 dilution aliquots of the original suspension to results if different exposure conditions were used. State that
inoculate carrier substrates that are most likely to be used by the user should determine the suitability of the biological
the purchaser of the spore suspensions for a given mode of indicator spore suspension for the users particular purpose
sterilization. Following a total viable count analysis, the and exposure conditions.
inoculated substrates are subjected to sterilization exposure
conditions intended to indicate survival.
Acceptance criteria: The inoculated carriers must show
evidence of growth among the exposed carriers. Biological Indicators for Moist Heat,
Analysis: A second study is conducted to demonstrate the
conditions necessary to result in total kill of the carriers.
Dry Heat, and Gaseous Modes of
Acceptance criteria: None of the carriers subjected to Sterilization, Nonpaper Carriers
conditions designed to induce total kill should show growth. (Comment on this Monograph)id=m663=Biological Indicators
If for either the survival-time test or the kill-time test, NMT for Moist Heat, Dry Heat, and Gaseous Modes of Sterilization,
one carrier out of both groups fails the survival or kill Nonpaper Carriers=B-Monos.pdf)
requirements, continue the corresponding test with four
additional groups, each consisting of 10 carriers, according DEFINITION
to the procedure described. For biological indicators for use Biological indicators for moist heat, dry heat, and gaseous
with moist-heat or dry-heat sterilization, if all of the modes of sterilization may be nonpaper carriers inoculated
additional specimens subjected to the specific sterilization with a culture of viable spores derived from one of several
process either meet the survival requirements for the sterilization-resistant microorganisms, based on the intended
survival-time test or meet the kill requirement for the kill- sterilization use. Cultures used for inoculation of carriers
time test, whichever is applicable, the requirements are met. include, among others, Clostridium sporogenes, Geobacillus
TOTAL VIABLE SPORE COUNT: stearothermophilus (formerly B. stearothermophilus), Bacillus
Analysis: Proceed as directed for Biological Indicators for atrophaeus (formerly B. subtilis), or Bacillus coagulans. The
Moist Heat, Dry Heat, and Gaseous Modes of Sterilization, carriers should be individually packaged for use either within
Liquid Spore Suspensions in Total Viable Spore Count the package or for use upon removal from the package as an
Acceptance criteria The requirements for this test are met unpackaged biological indicator. The packaged biological
if the total viable spore count within the suspension is within indicator on the carrier has a particular labeled spore count of
1 log of the value stipulated by the manufacturer. 103 - 109 NLT 103 and NMT 109 spores/carrier. It has a survival time and
spores/mL of suspension. kill time appropriate to the labeled spore count and to the
decimal reduction value (D value, in min), specified by:
SPECIFIC TESTS Survival time (in min) = NLT (labeled D value) (log of labeled
PURITY: There is no evidence of contamination with other spore count per carrier 2), and
microorganisms following examination of spores recovered Kill time (in min) = NMT (labeled D value) (log of labeled
from the metal carriers using suitable plate-culture medium. spore count per carrier + 4).
ADDITIONAL REQUIREMENTS IDENTIFICATION
PACKAGING AND STORAGE: Preserve in the original tube or Identification for the biological indicator is of less importance
container under the conditions recommended on the label, than the more relevant concerns of population and
and protect the contents of the tube or container from light,
USP 32 Official Monographs / Biological 83
resistance to the sterilization processes. The manufacturer biological indicator carrier under specified conditions on the
should identify the species used. label. The total viable spore count per carrier following heat
shock treatment must also appear on the label or package
PERFORMANCE TESTS insert. State in the labeling the strain and ATCC number of
[NOTESee Biological IndicatorsResistance Performance Tests the spore suspension used to inoculate the carriers and
55] instructions for spore recovery and for safe disposal of the
D VALUE carriers. Indicate in the labeling that the stated D value is
Analysis: Proceed as directed in the relevant procedure for reproducible only under the exact conditions determined by
D Value Determination. the manufacturer and that the user would not necessarily
Acceptance criteria: The requirements of the test are met obtain the same results if different exposure conditions were
if the determined D value is within 20% of the labeled D used. State that the user should determine the suitability of
value for the selected sterilizing conditions, and if the the carrier biological indicator for the users particular
confidence limits of the estimate are within 10% of the purpose and exposure conditions.
determined D value. The D value determination method
used should be that identified by the biological indicator
manufacturer.
SURVIVAL TIME and KILL TIME Biological Indicator for Steam
Analysis: Follow the procedure in the subsection Survival Sterilization, Paper Carrier
Time and Kill Time in D Value Determination. (Comment on this Monograph)id=m9510=Biological Indicator
Acceptance criteria: The requirements of the test are met for Steam Sterilization, Paper Carrier=B-Monos.pdf)
if all of the carriers subjected to sterilization exposure
conditions intended to indicate survival show evidence of DEFINITION
growth among the exposed carriers, while none of the Biological Indicator for Steam Sterilization, Paper Carrier, is a
carriers subjected to conditions designed to induce total kill defined solution of viable spores made from a culture derived
show growth. If for either the survival test or the kill time from a specified strain of Bacillus stearothermophilus, on a
test, NMT one carrier out of both groups fails the survival or suitable grade of paper carrier, individually packaged in a
kill requirements, continue the corresponding test with four suitable container readily penetrable by steam, and
additional groups, each consisting of 10 carriers, according characterized for predictable resistance to steam sterilization.
to the procedure described. If all of the additional specimens The packaged Biological Indicator for Steam Sterilization, Paper
subjected to the specific sterilization process either meet the Carrier, has a particular labeled spore count per carrier of NLT
survival requirements for the survival test time or meet the 104 and NMT 109 spores. When labeled for and subjected to
kill requirement for the kill test, whichever is applicable, the steam sterilization conditions at a particular temperature, it has
requirements are met. a survival time and kill time appropriate to the labeled spore
TOTAL VIABLE SPORE COUNT count and to the decimal reduction value (D Value, in min) of
Analysis: Proceed as directed in the subsection Biological the solution, specified by:
Indicators for Moist Heat, Dry Heat, and Gaseous Modes of Survival time (in min) = NLT (labeled D Value) (log labeled
Sterilization, Nonpaper Carriers in Total Viable Spore Count. spore count per carrier 2); and
Acceptance criteria: The requirements of the test are met if Kill time (in min) = NMT (labeled D Value) (log labeled spore
the average number of viable spores/carrier are within 50% count per carrier + 4)
and +300% of the labeled count/carrier or within a lesser
range that may be stated by the manufacturer. 103109 IDENTIFICATION
spores/carrier of labeled spore count. The biological indicator organism complies substantially with
the morphological, cultural, and biochemical characteristics
SPECIFIC TESTS of the strain of Bacillus stearothermophilus, ATCC No. 7953
PURITY: There is no evidence of contamination with other or 12980, whichever is stated in the labeling: under
microorganisms following examination of spores recovered microscopic examination it consists of Gram-positive rods
from the carriers using a suitable plate culture medium. with oval endospores in subterminally swollen cells; when
incubated in nutrient broth for 17 h and used to inoculate
ADDITIONAL REQUIREMENTS appropriate solid media, growth occurs when the inoculated
PACKAGING AND STORAGE: Preserve in the original package media are incubated aerobically for 24 h at 55 to 60, and
under the conditions recommended on the label, and similar inoculated media incubated concomitantly at 30 to
protect the package from light, toxic substances, excessive 35 show no evidence of growth in the same period. When
heat, and high relative humidity or moisture. The packaging examined under conventional biochemical tests for microbial
or container materials do not adversely affect the characterization, it shows a delayed weak positive catalase
performance of the article used as directed in the labeling. reaction, it does not utilize citrate, propionate or hippurate,
EXPIRATION DATE: The expiration date is determined on the it reduces nitrate, but it does not liquefy gelatin, and it gives
basis of stability studies and is NMT 18 months from the a negative result with the Voges-Proskauer test. Organisms
date of manufacture. The date of manufacture is the date on derived from ATCC strain No. 7953 show negative egg yolk
which the first determination of the total viable count was and starch hydrolysis reactions, while those derived from
made. ATCC strain No. 12980 show positive reactions in both tests.
DISPOSAL: Prior to destruction or discarding the carriers,
sterilize by moist heat sterilization to ensure that the carrier PERFORMANCE TESTS
surface is exposed to 121 for NLT 30 min, or by an [NOTESee Biological IndicatorsResistance Performance Tests
equivalent method recommended by the manufacturer. 55.]
LABELING: Label the package or package insert to state that D VALUE
it is a biological indicator prepared on a carrier for use in Analysis: Proceed as directed for D Value, using the
label-specified applications for moist heat, dry heat, and/or procedure for Biological Indicator for Steam Sterilization, Paper
gaseous sterilization. State the biological indicator D value Carrier.
obtained under defined exposures to stated sterilization Acceptance criteria: The determined D Value is within 20%
conditions using the Survival Curve method, Spearman- of the labeled D Value for the selected sterilizing
Karber method, or Stumbo-Murphy-Cochran method of D
value analysis. State the survival time and kill time for the
84 Biological / Official Monographs USP 32
temperature, and the confidence limits of the estimate are Biological Indicator for Steam
within 10% of the determined D Value. Sterilization, Self-Contained
SURVIVAL TIME and KILL TIME
Analysis: Proceed as directed for Survival Time and Kill Time, (Comment on this Monograph)id=m9512=Biological Indicator
using the procedure for Biological Indicator for Steam for Steam Sterilization, Self-Contained=B-Monos.pdf)
Sterilization, Paper Carrier. DEFINITION
Acceptance criteria Biological Indicator for Steam Sterilization, Self-Contained, is a
Survival time: All specimens show evidence of growth. Biological Indicator for Steam Sterilization, Paper Carrier
Kill time: No specimen shows growth. If for either the individually packaged in a suitable container readily penetrable
Survival time test or the Kill time test, NMT 1 specimen out by steam and designed to hold an appropriate bacteriological
of both groups fails the survival requirement or the kill culture medium, so as to enable the packaged carrier, after
requirement (whichever is applicable), continue the subjection to saturated steam sterilization conditions, to be
corresponding test with four additional groups, each incubated in the supplied medium in a self-contained system.
consisting of 20 specimens, according to the procedure The supplied medium may contain a suitable indicator as a
described. If all of the additional specimens subjected to convenience for determining by a color change whether or
steam sterilization either meet the survival requirement for not spores have survived. The design of the self-contained
the Survival time test or meet the kill requirement for the system is such that, after exposure to the specified sterilization
Kill time test, whichever is applicable, the requirements of conditions and inoculation of the medium under closed
the test are met. conditions as stated in the labeling, there is no loss of medium
TOTAL VIABLE SPORE COUNT and inoculum during subsequent transport and handling, if
Analysis: Proceed as directed for Total Viable Spore Count, done according to the provided instructions. The materials of
using the procedure for Biological Indicator for Steam which the self-contained system are made are such that there
Sterilization, Paper Carrier. is no retention or release of any substance that may cause
Acceptance criteria: The log average number of viable inhibition of growth of surviving spores under the incubation
spores per carrier is NLT 0.3 log of the labeled spore count conditions stated in the labeling.
per carrier and does not exceed the log labeled spore count
per carrier by 0.48. IDENTIFICATION
The biological indicator organism complies substantially with
SPECIFIC TESTS the morphological, cultural, and biochemical characteristics
PURITY of the strain of Bacillus stearothermophilus, ATCC No. 7953
Presence of contamination by other microorganisms: By or 12980, whichever is stated in the labeling: under
examination of the spores on a suitable plate culture microscopic examination it consists of Gram-positive rods
medium, there is no evidence of contamination with other with oval endospores in subterminally swollen cells; when
microorganisms. incubated in nutrient broth for 17 h and used to inoculate
ADDITIONAL REQUIREMENTS appropriate solid media, growth occurs when the inoculated
PACKAGING AND STORAGE: Preserve in the original package media are incubated aerobically for 24 h at 5560, and
under the conditions recommended on the label, and similar inoculated media incubated concomitantly at
protect it from light, toxic substances, excessive heat, and 3035 show no evidence of growth in the same period.
moisture. The packaging and container materials do not When examined under conventional biochemical tests for
adversely affect the performance of the article used as microbial characterization, it shows a delayed weak positive
directed in the labeling. catalase reaction; it does not utilize citrate, propionate, or
EXPIRATION DATE: The expiration date is determined on the hippurate; it reduces nitrate, but it does not liquefy gelatin;
basis of stability studies and is NLT 18 months from the date and it gives a negative result with the Voges-Proskauer test.
of manufacture, the date of manufacture being the date on Organisms derived from ATCC strain No. 7953 show
which the first determination of the total viable spore count negative egg yolk and starch hydrolysis reactions, while
was made. those derived from ATCC strain No. 12980 show positive
DISPOSAL: Prior to destruction or discard, sterilize it by reactions in both tests.
steam at 121 for NLT 30 min, or by no less than an PERFORMANCE TESTS
equivalent method recommended by the manufacturer. This [NOTESee Biological IndicatorsResistance Performance Tests
includes a test strip employed in any test procedures for the 55.]
strips themselves. D VALUE
LABELING: Label it to state that it is a Biological Indicator for Analysis: Proceed as directed for the relevant procedure for
Steam Sterilization, Paper Carrier; to indicate its D Value, the D Value Determination.
method used to determine such D Value, i.e., by spore count Acceptance criteria: The determined D value is within 20%
or fraction negative procedure after graded exposures to the of the labeled D value for the selected sterilizing
sterilization conditions; the Survival time and Kill time under temperature, and the confidence limits of the estimate are
specified sterilization conditions stated on the label; its within 10% of the determined D value.
particular total viable spore count with a statement that such SURVIVAL TIME and KILL TIME
count has been determined after preliminary heat treatment; Analysis: Proceed as directed for Survival Time and Kill Time,
and its recommended storage conditions. State in the using the procedure for Biological Indicator for Steam
labeling the size of the paper carrier, the strain and ATCC Sterilization, Self-Contained.
number from which the spores were derived, and Acceptance criteria
instructions for spore recovery and for safe disposal of the Survival time: All specimens show evidence of growth.
indicator. Indicate in the labeling that the stated D Value is Kill time: No specimen shows growth. If for either the
reproducible only under the exact conditions under which it Survival time test or the Kill time test, NMT 1 specimen out
was determined, that the user would not necessarily obtain of both groups fails the survival requirement or the kill
the same result and that the user should determine the requirement (whichever is applicable), continue the
suitability of the biological indicator for the particular use.
USP 32 Official Monographs / Biotin 85
corresponding test with 4 additional groups, each microorganisms in 1 mL. Inoculate the pooled medium
consisting of 20 specimens, according to the procedure with enough suspension to contain a total of 1001000
described. If all of the additional specimens subjected to microorganisms in a 10 mL aliquot of NMT the volume
steam sterilization either meet the survival requirement for from 10 units of the pooled medium. Incubate the
the Survival time test or meet the kill requirement for the inoculated pooled medium as directed for Total Viable Spore
Kill time test, whichever is applicable, the requirements of Count.
the test are met. Acceptance criteria: Clear evidence of growth is obtained
TOTAL VIABLE SPORE COUNT within 7 days.
Analysis: Proceed as directed for Total Viable Spore Count,
using the procedure for Biological Indicator for Steam ADDITIONAL REQUIREMENTS
Sterilization, Paper Carrier. PACKAGING AND STORAGE: Preserve in the original package
Acceptance criteria: The average number of viable spores under the conditions recommended on the label, and
per carrier is NLT 0.3 log of the labeled spore count per protect from light, from substances that may adversely affect
carrier and does not exceed the log labeled spore count per the contained microorganisms, from excessive heat, and
carrier by 0.48. from moisture.
EXPIRATION DATE: The expiration date is determined on the
SPECIFIC TESTS basis of stability studies and is NLT 18 months from the date
MEDIUM SUITABILITY of manufacture, the date of manufacture being the date on
Sterility: Incubate 10 self-contained biological indicator which the first determination of the total viable spore count
systems at 5560, or at the optimal recovery temperature was made.
specified by the manufacturer, for 48 h, making sure that DISPOSAL: Prior to destruction or discard, sterilize it by
there is no contact between the individual spore strips and steam at 121 for NLT 30 min, or by NLT an equivalent
the supplied medium. Examine the incubated medium method recommended by the manufacturer. This includes
visually (for change in color indicator or turbidity) and test strips employed in any test procedures for the strips
microscopically (for absence of microbial growth). themselves.
Growth promotion of medium prior to sterilization LABELING: Label it to state that it is a Biological Indicator for
treatment Steam Sterilization, Self-Contained; to indicate the D value of
Analysis: Submerge 10 self-contained units in a water bath the self-contained system, the method used to determine
maintained at 95100 for 15 min. Start timing when the such D value (i.e., by spore count or fraction negative
temperature of the container contents reach 95. Cool procedure after graded exposures to the sterilization
rapidly in an ice-water bath (04). Remove the units from conditions); the Survival time and Kill time under the specified
the ice-water bath, submerge each spore strip with the self- conditions stated on the label; its particular total viable spore
contained medium, incubate at 5560, or at the optimal count, with a statement that such count has been
recovery temperature specified by the manufacturer, and determined after preliminary heat treatment; and its
examine visually after 48 h for growth (for turbidity or recommended storage conditions. State on the labeling that
change in color), and microscopically (for microbial the supplied bacteriological medium will meet requirements
growth). for growth-promoting ability, the strain and ATCC number
Acceptance criteria: All the specimens under test show from which the spores were derived, and the instructions for
growth. If one or more of the specimens do not show spore recovery and for safe disposal of the indicator unit.
growth, repeat the test with 20 additional units. The Also indicate in the labeling that the stated resistance
additional units all show growth. characteristics are reproducible only under steam sterilization
Growth promotion of medium after exposure to conditions at the stated temperature and only under the
sterilization conditions: Expose the specified number of exact conditions under which it was determined, that the
units for both the Survival time and Kill time stated in the user would not necessarily obtain the same result, and that
labeling, as described in the section Biological Indicator for the user should determine the suitability of the biological
Steam Sterilization, Self-Contained under Biological indicator for the particular use.
IndicatorsResistance Performance Tests 55. Incubate the
spore strips submerged in the self-contained medium
according to the instructions of the manufacturer. At the
end of the incubation period confirm the existence of Biotin
growth in each of the specimens that were exposed for each (Comment on this Monograph)id=m9515=Biotin=B-Monos.pdf)
Survival time and the absence of growth in each of the
specimens that were exposed for each Kill time by visual
inspection (turbidity or color indicator change) and by
separate microscopic examination of each specimen and
confirm, where applicable, correspondence of the labeled
color to the appearance of growth in the supplied medium.
Ability of medium to support growth after exposure to
the sterilization conditions
Analysis: Take a stated number of units (e.g., 10) after C10H16N2O3S 244.31
they have been exposed for each Kill time stated in the 1H-Thieno3,4-dimidazole-4-pentanoic acid, hexahydro-2-oxo-,
labeling as directed in the preceding section. Aseptically 3aS-(3a,4,6a)-;
remove and pool the medium from each unit. Prepare a (3aS,4S,6aR)-Hexahydro-2-oxo-1H-thieno3,4-dimidazole-4-valeric
suspension of the indicator microorganism as directed for acid [58-85-5].
Total Viable Spore Counts under Biological Indicator for Steam
Sterilization, Paper Carrier. Prepare a dilution of that DEFINITION
suspension so as to contain 1001000 viable Biotin contains NLT 97.5% and NMT 100.5% of C10H16N2O3S.
86 Biotin / Official Monographs USP 32
IDENTIFICATION ASSAY
INFRARED ABSORPTION 197K PROCEDURE
Sample: 500 mg
ASSAY Analysis: Dissolve the Sample in 20 mL of benzene, add 2
PROCEDURE drops of crystal violet TS, and titrate with 0.1 N perchloric
Sample: 500 mg of Biotin acid VS to a blue endpoint. Perform a blank determination,
Analysis: Mix the sample with 100 mL of water. Add and make any necessary correction (see Titrimetry 541).
phenolphthalein TS and titrate the suspension slowly with Each mL of 0.1 N perchloric acid is equivalent to 31.15 mg
0.1 N sodium hydroxide VS, while heating and stirring of C21H29NO.
continuously. Each mL of 0.1 N sodium hydroxide is Acceptance criteria: NLT 98.0% and NMT 101.0%
equivalent to 24.43 mg of C10H16N2O3S.
Acceptance criteria: NLT 97.5% and NMT 100.5% IMPURITIES
OPTICAL ROTATION, Specific Rotation 781S Organic Impurities
Sample solution: 20 mg/mL in 0.1 N sodium hydroxide PROCEDURE: ORDINARY IMPURITIES 466
Acceptance criteria: Between +89 and +93 Standard solution: Prepare in methanol as directed.
Sample solution: Prepare in methanol as directed.
ADDITIONAL REQUIREMENTS Eluant: A mixture of methanol and ammonium hydroxide
PACKAGING AND STORAGE: Store in tight containers. (100:1.5)
USP REFERENCE STANDARDS 11 Visualization: 17
USP Biotin RS Analysis: Proceed as directed.
SPECIFIC TESTS
MELTING RANGE OR TEMPERATURE, Class I 741: Between
Biperiden 112 and 116
(Comment on this Monograph)id=m9530=Biperiden=B- LOSS ON DRYING 731
Monos.pdf) Analysis: Dry a sample at 105 for 3 h.
Acceptance criteria: The sample loses NMT 1.0% of its
weight.
USP Biperiden RS
C21H29NO 311.46
1-Piperidinepropanol, -bicyclo[2.2.1]hept-5-en-2-yl--phenyl-;
-5-Norbornen-2-yl--phenyl-1-piperidinepropanol [514-65-8].
Biperiden Hydrochloride
DEFINITION (Comment on this Monograph)id=m9550=Biperiden
Biperiden contains NLT 98.0% and NMT 101.0% of C21H29NO, Hydrochloride=B-Monos.pdf)
calculated on the dried basis.
C21H29NO HCl 347.92
IDENTIFICATION 1-Piperidinepropanol, -bicyclo[2.2.1]hept-5-en-2-yl--phenyl-,
A. INFRARED ABSORPTION 197K hydrochloride;
B. ULTRAVIOLET ABSORPTION 197U -5-Norbornen-2-yl--phenyl-1-piperidinepropanol
Solution: Transfer 180 mg of biperiden to a 200-mL hydrochloride [1235-82-1].
volumetric flask, add 1 mL of lactic acid and dilute with
water to volume (0.9 mg/mL). DEFINITION
Acceptance criteria: Absorptivities at 257 nm, calculated Biperiden Hydrochloride contains NLT 98.0% and NMT 101.0%
on the dried basis, do not differ by more than 3.0%. of C21H29NO HCl, calculated on the dried basis.
C. PROCEDURE IDENTIFICATION
Sample: 20 mg A. INFRARED ABSORPTION 197K
Analysis: Dissolve the Sample in 5 mL of phosphoric acid. B. ULTRAVIOLET ABSORPTION 197U
Acceptance criteria: A green color is produced. Solution: 1 mg/mL
D. PROCEDURE Medium: Methanol
Sample solution: Dissolve 200 mg in 80 mL of water with Acceptance criteria: Absorptivities at 257 nm do not differ
the aid of 0.5 mL of 3 N hydrochloric acid, warming, if by more than 3.0%.
necessary, to effect solution, and then cool. C. PROCEDURE
Analysis: To 5 mL of Sample solution, add 1 drop of Sample: 20 mg
hydrochloric acid and several drops of mercuric chloride TS. Analysis: Dissolve the Sample in 5 mL of phosphoric acid.
Acceptance criteria: A white precipitate is formed. Acceptance criteria: A green color is produced.
E. PROCEDURE D. PROCEDURE
Sample solution: Use Sample solution from Procedure D Sample solution: 1 in 500
Analysis: To a second 5-mL portion of the Sample solution, Analysis: To a 5-mL portion of the Sample solution add
add bromine TS dropwise. bromine TS dropwise.
Acceptance criteria: A yellow precipitate forms, which
redissolves on shaking, and finally, upon the addition of
more bromine TS, a permanent precipitate is formed.
USP 32 Official Monographs / Biperiden 87
Acceptance criteria: A yellow precipitate, which dissolves Visualization: Iodine vapor, 10 min
on shaking, is formed. Addition of more bromine TS Analysis: Separately apply the Sample solution and the
produces a precipitate that does not dissolve on shaking. Standard solution to the chromatographic plate. Allow the
E. IDENTIFICATION TESTSGENERAL, Chloride 191 applications to dry, and develop the chromatogram in the
Sample solution: 1 in 500 Developing solvent system until the solvent front has moved
Acceptance criteria: Meets the requirements of the tests about three-fourths of the length of the plate. Remove the
plate from the developing chamber, mark the solvent front,
ASSAY and allow the solvent to evaporate. Locate the spots on the
PROCEDURE plate by exposing the plate for 10 min to iodine vapors in a
Sample: 500 mg pre-equilibrated closed chamber, on the bottom of which
Analysis: Dissolve the Sample in 80 mL of glacial acetic there are iodine crystals.
acid, warming slightly, if necessary, to effect solution. Cool, Acceptance criteria: The RF value of the principal spot of
add 1 drop of crystal violet TS and 10 mL of mercuric the Sample solution corresponds to that of the Standard
acetate TS, and titrate with 0.1 N perchloric acid VS to a solution.
blue endpoint. Perform a blank determination, and make
any necessary correction (see Titrimetry 541). Each mL of ASSAY
0.1 N perchloric acid is equivalent to 34.79 mg of C21H29NO PROCEDURE
HCl. Solution A: 38 mg/mL of monobasic sodium phosphate
Acceptance criteria: NLT 98.0% and NMT 101.0% and 2 mg/mL of anhydrous dibasic sodium phosphate in
water. Adjust to a pH of 5.3 0.1, if necessary.
IMPURITIES Solution B: Dissolve 400 mg of bromocresol purple in 30
Organic Impurities mL of water, add 6.3 mL of 0.1 N sodium hydroxide, and
PROCEDURE: ORDINARY IMPURITIES 466 dilute with water to 500 mL.
Standard solution: Prepare in methanol as directed. Phosphate bufferbromocresol purple solution: Mix equal
Sample solution: Prepare in methanol as directed. volumes of Solution A, Solution B, and chloroform, shake in a
Eluant: A mixture of methanol and ammonium hydroxide separator, and discard the chloroform. If appreciable color is
(100:1.5) extracted, repeat with additional portions of chloroform until
Visualization: 17 no color is extracted.
Analysis: Proceed as directed. Standard stock solution: 0.8 mg/mL of USP Biperiden
Hydrochloride RS in methanol
SPECIFIC TESTS Standard solution: 40 g/mL of USP Biperiden
LOSS ON DRYING 731 Hydrochloride RS from Standard stock solution: transfer 5.0
Analysis: Dry a sample at 105 for 3 h. mL of Standard stock solution to a 100-mL volumetric flask,
Acceptance criteria: The sample loses NMT 0.5% of its add 25 mL of water, and dilute with methanol to volume.
weight. Sample solution: Transfer a portion of finely powdered
ADDITIONAL REQUIREMENTS Tablets, equivalent to 2 mg of biperiden hydrochloride, from
PACKAGING AND STORAGE: Preserve in well-closed, light- NLT 20 Tablets, to a 50-mL volumetric flask, add 12.5 mL of
resistant containers. water, and heat on a steam bath for 15 min. Cool, and
USP REFERENCE STANDARDS 11 dilute with methanol to volume.
USP Biperiden Hydrochloride RS Blank: Methanol and water (3:1)
Analysis
Samples: Standard solution, Sample solution, and Blank
Transfer 5 mL each of the Standard solution, the Sample
Biperiden Hydrochloride Tablets solution, and the Blank to individual separators, each
(Comment on this Monograph)id=m9580=Biperiden containing 10 mL of Phosphate bufferbromocresol purple
Hydrochloride Tablets=B-Monos.pdf) solution. Extract the solution in each separator with 20 mL
of chloroform for 2 min. After the layers have separated,
DEFINITION pass each chloroform extract through filter paper
Biperiden Hydrochloride Tablets contain NLT 93.0% and NMT (Whatman No. 31 or equivalent) into separate glass-
107.0% of the labeled amount of C21H29NO HCl. stoppered, 50-mL volumetric flasks. In the same manner,
extract the solution in each separator with another 20-mL
IDENTIFICATION portion of chloroform, filter, and wash each filter with 8
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 mL of chloroform, collecting each combined filtrate and
Adsorbent: 0.25-mm layer of chromatographic silica gel washing, respectively, in the 50-mL volumetric flask
mixture. Condition by heating the plate at 105 for 1 h and containing the first extract. Dilute each with chloroform to
allowing to cool. volume. Concomitantly determine the absorbances of the
Standard solution: Proceed as directed for the Sample solutions in 1-cm cells at the wavelength of maximum
solution using 10 mg of USP Biperiden Hydrochloride RS in absorbance at about 408 nm, with a suitable
place of the powdered Tablets. spectrophotometer, using the Blank to set the instrument.
Sample solution: To a quantity of finely powdered Tablets, Calculate the percentage of C21H29NO HCl in the portion of
equivalent to 10 mg of biperiden hydrochloride, add 5 mL Tablets taken:
of water, mix, and sonicate to disperse the powder. Add 5
mL of methanol to the flask, mix, and sonicate for 15 min. Result = (AU/AS) (CS/CU) F 100/L
Filter the solution into a separator, add 2 mL of 1 N sodium
hydroxide and 10 mL of chloroform, and shake for 3 min. AU = absorbance of the Sample solution
Filter the chloroform layer into a stoppered flask, and use AS = absorbance of the Standard solution
the chloroform filtrate as the Sample solution. CS = concentration of USP Biperiden Hydrochloride RS
Application volume: 20 L in the Standard solution (g/mL)
Developing solvent system: Methanol and ammonium CU = nominal concentration of the Sample solution
hydroxide (100:1.5) (g/mL)
88 Biperiden / Official Monographs USP 32
F = conversion factor, 0.001 (g to mg) IDENTIFICATION

Acceptance criteria: NLT 93.0% and NMT 107.0% IDENTIFICATIONORGANIC NITROGENOUS BASES 181
Sample solution: A volume of Injection equivalent to 50
PERFORMANCE TESTS mg of biperiden lactate
DISSOLUTION 711 Standard solution: 50 mg of USP Biperiden RS in 25 mL of
Medium: 0.01 N hydrochloric acid; 500 mL 0.01 N hydrochloric acid
Apparatus 2: 50 rpm Analysis: Proceed as directed under IdentificationOrganic
Time: 45 min Nitrogenous Bases 181, beginning with Transfer the liquid
[NOTEDetermine the amount of C21H29NO HCl dissolved to a separator.
by employing the following method.] Acceptance criteria: Meets the requirements
Phosphate bufferbromocresol purple solution: Prepare
as directed for Assay. ASSAY
Standard stock solution: 0.8 mg/mL of USP Biperiden PROCEDURE
Hydrochloride RS in methanol Solution A: 38 mg/mL of monobasic sodium phosphate
Standard solution: 2 g/mL of USP Biperiden and 2 mg/mL of anhydrous dibasic sodium phosphate in
Hydrochloride RS, prepared as follows: Pipet 5 mL of water
Standard stock solution into a 500-mL volumetric flask, and Adjust the pH of the solution to 5.3 0.1, if necessary.
add 0.01 N hydrochloric acid to volume. Pipet 25 mL of this Solution B: Dissolve 400 mg of bromocresol purple in 30
solution into a suitable beaker, and adjust with 0.01 N mL of water. Add 6.3 mL of 0.1 N sodium hydroxide, and
sodium hydroxide to a pH of 5.3. Transfer this solution to a dilute with water to 500 mL.
100-mL volumetric flask with the aid of water, and dilute Phosphate bufferbromocresol purple solution: Mix equal
with water to volume. volumes of Solution A, Solution B, and chloroform. Shake in a
Sample solution: Sample per Dissolution 711. Filter 75 mL separator, and discard the chloroform. If appreciable color is
of the solution under test, pipet 50 mL of the clear filtrate extracted, repeat with additional portions of chloroform until
into a suitable beaker, and adjust with 0.01 N sodium no color is extracted.
hydroxide to a pH of 5.3. Transfer this solution to a 100-mL Standard stock solution: 0.8 mg/mL of USP Biperiden RS
volumetric flask with the aid of water, and dilute with water in methanol
to volume. Standard solution: 40 g/mL of USP Biperiden RS from
Blank: Water Standard stock solution
Analysis Transfer 5.0 mL of Standard stock solution to a 100-mL
Samples: Standard solution, Sample solution, and Blank volumetric flask, add 25 mL of water, and dilute with
Pipet 20 mL each of the Standard solution, the Sample methanol to volume.
solution, and the Blank into individual separators, each Sample solution: Transfer an amount of the Injection,
containing 10.0 mL of Phosphate bufferbromocresol purple equivalent to 5 mg of biperiden lactate, to a 100-mL
solution. Extract the solution in each separator with 40.0 volumetric flask. Add 25 mL of water, and dilute to volume
mL of chloroform for 10 min. After the layers have with methanol.
separated, pass each chloroform extract through filter Blank: Methanol and water (3:1)
paper into separate, glass-stoppered containers, discarding Analysis
the first 10 mL of each filtrate. Determine the amount of Samples: Standard solution, Sample solution, and Blank
C21H29NO HCl dissolved from absorbances at the Transfer 5.0 mL each of the Standard solution, the Sample
wavelength of maximum absorbance at about 408 nm solution, and the Blank to individual separators, each
(10-cm cells) of the extract from the Sample solution in containing 10.0 mL of Phosphate bufferbromocresol purple
comparison with that of the extract from the Standard solution. Extract the solution in each separator with 20.0
solution, using the Blank to set the instrument. mL of chloroform for 2 min. After the layers have
Acceptance criteria: NLT 75% (Q) separated, pass each chloroform extract through filter
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements paper (Whatman No. 31 or equivalent) into separate
glass-stoppered, 50-mL volumetric flasks. In the same
ADDITIONAL REQUIREMENTS manner, extract the solution in each separator with
PACKAGING AND STORAGE: Preserve in tight containers. another 20.0-mL portion of chloroform, filter, and wash
USP REFERENCE STANDARDS 11 each filter with 8 mL of chloroform, collecting each
USP Biperiden Hydrochloride RS combined filtrate and washing, respectively, in the 50-mL
volumetric flask containing the first extract. Dilute each
with chloroform to volume. Concomitantly determine the
Biperiden Lactate Injection absorbances of the solutions in 1-cm cells at the
wavelength of maximum absorbance at about 408 nm,
(Comment on this Monograph)id=m9610=Biperiden Lactate with a suitable spectrophotometer, using the blank to set
Injection=B-Monos.pdf) the instrument.
C21H29NO C3H6O3 401.54 Calculate the percentage of C21H29NO C3H6O3 in the
1-Piperidinepropanol, -bicyclo[2.2.1]hept-5-en-2-yl--phenyl-, portion of Injection taken:
compd. with 2-hydroxypropanoic acid (1:1);
-5-Norbornen-2-yl--phenyl-1-piperidinepropanol lactate Result = (AU/AS) (CS/CU) (Mr1/Mr2) F 100
(salt) [7085-45-2].
AU = absorbance from the Sample solution
DEFINITION AS = absorbance from the Standard solution
Biperiden Lactate Injection is a sterile solution of biperiden CS = concentration of USP Biperiden Hydrochloride RS
lactate (C21H29NO C3H6O3) in Water for Injection, prepared in the Standard solution (g/mL)
from Biperiden with the aid of Lactic Acid. It contains NLT CU = nominal concentration of biperiden lactate in the
95.0% and NMT 105.0% of the labeled amount of C21H29NO Sample solution (mg/mL)
C3H6O3. Mr1 = molecular weight of biperiden lactate, 401.55
USP 32 Official Monographs / Bisacodyl 89
Mr2 = molecular weight of biperiden, 311.47 ADDITIONAL REQUIREMENTS

F = conversion factor, 0.001 (g to mg) PACKAGING AND STORAGE: Preserve in well-closed containers.
USP Bisacodyl RS
SPECIFIC TESTS
BACTERIAL ENDOTOXINS TEST 85: NMT 83.3 USP Endotoxin
Units/mg of biperiden lactate
PH 791: 4.85.8 Bisacodyl Suppositories
OTHER REQUIREMENTS: Meets the requirements under (Comment on this Monograph)id=m9685=Bisacodyl
Injections 1 Suppositories=B-Monos.pdf)
ADDITIONAL REQUIREMENTS DEFINITION
PACKAGING AND STORAGE: Preserve in single-dose containers, Bisacodyl Suppositories contain NLT 90.0% and NMT 110.0%
preferably of Type I glass, protected from light. of the labeled amount of C22H19NO4.
USP Biperiden RS IDENTIFICATION
USP Endotoxin RS A. PROCEDURE
Sample: Transfer the equivalent to 150 mg of bisacodyl
from Suppositories, to a 500-mL conical flask, add 75 mL of
solvent hexane, and heat on a steam bath until melted.
Bisacodyl Filter the solution, with the aid of a vacuum, through a
(Comment on this Monograph)id=m9660=Bisacodyl=B- medium-porosity, sintered-glass funnel, and wash the
Monos.pdf) residue with about 100 mL of warm solvent hexane until it
is free from fat. Continue the vacuum until the residue
appears dry. Dissolve the residue by rinsing the filter with
about 50 mL of warm acetone, collecting the filtrate in a
150-mL beaker, and evaporating the filtrate on a steam
bath to a volume of about 5 mL. To the residual liquid add
about 75 mL of water, heat on a steam bath for 15 min,
and cool. Scratch the sides of the beaker to induce
crystallization, filter the crystals, and dry at 100 for about
C22H19NO4 361.39 15 min.
Phenol, 4,4-(2-pyridinylmethylene)bis-, diacetate (ester); Analysis 1: The bisacodyl so obtained melts between 129
4,4-(2-Pyridylmethylene)diphenol diacetate (ester). [603-50-9]. and 135.
Analysis 2: Infrared Absorption 197S
DEFINITION Cell: 1.0 mm
Bisacodyl contains NLT 98.0% and NMT 101.0% of C22H19NO4, Sample solution: Dissolve a portion of the crystalline
calculated on the dried basis. Sample in chloroform (1 in 200).
[CAUTIONAvoid inhalation and contact with the eyes, skin, and solution corresponds to that of the Standard solution, as
mucous membranes.]
IDENTIFICATION
A. INFRARED ABSORPTION 197S ASSAY
Cell: 1.0 mm PROCEDURE
Solution: 1 in 200 solution in chloroform, previously dried Solution A: 0.074 M sodium acetate in water, adjusted with
B. ULTRAVIOLET ABSORPTION 197U 2.5% acetic acid to a pH of 7.4
Analytical wavelength: 263 nm Mobile phase: Acetonitrile and Solution A (9:11)
Solution: 20 g/mL in 0.05 N hydrochloric acid Standard solution: 0.5 mg/mL of USP Bisacodyl RS in
Acceptance criteria: Absorptivities, calculated on the dried acetonitrile
basis, do not differ by more than 3.0%. Sample solution: To the equivalent to 100 mg of bisacodyl
from Suppositories, in a 500-mL separator, add 150 mL of
ASSAY n-hexane, and shake until all the suppositories are dissolved.
PROCEDURE Add 50 mL of acetonitrile, shake for 1 min, and allow the
Sample solution: Dissolve 250 mg of Bisacodyl in 70 mL of layers to separate. Drain the lower layer into a 200-mL
glacial acetic acid, and add 3 drops of p-naphtholbenzein volumetric flask, and extract the n-hexane layer remaining in
TS. the separator with two 50-mL portions of acetonitrile,
Titrate with 0.1 N perchloric acid VS. Perform a blank combining the lower layers in the volumetric flask. Dilute
determination, and make any necessary correction (see the combined extracts in the volumetric flask with
Titrimetry 541). Each mL of 0.1 N perchloric acid is acetonitrile to volume, mix, and filter.
equivalent to 36.14 mg of C22H19NO4. Chromatographic system
Acceptance criteria: 98.0%101.0% (See Chromatography 621, System Suitability.)
IMPURITIES
SPECIFIC TESTS
0.5% of its weight.
90 Bisacodyl / Official Monographs USP 32
Mode: LC Mode: LC
Detector: UV 265 nm Detector: UV 254 nm
Guard column: Packing L2 Column: 3.9-mm 30-cm; packing L1
Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min
Injection size: 10 L System suitability
System suitability Sample: Standard solution
Sample: Standard solution [NOTEThe relative retention times for bisacodyl and
Suitability requirements ethylparaben are about 2.0 and 1.0, respectively.]
Tailing factor: NMT 2.0 Suitability requirements
Relative standard deviation: NMT 2.0% of replicate Resolution: NLT 7.0 between bisacodyl and the internal
injections standard
Samples: Standard solution and Sample solution Tailing factor: NMT 1.2
Calculate the percentage of C22H19NO4 taken: Relative standard deviation: NMT 2.0%
Analysis
Result = (rU/rS) (CS/CU) 100 Samples: Standard solution and Sample solution
Calculate the percent label claim of C22H19NO4:
rS = peak response from the Standard solution Result = (RU/RS) (CS/CU) 100
CS = concentration of USP Bisacodyl RS in the
Standard solution (mg/mL) RU = peak response ratio of the bisacodyl peak to the
CU = concentration of bisacodyl in the Sample solution internal standard peak from the Sample solution
(mg/mL) RS = peak response ratio of the bisacodyl peak to the
Acceptance criteria: 90.0%110.0% internal standard peak from the Standard
solution
ADDITIONAL REQUIREMENTS CS = concentration of USP Bisacodyl RS in the
PACKAGING AND STORAGE: Preserve in well-closed containers Standard solution (g/mL)
at a temperature not exceeding 30. CU = concentration of bisacodyl in the Sample solution
USP REFERENCE STANDARDS 11 (g/mL)
USP Bisacodyl RS Acceptance criteria: 90.0%115.0%
SPECIFIC TESTS
PH 791: 5.06.8
Bisacodyl Rectal Suspension
(Comment on this Monograph)id=m9680=Bisacodyl Rectal ADDITIONAL REQUIREMENTS
Suspension=B-Monos.pdf) PACKAGING AND STORAGE: Preserve in unit-dose containers at
a temperature not exceeding 30.
DEFINITION USP REFERENCE STANDARDS 11
Bisacodyl Rectal Suspension is a suspension of Bisacodyl in a USP Bisacodyl RS
suitable aqueous medium. It contains NLT 90.0% and NMT
115.0% of the labeled amount of C22H19NO4.
IDENTIFICATION Bisacodyl Delayed-Release Tablets
The retention time of the major peak of the Sample solution (Comment on this Monograph)id=m9695=Bisacodyl Delayed-
corresponds to that of the Standard solution, as obtained in the Release Tablets=B-Monos.pdf)
Assay.
DEFINITION
ASSAY Bisacodyl Delayed-Release Tablets contain NLT 90.0% and NMT
PROCEDURE 110.0% of the labeled amount of C22H19NO4. Bisacodyl
Mobile phase: Methanol and 0.01 M monobasic potassium Delayed-Release Tablets are enteric coated.
phosphate (3:2)
Internal standard solution: Dissolve ethylparaben in IDENTIFICATION
methanol and dilute with an equal volume of water (5.0 A. INFRARED ABSORPTION 197S
mg/mL). Cell: 1.0 mm
Standard solution: Dissolve a quantity of USP Bisacodyl RS Sample solution: Macerate the equivalent of 300 mg of
in methanol, add a volume of Internal standard solution, and bisacodyl from powdered Tablets, with 100 mL of acetone.
dilute with methanol to obtain a solution of 67 g/mL of Heat on a steam bath to boiling, filter, and evaporate to
bisacodyl and 250 g/mL of ethylparaben. about 20 mL. Add 200 mL of water, and warm the mixture
Sample solution: Transfer a measured volume of Rectal on the steam bath, passing a stream of nitrogen over the
Suspension equivalent to 6.7 mg of bisacodyl to a 100-mL surface to evaporate the acetone. After 30 min, cool the
volumetric flask. Add 5.0 mL Internal standard solution and mixture, and filter through a sintered-glass funnel. Discard
dilute with methanol to volume. the filtrate, and dissolve the crystals in 50 mL of acetone.
Chromatographic system Evaporate the solution to about 15 mL, add about 75 mL of
(See Chromatography 621, System Suitability.) water, heat on a steam bath for 15 min, and then cool.
USP 32 Official Monographs / Bismuth 91
Scratch the sides of the beaker to induce crystallization, filter Milk of Bismuth
the crystals, and dry at 100 for about 15 min. Using the (Comment on this Monograph)id=m9750=Milk of Bismuth=B-
crystals so obtained prepare a solution (1 in 200) in Monos.pdf)
chloroform.
B. The retention time of the major peak for bisacodyl of the DEFINITION
Sample solution corresponds to that of the Standard solution, Milk of Bismuth contains bismuth hydroxide and bismuth
as obtained in the Assay. subcarbonate in suspension in water, and yields NLT 5.2% and
NMT 5.8% (w/w) of bismuth trioxide (Bi2O3).
ASSAY
PROCEDURE
Solution A: 0.074 M sodium acetate in water, adjusted with Bismuth Subnitrate 80 g
2.5% acetic acid to a pH of 7.4 Nitric Acid 120 mL
Mobile phase: Acetonitrile and Solution A (9:11) Ammonium Carbonate 10 g
Standard solution: 0.5 mg/mL of USP Bisacodyl RS in
acetonitrile Strong Ammonia Solution
Sample solution: Equivalent to 100 mg of bisacodyl from Purified Water, each A sufficient quantity
powdered Tablets, in a 200-mL volumetric flask, add 25 mL To make 1000 mL
of water, and shake by mechanical means for 15 min
followed by sonication for 15 min. Add 100 mL of Mix the Bismuth Subnitrate with 60 mL of Purified Water and
acetonitrile, and shake by mechanical means for 15 min 60 mL of the Nitric Acid in a suitable container, and agitate,
followed by sonication for 15 min. Dilute with acetonitrile to warming gently until solution is effected. Pour this solution,
volume, mix, and filter. with constant stirring, into 5000 mL of Purified Water
Chromatographic system containing 60 mL of the Nitric Acid. Dilute 160 mL of Strong
(See Chromatography 621, System Suitability.) Ammonia Solution with 4300 mL of Purified Water in a glazed
Mode: LC or glass vessel of at least 12-L capacity. Dissolve the
Detector: UV 265 nm Ammonium Carbonate in this solution, and then pour the
Guard column: Packing L2 bismuth solution quickly into it with constant stirring. Add
Column: 3.9-mm 30-cm; packing L1 sufficient 6 N ammonium hydroxide, if necessary, to render
Flow rate: 2 mL/min the mixture distinctly alkaline, allow to stand until the
Injection size: 10 L precipitate has settled, then pour or siphon off the
System suitability supernatant, and wash the precipitate twice with Purified
Sample: Standard solution Water, by decantation. Transfer the magma to a strainer of
Suitability requirements close texture, so as to provide continuous washing with
Tailing factor: NMT 2.0 Purified Water, the outlet tube being elevated to prevent the
Relative standard deviation: NMT 2.0% surface of the magma from becoming dry. When the washings
Analysis no longer yield a pink color with phenolphthalein TS, drain
Samples: Standard solution and Sample solution the moist solution, transfer to a graduated vessel, add
Calculate the percent label claim of C22H19NO4: sufficient Purified Water to make 1000 mL, and mix.
[NOTEThis method of solution may be varied, provided the
Result = (rU/rs) (CS/CU) 100
product meets the following requirements.]
rU = peak response from the Sample solution IDENTIFICATION
rS = peak response from the Standard solution A. IDENTIFICATION TESTSGENERAL, Bismuth 191 and
CS = concentration of USP Bisacodyl RS in the Carbonate 191: It meets the requirements.
Standard solution (mg/mL) B. Add 1 mL of 3 N hydrochloric acid to 1 mL of Milk of
CU = concentration of bisacodyl in the Sample solution Bismuth: a clear solution is produced. Pour the clear solution
(mg/mL) into 10 volumes of water: a white precipitate is formed.
ASSAY
DISINTEGRATION 701: Proceed as directed for Delayed- Sample solution: Evaporate a quantity of Milk of Bismuth
Release (Enteric Coated) Tablets: the tablets do not to dryness, and ignite the residue to constant weight. From
disintegrate after 1 h of agitation in simulated gastric fluid the weight of the Bi2O3 so obtained determine the
TS, but then disintegrate within 45 min in simulated percentage in the Assay sample.
intestinal fluid TS. Acceptance criteria: 5.2%5.8%
IMPURITIES
ADDITIONAL REQUIREMENTS Inorganic Impurities
PACKAGING AND STORAGE Preserve in well-closed containers ARSENIC, Method I 211
at a temperature not exceeding 30. Sample solution: Evaporate 3.75 mL on a steam bath to
LABELING: Label the Tablets to indicate that they are enteric dryness, add 2 mL of sulfuric acid, and heat until copious
coated. fumes of sulfur trioxide are evolved.
USP Bisacodyl RS
92 Bismuth / Official Monographs USP 32
Acceptance criteria: NMT 0.8 ppm Allow to cool, add 2 mL of nitric acid to the residue,
LEAD: To 5 mL add warm nitric acid, dropwise, until it is dropwise, and warm until complete solution has been
just dissolved, and pour the solution into 50 mL of water: a effected. Add about 60 mL of water and 0.3 mL of xylenol
white precipitate may form. Filter, if necessary, evaporate the orange TS.
filtrate on a steam bath to 15 mL, again filter, and to 10 mL Analysis
of the filtrate add an equal volume of 2 N sulfuric acid. Sample: Sample solution
Acceptance criteria: No precipitate is formed. Titrate with 0.05 N edetate disodium VS to a yellow
LIMIT OF ALKALIES AND ALKALINE EARTHS endpoint. Each mL of 0.05 N edetate disodium is
Sample solution: 2.0 mL in 5 mL of hydrochloric acid, equivalent to 10.45 mg of Bi.
dilute with water to 100 mL, add hydrogen sulfide to Acceptance criteria: 49%54%
precipitate the bismuth completely, and filter
Analysis: To 50 mL of the clear filtrate add 5 drops of IMPURITIES
sulfuric acid, evaporate to dryness, and ignite. Inorganic Impurities
Acceptance criteria: The weight of the residue does not ARSENIC, Method I 211
exceed 3 mg (0.3%). Sample: 300 mg of Bismuth Citrate
WATER-SOLUBLE SUBSTANCES: Boil 10 mL with 90 mL of Analysis: Triturate Sample with an equal weight of calcium
water for 10 min, cool, add water to make the total volume hydroxide, and ignite. Dissolve the residue in 5 mL of 3 N
100 mL, mix, and filter. Evaporate 50 mL of the filtrate to hydrochloric acid.
dryness, and ignite it gently: the weight of the residue does Acceptance criteria: NMT 10 ppm
not exceed 5 mg (0.1%). LIMIT OF NITRATE
Sample solution: The second portion of the cooled
SPECIFIC TESTS solution reserved from Identification test C
MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED Analysis: To the Sample solution, add an equal volume of
MICROORGANISMS 62: The total bacterial count does not sulfuric acid, and allow to cool. Into the liquid, drop a
exceed 100 cfu/mL and the test for Escherichia coli is crystal of ferrous sulfate, and allow to stand for 30 min.
negative. Acceptance criteria: No brown or brownish black color
appears around the crystal.
ADDITIONAL REQUIREMENTS LIMIT OF COPPER, LEAD, AND SILVER
PACKAGING AND STORAGE: Preserve in tight containers, and Standard solution: Prepare a solution containing 1000
protect from freezing. g/mL of copper, a solution containing 1000 g/mL of
lead, and a solution containing 1000 g/mL of silver.
Transfer 3.0 mL of each solution to a 2000-mL volumetric
Bismuth Citrate flask, dilute with 1 N nitric acid to volume.
[NOTEThe concentrations of copper, lead, and silver in
(Comment on this Monograph)id=m9755=Bismuth Citrate=B- this solution may be modified by using a different
Monos.pdf) quantity or by further dilution to bring the absorption
responses within the working range of the atomic
absorption spectrophotometer.]
Sample solution: Ignite 3 g of Bismuth Citrate, in a
porcelain crucible, cool, and cautiously add 6 N nitric acid
to dissolve the residue. Add 100 mL of water. A white
precipitate forms. Filter this mixture, evaporate on a steam
bath to obtain about 15 mL of solution, and filter again.
Dilute the filtrate with water to 20.0 mL.
BiC6H5O7 398.08 Analysis
[813-93-4]. Samples: Standard solution and Sample solution
DEFINITION Concomitantly determine the absorbances of the Standard
Bismuth Citrate contains NLT 49% and NMT 54% of bismuth solution and the Sample solution at the emission lines of
(Bi). 324.7, 217, and 328.1 nm for copper, lead, and silver,
respectively, with an atomic absorption
IDENTIFICATION spectrophotometer (see Spectrophotometry and Light-
A. INFRARED ABSORPTION 197K: On the undried specimen Scattering 851) equipped with copper, lead, and silver
B. When strongly heated, the salt chars, and on ignition hollow-cathode lamps and an oxidizing flame.
leaves a more or less blackened residue having a yellow Acceptance criteria: The absorbances of the Sample
surface. The residue is soluble in warm nitric acid, and this solution do not exceed those of the Standard solution for
solution, when dropped into a large excess of water, each element (NMT 10 ppm).
produces a white turbidity. LIMIT OF SOLUBLE BISMUTH
C. PROCEDURE Standard solution: 242.0 mg of bismuth nitrate
Sample solution: 1 g in ammonia TS pentahydrate to a 100-mL volumetric flask. Add 3 mL of
Analysis: When treated with hydrogen sulfide in excess, a 1.5 N nitric acid, swirl to dissolve, and dilute with water to
black precipitate is obtained. Filter this mixture, drive off the volume. Transfer 1.0 mL of this solution to a 500-mL
excess hydrogen sulfide by heating, and allow to cool. To a volumetric flask, add 250 mL of 1.5 N nitric acid, and
portion of this cooled solution add an excess of calcium dilute with water to volume. This solution contains 2.0
hydroxide TS, and boil. g/mL of Bi.
Acceptance criteria: A white precipitate is formed. Reserve [NOTEThe concentration of bismuth in this solution may
a second portion of the cooled solution for the test for Limit be modified by using a different quantity or by further
of nitrate. dilution to bring the absorption responses within the
working range of the atomic absorption
ASSAY spectrophotometer.]
PROCEDURE Sample solution: Prepare a mixture of 5.0 g of Bismuth
Sample solution: Transfer 300 mg of Bismuth Citrate to a Citrate in 100 mL of water, and stir by mechanical means
porcelain crucible, and ignite.
the suspension thus obtained for 2 h. Pass through filter Analysis: To the Standard solution and the Sample solution,
paper. Pass the filtrate thus obtained through a filter add 0.05 mL of Indigo carmine titrant. Carefully add 30 mL
having a 0.1-m or finer porosity. To 10.0 mL of the filtrate of sulfuric acid, and immediately titrate with Indigo carmine
add 0.1 mL of nitric acid. titrant to a stable blue endpoint.
Analysis Acceptance criteria: The volume of Indigo carmine titrant
Samples: Standard solution and Sample solution consumed by the Sample solution does not exceed that
Concomitantly determine the absorbances of the Standard consumed by the Standard solution (0.4%).
solution and the Sample solution at the emission line of LIMIT OF SILVER
223.06 nm for bismuth with an atomic absorption Standard solution: 7.87 g/mL of silver nitrate
spectrophotometer (see Spectrophotometry and Light- Sample solution: 2.0 g of Bismuth Subcarbonate, add 1
Scattering 851) equipped with a bismuth hollow- mL of water and 4 mL of nitric acid
cathode lamp and an oxidizing flame. Analysis: Heat gently to achieve dissolution, add water to
Acceptance criteria: The absorbances of the Sample obtain 11 mL of solution, and cool. Add 2 mL of 1 N
solution do not exceed those of the Standard solution (NMT hydrochloric acid, and allow to stand in a dark place for 5
40 ppm). min.
Acceptance criteria: No more turbidity is produced than
ADDITIONAL REQUIREMENTS corresponds to that produced with 10 mL of the Standard
PACKAGING AND STORAGE: Preserve in tight, light-resistant solution concomitantly treated with 1 mL of nitric acid and
containers, store at controlled room temperature, and 2 mL of 1 N hydrochloric acid (0.0025%).
prevent exposure to excessive heat. LIMIT OF COPPER
USP REFERENCE STANDARDS 11 Standard stock solution: 1.34 g of cupric chloride, 10 g
USP Bismuth Citrate RS of ammonium chloride, and 3 mL of sodium metabisulfite
solution (275 mg/mL) to 100.0 mL. This stock solution
contains the equivalent of 5 mg/mL of copper.
Bismuth Subcarbonate Standard solution: equivalent of 10 g/mL of copper
made from the Standard stock solution in 2 N nitric acid.
(Comment on this Monograph)id=m9760=Bismuth Mix 0.25 mL of this solution and 9.75 mL of water.
Subcarbonate=B-Monos.pdf) Sample solution: 5 mL of the Sample stock solution
retained from the test for Chloride, add 2 mL of 6 N
DEFINITION ammonium hydroxide, dilute with water to 50 mL, mix,
Bismuth Subcarbonate contains NLT 97.6% and NMT 100.7% and filter.
of (BiO)2CO3, calculated on the dried basis. Analysis: To 10 mL each of the Standard solution and the
IDENTIFICATION Sample solution, add 1 mL of a solution of sodium
IDENTIFICATION TESTSGENERAL, Bismuth 191 and Carbonate diethyldithiocarbamate (1 in 1000).
Acceptance criteria: No more color is obtained from the
191 Sample solution than is obtained from the Standard solution
ASSAY (0.005%).
PROCEDURE LIMIT OF LEAD
Sample solution: 500 mg of Bismuth Subcarbonate in 3 Diluent: 6 N nitric acid, lead-free
mL of nitric acid. Dilute with water to 250 mL, add 0.3 mL Standard stock solution: 0.1598 mg/mL of lead nitrate in
of xylenol orange TS Diluent [NOTEThis solution contains 100 g/mL of lead.]
Analysis: Titrate with 0.05 M edetate disodium VS to a Standard solution: 1.0 g/mL, 2.0 g/mL, and 3.0
yellow endpoint. Each mL of 0.05 M edetate disodium is g/mL of lead from Standard stock solution in Diluent
equivalent to 12.75 mg of (BiO)2CO3. Sample solution: 12.5 g of Bismuth Subcarbonate in 75
Acceptance criteria: 97.6%100.7% mL of Diluent. Heat to boiling for 1 min, cool, and dilute
with water to 100 mL.
IMPURITIES Analysis: Concomitantly determine the absorbances of the
Inorganic Impurities Standard solutions and the Sample solution at the lead
CHLORIDE AND SULFATE, Chloride 221 emission line of 283.3 nm with an atomic absorption
Sample stock solution: 5.0 g in 10 mL of water. Add 20 spectrophotometer (See Spectrophotometry and Light-
mL of nitric acid, warm to achieve dissolution, allow to scattering 851) equipped with a lead hollow-cathode lamp
cool, and dilute with water to obtain 100 mL of solution. and an airacetylene flame, using a 1:5 dilution of the
Sample solution: To 6.6 mL of the Sample stock solution Diluent as the blank. Plot the absorbances of the Standard
add 4 mL of nitric acid, and dilute with water to 50 mL. solutions versus concentration, in g/mL, of lead, and draw
Acceptance criteria: A 15.0-mL portion of the Sample the straight line best fitting the three plotted points. From
solution shows no more chloride than corresponds to 70 L the graph so obtained, determine the concentration, C, in
of 0.020 N hydrochloric acid (0.05%). g/mL, of lead in the Sample solution.
LIMIT OF ARSENIC, Method I 211 Calculate the percentage of lead (Pb) in the portion of
Sample solution: 600 mg in 35 mL of 3 N hydrochloric Bismuth Subcarbonate taken:
acid
Acceptance criteria: NMT 5 ppm Result = C/12,500
LIMIT OF NITRATE
Indigo carmine titrant: 4 g of indigo carmine in 900 mL C = concentration of lead in the Sample solution
of water, add 2 mL of sulfuric acid, and dilute with water LIMIT OF ALKALIES AND ALKALINE EARTHS
to 1000 mL Sample solution: Boil 1.0 g with 20 mL of a mixture of
Standard solution: 0.0815 mg/mL of potassium nitrate acetic acid and water (1:1). After 2 min, cool and filter.
(equivalent to 0.05 mg/mL of nitrate). Place 20.0 mL in a Analysis: Collect the filtrate, wash the residue with 20 mL
125-mL conical flask. of water, and add the washing to the filtrate. To this
Sample solution: 250 mg of Bismuth Subcarbonate in a solution add 2 mL of 2 N hydrochloric acid and 20 mL of
125-mL conical flask, add 20 mL of water, and swirl to water. Heat to boiling and precipitate the bismuth by
suspend adding hydrogen sulfide. Cool the mixture, and filter.
Collect the filtrate, wash the residue with water, and add Acceptance criteria: No reddish brown color appears at
the washing to the filtrate. Evaporate this solution to the zone of contact of the two liquids.
dryness on a water bath. To the residue add 0.5 mL of COPPER, LEAD, AND SILVER
sulfuric acid, dry slowly, and cool. Sample: Ignite 3 g in a porcelain crucible, cool, and
Acceptance criteria: The weight of the residue does not cautiously add, dropwise, just sufficient nitric acid to
exceed 10 mg (1.0%). dissolve the residue upon warming. Evaporate the solution
to dryness, again ignite, and cool. Cautiously dissolve the
SPECIFIC TESTS residue in just sufficient nitric acid with the aid of gentle
LOSS ON DRYING 731: Dry at 105 to constant weight: it heat, concentrate the solution to about 4 mL, pour it into
loses NMT 1.0% of its weight. 100 mL of water, filter, evaporate the filtrate on a steam
bath to 20 mL, again filter, and divide this filtrate into
ADDITIONAL REQUIREMENTS portions of 5 mL each.
PACKAGING AND STORAGE: Preserve in well-closed containers, Acceptance criteria
protected from light. Copper: Add a slight excess of 6 N ammonium
hydroxide: the liquid does not exhibit a bluish color.
Lead: Add 5 mL of 2 N sulfuric acid: the liquid does not
Bismuth Subgallate become cloudy.
Silver: Add hydrochloric acid, dropwise: no precipitate is
(Comment on this Monograph)id=m9770=Bismuth formed that is insoluble in a slight excess of hydrochloric
Subgallate=B-Monos.pdf) acid.
LIMIT OF ALKALIES AND ALKALINE EARTHS
Sample solution: Boil 1.0 g with 20 mL of a mixture of
equal volumes of 6 N acetic acid and water, cool, and
filter.
Analysis: Precipitate the bismuth from the filtrate by the
addition of hydrogen sulfide, boil the mixture, and filter.
Add 5 drops of sulfuric acid to the filtrate, evaporate to
C7H5BiO6 394.09 dryness, and ignite to constant weight.
Gallic acid bismuth basic salt [99-26-3]. Acceptance criteria: The weight of the residue does not
exceed 5 mg (0.5%).
DEFINITION Organic Impurities
Bismuth Subgallate is a basic salt which, when dried at 105 for FREE GALLIC ACID
3 h, contains the equivalent of NLT 52.0% and NMT 57.0% of Analysis: Shake 1.0 g with 20 mL of alcohol for 1 min,
Bi2O3. filter and evaporate the filtrate to dryness on a steam bath,
and dry the residue at 105 for 1 h.
IDENTIFICATION Acceptance criteria: The weight of the residue does not
A. IDENTIFICATION TESTSGENERAL, Bismuth 191: Meets the exceed 5 mg (0.5%).
requirements
Sample: When heated to redness, it at first chars, leaving SPECIFIC TESTS
finally a yellow residue. LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT
B. PROCEDURE 7.0% of its weight.
Analysis: Agitate thoroughly 100 mg with an excess of
hydrogen sulfide TS, filter, and boil the filtrate to expel the ADDITIONAL REQUIREMENTS
dissolved gas. Cool, and add 1 drop of ferric chloride TS. PACKAGING AND STORAGE: Preserve in tight, light-resistant
Acceptance criteria: A purplish blue mixture is produced. containers.
ASSAY
PROCEDURE
Sample solution: Dry 1 g of Bismuth Subgallate at 105 for Bismuth Subnitrate
3 h, then weigh accurately and ignite in a porcelain crucible. (Comment on this Monograph)id=m9780=Bismuth
Allow it to cool and add nitric acid to the residue, dropwise, Subnitrate=B-Monos.pdf)
warming until complete solution has been effected. Bi5O(OH)9(NO3)4 1461.99
Analysis: Evaporate Sample solution to dryness and carefully Bismuth hydroxide nitrate oxide Bi5O(OH)9(NO3)4 [1304-85-4].
ignite the residue to constant weight. From the weight of
the residue so obtained, determine the percentage of Bi2O3 DEFINITION
in the portion of Bismuth Subgallate taken. Bismuth Subnitrate is a basic salt that contains the equivalent of
Acceptance criteria: 52.0%57.0% NLT 79.0% of bismuth trioxide (Bi2O3), calculated on the dried
basis.
IMPURITIES
Inorganic Impurities IDENTIFICATION
ARSENIC 211 IDENTIFICATION TESTSGENERAL, Bismuth 191 and Nitrate
Sample: Triturate 400 mg with an equal weight of calcium 191: Meets the requirements
hydroxide, and ignite. Dissolve the residue in 5 mL of 3 N
hydrochloric acid. ASSAY
Acceptance criteria: The solution, without further PROCEDURE
treatment, meets the requirements (NMT 7.5 ppm). Sample solution: 400 mg of Bismuth Subnitrate to a 250-
LIMIT OF NITRATE mL beaker. Add 5 mL of water, then add 2 mL of nitric acid,
Sample: Mix 100 mg with 5 mL of 2 N sulfuric acid and 5 and warm, if necessary, to effect solution. Dilute to 100 mL,
mL of ferrous sulfate TS, filter the mixture, and carefully and add 0.3 mL of xylenol orange TS.
superimpose the filtrate, without mixing, on 5 mL of
sulfuric acid, in a test tube.
Analysis: Titrate with 0.05 M edetate disodium VS to a ADDITIONAL REQUIREMENTS

yellow endpoint. Each mL of 0.05 M edetate disodium is PACKAGING AND STORAGE: Preserve in well-closed containers.
equivalent to 11.65 mg of Bi2O3.
Acceptance criteria: NLT 79.0% of Bi2O3, calculated on the
dried basis.
Bismuth Subsalicylate
IMPURITIES (Comment on this Monograph)id=m9785=Bismuth
Inorganic Impurities Subsalicylate=B-Monos.pdf)
CARBONATE: Add 3 g to 3 mL of warm nitric acid: no
effervescence occurs. Pour the solution into 100 mL of C7H5BiO4 362.09
water: a white precipitate forms. Filter, evaporate the filtrate (2-Hydroxybenzoato-O1)-oxobismuth;
on a steam bath to 30 mL, again filter the liquid, divide the 2-Hydroxybenzoic acid bismuth (3+) salt, basic [14882-18-9].
latter filtrate into portions of 5 mL each, and use these DEFINITION
several portions in the tests for Chloride, Sulfate, Copper, Bismuth Subsalicylate is a basic salt that when dried at 105 for
Lead, and Silver. 3 h contains NLT 56.0% and NMT 59.4% of bismuth (Bi) and
CHLORIDE AND SULFATE, Chloride 221: A 10-mL portion of NLT 36.5% and NMT 39.3% of total salicylates.
the test liquid retained in the test for Carbonate shows no
more chloride than corresponds to 0.50 mL of 0.020 N IDENTIFICATION
hydrochloric acid (0.035%). A. INFRARED ABSORPTION 197M
CHLORIDE AND SULFATE, Sulfate 221: To a 5-mL portion of B. IDENTIFICATION TESTSGENERAL, Bismuth 191: Meets the
the test liquid retained in the test for Carbonate, add 5 requirements
drops of barium nitrate TS.
Acceptance criteria: No turbidity is produced ASSAY
immediately. BISMUTH
ARSENIC, Method I 211: Mix 375 mg with 5 mL of water, Sample solution: 300 mg of Bismuth Subsalicylate,
cautiously add 2 mL of sulfuric acid, and heat the mixture previously dried at 105 for 3 h in a porcelain crucible
until fumes of sulfur trioxide are copiously evolved. Cool, Ignite, allow it to cool, and add about 2 mL of nitric acid to
cautiously add 10 mL of water, and again evaporate to the residue, dropwise, warming until complete solution has
strong fuming, repeating, if necessary, to remove any trace been effected. Add about 60 mL of water and 0.3 mL of
of nitric acid. xylenol orange TS.
Acceptance criteria: NMT 8 ppm Analysis: Titrate with 0.05 M edetate disodium VS to a
COPPER: To a 5-mL portion of the test liquid retained in the yellow endpoint. Each mL of 0.05 M edetate disodium is
test for Carbonate, add a slight excess of 6 N ammonium equivalent to 10.45 mg of Bi.
hydroxide. Acceptance criteria: 56.0%59.4%
Acceptance criteria: The liquid does not exhibit a bluish TOTAL SALICYLATES
color. Ferric ammonium sulfate solution: Ferric ammonium
LEAD: Mix a 5-mL portion of the test liquid retained in the sulfate TS, 1 N hydrochloric acid, and water (4:1:15)
test for Carbonate with an equal volume of 2 N sulfuric acid. Standard stock solution: 0.2 mg/mL of USP Salicylic Acid
Acceptance criteria: The liquid does not become cloudy. RS in water
SILVER: To a 5-mL portion of the test liquid retained in the Standard solution: 0.05 mg/mL salicylic acid in water
test for Carbonate, add hydrochloric acid, dropwise. [NOTEAdjust with 0.5 N sodium hydroxide or 1 N
Acceptance criteria: No precipitate is formed that is hydrochloric acid to a pH of 4.5, prior to final dilution.]
insoluble in a slight excess of hydrochloric acid, but that is Sample solution: 52 mg of Bismuth Subsalicylate,
soluble in 6 N ammonium hydroxide. previously dried at 105 for 3 h, in a 200-mL volumetric
LIMIT OF ALKALIES AND ALKALINE EARTHS: Boil 1.0 g with 20 flask
mL of a mixture of equal volumes of 6 N acetic acid and Add 10 mL of 0.5 N sodium hydroxide, heat on a steam
water, cool, and filter. Add 2 mL of 3 N hydrochloric acid, bath for 15 min, allow to cool, and dilute with water to
precipitate the bismuth by the addition of hydrogen sulfide, volume. Centrifuge about 70 mL of this solution, then
boil the mixture, and filter it. Add 5 drops of sulfuric acid to transfer 50.0 mL of the clear supernatant to a beaker. Add
the filtrate, evaporate to dryness, and ignite to constant about 40 mL of water, and adjust with 0.5 N sodium
weight. hydroxide or 1 N hydrochloric acid to a pH of 4.5. Transfer
Acceptance criteria: The weight of the residue does not this solution to a 100-mL volumetric flask with the aid of
exceed 5 mg (0.5%). water, and dilute with water to volume.
LIMIT OF AMMONIUM SALTS: Boil about 100 mg with 5 mL of Blank: Water, adjusted with 0.5 N sodium hydroxide or 1 N
1 N sodium hydroxide. hydrochloric acid to a pH of 4.5
Acceptance criteria: The vapor does not turn moistened Analysis
red litmus paper blue. Samples: Standard solution, Sample solution, and Blank
Prepare the following solutions in separate 50-mL conical
SPECIFIC TESTS flasks.
3.0% of its weight.
Reacted standard solution: To 25.0 mL of Standard Scattering 851) equipped with copper, lead, and silver
solution, add 1.0 mL of Ferric ammonium sulfate solution. hollow-cathode lamps and an oxidizing flame.
Reacted sample solution: To 25.0 mL of Sample solution, Acceptance criteria: NMT 10 ppm
add 1.0 mL of Ferric ammonium sulfate solution. LIMIT OF SOLUBLE BISMUTH
Reacted blank solution: To 25.0 mL of Blank, add 1.0 mL Standard solution: 242.0 mg of bismuth nitrate
of Ferric ammonium sulfate solution. pentahydrate in a 100-mL volumetric flask
Unreacted standard solution: To 25.0 mL of the Standard Add 3 mL of 1.5 M nitric acid, swirl to dissolve, and dilute
solution, add 1.0 mL of 0.05 N hydrochloric acid. with water to volume. Transfer 1.0 mL of this solution to
Unreacted sample solution: To 25.0 mL of the Sample a 500-mL volumetric flask, add 250 mL of 1.5 M nitric
solution, add 1.0 mL of 0.05 N hydrochloric acid. acid, dilute with water to volume, and mix. This solution
Unreacted blank solution: To 25.0 mL of Blank, add 1.0 contains 2 g/mL of bismuth.
mL of 0.05 N hydrochloric acid. [NOTEThe concentration of bismuth in this solution
Concomitantly determine the absorbances of these six may be modified by using a lesser dilution or by further
solutions at the wavelength of maximum absorption at dilution to bring the absorption response within the
about 525 nm, using water to zero the working range of the atomic absorption
spectrophotometer. spectrophotometer.]
Calculate the percentage of total salicylates in the portion Sample solution: 5.0 g of Bismuth Subsalicylate in 100
of C7H5BiO4 taken: mL of water
Stir the suspension thus obtained for 2 h at 2023. Filter
Result = [(AUR AUU B)/(ASR ASU B)] (CS/W) 10,000 through filter paper. Filter the filtrate thus obtained
through a filter having a porosity of 0.1 m or less. To
AUR = absorbance of the Reacted sample solution 10.0 mL of the filtrate add 0.1 mL of nitric acid.
AUU = absorbance of the Unreacted sample solution [NOTEThe concentrate of Bismuth Subsalicyclate may
B = difference in the absorption of the Reacted blank be modified by using the same proportions used for
solution and the absorption of the Unreacted modifying the Standard Solution, by using a different
blank solution quantity, or by further dilution.]
ASR = absorbance of the Reacted standard solution Analysis
ASU = absorbance of the Unreacted standard solution Samples: Standard solution and Sample solution
CS = concentration of USP Salicylic Acid RS in the Concomitantly determine the absorbances of the
Standard stock solution (mg/mL) Standard solution and the Sample solution at the
W = weight, in mg, of Bismuth Subsalicylate taken emission line of 223.06 nm for bismuth with an atomic
Acceptance criteria: 36.5%39.3% absorption spectrophotometer (see Spectrophotometry
and Light-Scattering 851) equipped with a bismuth
IMPURITIES hollow-cathode lamp and an oxidizing flame.
Inorganic Impurities Acceptance criteria: NMT 40 ppm
ARSENIC, Method I 211: Triturate 300 mg of bismuth LIMIT OF NITRATE
subsalicylate with 300 mg of calcium hydroxide, and ignite. Sample solution: To 0.1 g add 10 mL of water, and
Dissolve the residue in 5 mL of 3 N hydrochloric acid. carefully add 20 mL of sulfuric acid.
Acceptance criteria: NMT 10 ppm Standard solution: Add to 0.1 g of salicylic acid, 6 mL of
LIMIT OF COPPER, LEAD, AND SILVER water, 4.0 mL of a solution containing 100 g of nitrate
Standard solution: 3.0 mL each of solutions containing per mL, and 20 mL of sulfuric acid.
1000 g/mL of copper, lead, and silver, respectively, to a Acceptance criteria: The Sample solution should not be
2000-mL volumetric flask more yellow than the Standard solution (0.4%), prepared
Dilute with 1 M nitric acid to volume. [NOTEThe concomitantly.
concentrations of copper, lead, and silver may be Organic Impurities
modified by using different volumes or concentrations to LIMIT OF FREE SALICYLIC ACID
bring the absorption response within the working range Mobile phase: Methanol and 0.06 M acetic acid (11:9)
of the atomic absorption spectrophotometer.] Diluent: Acetonitrile and water (1:1)
Sample solution: Ignite 3 g of sample in a porcelain Standard solution: 0.02 mg/mL of USP Salicylic Acid RS in
crucible, cool, cautiously add 6 M nitric acid to dissolve Diluent
the residue, and evaporate on a steam bath. Ignite the Sample solution: 260 mg of Bismuth Subsalicylate in a
residue, cool, and transfer the residue to a tared conical glass centrifuge tube
flask. Wash the flask with about 5 mL of 6 M nitric acid, Add about 12 mL of acetonitrile, shake by mechanical
adding the wash to the conical flask. Dissolve the residue means for 20 min, and centrifuge. Decant the supernatant
with the aid of heat, and add water to obtain a solution into a suitable container. Repeat the acetonitrile addition,
weighing 20.0 g. [NOTEThe concentrate of Bismuth shaking, centrifuging, and decanting, combining the
Subsalicyclate may be modified by using the same decanted liquid with the first decantate. Pass the
proportions used for modifying the Standard solution, by combined liquid through a filter having a 0.5-m or finer
using a different quantity, or by further dilution.] porosity, collecting the filtrate in a 50-mL volumetric flask.
Analysis Wash the container with 5 mL of acetonitrile, and filter
Samples: Standard solution and Sample solution the wash, collecting the filtrate in the volumetric flask.
Concomitantly determine the absorbances of the Standard Dilute with water to volume.
solution and the Sample solution at the emission lines of Chromatographic system
324.7 nm, 217 nm, and 328.1 nm for copper, lead, and (See Chromatography 621, System Suitability.)
silver, respectively, with an atomic absorption
spectrophotometer (see Spectrophotometry and Light-
Mode: LC Acceptance criteria: 56.0%59.4% of Bi

Detector: UV 300 nm TOTAL SALICYLATES
Column: 4.6-mm 30-cm; 5-m packing L1 Solution A: Ferric ammonium sulfate TS, 1 N hydrochloric
Guard column: 3.2-mm 1.5-cm; 5-m packing L1 acid, and water (4:1:15)
Flow rate: 1 mL/min Standard stock solution: 0.2 mg/mL of USP Salicylic Acid
Injection size: 20 L RS in water
System suitability Standard solution: 0.05 mg/mL salicylic acid in water,
Sample: Standard solution adjust with 0.5 N sodium hydroxide or 1 N hydrochloric
Suitability requirements acid to a pH of 4.5, prior to final dilution
Tailing factor: NMT 2.0 Reacted standard solution: To 25.0 mL of Standard
Relative standard deviation: NMT 2.0% solution, add 1.0 mL of Solution A.
Analysis Unreacted standard solution: To 25.0 mL of the Standard
Samples: Standard solution and Sample solution solution, add 1.0 mL of 0.05 N hydrochloric acid.
Calculate the percentage of free salicylic acid in C7H5BiO4: Sample solution: Equivalent to 52 mg of bismuth
subsalicylate from dried magma in a 200-mL volumetric
Result = (rU/rS) (CS/CU) 100 flask. Add 10 mL of 0.5 N sodium hydroxide, heat on a
steam bath for 15 min, allow to cool, and dilute with water
rU = peak area response of salicylic acid in the to volume. Centrifuge and then transfer 50.0 mL of the clear
Sample solution supernatant to a beaker. Add about 40 mL of water, and
rS = peak area response of the Standard solution adjust with 0.5 N sodium hydroxide or 1 N hydrochloric
CS = concentration of USP Salicylic Acid RS in the acid to a pH of 4.5. Transfer this solution to a 100-mL
Standard solution (mg/mL) volumetric flask with the aid of water, and dilute with water
CU = concentration of the Bismuth Subsalicylate in to volume.
the Sample solution (mg/mL) Reacted sample solution: To 25.0 mL of Sample solution,
Acceptance criteria: NMT 0.2% add 1.0 mL of Solution A.
Unreacted sample solution: To 25.0 mL of the Sample
SPECIFIC TESTS solution, add 1.0 mL of 0.05 N hydrochloric acid.
PH 791 Blank: Water, adjusted with 0.5 N sodium hydroxide or 1 N
Analysis: Place 10 g of Bismuth Subsalicylate in 90 mL of hydrochloric acid to a pH of 4.5
water, shake by mechanical means for 10 min, and filter. Reacted blank solution: To 25.0 mL of Blank, add 1.0 mL
Acceptance criteria: 2.75.0 of Solution A.
LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT Unreacted blank: To 25.0 mL of Blank, add 1.0 mL of 0.05
1.0% of its weight. N hydrochloric acid.
PACKAGING AND STORAGE: Preserve in tight, light-resistant Samples: Reacted standard solution, Unreacted standard
containers. solution, Reacted sample solution, Unreacted sample solution,
USP REFERENCE STANDARDS 11 Reacted blank solution, and Unreacted blank
USP Bismuth Subsalicylate RS Concomitantly determine the absorbances of the Samples
USP Salicylic Acid RS at the wavelength of maximum absorption at about 525
nm, using water to zero the spectrophotometer.
Calculate the percentage of total salicylates in the portion
of dried magma taken:
Bismuth Subsalicylate Magma
(Comment on this Monograph)id=m9786=Bismuth Result = [(AUR AUU B)/(ASR ASU B)] (CS/W) 10,000
Subsalicylate Magma=B-Monos.pdf) AUR = absorbance of the Reacted sample solution
DEFINITION AUU = absorbance of the Unreacted sample solution
Bismuth Subsalicylate Magma is a suspension of Bismuth B = difference in the absorption of the Reacted blank
Subsalicylate in water that contains NLT 90.0% and NMT solution and the absorption of the Unreacted
110.0% of the labeled amount of C7H5O4Bi. Bismuth blank
Subsalicylate is a basic salt that when dried at 105 for 3 h ASR = absorbance of the Reacted standard solution
contains NLT 56.0% and NMT 59.4% bismuth and NLT ASU = absorbance of the Unreacted standard solution
36.5% and NMT 39.3% of total salicylates. CS = concentration of USP Salicylic Acid RS in the
[NOTEDry at 105 for 3 h to determine the solids content Standard stock solution (mg/mL)
and, after determining the solids content, perform all tests on W = weight, in mg, of Bismuth Subsalicylate taken for
a portion of the dried magma.] the Sample solution
Acceptance criteria: 36.5%39.3% of total salicylates
IDENTIFICATION
A. INFRARED ABSORPTION 197M IMPURITIES
B. IDENTIFICATION TESTSGENERAL, Bismuth 191: Meets the Inorganic Impurities
requirements LIMIT OF COPPER, LEAD, AND SILVER
Standard solution: 1.5 g/mL of copper, 1.5 g/mL of
ASSAY lead, and 1.5 g/mL of silver, in 1 M nitric acid
BISMUTH [NOTEThe concentrations of copper, lead, and silver may
Sample solution: Ignite an equivalent to 300 mg of be modified by using different volumes or concentrations
bismuth subsalicylate from dried magma. Allow it to cool, to bring the absorption response within the working
and add about 2 mL of nitric acid to the residue, dropwise, range of the atomic absorption spectrophotometer.]
warming until complete solution has been effected. Add Sample solution: Ignite 3 g of sample in a porcelain
about 60 mL of water and 0.3 mL of xylenol orange TS. crucible, cool, and cautiously add 6 M nitric acid to
Analysis: Titrate the Sample solution with 0.05 M edetate dissolve the residue, and evaporate on a steam bath. Ignite
disodium VS to a yellow endpoint. Each mL of 0.05 M the residue, cool, and transfer the residue to a tared conical
edetate disodium is equivalent to 10.45 mg of bismuth (Bi). flask, and wash the flask with about 5 mL of 6 M nitric
acid, adding the wash to the conical flask. Dissolve the decanting, combining the decanted liquid with the first
residue with the aid of heat, and add water to obtain a decantate. Pass the combined liquid through a filter having
solution weighing 20.0 g. a 0.5-m or finer porosity, collecting the filtrate in a 50-mL
[NOTEThe concentrate of bismuth subsalicyclate may be volumetric flask. Wash the container with 5 mL of
modified by using the same proportions used for acetonitrile, and filter the wash, collecting the filtrate in the
modifying the Standard solution, by using a different volumetric flask. Dilute with water to volume.
quantity, or by further dilution.] Chromatographic system
Samples: Standard solution and Sample solution Mode: LC
Concomitantly determine the absorbances of the Standard Detector: UV 300 nm
solution and the Sample solution at the emission lines of Guard column: 3.2-mm 1.5-cm; 5-m packing L1
324.7 nm, 217 nm, and 328.1 nm, for copper, lead, and Analytical column: 4.6-mm 30-cm; 5-m packing L1
silver, respectively, with an atomic absorption Flow rate: 1 mL/min
spectrophotometer (see Spectrophotometry and Light- Injection size: 20 L
scattering 851) equipped with copper, lead, and silver System suitability
hollow-cathode lamps and an oxidizing flame. Sample: Standard solution
Acceptance criteria: The absorbances of the Sample Suitability requirements
solution do not exceed those of the Standard solution for Tailing factor: NMT 2.0
each element (NMT 10 ppm). Relative standard deviation: NMT 2.0%
LIMIT OF SOLUBLE BISMUTH Analysis
Standard solution: 242.0 mg of bismuth nitrate Samples: Standard solution and Sample solution
pentahydrate in a 100-mL volumetric flask, add 3 mL of Calculate the percentage of free salicylic acid in the
1.5 M nitric acid and swirl to dissolve, dilute with water to Bismuth Subsalicylate taken:
volume, and mix. Transfer 1.0 mL of this solution to a 500-
mL volumetric flask, add 250 mL of 1.5 M nitric acid, dilute Result = (rU/rS) (CS/CU) 100
with water to volume, and mix (2 g/mL of bismuth).
[NOTEThe concentration of bismuth in this solution may rU = peak area of salicylic acid in the Sample solution
be modified by using a lesser dilution or by further rS = peak area of the Standard solution
dilution to bring the absorption response within the CS = concentration of USP Salicylic Acid RS in the
working range of the atomic absorption Standard solution (mg/mL)
spectrophotometer.] CU = concentration of the bismuth subsalicylate in
Sample solution: 5.0 g of bismuth subsalicylate in 100 mL the Sample solution (mg/mL)
of water, and stir the suspension thus obtained for 2 h at Acceptance criteria: NMT 0.2%
2023. Pass through filter paper. Pass the filtrate thus
obtained through a filter having a porosity of 0.1 m or ADDITIONAL REQUIREMENTS
less. To 10.0 mL of the filtrate add 0.1 mL of nitric acid. PACKAGING AND STORAGE: Preserve in tight, light-resistant
[NOTEThe concentrate of bismuth subsalicyclate may be containers.
modified by using the same proportions used for LABELING: The label states that this article is not intended for
modifying the Standard solution, by using a different direct administration to humans or animals.
quantity, or by further dilution.] USP REFERENCE STANDARDS 11
Analysis USP Bismuth Subsalicylate RS
Samples: Standard solution and Sample solution USP Salicylic Acid RS
Concomitantly determine the absorbances of the Standard
solution and the Sample solution at the emission line of
223.06 nm for bismuth with an atomic absorption Bismuth Subsalicylate Oral Suspension
spectrophotometer (see Spectrophotometry and Light-
scattering 851) equipped with a bismuth hollow- (Comment on this Monograph)id=m9787=Bismuth
cathode lamp and an oxidizing flame. Subsalicylate Oral Suspension=B-Monos.pdf)
Acceptance criteria: The absorbances of the Sample DEFINITION
solution do not exceed those of the Standard solution (NMT Bismuth Subsalicylate Oral Suspension is a suspension that
40 ppm). contains NLT 90.0% and NMT 110.0% of the labeled amount
LIMIT OF NITRATE of C7H5BiO4. It may contain one or more suitable buffers,
Sample solution: To 0.1 g add 10 mL of water, carefully coloring agents, flavors, preservatives, stabilizers, sweeteners,
add 20 mL of sulfuric acid, and mix. and suspending agents.
Standard solution: Add to 0.1 g of salicylic acid, 6 mL of
water, 4.0 mL of a solution containing 100 g of nitrate IDENTIFICATION
(NO3) per mL, and 20 mL of sulfuric acid A. IDENTIFICATION TESTSGENERAL, Bismuth 191
Acceptance criteria: The Sample solution should not be B. IDENTIFICATION TESTSGENERAL, Salicylate 191: After
more yellow than the Standard solution (0.4%), prepared acidifying with nitric acid
concomitantly.
LIMIT OF FREE SALICYLIC ACID PROCEDURE
Mobile phase: Methanol and 0.06 M acetic acid (11:9) Standard stock solution: 500 mg of bismuth metal, in a
Diluent: Acetonitrile and water (1:1) 200-mL volumetric flask, dissolve in 12 mL of nitric acid,
Standard solution: 0.02 mg/mL of USP Salicylic Acid RS in and dilute with 0.01 N nitric acid to volume
Diluent Standard solution: 50 g/mL from Standard stock solution
Sample solution: 260 mg of bismuth subsalicylate, in a in 1 N nitric acid
glass centrifuge tube, add about 12 mL of acetonitrile, Sample solution: 10 g of Oral Suspension, previously well-
shake by mechanical means for 20 min, and centrifuge. shaken in its original container to ensure homogeneity, in a
Decant the supernatant into a suitable container. Repeat 200-mL volumetric flask. Add about 100 mL of 1 N nitric
the acetonitrile addition, shaking, centrifuging, and acid, mix, and dilute with 1 N nitric acid to volume. Mix
USP 32 Official Monographs / Bisoctrizole 99
well without shaking, transfer 10.0 mL of this mixture to a Sample stock solution: Equivalent to 90 mg of bismuth
100-mL volumetric flask, and dilute with 1 N nitric acid to subsalicylate from powdered Tablets, in a 200-mL volumetric
volume. Centrifuge about 20 mL at 4500 rpm for at least 10 flask, add 150 mL of 1 N nitric acid, and sonicate for 2 min.
min. Dilute with 1 N nitric acid to volume.
Spectrometric conditions Sample solution: Sample stock solution and 1 N nitric acid
Mode: UV-Vis (1:4). Centrifuge a portion at 4500 rpm for at least 10 min.
Analytical wavelength: 463 nm Spectrometric conditions
Cell: 1 cm Mode: Spectrophotometry
Analysis Analytical wavelength: 463 nm
Samples: Standard solution and Sample solution Cell: 1 cm
Transfer an accurately measured volume of the Sample Analysis
solution that contains 0.9 mg of bismuth subsalicylate and Samples: Standard solution and Sample solution
10 mL of the Standard solution to separate 50-mL Analysis: Transfer 10.0 mL of the Sample solution and the
volumetric flasks. Add 10.0 mL of 10% ascorbic acid Standard solution to separate 50.0-mL volumetric flasks.
solution and 25.0 mL of 20% potassium iodide solution Add 10.0 mL of 10% ascorbic acid solution and 25.0 mL of
into each volumetric flask, dilute with water to volume, 20% potassium iodide solution into each volumetric flask,
and mix well. Concomitantly determine the absorbances and dilute with 1 N nitric acid to volume. Concomitantly
of both solution, using the reagent blank to set the determine the absorbance of the solutions at the
spectrophotometer. wavelength of maximum absorbance at 463 nm with a
Calculate the percentage of bismuth subsalicylate suitable spectrophotometer using the combined reagent
(C7H5BiO4) in the portion of Oral Suspension: solutions as the blank.
Calculate the percentage of C7H5BiO4 in the portion of
Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 Tablets:
AU = absorbance of the Sample solution Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100
CS = concentration of bismuth in the Standard AU = absorbance of the Sample solution
solution (g/mL) AS = absorbance of the Standard solution
CU = nominal concentration of the Sample solution CS = concentration of bismuth in the Standard
(g/mL) solution (g/mL)
Mr1 = molecular weight of bismuth subsalicylate, CU = nominal concentration of bismuth in the Sample
362.09 solution (g/mL)
Mr2 = molecular weight of bismuth, 208.98 Mr1 = molecular weight of bismuth subsalicylate,
Acceptance criteria: 90.0%110.0% 362.09
Mr2 = molecular weight of bismuth, 208.98
MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED
MICROORGANISMS 62 PERFORMANCE TESTS
Total aerobic microbial count: NMT 100 cfu/g DISINTEGRATION 701
Total combined yeasts and molds count: NMT 50 cfu/g Time: 10 min
It meets the requirements of the tests for absence of [NOTEThis test does not apply for Tablets labeled as
Escherichia coli. chewable.]
PH 791: 3.05.0
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Avoid
PACKAGING AND STORAGE: Preserve in tight containers. excessive heat (over 40).
Protect from freezing. Avoid excessive heat (over 40). LABELING: Label chewable Tablets to indicate that they are
to be chewed before swallowing.
Bismuth Subsalicylate Tablets

(Comment on this Monograph)id=m9788=Bismuth Bisoctrizole
Subsalicylate Tablets=B-Monos.pdf) (Comment on this Monograph)id=m1129=Bisoctrizole=B-
Monos.pdf)
DEFINITION
Bismuth Subsalicylate Tablets contain NLT 90.0% and NMT
110.0% of the labeled amount of bismuth subsalicylate
(C7H5BiO4).
IDENTIFICATION
A. IDENTIFICATION TESTSGENERAL, Bismuth 191
B. IDENTIFICATION TESTSGENERAL, Salicylate 191: After
acidifying with nitric acid, it meets the requirements of the C41H50N6O2 658.90
test with ferric chloride TS. Phenol, 2,2-methylenebis[6-(2H-benzotriazol-2-yl)-4-(1,1,3,3-
tetramethylbutyl)]-;
ASSAY 2,2-Methylenebis[6-(2H-benzotriazol-2-yl)-4-(1,1,3,3-
PROCEDURE tetramethylbutyl)phenol] [103597-45-1].
Standard stock solution: 500 mg of bismuth, in a 200-mL
volumetric flask, dissolve in 12 mL of nitric acid, and dilute DEFINITION
with 0.01 N nitric acid to volume Bisoctrizole contains NLT 96.0% and NMT 102.0% of
Standard solution: 50 g/mL of bismuth from Standard C41H50N6O2, calculated on the as-is basis.
stock solution and 1 N nitric acid (1:49)
100 Bisoctrizole / Official Monographs USP 32

B. The retention time of the major peak of the Sample IMPURITIES
solution corresponds to that of the Standard solution, as Inorganic Impurities
obtained in the Assay. HEAVY METALS, Method II 231: NMT 20 ppm
Organic Impurities
ASSAY PROCEDURE 1
PROCEDURE Diluent, Solution A, Solution B, Mobile phase, Standard
Diluent: Tetrahydrofuran and a 0.2% (w/v) aqueous solution, Sample solution, Chromatographic system, and
solution of 1-pentane sulfonic acid sodium salt (3:2) System suitability: Proceed as directed in the Assay.
Solution A: 0.4 g of 1-pentane sulfonic acid sodium salt, Analysis
800 mL of methanol, 200 mL of water, and 0.5 mL of Samples: Standard solution and Sample solution
phosphoric acid Calculate the percentage of each impurity in the portion
Solution B: 0.4 g of 1-pentane sulfonic acid sodium salt, of Bisoctrizole taken:
1000 mL of methanol, and 0.5 mL of phosphoric acid
Mobile phase: See gradient table below. Result = (ru/rT) 100
ru = peak response for each individual impurity
rT = sum of the responses of all the peaks
(min) (%) (%)
Acceptance criteria
0 70 30 Individual impurities: NMT 0.1% of any individual
1 70 30 impurity, excluding bisoctrizole related compound A and
the bisoctrizole isomer
11 3 97
Total impurities: NMT 4.0% of total impurities, including
27 3 97 bisoctrizole related compound A, and the bisoctrizole
28 70 30 isomer, determined in the test for Limit of bisoctrizole
related compound A and the bisoctrizole isomer
System suitability solution: 0.8 mg/mL of bisoctrizole from PROCEDURE 2: LIMIT OF BISOCTRIZOLE RELATED COMPOUND A
USP Bisoctrizole Resolution Mixture RS dissolved in AND THE BISOCTRIZOLE ISOMER
tetrahydrofuran, and dilute with Diluent Diluent, Solution A, Solution B, Mobile phase, Sample
Standard solution: 80 mg of USP Bisoctrizole RS to a 100- solution, and Chromatographic system: Proceed as
mL volumetric flask. Dissolve in 60 mL of tetrahydrofuran, directed in the Assay.
and dilute with Diluent to volume. Standard stock solution A: 0.65 mg/mL of USP
Sample solution: 80 mg of Bisoctrizole to a 100-mL Bisoctrizole RS in tetrahydrofuran
volumetric flask. Dissolve in 60 mL of tetrahydrofuran, and Standard stock solution B: 0.40 mg/mL of USP
dilute with Diluent to volume. Bisoctrizole Related Compound A RS in tetrahydrofuran
Chromatographic system Standard solution: Standard stock solution A, Standard
(See Chromatography 621, System Suitability.) stock solution B, tetrahydrofuran, and Diluent (5:1:60:34)
Mode: LC System suitability
Detector: UV 346 nm Sample: Standard solution
Column: 3.0-mm 25-cm; packing L1 [NOTEThe relative retention times for bisoctrizole
Temperature: 40 related compound A and the bisoctrizole isomer are
Flow rate: 0.8 mL/min about 0.42 and about 1.1, respectively.]
Injection size: 10 L Suitability requirements
System suitability Resolution: NLT 1.5 between bisoctrizole and the
Sample: System suitability solution and Standard solution bisoctrizole isomer
[NOTEThe relative retention times for bisoctrizole and the Analysis
bisoctrizole isomer are about 1.0 and 1.1, respectively.] Samples: Standard solution and Sample solution
Suitability requirements Calculate the percentage of bisoctrizole related compound
Resolution: NLT 1.5 between bisoctrizole and the A taken:
bisoctrizole isomer, Standard solution
Relative standard deviation: NMT 2.0%, Standard Result = (rU/rS) (CS/CU) 100
solution
Analysis rU = peak response for the bisoctrizole related
Samples: Standard solution and Sample solution compound A in the Sample solution
Calculate the percentage of C41H50N6O2 in the portion of rS = peak response for bisoctrizole related compound
Bisoctrizole taken: A in the Standard solution
CS = concentration of USP Bisoctrizole Related
Result = (rU/rS) (CS/CU) 100 Compound A RS in the Standard solution
(mg/mL)
rU = peak response from the Sample solution CU = nominal concentration of bisoctrizole related
rS = peak response from the Standard solution compound A in the Sample solution (mg/mL)
CS = concentration of USP Bisoctrizole RS in the Acceptance criteria: NMT 0.5% of the bisoctrizole related
Standard solution (mg/mL) compound A
CU = nominal concentration of bisoctrizole in the
USP 32 Official Monographs / Bisoprolol 101
Calculate the percentage of bisoctrizole isomer taken: Sample solution: 1 mg/mL of Bisoprolol Fumarate in
Diluent
Result = (rU/rS) (CS/CU) 100 Chromatographic system
rU = peak response for the bisoctrizole isomer in the Mode: LC
Sample solution Detector: UV 273 nm
rS = peak response for Bisoctrizole in the Standard Column: 4.6-mm 12.5-cm; packing L7
solution Flow rate: 1 mL/min
CS = concentration of USP Bisoctrizole RS in the Injection size: 10 L
Standard solution (mg/mL) System suitability
CU = nominal concentration of bisoctrizole in the Samples: System suitability solution and Standard solution
Sample solution (mg/mL) Suitability requirements
Acceptance criteria: NMT 4.0% of the bisoctrizole isomer Resolution: NLT 7.0 between bisoprolol and propranolol
in the System suitability solution
ADDITIONAL REQUIREMENTS Tailing factor: NMT 2.0 in the Standard solution
PACKAGING AND STORAGE: Preserve in well-closed containers, Relative standard deviation: NMT 2.0% in the Standard
and store at controlled room temperature. solution
USP Bisoctrizole RS Samples: Standard solution and Sample solution
USP Bisoctrizole Related Compound A RS Calculate the percentage of (C18H31NO4)2 C4H4O4 in the
USP Bisoctrizole Resolution Mixture RS portion of Bisoprolol Fumarate taken:
Bisoprolol Fumarate rU = peak response from the Sample solution
(Comment on this Monograph)id=m9792=Bisoprolol rS = peak response from the Standard solution
Fumarate=B-Monos.pdf) CS = concentration of USP Bisoprolol Fumarate RS in
CU = concentration of bisoprolol fumarate in the
IMPURITIES
(C18H31NO4)2 C4H4O4 766.96 HEAVY METALS, Method I 231: NMT 20 ppm
2-Propanol, 1-[4-[[2-(1-methylethoxy)ethoxy]methyl] Organic Impurities
phenoxy]-3-[(1-methylethyl)amino]-, ()-, (E)-2-butenedioate PROCEDURE
(2:1) (salt); Diluent, Mobile phase, System suitability solution,
()-1-[[-(2-Isoproproxyethoxy)-p-tolyl]oxy]-3- Standard solution, Sample solution, and
(isopropylamino)-2-propanol fumarate (2:1) (salt) Chromatographic system: Proceed as directed in the
[104344-23-2]. Assay.
System suitability
DEFINITION Samples: System suitability solution and Standard solution
Bisoprolol Fumarate contains NLT 97.5% and NMT 102.0% of Suitability requirements
(C18H31NO4)2 C4H4O4, calculated on the anhydrous basis. Resolution: NLT 7.0 between bisoprolol and propranolol
IDENTIFICATION in the System suitability solution
A. INFRARED ABSORPTION 197K Tailing factor: NMT 2.0 in the Standard solution
B. LIQUID CHROMATOGRAPHY: The retention time of the Relative standard deviation: NMT 2.0% in the
major peak of the Sample solution corresponds to that of the Standard solution
Standard solution, as obtained in the Assay. Analysis
ASSAY Calculate the percentage of total impurities in the portion
PROCEDURE of Bisoprolol Fumarate taken:
Diluent: Acetonitrile and water (7:13)
Mobile phase: To a 1-L portion of Diluent add 5 mL of Result = (ru/rT) 100
heptafluorobutyric acid, 5 mL of diethylamine, and 2.5 mL
of formic acid. Ru = sum of areas for all the peaks, excluding the
System suitability solution: 0.5 mg/mL of propranolol fumaric acid and bisoprolol peaks
hydrochloride and 1 mg/mL of Bisoprolol Fumarate in RT = sum of the areas of all the peaks in the
Diluent chromatogram
Standard solution: 1 mg/mL of USP Bisoprolol Fumarate RS
in Diluent
102 Bisoprolol / Official Monographs USP 32

Total impurities: NMT 0.5% (See Chromatography 621, System Suitability.)
Mode: LC
OPTICAL ROTATION, Specific Rotation 781 Column: 4.6-mm 12.5-cm; packing L7
Sample solution: 10 mg/mL, in methanol Flow rate: 1 mL/min
Acceptance criteria: 2 to +2 Injection size: 10 L
WATER DETERMINATION, Method I 921: NMT 0.5% System suitability
CONTENT OF FUMARIC ACID Samples: System suitability solution and Standard solution
Sample solution: 500 mg of Bisoprolol Fumarate in 70 mL Suitability requirements
of dehydrated alcohol Resolution: NLT 7.0 between bisoprolol and propranolol,
Analysis: Add 8.0 mL of 0.1 N tetrabutylammonium System suitability solution
hydroxide VS, and stir for 2 min. Titrate with 0.1 N Tailing factor: NMT 2.0 in the Standard solution
tetrabutylammonium hydroxide VS, using a glass-calomel Relative standard deviation: NMT 2.0%, Standard
electrode system. Perform a blank determination, and make solution
any necessary correction (see Titrimetry 541). Each mL of Analysis
0.1 N tetrabutylammonium hydroxide is equivalent to 5.804 Samples: Standard solution and Sample solution
mg of fumaric acid. Calculate the percentage of (C18H31NO4)2 C4H4O4 in the
Acceptance criteria: NLT 14.8% and NMT 15.4% of portion of Tablets taken:
fumaric acid is found, calculated on the anhydrous basis.
PACKAGING AND STORAGE: Preserve in tight, light-resistant rU = peak response from the Sample solution
containers. Store at controlled room temperature. rS = peak response from the Standard solution
USP REFERENCE STANDARDS 11 CS = concentration of USP Bisoprolol Fumarate RS in
USP Bisoprolol Fumarate RS the Standard solution (mg/mL)
CU = nominal concentration of bisoprolol fumarate in
the Sample solution (mg/mL)
Bisoprolol Fumarate Tablets Acceptance criteria: 90.0%105.0%
(Comment on this Monograph)id=m9794=Bisoprolol Fumarate PERFORMANCE TESTS
Tablets=B-Monos.pdf) DISSOLUTION 711
Test 1
DEFINITION Medium: Water; 900 mL
Bisoprolol Fumarate Tablets contain NLT 90.0% and NMT Apparatus 2: 75 rpm
105.0% of the labeled amount of bisoprolol fumarate Time: 20 min
[(C18H31NO4)2 C4H4O4]. Analysis: Determine the amount of (C18H31NO4)2 C4H4O4
dissolved by employing the following method.
IDENTIFICATION Diluent: Methanol, triethylamine, phosphoric acid, and
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 water (160:5:2.5:35)
Sample solution: Equivalent to 40 mg of bisoprolol Mobile phase: Methanol, triethylamine, and water
fumarate, from powdered Tablets (NLT 5), in a 50-mL flask. (34:1:50), adjust with phosphoric acid to a pH of 4.0 0.1.
Add about 20 mL of a mixture of dichloromethane and Standard stock solution: USP Bisoprolol Fumarate RS in
methanol (7:3), shake for 30 min, centrifuge, and use the water to obtain a solution having a known concentration of
clear solution. about twice the concentration of bisoprolol fumarate in the
Application volume: 20 L Sample solution
Developing solvent system: Dichloromethane, methanol, Standard solution: Standard stock solution and Diluent
and ammonia TS, stronger (70:10:0.8) (1:1)
Analysis Sample solution: Sample per Dissolution 711. Withdraw a
Sample: Sample solution portion of the solution under test, filter, and dilute with an
Proceed as directed in the chapter, except to develop the equal volume of Diluent.
chromatogram until the solvent front has moved about Chromatographic system
two-thirds of the length of the plate and to dry the plate in (See Chromatography 621, System Suitability.)
a current of cold air. Mode: LC
ASSAY Detector: UV 227 nm
PROCEDURE Column: 4.6-mm 33-cm; packing L7
Diluent: Acetonitrile and water (7:13) Flow rate: 1 mL/min
Mobile phase: 1-L portion of Diluent, add 5 mL of Injection size: 50 L
heptafluorobutyric acid, 5 mL of diethylamine, and 2.5 mL System suitability
of formic acid Sample: Standard solution
System suitability solution: 0.5 mg/mL of propranolol Suitability requirements
hydrochloride and 1 mg/mL of bisoprolol fumarate in Relative standard deviation: NMT 2.0%
Diluent Analysis
Standard solution: 1 mg/mL of USP Bisoprolol Fumarate RS Samples: Standard solution and Sample solution
in Diluent Calculate the percentage of (C18H31NO4)2 C4H4O4
Sample solution: Transfer an equivalent of 25 mg of dissolved.
bisoprolol fumarate, from powdered Tablets (NLT 20), to a Tolerances: NLT 80% (Q) of the labeled amount of
25-mL volumetric flask. Add 10 mL of Diluent, and sonicate (C18H31NO4)2 C4H4O4 is dissolved.
for 10 min. Cool, dilute with Diluent to volume, and mix. Test 2: If the product complies with this test, the labeling
Centrifuge for 20 min, and use the clear supernatant. indicates that it meets USP Dissolution Test 2.
USP 32 Official Monographs / Bisoprolol 103
Medium: 0.5 M sodium chloride; 900 mL Mobile phase: See the gradient table below
Apparatus 2: 75 rpm
Time: 20 min Time Solution A Solution B
Analysis: Proceed as directed in Performance Tests, Test 1, (min) (%) (%)
Analysis.
Tolerances: NLT 80% (Q) of the labeled amount of 0 100 0
(C18H31NO4)2 C4H4O4 is dissolved. 9.0 40 60
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements 9.1 100 0
ADDITIONAL REQUIREMENTS 12.0 100 0
containers, and store at controlled room temperature. System suitability solution: 40 g/mL of USP
LABELING: When more than one Dissolution test is given, the Chlorothiazide RS and 40 g/mL of USP Hydrochlorothiazide
labeling states the Dissolution test used only if Test 1 is not RS in Diluent
used. Standard solution: 100 g/mL of USP Bisoprolol Fumarate
USP REFERENCE STANDARDS 11 RS and 100 g/mL of USP Hydrochlorothiazide RS in Diluent.
USP Bisoprolol Fumarate RS Stir by mechanical means for 1 h.
Sample stock solution: Weigh 10 Tablets, and transfer to a
100-mL volumetric flask. Add about 50 mL of Diluent,
sonicate for 10 min, and cool. Dilute with Diluent to volume,
Bisoprolol Fumarate and stir by mechanical means for 1 h, and centrifuge.
Hydrochlorothiazide Tablets Sample solution A: 100 g/mL of bisoprolol fumarate from
(Comment on this Monograph)id=m9796=Bisoprolol Fumarate Sample stock solution in Diluent
and Hydrochlorothiazide Tablets=B-Monos.pdf) Sample solution B: 62.5 g/mL of hydrochlorothiazide
from Sample stock solution in Diluent
DEFINITION Chromatographic system
Bisoprolol Fumarate and Hydrochlorothiazide Tablets contain (See Chromatography 621, System Suitability.)
NLT 90.0% and NMT 110.0% of the labeled amounts of Mode: LC
bisoprolol fumarate [(C18H31NO4)2 C4H4O4] and Detector: UV 225 nm
hydrochlorothiazide (C7H8ClN3O4S2). Column: 8-mm 10-cm; packing L11
Flow rate: 3 mL/min
A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 System suitability
Standard solution A: 1 mg/mL of USP Bisoprolol Fumarate Samples: System suitability solution and Standard solution
RS in methanol Suitability requirements
Standard solution B: 1 mg/mL of USP Hydrochlorothiazide Resolution: NLT 1.5 between chlorothiazide and
RS in methanol hydrochlorothiazide in the System suitability solution
Sample solution: Finely powder 1 Tablet, and transfer the Tailing factor: NMT 1.3 for the hydrochlorothiazide peak
powder to a 5-mL volumetric flask. Dilute with methanol to in the Standard solution
volume, sonicate for 5 min, centrifuge, and use the Relative standard deviation: NMT 2.0% in the Standard
supernatant. solution
Developing solvent system: Methylene chloride, methanol, Samples: Standard solution, Sample solution A, and Sample
and 14.5 M ammonium hydroxide solution (43:20:8) solution B
Analysis Calculate the percentage of (C18H31NO4)2 C4H4O4 in the
Samples: Standard solution A, Standard solution B, and portion of Tablets taken:
Sample solution
Locate the spots on the plate under short-wavelength UV Result = (rU/rS) (CS/CU) 100
light and by exposure to iodine vapors.
Acceptance criteria: The RF values of the principal spots of rU = peak response for bisoprolol fumarate in Sample
the Sample solution correspond to the principal spots of solution A
Standard solution A and Standard solution B. rS = peak response for bisoprolol fumarate in the
B. LIQUID CHROMATOGRAPHY: The retention times of the Standard solution
major peaks of the Sample solution A and Sample solution B CS = concentration of USP Bisoprolol Fumarate RS in
corresponds to those of the Standard solution, as obtained in the Standard solution (mg/mL)
the Assay. CU = nominal concentration of bisoprolol fumarate in
Sample solution A (mg/mL)
ASSAY Calculate the percentage of C7H8ClN3O4S2 in the portion of
PROCEDURE Tablets taken:
Diluent: 10 mL of 1 M dibutylammonium phosphate per L
of acetronitrile and water (1:1) Result = (rU/rS) (CS/CU) 100
Solution A: 1 M dibutylammonium phosphate and water
(1:100) rU = peak response for hydrochlorothiazide in Sample
Solution B: 10 mL of 1 M dibutylammonium phosphate per solution B
L of acetronitrile and water (3:2), stir vigorously for 2 min, rS = peak response for hydrochlorothiazide in the
filter Standard solution
104 Bisoprolol / Official Monographs USP 32
CS = concentration of USP Hydrochlorothiazide RS in Tailing factor: NMT 1.3 in Standard solution

the Standard solution (mg/mL) Relative standard deviation: NMT 2.0% in Standard
CU = nominal concentration of hydrochlorothiazide in solution
the Sample solution B (mg/mL) Analysis
Acceptance criteria: 90.0%110.0% Samples: Standard solution and Sample solution
PERFORMANCE TESTS of Tablets taken:
DISSOLUTION 711
Medium: 0.1 N hydrochloric acid; 900 mL Result = (ru/rS) (WB/WH) (CS/CU) 1/F 100
Apparatus 2: 75 rpm
Times: 20 min for bisoprolol fumarate; 30 min for ru = peak response of each of the two impurities
hydrochlorothiazide from the Sample solution
Solution A: Triethylamine and water (1:499), adjust with rS = peak response for hydrochlorothiazide in the
phosphoric acid to a pH of 3.0 Standard solution
Mobile phase: Acetonitrile and Solution A (1:4) WB = labeled quantity of bisoprolol fumarate in each
Standard stock solution A: 0.5 mg/mL of USP Bisoprolol Tablet (mg)
Fumarate RS in Medium WH = labeled quantity of hydrochlorothiazide in each
Standard stock solution B: Transfer 30 mg of USP Tablet (mg)
Hydrochrolothiazide RS to a 50-mL volumetric flask, dissolve CS = concentration of USP Hydrochlorothiazide RS in
with 5 mL of methanol, and dilute with Medium to volume the Standard solution (mg/mL)
(0.6 mg/mL). CU = concentration of bisoprolol fumarate in the
Standard solution: Dilute Standard stock solution A and Sample solution (mg/mL)
Standard stock solution B in Medium to obtain a solution F = relative response factor, see Impurity Table 1.
having known concentrations of bisoprolol fumarate and Acceptance criteria
hydrochlorothiazide corresponding to those of the solution Individual impurities: See Impurity Table 1.
under test.
Sample solution: Sample per Dissolution 711 Impurity Table
(See Chromatography 621, System Suitability.) Relative Relative Acceptance
Mode: LC Retention Response Criteria,
Detector: UV 227 nm for bisoprolol and 272 nm for Impurity Name Time Factor NMT (%)
hydrochlorothiazide Compound 1 0.69 1.2 1.0
Column: 3.9-mm 15-cm; packing L11 Compound 2 1.2 1.4 2.0
Injection size: 20 L Hydrochlorothiazide 1.0
System suitability
Relative standard deviation: NMT 2.0% PACKAGING AND STORAGE: Preserve in tight, light-resistant
Analysis containers. Store at controlled room temperature.
Samples: Standard solution and Sample solution USP REFERENCE STANDARDS 11
Calculate the quantities, in mg, of (C18H31NO4)2 C4H4O4 USP Bisoprolol Fumarate RS
and C7H8ClN3O4S2 dissolved. USP Chlorothiazide RS
Tolerances: NLT 80% (Q) of the labeled amount of USP Hydrochlorothiazide RS
(C18H31NO4)2 C4H4O4 and NLT 80% (Q) of the labeled
amount of C7H8ClN3O4S2
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Bleomycin Sulfate
with respect to bisoprolol fumarate and to
hydrochlorothiazide (Comment on this Monograph)id=m9835=Bleomycin Sulfate=B-
Monos.pdf)
IMPURITIES Bleomycin sulfate (salt) [9041-93-4].
Organic Impurities
Diluent, Solution A, Solution B, Mobile phase, System Bleomycin Sulfate is the sulfate salt of bleomycin, a mixture of
suitability solution, and Sample stock solution: Proceed basic cytotoxic glycopeptides produced by the growth of
as directed in the Assay. Streptomyces verticillus, or produced by other means. It has a
Standard solution: 2 g/mL of USP Hydrochlorothiazide potency of NLT 1.5 Bleomycin Units/mg and NMT 2.0
RS in Diluent Bleomycin Units/mg.
Sample solution: 100 g/mL of bisoprolol fumarate from
Sample stock solution in Diluent IDENTIFICATION
Chromatographic system A. INFRARED ABSORPTION 197K
(See Chromatography 621, System Suitability.) B. IDENTIFICATION TESTSGENERAL, Sulfate 191
Mode: LC
Column: 8-mm 10-cm; packing L11 PROCEDURE
Flow rate: 3 mL/min Sample solution: Dissolve a suitable quantity of Bleomycin
Injection size: 10 L Sulfate, in Buffer No. 16, and dilute with Buffer No. 16 to
System suitability obtain a solution having a convenient concentration.
Samples: System suitability solution and Standard solution Analysis: Proceed as directed under AntibioticsMicrobial
Suitability requirements Assays 81, using an measured volume of Sample solution
Resolution: NLT 1.5 between chlorothiazide and diluted with Buffer No. 16 to yield a Sample Dilution having
hydrochlorothiazide in the System suitability solution
USP 32 Official Monographs / Bleomycin 105
a concentration assumed to be equal to the median dose Blank: Solution A

level of the Standard. Analysis
Acceptance criteria: 1.52.0 Bleomycin Units/mg Samples: Standard solutions, Sample solution, and Blank
Analysis: Plot the absorbances of the Standard solutions
OTHER COMPONENTS versus concentration, in g/mL, of copper, and draw the
CONTENT OF BLEOMYCINS straight line best fitting the three plotted points.
Mobile phase: 960 mg of sodium 1-pentanesulfonate in Calculate the percentage of copper in the portion of
1000 mL of deaerated 0.08 N acetic acid, adjust with Bleomycin Sulfate:
ammonium hydroxide to a pH of 4.3, filter, and degas.
[NOTE1.86 g of edetate disodium may be included if Result = C/W 100
needed to obtain satisfactory chromatography.] Use a linear
gradient of 10%40% methanol mixed with this solution, C = concentration of copper from the graph
with a gradient mixing time of 60 min, and allow obtained, in the Sample solution (g/mL)
chromatography to proceed with the final gradient mixture W = weight of Bleomycin Sulfate taken to prepare
for a further 20 min or until demethylbleomycin A2 has been the Sample solution (mg)
eluted.
Sample solution: 2.5 Bleomycin Units/mL from Bleomycin SPECIFIC TESTS
Sulfate in deaerated water [NOTEStore this solution in a PH 791: 4.56.0, in a solution containing 10 Bleomycin
refrigerator until just prior to use.] Units/mL
Chromatographic system LOSS ON DRYING 731: Dry it in a vacuum at a pressure not
(See Chromatography 621, System Suitability.) exceeding 5 mm of mercury at 60 for 3 h: it loses NMT
Mode: LC 6.0% of its weight.
Detector: UV 254 nm OTHER REQUIREMENTS
Column: 4.6-mm 250-mm stainless steel; packing L1 Where the label states that Bleomycin Sulfate is sterile, it
Flow rate: 1.2 mL/min meets the requirements for the following tests:
Injection size: 10 L Bacterial Endotoxins Test 85: NMT 10.0 USP Endotoxin
Analysis Units/Bleomycin Unit
Sample: Sample solution Sterility Tests 71: It meets the requirements when tested
Analysis: The elution order is bleomycinic acid, bleomycin as directed for Test for Sterility of the Product to be Examined,
A2 (major peak), bleomycin A5, bleomycin B2 (major peak), Membrane Filtration.
bleomycin B4, and demethylbleomycin A2. Where the label states that Bleomycin Sulfate must be
Calculate the percentage contents of bleomycin A2, subjected to further processing during the preparation of
bleomycin B2, and bleomycin B4 taken: injectable dosage forms, it meets the requirements for:
Bacterial Endotoxins Test 85: NMT 10.0 USP Endotoxin
Result = (ru/rT) 100 Units/Bleomycin Unit
ru = peak response corresponding to the particular ADDITIONAL REQUIREMENTS

bleomycin PACKAGING AND STORAGE: Preserve in tight containers.
rT = total of the responses of all peaks LABELING: Where it is intended for use in preparing
Acceptance criteria: The content of bleomycin A2 is injectable dosage forms, the label states that it is sterile or
55%70%; the content of bleomycin B2 is 25%32%; the must be subjected to further processing during the
content of bleomycin B4 is NMT 1%; and the combined preparation of injectable dosage forms.
percentage of bleomycin A2 and bleomycin B2 is NLT 90%. USP REFERENCE STANDARDS 11
USP Bleomycin Sulfate RS
IMPURITIES USP Endotoxin RS
COPPER: NMT 0.1%
Solution A: Nitric acid and water (1:99)
Standard stock solution: 1.000 g of copper to a 1000-mL Bleomycin for Injection
volumetric flask, dissolve in 20 mL of nitric acid, and dilute (Comment on this Monograph)id=m9840=Bleomycin for
with Solution A to volume. Store in a polyethylene bottle. Injection=B-Monos.pdf)
This solution contains 1000 g/mL of copper.
Standard solutions: Transfer 5.0 mL of Standard stock DEFINITION
solution to a 100-mL volumetric flask, and dilute with Bleomycin for Injection contains an amount of Bleomycin
Solution A to volume. Transfer 3.0, 9.0, and 15.0 mL, Sulfate equivalent to NLT 90.0% and NMT 120.0% of the
respectively, of Standard stock solution to separate 100-mL labeled amount of bleomycin.
volumetric flasks, dilute the contents of each flask with
Solution A to volume, and mix. These Standard solutions ASSAY
contain, respectively, 1.5, 4.5, and 7.5 g/mL of copper. PROCEDURE
Sample solution: 7.5 mg/mL of Bleomycin Sulfate in Sample solution: Constitute Bleomycin for Injection as
Solution A directed in the labeling. Withdraw all of the withdrawable
Spectrometric conditions contents, using a suitable hypodermic needle and syringe,
Mode: Atomic absorption spectrophotometry (see and quantitatively dilute with Buffer No. 16 to obtain a
Spectrophotometry and Light-Scattering 851), equipped solution having a convenient concentration.
with a copper hollow-cathode lamp and an airacetylene Analysis: Proceed as directed under AntibioticsMicrobial
flame Assays 81, using the Sample solution diluted quantitatively
Analytical wavelength: Copper emission line at 324.8 and stepwise with Buffer No. 16 to yield a dilution having a
nm
106 Bleomycin / Official Monographs USP 32
concentration assumed to be equal to the median dose level Adjust with ammonium hydroxide to a pH of 4.3.
of the Standard. [NOTE1.86 g of edetate disodium may be included if
Acceptance criteria: 90.0%120.0% needed to obtain satisfactory chromatography.]
Use a linear gradient of 10% to 40% methanol mixed with
PERFORMANCE TESTS this solution, with a gradient mixing time of 60 min, and
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements allow chromatography to proceed with the final gradient
mixture for a further 20 min or until demethylbleomycin A2
IMPURITIES has been eluted.
Inorganic Impurities Sample solution: 2.5 Bleomycin Units/mL from Bleomycin
COPPER for Injection in water
Solution A: Nitric acid and water (1:100) [NOTEStore this solution in a refrigerator until just prior to
Standard stock solution: Transfer 1.000 g of copper to a use.]
1000-mL volumetric flask, dissolve in 20 mL of nitric acid, Chromatographic system
dilute with Solution A to volume, and mix. Store in a (See Chromatography 621, System Suitability.)
polyethylene bottle. This solution contains 1000 g of Mode: LC
copper per mL. Detector: UV 254 nm
Standard solutions: 1.5 g/mL, 4.5 g/mL, and 7.5 Column: 4.6-mm 250-mm stainless steel; packing L1
g/mL from Standard stock solution in Solution A Flow rate: 1.2 mL/min
Sample solution: 7.5 mg/mL of Bleomycin for Injection in Injection size: 10 L
Solution A Analysis
Spectrometric conditions Sample: Sample solution
Mode: Atomic absorption spectrophotometry equipped The elution order is bleomycinic acid, bleomycin A2 (major
with a copper hollow-cathode lamp and an airacetylene peak), bleomycin A5, bleomycin B2 (major peak),
flame (see Spectrophotometry and Light-Scattering 851) bleomycin B4, and demethylbleomycin A2.
Analytical wavelength: The copper emission line at Calculate the percentage contents of bleomycin A2,
324.8 nm bleomycin B2, and bleomycin B4:
Analysis
Samples: Blank (Solution A), Sample solution, and Result = (ru/rT) 100
Standard solution
Plot the absorbances of the Standard solutions versus ru = peak response corresponding to the particular
concentration of copper, in g/mL, and draw the straight bleomycin
line best fitting the three plotted points. From the graph rT = sum of the responses of all peaks
so obtained, determine the concentration (C) of copper, Acceptance criteria
in g/mL, in the Sample solution. The content of bleomycin A2 is 55%70%; the content of
Calculate the percentage of copper in the portion of bleomycin B2 is 25%32%.
Bleomycin Sulfate taken: The combined percentage of bleomycin A2 and bleomycin
B2 is NLT 90%.
Result = (C/CU) F 100 The content of bleomycin B4 is NMT 1%.
OTHER REQUIREMENTS: Meets the requirements for
C = concentration of copper measured in the Sample Identification tests, under Bleomycin Sulfate and for Injections
solution (g/mL) 1, Labeling.
CU = concentration of Bleomycin Sulfate for injection
in the Sample solution (mg/mL) ADDITIONAL REQUIREMENTS
F = unit conversion factor (0.001 [mg/g]) PACKAGING AND STORAGE: Preserve as described under
Acceptance criteria: NMT 0.1% Injections 1, Containers for Sterile Solids.
SPECIFIC TESTS USP Bleomycin Sulfate RS
CONSTITUTED SOLUTION: At the time of use, it meets the USP Endotoxin RS
requirements for Injections 1, Constituted Solutions.
BACTERIAL ENDOTOXINS TEST 85: NMT 10.0 USP Endotoxin
Units/Bleomycin Unit
STERILITY TESTS 71: Meets the requirements when tested as Anti-A Blood Grouping Serum
directed for Test for Sterility of the Product to be Examined, (Comment on this Monograph)id=m9880=Anti-A Blood
Membrane Filtration, the entire contents of each container Grouping Serum=B-Monos.pdf)
being used.
WATER DETERMINATION, Method Ic 921 DEFINITION
Sample solution: Use a dry syringe to inject 4 mL of Anti-A Blood Grouping Serum conforms to the regulations of
anhydrous methanol through the stoppers of each of two the federal Food and Drug Administration concerning biologics
tared containers, respectively, and shake to dissolve. Using (660.20 to 660.29) (see Biologics 1041). It is a sterile, liquid
the same syringe, aspirate the contents of the two or dried preparation containing the particular blood group
containers, transfer to the titration vessel, and titrate. antibodies derived from high-titered blood plasma or serum of
Perform a blank determination on 8 mL of the anhydrous human subjects, with or without stimulation by the injection
methanol. Determine the weights of the empty containers, of Blood Group Specific Substance A (or AB). It agglutinates
and calculate the percentage of water. human red cells containing A-antigens, i.e., blood groups A
Acceptance criteria: NMT 6.0% and AB (including subgroups A1, A2, A1B, and A2B but not
PH 791: 4.56.0 in a solution containing 10 Bleomycin necessarily weaker subgroups). It contains a suitable
Units/mL antimicrobial preservative. It meets the requirements to the
CONTENT OF BLEOMYCINS test for potency, in parallel with, and not less than equivalent
Mobile phase: 960 mg of sodium 1-pentanesulfonate in to, the U.S. Reference Blood Grouping Serum Anti-A, in
1000 mL of 0.08 N acetic acid
USP 32 Official Monographs / Blood 107
agglutinating red blood cells from Group A1 and Group A2B human blood will not transmit hepatitis. Label it also to state
donors. It meets the requirements of the tests for specificity that it is for in vitro diagnostic use. [NOTEThe labeling is in
with Group A1, A2B, B, and O cells and confirms the absence black lettering imprinted on paper that is white or is colored
of contaminating antibodies reactive with Mg, Wra antigens as completely or in part to match the specified yellow color
well as other antigens having an incidence of 1% or greater in standard.]
the general population (see Biologics 1041, Blood Grouping
Serums Anti-D, Anti-C, Anti-E, Anti-c, Anti-e). It meets the
requirements of the tests for avidity with Group A1 and A2B
cells. All fresh or frozen red blood cell suspensions used for Blood Grouping Serums
these tests are prepared under specified conditions and meet (Comment on this Monograph)id=m9900=Blood Grouping
specified criteria. Anti-A Blood Grouping Serum may be Serums=B-Monos.pdf)
artificially colored blue.
DEFINITION
ADDITIONAL REQUIREMENTS [NOTEThis monograph deals with those Blood Grouping
PACKAGING AND STORAGE: Preserve at a temperature Serums for which there are no individual monographs and
between 2 and 8. which are not routinely used or required for the testing of
EXPIRATION DATE: The expiration date for liquid Serum is blood or blood products for transfusion.]
not later than 1 year, and for dried Serum not later than 5 Blood Grouping Serums conform to the regulations of the
years after date of issue from manufacturers cold storage federal Food and Drug Administration concerning biologics
(5, 1 year; or 0, 2 years), provided that the expiration date (see Sections 660.20 to 660.29; see also Biologics 1041). Each
for dried Serum is not later than 1 year after constitution. is a sterile, liquid or dried preparation containing one or more
LABELING: Label it to state that the source material was not of the particular blood group antibodies derived from high-
reactive for hepatitis B surface antigen, but that no known titered blood plasma or serum of human subjects, with or
test method offers assurance that products derived from without stimulation by the injection of red cells or other
human blood will not transmit hepatitis. Label it also to state substances, or of animals after stimulation by substances that
that it is for in vitro diagnostic use. [NOTEThe labeling is in cause such antibody production. It causes, either directly or
black lettering imprinted on paper that is white or is colored indirectly, by the antiglobulin test, the visible agglutination of
completely or in part to match the specified blue color human red cells containing the particular antigen(s) for which
standard.] it is specific. It contains a suitable antimicrobial preservative. It
meets the requirements of the test for potency, (1) in the case
of tube test reagents, when tested by the specified method, of
agglutinating red blood cells containing the specified antigens
Anti-B Blood Grouping Serum with the specified degree of reactivity, as defined, as follows:
(Comment on this Monograph)id=m9890=Anti-B Blood not less than a 1+ reaction (i.e., agglutinated cells dislodged
Grouping Serum=B-Monos.pdf) into finely granular, but definite, small clumps) with a 1:8
dilution of Serum for Anti-K, Anti-k, Anti-Jka, Anti-Fya, Anti-Cw;
DEFINITION NLT a 1+ reaction with a 1:4 dilution of Serum for Anti-S, Anti-
Anti-B Blood Grouping Serum conforms to the regulations of s, Anti-P1, Anti-M, Anti-I, Anti-e (saline), Anti-c (saline) and
the federal Food and Drug Administration concerning biologics Anti-A1; and not less than a 2+ reaction (i.e., agglutinated cells
(660.20 to 660.29) (see Biologics 1041). It is a sterile, liquid dislodged into many small clumps of equal size) with
or dried preparation containing the particular blood group undiluted Serum for Anti-U, Anti-Kpa, Anti-Kpb, Anti-Jsa, Anti-
antibodies derived from high-titered blood plasma or serum of Fyb, Anti-N, Anti-Lea, Anti-Leb, Anti-Dia, Anti-Mg, Anti-Jkb, and
human subjects, with or without stimulation by the injection Anti-Xga; and (2) in the case of reagents recommended for
of Blood Group Specific Substance B (or AB). It agglutinates slide test methods, of agglutinating red blood cells, with the
human red cells containing B-antigens, i.e., blood groups B specified degree of reactivity, as defined, with both undiluted
and AB (including subgroups A1B and A2B). It contains a Serum and with a 1:2 dilution of Serum, when tested by the
suitable antimicrobial preservative. It meets the requirements manufacturers recommended method(s) using cells
of the test for potency, in parallel with, and not less than heterozygous for the corresponding antigen(s). It meets the
equivalent to, the U.S. Reference Blood Grouping Serum Anti- requirements of the tests for specificity by the most sensitive
B, in agglutinating red blood cells from Group B donors. It method recommended by the manufacturer, in which not less
meets the requirements of the tests for specificity with Group than 4 positive and 4 negative phenotypes are included, and
A1, B, and O cells and confirms the absence of contaminating confirms the absence of contaminating antibodies reactive
antibodies reactive with Mg, Wra antigens as well as other with Mg, Wra antigens as well as other antigens having an
antigens having an incidence of 1% or greater in the general incidence of 1% or greater in the general population (see
population (see under Blood Grouping Serums Anti-D, Anti-C, Blood Grouping Serums Anti-D, Anti-C, Anti-E, Anti-c, Anti-e). It
Anti-E, Anti-c, Anti-e). It meets the requirements of the test for meets the requirements of the tests for avidity with the
avidity with Group B cells. All fresh or frozen red blood cell manufacturers recommended method, red blood cells
suspensions used for these tests are prepared under specified heterozygous for the corresponding antigen(s) being used. All
conditions and meet specified criteria. Anti-B Blood Grouping fresh or frozen red blood cell suspensions used for these tests
Serum may be artificially colored yellow. are prepared under specified conditions and meet specified
criteria.
PACKAGING AND STORAGE: Preserve at a temperature ADDITIONAL REQUIREMENTS
between 2 and 8. PACKAGING AND STORAGE: Preserve at a temperature
EXPIRATION DATE: The expiration date for liquid Serum is between 2 and 8.
not later than 1 year and for dried Serum not later than 5 EXPIRATION DATE: The expiration date for liquid Serum is
years after date of issue from manufacturers cold storage not later than 1 year and for dried Serum not later than 5
(5, 1 year; or 0, 2 years), provided that the expiration date years after date of issue from manufacturers cold storage
for dried Serum is not later than 1 year after constitution. (5, 1 year; or 0, 2 years), provided that the expiration date
LABELING: Label it to state that the source material was not for dried Serum is not later than 1 year after constitution.
reactive for hepatitis B surface antigen, but that no known
test method offers assurance that products derived from
108 Blood / Official Monographs USP 32
LABELING: Label each to state the source of the product if Table 2

other than human and, if of human origin, to state that the Serum Phenotype of Cells
source material was not reactive for hepatitis B surface
antigen, but that no known test method offers assurance Anti-D cDe
that products derived from human blood will not transmit Anti-C Ccde
hepatitis. Label each also to state that it is for in vitro Anti-E cdEe
diagnostic use.
Anti-CD cDe, Ccde
Anti-DE cDe, cdEe
Blood Grouping Serums Anti-D, Anti-C, Anti-CDE cdEe, cDe, Ccde
Anti-E, Anti-c, Anti-e Anti-c CcDEe
(Comment on this Monograph)id=m9910=Blood Grouping Anti-e cdEe
Serums Anti-D, Anti-C, Anti-E, Anti-c, Anti-e=B-Monos.pdf)
Anti-Rh Blood Grouping Serums Each serum meets the requirements of the tests for specificity
by the most sensitive method recommended by the
DEFINITION manufacturer, in which NLT 4 positive and 4 negative
Blood Grouping Serums Anti-D, Anti-C, Anti-E, Anti-c, Anti-e phenotypes are included (see Table 3), and confirms the
(Anti-Rh Group) conform to the regulations of the federal absence of contaminating antibodies reactive with Mg, Wra
Food and Drug Administration concerning biologics (660.20 to antigens as well as other antigens having an incidence of 1%
660.29) (see Biologics 1041). They are sterile, liquid, or dried or greater in the general population, except where some of
preparations derived from the blood plasma or serum of these confirmatory tests cannot be done by the manufacturer,
human subjects who have developed specific Rh antibodies. in which event such omissions are noted. Antigens having
They are free from agglutinins for the A or B antigens and such population incidence in the United States [other than
from alloantibodies other than those for which claims are low-incidence antigens, serum proteins (e.g., Gm, Km),
made in the labeling. They contain a suitable antimicrobial leukocyte factors, drugs, chemicals, or polyagglutinable cells,
preservative. Liquid serums are not artificially colored. Two which are not necessarily excluded] are the following: A, B, H,
varieties of Anti-Rh Blood Grouping Serums are recognized, Lea, Leb, Lec, Led, I, K, k, Kpa, Kpb, Jsb, P1, D, C, E, c, e, Cw, M,
i.e., (1) saline agglutinating complete antiserums, which N, S, s, U, Lua, Lub, Jka, Jkb, Fya, Fyb, Xga, Doa, Dob, Yta, Ytb,
specifically agglutinate human red blood cells suspended in Lan, Coa, Cob, Mg, Wra, and Sda.
saline TS, and (2) blocking or incomplete antiserums, which
contain protein or other macromolecular substances, usually Table 3
require the cells to be suspended in serum or plasma, and
generally are for slide or rapid tube tests. The most commonly Serum Cells
used of these blood grouping serums are listed in Table 1, Anti-D CcDe, cDe, Ccde, cdEe, A1 cde, B cde, O cde,
each reacting with the antigen(s) designated by the and where recommended for use by indirect
corresponding letter(s) with the alternative nomenclature antiglobulin technique, cde Bg(a+) cells from
indicated parenthetically. 3 different donors
Anti-C cDe, Ccde, cdEe, C + rhi neg. cells, A1 cde, B
Table 1 cde, O cde
Serum Antigen(s) Reacting Anti-E cDe, Ccde, cdEe, A1 cde, B cde, O cde
Anti-D (Anti-Rho) D (Rho) Anti-CD cDe, Ccde, cdEe, A1 cde, B cde, O cde, and
Anti-C (Anti-rh) C (rh) where recommended for detection of the G
antigen, rGr
Anti-E (Anti-rh) E (rh)
Anti-DE cDe, Ccde, cdEe, A1 cde, B cde, O cde
Anti-CD (Anti-Rho) D (Rho) and C (rh)
Anti-CDE cDe, Ccde, cdEe, A1 cde, B cde, O cde, and
Anti-DE (Anti-Rho) D (Rho) and E (rh)
where recommended for detection of the G
Anti-CDE (Anti-Rho) D (Rho), C (rh), and E (rh) antigen, rGr
Anti-c (Anti-hr) c (hr) Anti-c Ccde, A1 CDe, B CDe, O CDe, and CDEe or
Anti-e (Anti-hr) e (hr) CDE or CdE
Anti-e cdEe, A1 cDE, B cDE, O cDE, and CcDE or CDE
Each Serum meets the requirements of the test for potency in or CdE
the case of serums for saline tube test in parallel with, and NLT
equivalent to, the U.S. Reference Blood Grouping Serum for All fresh or frozen red blood cell suspensions used for these
Anti-D, Anti-C, or Anti-E, whichever is applicable, or, in the tests are prepared under specified conditions and meet
case of Anti-c and Anti-e for saline tube test which have no specified criteria.
reference preparations, the test for minimum agglutination
reactivity at a specified dilution; and in the case of serums for ADDITIONAL REQUIREMENTS
slide or rapid tube test in parallel with, and NLT than PACKAGING AND STORAGE: Preserve at a temperature
equivalent to, the U.S. Reference Blood Grouping Serum for between 2 and 8.
Anti-D, Anti-C, Anti-E, Anti-c, or Anti-e, whichever is EXPIRATION DATE: The expiration date for liquid Serums is
applicable, in agglutinating as a minimum red blood cells from not later than 1 year and for dried Serums not later than 5
the donors indicated in Table 2 (which may be from Group A, years after date of issue from manufacturers cold storage
B, AB, or O). (5, 1 year; or 0, 2 years), provided that the expiration date
Each serum for slide or rapid tube test meets the requirements for dried Serums is not later than 1 year after constitution.
of the tests for avidity with the cells as indicated under tests LABELING: Label each to state that the source material was
for potency above. not reactive for hepatitis B surface antigen, but that no
known test method offers assurance that products derived Analysis

from human blood will not transmit hepatitis. Label each to Samples: Control solutions and Sample solution
state that it is for in vitro diagnostic use. On a suitable U-bottomed microtiter plate, place 1 drop of
0.9% saline in each of three different wells in a row (Blank
Row). Place 1 drop from one of the lots of Anti-A reagent
in each of three different wells in a second row. Place 1
Whole Blood drop from the second lot of Anti-A reagent in each of
(Comment on this Monograph)id=m9950=Whole Blood=B- three different wells in a third row. Repeat the same with
Monos.pdf) two different lots of Anti-B reagent and one lot of Anti-AB
ACD Whole Blood reagent in separate rows. To each row, add 1 drop of
CPD Whole Blood Control solution A1, Control solution B, and the Sample
CPDA-1 Whole Blood solution in the first, second, and the third well,
Heparin Whole Blood respectively, of each row. Mix the contents of the wells by
gently tapping the sides of the plate. Centrifuge the plate
DEFINITION at the appropriate conditions established for the
Whole Blood conforms to the regulations of the federal Food centrifuge. Resuspend the cell buttons by manually
and Drug Administration concerning biologics (21 CFR 640.1 tapping the plate, or with the aid of a suitable mechanical
to 640.6) (see Biologics 1041). It is blood that has been shaker. Read the optical densities at different wells using a
collected from suitable human donors under rigid aseptic suitable automated photometric microtiter plate reader.
precautions, for transfusion to human recipients or for further Compare the optical density of each well in the Blank Row
processing into one or more of its components for transfusion. with the optical density of the wells to which the
It contains a citrate-based anticoagulant. Whole Blood must be corresponding Control solution A1, Control solution B, or
tested for syphilis, hepatitis B virus, Human T-Cell Sample solution were added. [NOTEA high optical density
Lymphotropic Virus (HTLV) type I and type II, and tested for comparable to those obtained for the wells in the Blank
blood group and Rh factors and unexpected antibodies to red Row indicates negative results (no hemagglutination),
cell antigens using FDA-licensed commercially available test which can be corroborated by visual observation of
kits, the results of which must be below the approved upper smooth suspensions. A low optical density indicates
limit of detection specified in the respective test kits. In positive results (hemagglutination), which can be
addition, Whole Blood must also be tested for hepatitis C and corroborated by visual observation of formation of
HIV using FDA-approved nucleic acid assays, the results of clumps.] The blood group of Red Blood Cells is A, B, AB,
which must be below the approved upper limit of detection or O, accordingly, as the Sample solution is
for the method. A unit (dose) of Whole Blood contains a hemagglutinated by Anti-A reagent, Anti-B reagent, both
minimum of 50 g (based on a minimum donor hematocrit of reagents, or neither, respectively. The blood group of the
38%) of hemoglobin in a total volume of 450 mL 10% or Sample solution conforms to the blood group indicated on
500 mL 10% as indicated on the label. Whole Blood may be the label. The test is not valid if Control solutions for Blood
further processed. When filtered for removal of leukocytes, the Group A and Blood Group B red blood cells are not
quantity of residual leukocytes in the unit of Whole Blood agglutinated by Anti-A reagent and Anti-B reagent,
must be less than 5 106. respectively, or if both Control solutions are not
agglutinated by Anti-AB reagent. The test is also not valid
IDENTIFICATION if the Sample solution is not agglutinated by Anti-AB
A. ABO BLOOD GROUP reagent but is agglutinated by either Anti-A reagent or
Anti-A reagent: Use approved commercially available Anti-B reagent, or is agglutinated by Anti-AB reagent but
monoclonal or polyclonal anti-A blood grouping reagent, not by either Anti-A reagent or Anti-B reagent.
two different lots from the same or different manufacturers. B. RH TYPE
Use in accordance with manufacturers instructions. Test 1
Anti-B reagent: Use approved commercially available Anti-D (Rho) reagent: Use anti-D (Rho) blood grouping
monoclonal or polyclonal anti-B blood grouping reagent, reagent approved for use in microtiter plate tests. Dilute, if
two different lots from the same or different manufacturers. necessary, following the manufacturers instructions.
Use in accordance with manufacturers instructions. Sample solution: Prepare as directed for Identification test
Anti-AB reagent: Use approved commercially available anti- A.
AB blood grouping reagent. Use in accordance with Analysis: On a suitable U-bottom microtiter plate, place 1
manufacturers instructions. drop each of 0.9% saline and Anti-D (Rho) reagent in two
Control solutions: On the day of use, dilute Blood Group separate wells. Label them as the B well (Blank) and the T
A1 (Control solution A1) and Blood Group B (Control well, respectively. Add 1 drop of Sample solution to each
solution B) red blood cells, obtained from an approved well, and mix by gently tapping the side of the plate.
commercial source or prepared by the testing laboratory, Centrifuge the plate at appropriate conditions established
with 0.9% saline to suspensions of approximately the same for the centrifuge. Resuspend the cell buttons by manually
concentration between 2% to 5% (v/v). [NOTEIf the Blood tapping the plate, or with the aid of a suitable mechanical
Group A1 and Blood Group B red blood cells are prepared in shaker. Read the optical densities of the wells using a
the testing laboratory from whole blood of a known blood suitable automated photometric microtiter plate reader,
group, it must be prepared on the day of use following the and determine if the Red Blood Cells in the T well are
procedure for the Sample solution under Whole Blood.] agglutinated as described under Identification test A. If the
Sample solution: Centrifuge at 4 a suitable volume of T well indicates negative results (no hemagglutination),
Whole Blood at 5000 g for 5 min. Remove the plasma from incubate the plate at 37 for 15 min, centrifuge, resuspend
the top, taking care not to disturb the pellet of red blood the cells, and read the optical densities of the wells as
cells at the bottom. Add 0.9% saline to obtain a final above. Agglutination of cells after immediate-spin or after
volume that is equal to the volume of Whole Blood. 37 incubation indicates Rh positive typing of Red Blood
Resuspend the pellet, and centrifuge as above. Repeat the Cells. The test is valid if cells in the B well are not
procedure once more. Dilute the red blood cells with 0.9% agglutinated. If the cells are not agglutinated in the T well,
saline to a suspension to obtain a concentration of red proceed as directed in Test 2.
blood cells that is the same as those of the Control solutions.
Test 2 Sample solution to each of T1 and T2, rinsing the pipet tip
Anti-D reagent: Use anti-D blood grouping reagent three to four times with Drabkins solution, and mix. Allow
approved for use for a weak D blood group test. to stand for at least 15 min at room temperature. [NOTE
Antihuman globulin reagent: Use polyspecific or anti-IgG Red Blood cells with appreciable carboxyhemoglobin
antihuman globulin reagent. Dilute, if necessary, following content, such as those obtained from smokers, may require
the manufacturers instructions. longer reaction time. If the donor characteristics are not
Control solution: Use IgG-coated red cells approved for known, the incubation time should be optimized prior to
use as a control for Rh typing. Dilute with 0.9% saline to testing]. Read the absorbances of the solutions against the
obtain a 2% solution. solution in tube B1 at 540 nm. The absorbance of the
Sample solution: Prepare as directed for Test 1. solution in tube B2 is recorded at the end. The test is not
Analysis: Place 1 drop of 0.9% saline into a suitable test valid if the absorbance of the solution in tube B2 is not
tube and 1 drop of Anti-D reagent in another, and mark within 0.005.
them as the Blank and Anti-D, respectively. To each tube, Calculations: Calculate the concentrations, in mg/mL, of
add 1 drop of the Sample solution, mix, and incubate at hemoglobin in Standard solutions A, B, and C. Plot a
37 for 15 to 30 min. Centrifuge the tubes at 1000 g for calibration curve of absorbance values against the
15 to 30 s. Gently resuspend the cell buttons, and examine hemoglobin concentration by drawing a best-fit straight line
them for hemagglutination (formation of clump) by visual using the least-square linear regression analysis. From the
examination. The Rh typing of Red Blood Cells is positive absorbance value of the Sample solution, obtain the
or negative according to whether the cells in Anti-D tube concentration, in mg/mL, of hemoglobin in the Sample
are agglutinated or not. If the cells are not agglutinated, solution.
add 1 mL of 0.9% saline to the Anti-D tube, and resuspend Calculate the total hemoglobin content in the Red Blood
the cells. Centrifuge the tube at 1000 g for 1 min, and Cells unit, in g, by the formula:
remove the saline completely. Repeat the step two to three Conc. of hemoglobin (mg/mL) the volume of the Red
times more to wash the Red Blood Cells. Add 1 drop of Blood Cells unit (mL)/103
0.9% saline and 1 to 2 drops of Antihuman globulin Leukocyte count (for units labeled as Whole Blood,
reagent to the Anti-D tube. Mix gently, and centrifuge the Leukocytes Reduced)
tube at 1000 g for 15 to 30 s. Gently resuspend the cell Sample solution: Pipet 40 L of a suitable red cell-lysing
button, add 1 drop of Control solution, mix gently, agent into a clean test tube, add 100 L of Whole Blood
centrifuge as above, and examine for agglutination. The Rh diluted with 0.9% saline, if necessary, such that the
Type of the Sample solution conforms to the Rh Type hematocrit of Red Blood Cells is not greater than 60%.
indicated on the label. The test is not valid if the cells in Analysis: Mix by pipetting up and down several times. Add
the Anti-D tube are not agglutinated after addition of the 360 L of 0.01% (w/v) crystal violet in 15% (v/v) acetic acid
Control solution. Also, for the test to be valid, the cells in into the mixture, and mix thoroughly. Fit a hemocytometer
the Blank tube must not be agglutinated. with a 50-L counting volume and a bright background,
with a cover slip, and load the counting chamber with the
SPECIFIC TESTS mixture until the counting area is completely covered, but
VISUAL INSPECTION: Inspect visually during storage and not overfull. Cover the counting chamber with a suitable
immediately prior to use. If the color or physical appearance moist lid to prevent evaporation, and allow to settle
is abnormal or there is any indication or suspicion of undisturbed for 10 to 15 min. Remove the lid, place the
microbial contamination, the unit is unsuitable for chamber on the stage of a light microscope fitted with 10
transfusion. ocular lens and 20 objective. Count the leukocytes in the
HEMOGLOBIN CONTENT entire 50-L counting volume. Calculate the leukocyte count
Drabkins solution: Dissolve Drabkins reagent in a suitable in Red Blood Cells, expressed in leukocytes/L, by dividing
volume of water, and add a suitable volume of a 30% (w/v) the observed leukocyte count by 10.
polyoxyethylene (23) lauryl ether solution such that the final Calculate the total number of leukocytes in the Red Blood
concentrations of potassium cyanide, potassium Cells unit by using the following formula:
ferrocyanide, and polyoxyethylene (23) lauryl ether in the
solution are approximately 0.75 mM, 0.6 mM, and 0.015%, Total leukocytes = leukocytes/L 103 volume of Red
respectively. Store the solution in the dark between 1826. Blood Cells unit (mL)
[CautionDrabkins reagent and Drabkins solution contain
cyanide and are HIGHLY TOXIC. Do not inhale or swallow ADDITIONAL REQUIREMENTS
or allow contact with skin or eyes. Wear suitable PACKAGING AND STORAGE: Collect into an approved
protective clothing, gloves, and eye and face protection. container (see Transfusion and Infusion Assemblies and Similar
Do not mix with acids. Contact with acids liberates a very Medical Devices 161) containing a sterile, pyrogen-free
toxic gas. If ingested, perform gastric lavage, and approved anticoagulant (see USP monographs Anticoagulant
immediately call a physician.] Citrate Dextrose Solution, Anticoagulant Citrate Phosphate
Blank solution: Water Dextrose Solution, or Anticoagulant Citrate Phosphate Dextrose
Standard solution A: 300 mg of USP Hemoglobin RS in a Adenine Solution). Store Whole Blood in the original
2-mL volumetric tube, add 1 mL water, dissolve in and container, or transfer to an equivalent one using a technique
dilute with water to volume. that does not compromise sterility. Whole Blood is stored at
Standard solution B: Mix 50 L of Standard solution A with a temperature between 1 and 6 unless platelets are to be
25 L of water. prepared, in which case the blood is stored for no longer
Standard solution C: Mix 50 L of Standard solution A with than 8 h following collection at room temperature. (See
100 L of water. General Notices and Requirements.)
Sample solution: 20 L of Whole Blood (without dilution) EXPIRATION DATE: Whole Blood collected in Anticoagulant
Analysis: Label suitable tubes as B1, B2, SA1, SA2, SB1, Citrate Dextrose Solution, Anticoagulant Citrate Phosphate
SB2, SC1, SC2, T1, and T2. Add 5.0 mL of Drabkins Dextrose Solution, or in Anticoagulant Citrate Phosphate
solution to each tube. Add 20 L of water to each of B1 and DextroseDextrose Solution may be stored for up to 21 days
B2, 20 L of Standard solution A to each of SA1 and SA2, 20 at 16 after the blood has been drawn. Whole Blood
L of Standard solution B to each of SB1 and SB2, 20 L of collected in Anticoagulant Citrate Phosphate Dextrose
Standard solution C to each of SC1 and SC2, and 20 L of Adenine Solution may be stored for up to 35 days at 16.
If the hermetic seal of the container is broken during approved and licensed commercially available test kits, the
collection, solution, or further processing, the expiration date results of which must be below the limits of detection
is not later than 24 h after the seal is broken (when blood is specified in the respective test kits by the manufacturers. In
stored at 16), but not to exceed the original expiration addition, the source blood must also be tested for hepatitis C
date of the unit. and HIV using FDA-approved nucleic acid assays, the results of
LABELING: Label the container to indicate the volume of the which must be below the approved limits of detection for the
Whole Blood collected from the donor, the collection date, method. A unit (dose) of Red Blood Cells contains a minimum
the donation number or other coding means to uniquely of 50 g of hemoglobin in a total volume of about 180325
identify the unit and to provide traceability to the donor, the mL. A unit (dose) of Red Blood Cells, Leukocytes Reduced
storage temperature, and the expiration date (see below). contains a minimum of 42.5 g of hemoglobin in a total
Label to indicate the type of anticoagulant or other volume of about 150275 mL, and has a residual leukocyte
preservative solution used to collect it. Label also to identify count of less than 5 106. A unit (dose) of Red Blood Cells,
donor status (i.e., volunteer or paid). Label also with the Deglycerolized contains a minimum of 40 g of hemoglobin in
following statements: See Circular of Information for a total volume of about 180325 mL. A unit (dose) of Red
indications, contraindications, cautions, and methods of Blood Cells, Leukocytes Reduced and Deglycerolized contains a
infusion.; Properly identify recipient; and [CAUTIONRx minimum of 34 g of hemoglobin in a total volume of
only.] Label to indicate the ABO group and Rh type, as approximately 180325 mL and has a residual leukocyte count
indicated in Table 1. [NOTEEach Whole Blood product must of less than 5 106. A unit (dose) of Red Blood Cells, Pheresis
have a determination made as to its ABO group and Rh type contains a mean hemoglobin content of 60 g of hemoglobin.
and specificity of unexpected red cell antibodies, if any.] A unit (dose) of Red Blood Cells, Pheresis, Leucocytes Reduced
contains a mean hemoglobin content of 51 g (or 153 mL
Table 1 packed red cell volume) and has a residual leukocyte count of
less than 5 106.
ABO Group Rh Type
A Positive IDENTIFICATION
A. ABO BLOOD GROUP
A Negative Anti-A reagent: Use approved commercially available
B Positive monoclonal or polyclonal anti-A blood grouping reagent,
B Negative two different lots from the same or different manufacturers.
Use in accordance with manufacturers instructions.
AB Positive Anti-B reagent: Use approved commercially available
AB Negative monoclonal or polyclonal anti-B blood grouping reagent,
O Positive two different lots from the same or different manufacturers.
Use in accordance with manufacturers instructions.
O Negative Anti-AB reagent: Use approved commercially available anti-
AB blood grouping reagent. Use in accordance with
If an ABO blood group color scheme is used, use the manufacturers instructions.
following labeling color: Group A (yellow), Group B (pink), Control solutions: On the day of use, dilute Blood Group
Group O (blue), and Group AB (white). Label Whole Blood A1 (Control solution A1) and Blood Group B (Control solution
with the type and results of tests for adventitious agents. If B) red blood cells, obtained from an approved commercial
it has been issued prior to determination of the test results, source or prepared by the testing laboratory, with 0.9%
label also with the warning Donor Untested and to saline to suspensions of about the same concentration
further specify Uncrossmatched Blood, when appropriate. between 2% and 5%.
If the unit of Whole Blood was filtered to reduce [NOTEIf the Blood Group A1 and Blood Group B red
leukocytes, label as Whole Blood, Leukocytes Reduced. blood cells are prepared in the testing laboratory from
USP REFERENCE STANDARDS 11 whole blood of a known blood group, it must be
USP Hemoglobin RS prepared on the day of use according to the following
procedure: Centrifuge at 4 a suitable volume of Whole
Blood at 5000 g for 5 min. Remove the plasma from the
Red Blood Cells top, taking care not to disturb the pellet of red blood cells
at the bottom. Add 0.9% saline to obtain a final volume
(Comment on this Monograph)id=m9940=Red Blood Cells=B- that is about equal to the volume of Whole Blood.
Monos.pdf) Resuspend the pellet, and centrifuge as above. Repeat the
DEFINITION procedure once more. Dilute the red blood cells with
Red Blood Cells is the portion of blood that contains 0.9% saline to a suspension to obtain a concentration of
hemoglobin and is derived from human whole blood, from red blood cells that is about the same as those of the
which plasma and platelets are removed by centrifugation, Control solutions.]
sedimentation, or by apheresis. In the case of apheresis, the Sample solution: On the day of use, dilute Red Blood Cells
plasma is automatically removed and returned directly to the with 0.9% saline to a suspension of about the same
donor. Red Blood Cells derived from whole blood may be concentration as the Control solutions.
prepared at any time during the dating period of the whole Analysis
blood from which it is derived. Samples: Control solutions and Sample solution
Red Blood Cells may be further processed by addition of red On a suitable U-bottomed microtiter plate, place 1 drop of
cell preservatives, irradiation to inactivate lymphocytes, 0.9% saline in each of three different wells in a row (Blank
filtration for removal of leukocytes, washing to remove Row). Place 1 drop from one of the lots of Anti-A reagent
proteins, freezing and thawing, or rejuvenation using validated in each of three different wells in a second row. Place 1
and approved procedures. The source blood for Red Blood drop from the second lot of Anti-A reagent in each of
Cells must be tested for syphilis, hepatitis B virus, hepatitis C three different wells in a third row. Repeat the same with
virus, Human T-cell Lymphotropic Virus (HTLV) type I and type two different lots of Anti-B reagent and one lot of Anti-AB
II, human immunodeficiency virus (HIV) type 1 and type 2, reagent in separate rows. To each row, add 1 drop of
and unexpected antibodies to red cell antigens using FDA- Control solution A1, Control solution B, and the Sample
solution in the first, second, and the third well, them as the Blank and Anti-D, respectively. To each tube,
respectively, of each row. Mix the contents of the wells by add 1 drop of the Sample solution, and incubate at 37 for
gently tapping the sides of the plate. Centrifuge the plate 1530 min. Centrifuge the tubes at about 1000 g for
at the appropriate conditions established for the 1530 s. Gently resuspend the cell buttons, and examine
centrifuge. Resuspend the cell buttons by manually them for hemagglutination (formation of clump) by visual
tapping the plate, or with the aid of a suitable mechanical examination. The Rh typing of Red Blood Cells is positive
shaker. Read the optical densities at different wells using a or negative according to whether the cells in Anti-D tube
suitable automated photometric microtiter plate reader. are agglutinated or not. If the cells are not agglutinated,
Compare the optical density of each well in the Blank Row add 1 mL of 0.9% saline to the Anti-D tube, and resuspend
with the optical density of the wells to which the the cells. Centrifuge the tube at about 1000g for 1 min,
corresponding Control solution A1, Control solution B, or and remove the saline completely. Repeat the step two to
Sample solution were added. [NOTEA high optical density three times more to wash the Red Blood Cells. Add 1 drop
comparable to those obtained for the wells in the Blank of 0.9% saline and 1 to 2 drops of Antihuman globulin
Row indicates negative results (no hemagglutination), reagent to the Anti-D tube. Mix gently, and centrifuge the
which can be corroborated by visual observation of tube at about 1000 g for 1530 s. Gently resuspend the
smooth suspensions. A low optical density indicates cell button, add 1 drop of Control solution, mix gently,
positive results (hemagglutination), which can be centrifuge as above, and examine for agglutination. The Rh
corroborated by visual observation of formation of Type of the Sample solution conforms to the Rh Type
clumps.] The blood group of Red Blood Cells is A, B, AB, indicated on the label. The test is not valid if the cells in
or O, accordingly, as the Sample solution is the Anti-D tube are not agglutinated after adding the
hemagglutinated by Anti-A reagent, Anti-B reagent, both Control solution. Also, for the test to be valid, the cells in
reagents, or neither, respectively. The blood group of the the Blank tube must not be agglutinated.
Sample solution conforms to the blood group indicated on
the label. The test is not valid if Control solutions for Blood SPECIFIC TESTS
Group A and Blood Group B red blood cells are not VISUAL INSPECTION: Inspect visually during storage and
agglutinated by Anti-A reagent and Anti-B reagent, immediately prior to use. If the color or physical appearance
respectively, or if both Control solutions are not is abnormal or there is any indication or suspicion of
agglutinated by Anti-AB reagent. The test is also not valid microbial contamination, the unit is unsuitable for
if the Sample solution is not agglutinated by Anti-AB transfusion.
reagent but is agglutinated by either Anti-A reagent or HEMOGLOBIN CONTENT
Anti-B reagent, or is agglutinated by Anti-AB reagent but Drabkins solution: Dissolve Drabkins reagent in a suitable
not by either Anti-A reagent or Anti-B reagent. volume of water, and add a suitable volume of a 30% (w/v)
B. RH TYPE polyoxyethylene (23) lauryl ether solution such that the final
Test 1 concentrations of potassium cyanide, potassium
Anti-D (Rho) reagent: Use anti-D (Rho) blood grouping ferrocyanide, and polyoxyethylene (23) lauryl ether in the
reagent approved for use in microtiter plate tests. Dilute, if solution are approximately 0.75 mM, 0.6 mM, and 0.015%,
necessary, following the manufacturers instructions. respectively. Store the solution in the dark between 18 and
Sample solution: On the day of use, dilute Red Blood 26.
Cells with 0.9% saline to obtain a 2%5% suspension.
Analysis: On a suitable U-bottom microtiter plate, place 1 [CautionDrabkins reagent and Drabkins solution contain
drop each of 0.9% saline and Anti-D (Rho) reagent in two cyanide and are HIGHLY TOXIC. Do not inhale, swallow,
separate wells. Label them as the B well (Blank) and the T or allow contact with skin or with eyes. Wear suitable
well, respectively. Add 1 drop of Sample solution to each protective clothing, gloves, and eye and face protection.
well, and mix by gently tapping the side of the plate. Do not mix with acids. Contact with acids liberates a very
Centrifuge the plate at appropriate conditions established toxic gas. If ingested, perform gastric lavage, and
for the centrifuge. Resuspend the cell buttons by manually immediately call a physician.]
tapping the plate or with the aid of a suitable mechanical Blank solution: Water
shaker. Read the optical densities of the wells using a Standard solution A: 300 mg USP Hemoglobin RS in a 2-
suitable automated photometric microtiter plate reader, mL volumetric tube, add 1 mL water, then dissolve in and
and determine if the Red Blood Cells in the T well is dilute with water to volume
agglutinated as described under Identification test A. If the Standard solution B: Mix 50 L of Standard solution A with
T well indicates negative results (no hemagglutination), 25 L of water.
incubate the plate at 37 for 15 min, centrifuge, resuspend Standard solution C: Mix 50 L of Standard solution A with
the cells, and read the optical densities of the wells as 100 L of water.
above. Agglutination of cells after immediate-spin or after Sample solution: Mix 50 L of Red Blood Cells with 50 L
37 incubation indicates Rh positive typing of Red Blood of water.
Cells. The test is valid if cells in the B well are not Analysis: Label suitable tubes as B1, B2, SA1, SA2, SB1,
agglutinated. If the cells are not agglutinated in the T well, SB2, SC1, SC2, T1, and T2. Add 5.0 mL of Drabkins solution
proceed as directed in Test 2. to each tube. Add 20 L of water to each of B1 and B2, 20
Test 2 L of Standard solution A to each of SA1 and SA2, 20 L of
Anti-D reagent: Use an anti-D blood grouping reagent Standard solution B to each of SB1 and SB2, 20 L of
approved for use for a weak D blood group test. Standard solution C to each of SC1 and SC2, and 20 L of
Antihuman globulin reagent: Use a polyspecific or anti- Sample solution to each of T1 and T2, rinsing the pipet tip
IgG antihuman globulin reagent. Dilute, if necessary, three to four times with Drabkins solution. Allow to stand for
following the manufacturers instructions. at least 15 min at room temperature. [NOTERed Blood
Control solution: Use IgG-coated red cells approved for Cells with appreciable carboxyhemoglobin content, such as
use as a control for Rh typing. Dilute with 0.9% saline to those obtained from smokers, may require longer reaction
obtain a 2% solution. time. If the donor characteristics are not known, the
Sample solution: Prepare as directed for Test 1. incubation time should be optimized prior to testing.] Read
Analysis: Place 1 drop of 0.9% saline into a suitable test the absorbances of the solutions against the solution in tube
tube and 1 drop of Anti-D reagent in another, and mark B1 at 540 nm. The absorbance of the solution in tube B2 is
recorded at the end. The test is not valid if the absorbance after the blood has been drawn. Red Blood Cells in
of the solution in tube B2 is not within 0.005. Anticoagulant Citrate Phosphate Dextrose Adenine Solution may
Calculations: Calculate the concentrations, in mg/mL, of be stored for up to 35 days at 16. Red Blood Cells may
hemoglobin in Standard solutions A, B, and C. Plot a be stored in an approved additive solution (AS), for up to
calibration curve of absorbance values against the 42 day at 16. If the hermetic seal of the container is
hemoglobin concentration by drawing a best-fit straight line broken during collection, solution, or further processing, the
using the least-square linear regression analysis. From the expiration date is not later than 24 h after the seal is broken
absorbance value of the Sample solution, obtain the (when blood is stored at 16). The expiration date for
concentration, in mg/mL, of hemoglobin in the Sample frozen Red Blood Cells prepared with low glycerol content
solution. Multiply the value by 2 to obtain the concentration, (20% glycerol) is not later than 10 years from the date of
in mg/mL, of hemoglobin in Red Blood Cells. collection when stored at 120 or colder, except when Red
Calculate the total hemoglobin content in the Red Blood Blood Cells is prepared for freezing with high glycerol
Cells unit, in g: content (40% glycerol), in which case it may be stored at
65 or colder for no later than 10 years from date of
Result = Conc. of hemoglobin (mg/mL) the volume of the collection. If the frozen Red Blood Cells is processed for
Red Blood Cells unit (mL)/103 freezing or for thawing, in an open system, the expiration
date for the thawed Red Blood Cells is 24 h after removal
LEUKOCYTE COUNT from 65 storage, provided it is then stored at the
Sample solution: Pipet 40 L of a suitable red cell-lysing temperature of unfrozen Red Blood Cells.
agent into a clean test tube, add 100 L of Red Blood Cells LABELING: Label the container to indicate the volume of the
diluted with 0.9% saline, if necessary, such that the whole human blood collected from the donor, the collection
hematocrit of Red Blood Cells is not greater than 60%. date, the donation number or other coding means to
Analysis: Mix by pipetting up and down several times. Add uniquely identify the unit and to provide traceability to the
360 L of 0.01% (w/v) crystal violet in 15% acetic acid into donor, and the expiration date. Label it to indicate the type
the mixture, and mix thoroughly. Fit a hemocytometer with of anticoagulant used to collect whole human blood and any
a 50-L counting volume and a bright background, with a additive solutions added subsequent to collection. Label it
cover slip, and load the counting chamber with the mixture also to identify donor status (i.e., volunteer or paid). Label it
until the counting area is completely covered but not also with the following statements: See Circular of
overfull. Cover the counting chamber with a suitable moist Information for indications, contraindications, cautions, and
lid to prevent evaporation, and allow to settle undisturbed methods of infusion; Properly identify recipient; and
for 1015 min. Remove the lid, and place the chamber on Caution: Rx only. In addition, label it to indicate the
the stage of a light microscope fitted with 10 ocular lens product name as indicated in Table 1. [NOTEThe name is
and 20 objective. Count the leukocytes in the entire 50-L determined by the method of solution of the Red Blood
counting volume. Calculate the leukocyte count in Red Cells (derived from whole human blood or from apheresis)
Blood Cells, expressed in leukocytes/L, by dividing the and by performing the necessary testing to ensure that the
observed leukocyte count by 10. product meets the minimum requirements for the named
Calculate the total number of leukocytes in the Red Blood products, as indicated in Table 1.]
Cells unit:
Result = leukocytes/L 103 the volume of the Red Blood Table 1. Red Blood Cells Solutions
Cells unit (mL) Product Name Method of Solution
Red Blood Cells Prepared from whole human blood
ADEQUACY OF DEGLYCEROLIZATION (for Red Blood Cells,
Deglycerolized): Interrupt the last wash cycle of the Red Blood Cells, Pheresis Prepared using automated apheresis
deglycerolization process at a point where the wash fluid is systems
visible in the clear tubing segment leading to the waste Red Blood Cells, Leukocyte Prepared from Red Blood Cells or Red
receptacle. Hold the tubing against a well-lighted, white Reduced Blood Cells, Pheresis (total leukocytes
background. Note the coloration of the fluid in the tubing, count <5 106)
and compare it to a suitable hemolysis color comparator
Red Blood Cells, Frozen Prepared from Red Blood Cells or Red
standard. The color of the fluid should be no stronger than
Blood Cells, Pheresis suspended in
the block indicating 3% hemolysis. [NOTEIf the level of
cryoprotective solution (glycerol) and
hemolysis is more than 3%, continue the wash process, and
frozen at an appropriate temperature
repeat the test until the color is within acceptable limits.]
Red Blood Cells, Prepared from Red Blood Cells, Frozen,
ADDITIONAL REQUIREMENTS Deglycerolyzed from which glycerol is removed by
PACKAGING AND STORAGE: Collect into an approved washing by an approved procedure
container (see Transfusion and Infusion Assemblies and Similar
Medical Devices 161) containing a sterile, pyrogen-free, Label it to indicate, the ABO Group/Rh Type, as indicated in
approved anticoagulant (see Anticoagulant Citrate Dextrose Table 2. [NOTEEvery Red Blood Cell product must have a
Solution, Anticoagulant Citrate Phosphate Dextrose, or determination made as to its ABO Group and Rh Type and
Anticoagulant Citrate Phosphate Dextrose Adenine Solution). An specificity of unexpected red cell antibodies, if any]
approved additive solution may be added after removal of If an ABO blood group color scheme is used, use the
the plasma. Store Red Blood Cells in the original container, following labeling color: Group A (yellow), Group B (pink),
or transfer to an equivalent container using a technique that Group O (blue), and Group AB (white).
does not compromise sterility. Liquid Red Blood Cells is Label the Red Blood Cells with names of the adventitious
stored at a temperature between 1 and 6 (see General agents tested and the results of the tests. If it has been
Notices and Requirements). Frozen Red Blood Cells is stored issued prior to determination of the test results, label it also
at 65 or colder.
AS contains sodium chloride, dextrose, adenine, and other substances that
EXPIRATION DATE: Red Blood Cells in Anticoagulant Citrate
Dextrose Solution, in Anticoagulant Citrate Phosphate Dextrose support red cell survival and function. Examples of such solutions are AS-1,
Solution, or in Anticoagulant Citrate Phosphate Dextrose- AS-3, and AS-5.
Dextrose Solution may be stored for up to 21 days at 16
with a warning Donor Untested and to specify Analysis: Place a small drop of the diluted platelets onto
Uncrossmatched Blood, when appropriate. the end of a clean glass microscope slide. Obtain a second
clean glass microscope slide, and draw its edge across the
Table 2. Blood Group and Rh Type drop of platelets so that capillary action spreads the drop
across the first slide. Push the second slide in one smooth
ABO Group Rh Type motion across the first slide to make a smear of platelets.
A Positive Allow the smear to dry. Apply a liberal amount (1 to 2 mL)
A Negative
of Wrights stain to the platelet smear, and allow to stand
for 2 min. Dip the slide in deionized water, and gently blot
B Positive dry with absorbent paper. Examine the slide using a
B Negative microscope at 100 with bright illumination.
AB Positive
Acceptance criteria: Platelets appear as anuclear round or
oval shapes approximately 0.1 to 0.2 m in diameter, with
AB Negative some fine purple granulation. The platelets may occasionally
O Positive appear to have a purple center with clear cytoplasm at the
O Negative
periphery
SPECIFIC TESTS
USP REFERENCE STANDARDS 11 PLATELET COUNT: Use a commercially available, validated
USP Hemoglobin RS hematology analyzer to determine platelet count. Proceed as
directed in the instruments operating manual
RESIDUAL LEUKOCYTE COUNT:
Platelets Sample solution: Transfer 100 L of Platelets into a suitable
test tube and add 400 L of 0.01% (w/v) crystal violet in
(Comment on this Monograph)id=m763=Platelets=B- 15% (v/v) acetic acid, and mix thoroughly.
Monos.pdf) Analysis: Fit a hemocytometer with a 50-L counting
DEFINITION volume and a bright background with a cover slip. Load the
Platelets is the portion of blood that contains platelet cells. It is counting chamber with the mixture until the counting area
derived from human whole blood from which red blood cells is completely covered, but do not overfill. Cover the
and a portion of the plasma are removed by centrifugation, counting chamber with a suitable moist lid to prevent
sedimentation, or apheresis. In the apheresis removal method, evaporation, and allow to settle undisturbed for 10 to 15
the red blood cells and plasma are automatically removed and min. Remove the lid, and place the chamber on the stage of
returned directly to the donor. Platelets derived from whole a light microscope fitted with a 10 ocular lens and 20
blood must be prepared within 4 h after collecting the whole objective. Count the leukocytes in the entire 50-L counting
blood from which it is derived, or within the time frame volume. Calculate the leukocyte count in the platelets,
specified for the blood collecting, processing, and storage expressed in leukocytes/L, by dividing the observed
system used. leukocyte count by 10.
Platelets may be derived from whole blood collected in any Calculate the total number of leukocytes in the red blood
approved anticoagulant solution (see USP monographs for cell unit.
anticoagulant solutions). Platelets prepared by apheresis must Total leukocytes = leukocytes/L 103 volume of the
be collected using Anticoagulant Citrate Dextrose Solution A platelet unit in mL
as the anticoagulant solution. PH 791 (for stored Platelets): Transfer aseptically a volume
Platelets derived from whole blood should have a minimum of appropriate for the equipment: the pH must be greater than
5.5 1010 platelet cells suspended in a volume of 40 to 70 mL 6.2 throughout the storage period.
of original plasma. Platelets produced by apheresis should ADDITIONAL REQUIREMENTS
have a minimum of 3.0 1011 platelet cells, suspended in 100 PACKAGING AND STORAGE: Store Platelets in an approved
to 500 mL of original plasma or in an approved additive container. Platelets may be stored in plasma or in an
solution. approved additive solution at 20 to 24 with continuous
Platelets derived from whole blood or by apheresis may be gentle agitation for no more than 5 days after date of
further processed by filtration for removal of leukocytes, or by preparation.
irradiation to inactivate lymphocytes. Platelets derived from LABELING: Label the container to indicate the collection
whole blood may be pooled from multiple donors to form one date, the donation number or other coding means to
dose of platelets. uniquely identify the unit and to provide traceability to the
Leukocytes may be removed from platelets by filtration, using donor, approximate volume, storage temperature, and its
an approved platelet leukoreduction filter. Platelets derived expiration date. Indicate the anticoagulant solution used and
from whole blood must contain less than 8.3 105 leukocytes any additive solutions added subsequent to collection. Also
after filtration. label the container to identify donor status (for example,
The source blood for platelets must be tested for syphilis, volunteer or paid). Label it also with the following
hepatitis B, and human T-cell Lymphotropic Virus (HTLV) Type statements: See Circular of Information for the Use of
I and Type II, using FDA-approved and -licensed commercially Human Blood and Blood Components for indications,
available test kits. The test results must be below the limits of contraindications, cautions, and methods of infusion;
detection specified by the manufacturers of the respective test Properly identify intended recipient; This product may
kits. The source blood must also be tested for hepatitis C and transmit infectious agents; and Rx only. In addition, label
HIV Type 1 and Type 2, using FDA-approved nucleic acid the container to indicate the product name as indicated in
assays. The test results must be below the approved limits of Table 1. [NOTEThe name is determined by the method of
detection for the tests used. platelets solution (derived from whole blood or by apheresis)
IDENTIFICATION and by performing the necessary testing to ensure that the
PROCEDURE product meets the minimum requirements for the named
Sample solution: Dilute a small volume of Platelets 1:1000 products, as indicated in Table 1.].
with 0.9% sodium chloride solution
USP 32 Official Monographs / Bovine 115
Table 1. Names of Platelet Solutions plastic, and other reconstructive procedures to contribute to
Product Name Method of Solution
the repair, reinforcement, and generation of tissue. The sterile
material is surgically secured, onlayed, and/or packed into
Platelets Prepared from a single unit of whole human deficient soft tissues such as skin, tendon, muscle, and dura
blood within 8 h of collection. mater.
Platelets, Pooled Individual platelet units derived from whole The source fetal or neonatal bovine skin is mechanically and
human blood and pooled by aseptic chemically processed to isolate the dermis and remove cells
techniques. [NOTELabel this solution and cellular components. To prevent the transmission of
with a unique identifying number infectious disease, the manufacturing process has been
related to the number of individual validated to inactivate viruses potentially present in the source
units pooled, and with an expiration material. To prevent the spread of transmissible spongiform
date of 4 h after pooling of the encephalopathies, the source material is acquired from
individual units.] appropriate geographic locations in accordance with relevant
Platelets, Pheresis Prepared by apheresis from a single donor.
guidelines subject to governmental oversight. The product is
inspected and tested to assure the product meets
Platelets, Leukocyte Prepared from whole blood, either by specifications.
Reduced centrifugation or by sedimentation, and
filtered using an approved platelet IMPURITIES
leukoreduction filter to yield less than 8.3 Inorganic Impurities
105 white blood cells in the final container. ASH DETERMINATION
Platelets, Pheresis, Contains less than 5 106 white blood cells, Analysis: Place a sample of the final product, 5.0 g, in a
Leukocyte prepared by apheresis, with or without a kiln-dried, porcelain crucible. Record the weight to the
Reduced filter. nearest 0.1 mg. Place the crucible containing the sample
into an oven at 125 for 24 h. Then place the crucible
[NOTEPlatelets prepared by apheresis should be labeled containing the sample into a cool muffle furnace. Heat the
with the donors ABO blood group and Rh factors. Test the furnace to 350, and maintain the temperature until
donors whole blood or red blood cells as directed under smoking ceases (generally about 20 min). Heat the furnace
Whole Blood or Red Blood Cells, respectively.] to 550. Maintain the temperature for 2 h. Cool the
crucible in a desiccator. Weigh the crucible, and record the
weight to the nearest 0.1 mg.
Calculate the percentage of ash:
Botulism Antitoxin
(Comment on this Monograph)id=m10120=Botulism Result = [(W1 W2)/W3] 100
Antitoxin=B-Monos.pdf) W1 = weight of crucible and residue (g)
DEFINITION W2 = weight of crucible (g)
Botulism Antitoxin conforms to the regulations of the federal W3 = weight of sample (g)
Food and Drug Administration concerning biologics (See Acceptance criteria: NMT 0.3%
Biologics 1041). It is a sterile, nonpyrogenic solution of the SPECIFIC TESTS
refined and concentrated antitoxic antibodies, chiefly STERILITY TESTS 71: Meets the requirements
globulins, obtained from the blood of healthy horses that have BACTERIAL ENDOTOXINS TEST 85: It meets the requirements
been immunized against the toxins produced by the type A as directed under Transfusion and Infusion Assemblies and
and type B and/or type E strains of Clostridium botulinum. Its Similar Medical Devices 161.
potency is determined with the U.S. Standard Botulism HISTOLOGICAL EVALUATION
Antitoxin of the relevant type, tested by neutralizing activity in 1% Acid alcohol: 70% ethyl alcohol and hydrochloric acid
mice of the corresponding U.S. Control Botulism Test Toxin. It (37.5%) (99:1)
contains NMT 20.0% of solids, and contains a suitable Potassium alum solution: 100 mg/mL of potassium alum
antimicrobial agent. in distilled water, dissolve with the aid of heat and a
ADDITIONAL REQUIREMENTS magnetic stirrer
PACKAGING AND STORAGE: Preserve in single-dose containers Hematoxylinalcohol solution: 100 mg/mL of hematoxylin
only, at a temperature between 28. (see Reagents, Indicators, and SolutionsReagent
EXPIRATION DATE: The expiration date for Antitoxin Specifications) in 100% ethyl alcohol, dissolve at room
containing a 20% excess of potency is not later than 5 years temperature
after date of issue from manufacturers cold storage (5, 1 Hematoxylin solution: Slowly combine the 1000 mL of the
year; or 0, 2 years). Potassium alum solution with 50 mL of the
LABELING: Label it to state that it was prepared from horse Hematoxylinalcohol solution. Bring to a boil as rapidly as
blood. possible. Remove from heat and slowly add 2.5 g of
mercuric oxide. Return the solution to heat until it becomes
dark purple, remove from heat, and cool in a sink of cold
water.
Bovine Acellular Dermal Matrix Eosin solution: Dissolve 1.0 g of eosin Y, water soluble, in
(Comment on this Monograph)id=m1676=Bovine Acellular 100 mL of distilled water. Dissolve 1.0 g of phloxine B in
Dermal Matrix=B-Monos.pdf) 100.0 mL of distilled water. Combine 100 mL of eosin Y
solution with 10 mL of phloxine B solution, 780 mL of 95%
DEFINITION ethyl alcohol, and 4.0 mL of glacial acetic acid. [NOTEFilter
Bovine Acellular Dermal Matrix is a remodelable collagen daily before use.]
scaffold derived from fetal or neonatal bovine skin. It is Bluing agent: 15.4 mg/mL of lithium carbonate in distilled
presented to the physician as a flat white sheet that is cut to water
size and hydrated in room temperature sterile saline solution 10% Neutral buffered formalin: To 6.5 g of dibasic
prior to implantation. It is utilized as a structural scaffold in sodium phosphate (anhydrous) and 4.0 g of monobasic
orthopedic, neurosurgical, urogynecological, dermatological,
116 Bovine / Official Monographs USP 32
sodium phosphate, add 900 mL of distilled water and 100 LIPID ANALYSIS
mL of formaldehyde (37%40%). Analysis: A standard Soxhlet extraction apparatus is
Sample solution and staining: Remove a sample of required. Dry flasks in an oven/dessicator and weigh,
finished product with an 8.0-mm biopsy punch. Place the recording the weight to the nearest 0.0001 g. Grind or cut
sample in a labeled tissue cassette, and fix for 24 h in 10% into small pieces 3.04.0 g of test material and place into a
Neutral buffered formalin. Dehydrate the sample in sequential thimble. Record the weight of the test material to the
soaks of the following: 70% ethyl alcohol (45 min), 80% nearest 0.0001 g. Place the thimble of material and 8090
ethyl alcohol (45 min), 95% ethyl alcohol (90 min), 100% mL of petroleum ether into an extraction flask, and place
ethyl alcohol (180 min), and xylene (90 min). Embed the into the Soxhlet extraction tube. Reflux for 4 h. Collect all of
sample in melted paraffin, cool, and cut 5-m thick sections the ether into the flask, and evaporate. Weigh the flask,
with a microtome. Collect sections on microscope slides. recording weight to the nearest 0.0001 g.
Deparaffinize the slide with xylene and hydrate with distilled Calculation: For the weight of lipid, substract the weight of
water. Stain in Hematoxylin solution for 615 min. Wash in the clean flask from the final weight of the flask. Calculate
running tap water for 25 min. Dip two times in 1% Acid the percent of lipid based on the weight of the starting
alcohol. Wash briefly in tap water. Place in Bluing agent until material.
the sections are bright blue. Wash in running tap water for Acceptance criteria: The percentage of lipid in 3.04.0 g of
10 min. Place in 80% ethyl alcohol for 12 min. Dehydrate Dermal Matrix sample is between 0% and 1.5%.
and clear through two changes each of 95% ethyl alcohol, MOISTURE CONTENT
100% ethyl alcohol, and xylene, 2 min each. Affix a Analysis: Proceed as directed under Loss on Drying 731 to
coverslip over the tissue using an appropriate resinous calculate the moisture content, with the following specifics.
mounting media. The nuclei stains blue, the cytoplasm Mince approximately 5.0 g of Dermal Matrix; place it into
stains from pink to red, and the collagen fibers stain from an aluminum dish. Dry the sample in an air oven for 1618
pink to red. h at 100102.
Microscopic and morphological characteristics: The Calculate the percentage of moisture in the sample taken:
collagen fibers of the Dermal Matrix stain pink-red, and no
evidence of cell nuclei or cytoplasm are apparent in Result 1 = [(W1 W2)/W3] 100
prepared histological sections as shown in the USP Bovine
Acellular Dermal Matrix Reference Photomicrographs of W1 = weight of dried sample and pan (g)
products with acceptable histological appearance. W2 = weight of pan (g)
PROTEIN DETERMINATION: Use the Kjeldahl nitrogen (protein) W3 = weight of sample (g)
determination method to calculate the percent protein of
the final product as directed under Nitrogen Determination Result 2 = 100 Result 1
461 with the following specifics. Suitable equipment and
procedures are readily available1. Acceptance criteria: 10.0%12.0% of the original sample
Digestion: Prepare a rack of 1520 Kjeldahl digestion tubes. weight
In each, place 2.02.2 g of final product, 0.2 0.05 g of CARBOHYDRATES
ammonium sulfate, a metallic catalyst tablet,2 and boiling Analysis: The percentage of carbohydrates is determined:
chips.3 Prepare a blank tube with catalyst tablets and boiling Carbohydrate% = 100% (lipid% + protein% + moisture% +
chips (reagent blank). To each tube add 15 mL of ash%)
concentrated sulfuric acid, and then, very slowly, 3 mL of
hydrogen peroxide (30%35%). Place the digestion tubes Acceptance criteria: The percentage of carbohydrates is
on a digestion block, and heat to 410. Digest for 60 5 equal to or less than 0.0%. Because this is a calculated
min. The mixture in the tubes should be a clear green. value, influenced by the error inherent in the test methods
Distillation: Add excess base (50% sodium hydroxide). above (Lipid analysis, Moisture content, and Ash
Generally, for each 5 mL of concentrated sulfuric acid used determination), a calculated value less than 0.0% is
in the digestion, 20 mL of 40% (w/w) sodium hydroxide is acceptable.
required to make the digest strongly alkaline (pH > 11). Mix GEL ELECTROPHORESIS: Use the electrophoresis determination
each tube and let cool to room temperature. Distill each method as directed under Biotechnology-Derived Articles
tube to collect approximately 125 mL of total distillate in a Polyacrylamide Gel Electrophoresis 1056 with the following
flask containing 25 mL of 4% boric acid. A reagent blank is specifics.
run with each set. Collagen extraction solution: Prepare a 0.5M acetic acid
Titration: Titrate the collected distillate with standardized solution containing 2 mM ethylenediaminetetraacetic acid
0.2 N sulfuric acid to a neutral gray color endpoint. Record (EDTA).
the volume of sulfuric acid used. 2X Tris-glycine sample buffer: Prepare a 2X solution
Analysis: Calculate the percentage of protein: containing 63 mM Tris-HCl pH 6.8, 10% glycerol, 2%
sodium dodecyl sulfate (SDS), 0.05% 2-mercaptoethanol,
Result = [(S1 B) S2 W1 P]/W2 and 0.25% bromophenol blue4.
S1 = sulfuric acid (mL) 1X Sample buffer: 2X Tris-glycine sample buffer and water
B = blank (mL) (1: 1)
S2 = N of sulfuric acid SDS-PAGE running buffer: Prepare a solution containing
W1 = milliequivalent weight N 100 (%), 1.4007 25 mM Tris pH 8.3, 192 mM glycine, and 0.1% SDS5.
P = protein factor for meat, 6.25 Polyacrylamide gel: Prepare a Tris-HCl polyacrylamide gel
W2 = weight of sample (g) with a 4% to 20% gradient6.
Acceptance criteria: The percentage of protein in 2.02.2 g 4A suitable sample buffer can be obtained from Invitrogen Corporation, 1600
of Dermal Matrix sample is between 90.0% and 95.0%. Faraday Ave., P.O. Box 6482, Carlsbad, CA 92008.
5A suitable gel running buffer can be obtained from Bio-Rad Laboratories,
1A suitable device and associated procedures can be obtained from
1000 Alfred Nobel Dr., Hercules, CA 94547.
Labconoco, 8811 Prospect Ave., Kansas City, MO. 6A suitable precast acrylamide gel can be obtained from Bio-Rad Laboratories,
2A suitable catalyst is Pro-Pac CT-37, Alfie Packers, 8901 J St., Omaha, NE.
3Commonly referred to as Henger granules. 1000 Alfred Nobel Dr., Hercules, CA 94547.
USP 32 Official Monographs / Bovine 117
Molecular weight marker: Use a suitable molecular weight T = thickness (mm)

marker containing protein bands between 10 and 250 Acceptance criteria: The measured tensile strength for each
kilodaltons (kDa). lot is NLT 5 N/mm2.
Staining solution: Prepare a solution containing 0.25% SUTURE RETENTION FORCE
(w/v) Coomassie brilliant blue R-250 (see Reagents, Analysis: Cut representative 1 1 cm samples from final
Indicators, and SolutionsReagent Specifications) in 10% product lots. Using an appropriate suture material (e.g., 40
acetic acid and 10% npropanol. polypropylene suture), thread the suture 3 mm from the
Destain solution: Water, acetic acid, and npropanol (8:1:1) edge of the sample in the center and pull through. Clamp
Collagen preparations: Mince 0.5 g of Dermal Matrix final approximately 5 mm of the opposite, unsutured end of the
product. Weigh a sample of minced Dermal Matrix, and add sample in the upper pneumatic grip of a commercially
to a volume of Collagen extraction solution to obtain a available material test system.8 The suture tails are hanging
concentration of 5 mg/mL (dry weight of Dermal Matrix). freely. Clamp the suture tails to the lower grip. Pull the grips
Extract on a rocking platform at room temperature for 72 h. apart at 20 mm/min while concurrently measuring the force
Analysis: Dilute acid-extracted collagen samples in 2X exerted. Record the maximum force (N) measured.
Trisglycine sample buffer to a concentration of 0.5 mg/mL, Acceptance criteria: The suture retention force measured
and incubate for 5 min at 100. Load the Polyacrylamide gel for each lot is NLT 5N for a 1 1 cm sample of the Dermal
in the electrophoresis apparatus, and add SDS-PAGE running Matrix.
buffer to the top and bottom reservoirs. Load 10 L of THERMAL ANALYSIS
Molecular weight markers in the first well of the Analysis: A final product sample of approximately 1020
Polyacrylamide gel and 10 L of 1X Sample buffer in the mg is heated at 2/min from 3090, hydrated with water,
second well. Load 10 L (5 g) of each collagen sample into and the thermal characteristics of each processed skin is
subsequent gel wells. Attach the cathode and anode to the measured with a differential scanning calorimeter as directed
appropriate terminals, and apply 110 V to the gel. Run the under Thermal Analysis 891. Dermal Matrix displays a
gel until the bromophenol blue reaches the bottom of the single thermal transition peak between 58 and 67.
gel. Remove the gel from the electrophoresis apparatus, and VISUAL INSPECTION
place it in a tray containing enough Staining solution to Analysis: Each piece of final product is visually inspected
cover the gel. Incubate the gel for 3 h on a rocker at room under a white light at a distance of 3045 cm for color, the
temperature. Completely remove the Staining solution from presence of particulates, and holes. Dermal Matrix is white,
the tray, cover the gel with Destain solution, and slowly rock and neither particulates nor holes are visible.
the gel for 20 min. Remove the Destain solution, and repeat HYDRATION RATE
the destaining procedure three times. Inspect the gel for Analysis: Cut a sample of finished product lot
bands that have migrated from the origin. approximately 1 1 cm. The sample fully hydrates, as
System suitability: All bands between 20 and 200 kDa are indicated by a change in color from white to gray, in less
present. The lane containing 1X Sample buffer does not than 3 min when placed in room temperature saline
contain any bands. solution.
Data analysis: Where a protein band appears in the gel,
the molecular weight of this protein is determined by ADDITIONAL REQUIREMENTS
comparing the position of the band to that of the known PACKAGING AND STORAGE: The package is a sealed, foil
Molecular weight marker. pouch that provides an effective moisture, light, gas, and
Specificity: The lanes of the Polyacrylamide gel that sterility barrier. Store in clean, dry conditions between 15
correspond to Dermal Matrix show four major protein and 30.
bands. Two bands, when compared to the Molecular weight LABELING: Label it to indicate that it is derived from bovine
marker, appear at 96 and 94 kDa. These two bands origin. The product is labeled to indicate the products
correspond to the monomeric alpha 1 and alpha 2 chains of intended clinical use. It is labeled with the dimensions of the
collagen Type I, respectively. Another two bands appear product, the expiration date, the required storage
close together at 200 kDa, which correspond to alpha 1 and conditions, lot number, part number, and the manufacturers
alpha 1/alpha 2 collagen dimers. name and address. The label indicates that the product is
TENSILE STRENGTH sterile and nonpyrogenic and is designed for single patient,
Analysis: Cut samples 5-mm wide 50-mm long from one-time use. The labeling cautions the user to inspect the
representative pieces from final product lots. Measure the packaging for damage and to discard the product if the
thickness of the sample. Test the samples with a packaging has been compromised. The labeling also cautions
commercially available material test system.7 Mount and the user to hydrate the product only in room temperature
align the specimen, gripping 1 cm of the sample on both sterile saline solution.
ends to ensure a sample gauge length of 3 cm. Pull the USP REFERENCE STANDARDS 11
grips apart at 30 mm/min while concurrently measuring the USP Endotoxin RS
force exerted on the sample. Record the maximum force (N) USP AUTHENTIC VISUAL REFERENCES 11
measured during the test. USP Bovine Acellular Dermal Matrix Reference
Calculate the tensile strength (n/mm2): Photomicrographs. These Photomicrographs show the
histological appearance of failed, cell-containing source
Result = F/W T material (Photomicrographs 1 and 2) and of passing,
processed, decellularized material (Photomicrographs 3 and
F = maximum force (N) 4). The samples were prepared as directed in the test for
W = width, 5 mm Histological evaluation.
7Asuitable material test system is available from Instron Corporation, 825 8Asuitable material test system is available from Instron Corporation, 825
University Ave., Norwood, MA. University Ave., Norwood, MA.
118 Bretylium / Official Monographs USP 32
Bretylium Tosylate from the Sample solution is NMT two times the bretylium
(Comment on this Monograph)id=m10170=Bretylium response from the Standard solution (2%); and no
Tosylate=B-Monos.pdf) individual peak response is greater than that of the
bretylium peak from the Standard solution (1%).
SPECIFIC TESTS
LOSS ON DRYING 731: Dry it in a vacuum at 75 for 2 h: it
at 25, excursions permitted between 15 and 30.
C18H24BrNO3S 414.36 USP REFERENCE STANDARDS 11
Benzenemethanaminium, 2-bromo-N-ethyl-N,N-dimethyl-, salt USP Bretylium Tosylate RS
with 4-methylbenzenesulfonic acid (1:1);
(o-Bromobenzyl)ethyldimethylammonium p-toluenesulfonate
[61-75-6; 59-41-6].
DEFINITION
Bretylium Tosylate Injection
Bretylium Tosylate contains NLT 98.0% and NMT 101.0% of (Comment on this Monograph)id=m10180=Bretylium Tosylate
C18H24BrNO3S, calculated on the dried basis. Injection=B-Monos.pdf)
A. INFRARED ABSORPTION 197K Bretylium Tosylate Injection is a sterile solution of Bretylium
B. The retention time of the major peak of the Sample Tosylate in Water for Injection. It contains NLT 90.0% and
solution corresponds to that of the Standard solution, as NMT 110.0% of the labeled amount of C18H24BrNO3S.
obtained in the Organic ImpuritiesProcedure. IDENTIFICATION
ASSAY The retention time of the major peak of the Sample solution
PROCEDURE corresponds to that of the Standard solution, both relative to
Sample solution: 6 mg/mL of Bretylium Tosylate in dioxane the internal standard, as obtained in the Assay.
Titrant: 0.025 N perchloric acid in dioxane, standardized as ASSAY
described in perchloric acid, tenth-normal in dioxane VS PROCEDURE
Analysis: To 50 mL of Sample solution, add 2 drops of Solution A: 1.38 mg of monobasic sodium phosphate and
crystal violet TS, and titrate with Titrant to a blue-green 2.0 mL of 25% tetra-methylammonium hydroxide solution
endpoint. Perform a blank determination (see Titrimetry in methanol in 800 mL of water, adjust with phosphoric
541), and make any necessary correction. Each mL of acid to a pH of 3.1 0.1, dilute with water to 1000 mL
Titrant is equivalent to 10.36 mg of C18H24BrNO3S. Mobile phase: Acetonitrile, tetrahydrofuran, and Solution A
Acceptance criteria: 98.0%101.0% (15:3:182)
IMPURITIES Standard solution: 0.2 mg/mL of USP Bretylium Tosylate
Inorganic Impurities RS
RESIDUE ON IGNITION 281: NMT 0.1% Sample solution: Equivalent to 0.2 mg/mL of bretylium
HEAVY METALS, Method I 231: NMT 20 ppm tosylate from a volume of Injection in water
Organic Impurities Chromatographic system
PROCEDURE (See Chromatography 621, System Suitability.)
Solution A: 0.01 M 1-sodium octanesulfonate Mode: LC
Mobile phase: Acetonitrile, glacial acetic acid, Detector: UV 220 nm
triethylamine, and Solution A (19:2:0.5:81) Column: 3.9-mm 30-cm; packing L1
Standard solution: 0.02 mg/mL of USP Bretylium Tosylate Flow rate: 2 mL/min
RS in Mobile phase Injection size: 20 L
Sample solution: 2 mg/mL of Bretylium Tosylate in Mobile System suitability
phase Sample: Standard solution
Chromatographic system [NOTEThe relative retention times for tosylate and
(See Chromatography 621, System Suitability.) bretylium are about 0.7 and 1.0, respectively.]
Detector: UV 265 nm Resolution: NLT 3.0, between the bretylium and tosylate
Column: 4.6-mm 25-cm; packing L11 peaks
Flow rate: 1.9 mL/min Relative standard deviation: NMT 1.4%
Sample: Standard solution Calculate the percentage of C18H24BrNO3S in each mL of
[NOTEThe relative retention times for tosylate ion, the Injection:
o-bromobenzyldimethylamine, bretylium, m-bromo-
benzyldimethylamine, and p- Result = (rU/rS) (CS/CU) 100
bromobenzyldimethylamine are 0.25, 0.74, 1.0, 1.27, rU = peak response of bretylium in the Sample
and 1.40, respectively.] solution
Suitability requirements rS = peak response of bretylium in the Standard
Relative standard deviation: NMT 3.0% solution
Analysis CS = concentration of USP Bretylium Tosylate RS in
Acceptance criteria: The sum of the responses for all the
peaks, excluding those of the bretylium and tosylate peaks,
USP 32 Official Monographs / Bretylium 119
CU = concentration of bretylium tosylate taken to Suitability requirements

prepare the Sample solution (mg/mL) Resolution: NLT 3.0, between the bretylium and tosylate
Acceptance criteria: 90.0%110.0% peaks
BACTERIAL ENDOTOXINS TEST 85: NMT 0.20 USP Endotoxin Samples: Standard solution and Sample solution
Unit/mg of bretylium tosylate Calculate the percentage of C18H24BrNO3S in each mL of
PARTICULATE MATTER IN INJECTIONS 788: Meets the the Injection:
PH 791: 3.57.0 Result = (rU/rS) (CS/CU) 100
Injections 1. rU = peak response of bretylium from the Sample
solution
ADDITIONAL REQUIREMENTS rS = peak response of bretylium from the Standard
PACKAGING AND STORAGE: Preserve in single-dose containers, solution
preferably of Type I glass. CS = concentration of USP Bretylium Tosylate RS in
USP REFERENCE STANDARDS 11 the Standard solution (mg/mL)
USP Bretylium Tosylate RS CU = concentration of dextrose injection taken to
USP Endotoxin RS prepare the Sample solution (mg/mL)
DEXTROSE
Sample solution: Equivalent to 25 g of dextrose of the
Bretylium Tosylate in Dextrose Dextrose Injection, to a 100-mL volumetric flask. Add 0.2
mL of 6 N ammonium hydroxide, and dilute with water to
Injection volume.
(Comment on this Monograph)id=m10190=Bretylium Tosylate Analysis: Determine the angular rotation in a suitable
in Dextrose Injection=B-Monos.pdf) polarimeter tube (see Optical Rotation 781).
Calculate the percentage (g/100 mL) of C6H12O6 H2O in the
DEFINITION portion of Injection:
Bretylium Tosylate in Dextrose Injection is a sterile solution of
Bretylium Tosylate and Dextrose in Water for Injection. It (Mr1/Mr2) A R (100/Sr)
contains NLT 95.0% and NMT 105.0% of the labeled amounts
of bretylium tosylate (C18H24BrNO3S) and dextrose (C6H12O6 Mr1 = molecular weight for dextrose monohydrate,
H2O). It contains no antimicrobial agents. 198.17
Mr2 = molecular weight for anhydrous dextrose, 180.16
IDENTIFICATION A = 100 mm divided by the length of the polarimeter
A. The retention time of the major peak in the Sample tube (mm)
solution corresponds to that in the Standard solution, as R = observed rotation (degrees)
obtained in the Assay, Bretylium tosylate Sr = midpoint of the specific rotation range for
B. Add a few drops of a solution (1 in 20) to 5 mL of hot anhydrous dextrose (degrees), 52.9
alkaline cupric tartrate TS: a copious red precipitate of Acceptance criteria: 95.0%105.0%
cuprous oxide is formed.
SPECIFIC TESTS
ASSAY BACTERIAL ENDOTOXINS TEST 85: NMT 0.20 USP Endotoxin
BRETYLIUM TOSYLATE Unit/mg of bretylium tosylate
Solution A: 1.38 mg of monobasic sodium phosphate and PH 791: 3.06.5
2.0 mL of 25% tetra-methylammonium hydroxide solution OTHER REQUIREMENTS: It meets the requirements under
in methanol in 800 mL of water. Adjust with phosphoric Injections 1.
acid to a pH of 3.1 0.1, dilute with water to 1000 mL.
Mobile phase: Acetonitrile, tetrahydrofuran, and Solution A ADDITIONAL REQUIREMENTS
(15:3:182) PACKAGING AND STORAGE: Preserve in single-dose glass or
Standard solution: 0.2 mg/mL of USP Bretylium Tosylate plastic containers. Glass containers are preferably of Type I or
RS Type II glass.
Sample solution: Equivalent to 0.2 mg/mL of bretylium USP REFERENCE STANDARDS 11
tosylate from a volume of Injection in water USP Bretylium Tosylate RS
Chromatographic system USP Endotoxin RS
Mode: LC
Detector: UV 220 nm
Flow rate: 2 mL/min
System suitability
[NOTEThe relative retention times for tosylate and
bretylium are about 0.7 and 1.0, respectively.]
120 Brinzolamide / Official Monographs USP 32
Brinzolamide CU = nominal concentration of Brinzolamide in the

(Comment on this Monograph)id=m10206=Brinzolamide=B- Sample solution (mg/mL)
Monos.pdf) Acceptance criteria: 98.0%102.0%
IMPURITIES
Organic Impurities
PROCEDURE 1
Mobile phase: Dehydrated alcohol, chromatographic
C12H21N3O5S3 383.51 solvent hexane, methanol, and diethylamine (55:40:5:0.2)
2H-Thieno[3,2-e]-1,2-thiazine-6-sulfonamide, 4- System suitability solution: 0.4 mg/mL of USP
(ethylamino)-3,4-dihydro-2-(3-methoxypropyl)-, 1,1-dioxide, Brinzolamide RS and 0.02 mg/mL of USP Brinzolamide
(R)-; Related Compound A RS in dehydrated alcohol
(R)-4-(Ethylamino)-3,4-dihydro-2-(3-methoxypropyl)-2H-thieno Sample solution: 0.5 mg/mL of Brinzolamide in
[3,2-e]-1,2-thiazine-6-sulfonamide 1,1-dioxide dehydrated alcohol
[138890-62-7]. Chromatographic system
DEFINITION Mode: LC
Brinzolamide contains NLT 98.0% and NMT 102.0% of Detector: UV 254 nm
C12H21N3O5S3, calculated on the dried basis. Column: 4.6-mm 25-cm; packing L51
A. INFRARED ABSORPTION 197K System suitability
B. The retention time of the major peak of the Sample Sample: System suitability solution
solution corresponds to that of the System suitability solution, [NOTEThe relative retention times for brinzolamide and
as obtained in Organic Impurities, Procedure 1. brinzolamide related compound A are 1.0 and 1.2,
respectively.]
ASSAY Suitability requirements
PROCEDURE Resolution: NLT 1.8 between brinzolamide and
Buffer: Add 4.0 mL of triethylamine to 1000 mL of water, brinzolamide related compound A
and adjust with phosphoric acid to a pH of 3.0. Column efficiency: NLT 2000 theoretical plates from
Mobile phase: Acetonitrile and Buffer (1:3) brinzolamide
Standard solution: 0.1 mg/mL of USP Brinzolamide RS in Tailing factor: NMT 1.8 for the brinzolamide peak
Mobile phase Analysis
Sample solution: 0.1 mg/mL of Brinzolamide in Mobile Sample: Sample solution
phase Calculate the percentage of brinzolamide related
Chromatographic system compound A in the portion of Brinzolamide taken:
Mode: LC Result = (rU/rS) 100
Detector: UV 254 nm
Column: 4.6-mm 25-cm; 5-m packing L1 rU = peak response for brinzolamide related
Flow rate: 1 mL/min compound A
Injection size: 20 L rS = sum of the peak responses for brinzolamide and
System suitability brinzolamide related compound A
Sample: Standard solution Acceptance criteria: NMT 0.5% of brinzolamide related
Suitability requirements compound A is found
Column efficiency: NLT 1200 theoretical plates PROCEDURE 2
Tailing factor: NMT 2.0 Buffer: Prepare as directed in the Assay.
Relative standard deviation: NMT 2.0% Mobile phase A: Prepare as directed for Mobile phase in
Analysis the Assay.
Samples: Standard solution and Sample solution Mobile phase B: Acetonitrile and Buffer (7:13)
Calculate the percentage of C12H21N3O5S3 in the portion of System suitability solution: 0.1 mg/mL of each of USP
Brinzolamide taken: Brinzolamide RS and USP Brinzolamide Related Compound
B RS in Mobile phase A
Result = (rU/rS) (CS/CU) 100 Sample solution: 1 mg/mL of Brinzolamide in Mobile
phase A
rU = peak response of the Sample solution Chromatographic system
rS = peak response of the Standard solution (See Chromatography 621, System Suitability.)
CS = concentration of USP Brinzolamide RS in the
USP 32 Official Monographs / Brinzolamide 121
Detector: UV 230 nm The retention time of the major peak of the Sample solution
Column: 4.6-mm 25-cm; 5-m packing L1 corresponds to that of the Standard solution, as obtained in
Flow rate: 1 mL/min the Assay.
Sample: System suitability solution PROCEDURE
Equilibrate the system with Mobile phase A. Buffer: 11.75 g/L of ammonium acetate in water, adjust
[NOTEThe relative retention times for brinzolamide with acetic acid to a pH of 5.2
related compound B and brinzolamide are 0.8 and 1.0, Mobile phase: Methanol and Buffer (7:13)
respectively.] Standard solution: 0.2 mg/mL of USP Brinzolamide RS in
Suitability requirements Mobile phase
Resolution: NLT 2.0 between brinzolamide and Sample solution: Transfer a volume of Ophthalmic
brinzolamide related compound B Suspension, equivalent to about 10 mg of brinzolamide, to a
Column efficiency: NLT 1200 theoretical plates from 50-mL volumetric flask, and dilute with Mobile phase to
brinzolamide volume. Nominal concentration 0.2 mg/mL
Tailing factor: NMT 2.0 for the brinzolamide peak System suitability solution: 0.06 mg/mL of USP
Analysis A Brinzolamide Related Compound B RS in Standard solution
Samples: Mobile phase A and Sample solution Chromatographic system
Allow the elution to continue for 20 min, and measure the (See Chromatography 621, System Suitability.)
areas for all the peaks, excluding the peaks of Mobile Mode: LC
phase A. Detector: UV 254 nm
Calculate the percentage of each impurity in the portion Column: 4.6-mm 15-cm; 5-m packing L1
of Brinzolamide taken: Flow rate: 1 mL/min
Result = (ri/rS) 100 System suitability
ri = peak response for each impurity [NOTEThe relative retention times for brinzolamide related
rS = sum of the responses for all the peaks compound B and brinzolamide are about 0.48 and 0.61,
Acceptance criteria: NMT 0.3% of any individual impurity respectively.]
Analysis B Suitability requirements
Equilibrate the system with Mobile phase B. Resolution: NLT 4.5 between the brinzolamide and
Samples: Sample solution brinzolamide related compound B, System suitability
Again, record the chromatograms, allowing the elution to solution
continue for 20 min, and measure the areas for Column efficiency: NLT 2500 theoretical plates, System
brinzolamide and all the peaks having a relative retention suitability solution
time greater than 6. Tailing factor: NMT 2.0, System suitability solution
Calculate the percentage of each impurity in the portion of Relative standard deviation: NMT 2.0%, Standard
Brinzolamide taken: solution
Analysis
Result = (ri/rS) 100 Samples: Standard solution and Sample solution
ri = peak response for each impurity Ophthalmic Suspension taken:
rS = sum of the responses for all the peaks
Acceptance criteria: NMT 0.3% of any individual Result = (rU/rS) (CS/CU) 100
impurity; NMT 1.0% of total impurities from Analysis A and
Analysis B rU = peak response from the Sample solution
SPECIFIC TESTS CS = concentration of USP Brinzolamide RS in the
LOSS ON DRYING 731: Dry in vacuum at 100105 for 3 h: Standard solution (mg/mL)
it loses NMT 0.5% of its weight. CU = concentration of Brinzolamide in the Sample
solution
USP REFERENCE STANDARDS 11 IMPURITIES
USP Brinzolamide RS Organic Impurities
USP Brinzolamide Related Compound A RS PROCEDURE 1
USP Brinzolamide Related Compound B RS Mobile phase: Dehydrated alcohol, hexane, methanol,
and diethylamine (55:40:5:0.2)
Brinzolamide Ophthalmic Suspension Brinzolamide RS and 0.02 mg/mL of USP Brinzolamide
Related Compound A RS in dehydrated alcohol
(Comment on this Monograph)id=m10208=Brinzolamide Sample solution: Transfer volume of Ophthalmic
Ophthalmic Suspension=B-Monos.pdf) Suspension, equivalent to about 10 mg of brinzolamide, to
a 25-mL volumetric flask. Dilute with dehydrated alcohol to
DEFINITION volume. Nominal concentration 0.2 mg/mL
Brinzolamide Ophthalmic Suspension is a sterile, aqueous Chromatographic system
suspension of Brinzolamide containing a suitable antimicrobial (See Chromatography 621, System Suitability.)
preservative. It contains NLT 90.0% and NMT 110.0% of the
labeled amount of brinzolamide (C12H21N3O5S3).
122 Brinzolamide / Official Monographs USP 32
Mode: LC PH 791: 6.58.5

Detector: UV 254 nm
Column: 4.6-mm 25-cm; packing L51 ADDITIONAL REQUIREMENTS
Flow rate: 0.75 mL/min PACKAGING AND STORAGE: Preserve in tight containers. Store
Injection size: 5 L at a temperature between 4 and 30.
Sample: System suitability solution USP Brinzolamide RS
[NOTEThe relative retention times for brinzolamide and USP Brinzolamide Related Compound A RS
brinzolamide related compound A are about 1.0 and USP Brinzolamide Related Compound B RS
1.2, respectively.]
Resolution: NLT 1.8 between brinzolamide and Bromocriptine Mesylate
brinzolamide related compound A
Column efficiency: NLT 2000 theoretical plates from (Comment on this Monograph)id=m10230=Bromocriptine
brinzolamide Mesylate=B-Monos.pdf)
Tailing factor: NMT 1.8 for the brinzolamide peak
Analysis
Calculate the percentage of brinzolamide related
compound A in the portion of Brinzolamide taken:
Result = (rU/rS) 100
rU = peak response of brinzolamide related C32H40BrN5O5 CH4SO3 750.70

compound A in the Sample solution Ergotaman-3,6,18-trione, 2-bromo-12-hydroxy-;
rS = sum of the peak responses for brinzolamide and 2-(1-methylethyl)-5-(2-methylpropyl)-;
brinzolamide related compound A in the monomethanesulfonate (salt), (5)-;
Sample solution 2-Bromoergocryptine monomethanesulfonate (salt)
Acceptance criteria: NMT 1.5% of brinzolamide related [22260-51-1].
compound A is found
PROCEDURE 2 DEFINITION
Buffer and Mobile phase: Proceed as directed in the Bromocriptine Mesylate contains NLT 98.0% and NMT 102.0%
Assay. of C32H40BrN5 O5 CH4SO3, calculated on the dried basis.
Standard solution: 2.5 g/mL of USP Brinzolamide IDENTIFICATION
Related Compound B RS in Mobile phase A. INFRARED ABSORPTION 197M: Undried
Sample solution: Use the Sample solution as prepared in B. ULTRAVIOLET ABSORPTION 197U
the Assay. Solution: 50 g/mL in 0.1 M methanolic methanesulfonic
Chromatographic system: Proceed as directed in the acid
Assay.
System suitability Proceed as directed in Assay. ASSAY
Analysis: Proceed as directed in the Assay PROCEDURE
Calculate the percentage of each impurity in the portion of Sample solution: 600 mg of Bromocriptine Mesylate into a
Ophthalmic Suspension taken: titration vessel, dissolve in 80 mL of a mixture of acetic
anhydride and glacial acetic acid (7:1)
Result = (rU/rS) (CS/CU) (Mr1/Mr2) F 100 Analysis: Titrate with 0.1 N perchloric acid VS. Perform a
blank determination, and make any necessary correction
rU = peak response for each impurity from the Sample (see Titrimetry 541). Each mL of 0.1 N perchloric acid is
solution equivalent to 75.07 mg of C32H40BrN5O5 CH4SO3.
rS = peak response for USP Brinzolamide Related Acceptance criteria: 98.0%102.0%
Compound B RS from the Standard solution
CS = concentration of USP Brinzolamide Related IMPURITIES
Compound B RS in the Standard solution Inorganic Impurities
(mg/mL) RESIDUE ON IGNITION 281: NMT 0.1%
CU = nominal concentration of brinzolamide in the HEAVY METALS, Method II 231: NMT 20 ppm
Sample solution (mg/mL) Organic Impurities
Mr1 = molecular weight of des-ethyl brinzolamide, PROCEDURE 1: LIMIT OF METHANESULFONIC ACID CONTENT: NLT
356.46 12.5% and NMT 13.4% of CH3SO3H
Mr2 = molecular weight of des-ethyl brinzolamide Sample solution: 400 mg into a titration vessel, dissolve in
oxalate, 445.49 70 mL of methanol
F = unit conversion factor, 0.001 mg/g Analysis: Titrate under nitrogen with 0.1 N methanolic
Acceptance criteria: NMT 0.5% of any individual impurity potassium hydroxide VS. Perform a blank determination,
is found and make any necessary correction. Each mL of 0.1 N
Total impurities: NMT 2.0% methanolic potassium hydroxide is equivalent to 9.61 mg
of CH3SO3H.
SPECIFIC TESTS PROCEDURE 2
STERILITY TESTS 71: It meets the requirements when tested Solution A: 0.1 N citric acid solution, adjust with
as directed for Test for Sterility of the Product to be Examined, hydrochloric acid to a pH of 2.0 and mix.
USP 32 Official Monographs / Bromocriptine 123
Diluent: Methanol and Solution A (1:1) chloride CS, ferric chloride CS, cupric sulfate CS, and dilute
Solution B: Acetonitrile and 0.01 M phosphate buffer, pH hydrochloric acid (1 in 40):
7.0 (43:57) A: 3.0:3.0:2.4:31.6
Solution C: Acetonitrile and 0.01 M phosphate buffer, pH B: 1.0:2.4:0.4:36.2
7.0 (3:2) C: 0.6:2.4:0:37.0
Mobile phase: See the gradient table below. Sample solution: 10 mg/mL of Bromocriptine Mesylate in
methanol
Time (min) Solution B (%) Solution C (%) Procedure: Compare this solution with 10-mL portions of
the Matching solutions in suitable matched tubes.
0 100 0 Acceptance criteria: The solution is clear and not darker in
18 100 0 color than Matching solutions A, B, and C.
30 0 100 SPECIFIC ROTATION 781S: +95 to +105
Sample solution: 10 mg/mL, in a mixture of methylene
40 0 100 chloride and methanol (1:1)
41 100 0 LOSS ON DRYING :
(See Thermal Analysis 891.)
System suitability solution: 2.0 mg/mL of each of - Determine the percentage of volatile substances by
ergocryptine and Bromocriptine Mesylate in Diluent thermogravimetric analysis on an appropriately calibrated
Standard solution: Dissolve USP Bromocriptine Mesylate instrument, using 10 mg of Bromocriptine Mesylate. Heat
RS in methanol, dilute quantitatively with an equal volume the specimen under test at the rate of 10/min in an
of Solution A, and dilute quantitatively and stepwise if atmosphere of nitrogen at a flow rate of 45 mL/min.
necessary, with Diluent to obtain a solution having a Record the thermogram from ambient temperature to
concentration of 4.6 g per mL. 160: it loses NMT 4.0% of its weight
Sample solution: 46 mg of Bromocriptine Mesylate, in a
10-mL volumetric flask, dissolve in 5.0 mL of methanol, ADDITIONAL REQUIREMENTS
dilute with Solution A PACKAGING AND STORAGE: Preserve in tight, light-resistant
Chromatographic system containers, in a cold place.
Mode: LC USP Bromocriptine Mesylate RS
Detector: UV 300 nm
Flow rate: 2 mL/min Bromocriptine Mesylate Capsules
System suitability (Comment on this Monograph)id=m10234=Bromocriptine
Samples: System suitability solution and Standard solution Mesylate Capsules=B-Monos.pdf)
Resolution: NLT 15 between -ergocryptine and DEFINITION
bromocriptine mesylate Bromocriptine Mesylate Capsules contain C32H40BrN5O5 CH4SO3
Tailing factor: NMT 1.5 equivalent to NLT 90.0% and NMT 110.0% of the labeled
Relative standard deviation: NMT 10.0% amount of C32H40BrN5O5.
[NOTEThe Relative retention times are about 0.46 for IDENTIFICATION
-ergocryptine and 1.0 for bromocriptine mesylate.] The principal spot of the Sample solution corresponds, in RF
Analysis value and color, to that from the Standard solution, as
Samples: Standard solution and Sample solution obtained under the Procedure for Organic Impurities.
of Bromocriptine Mesylate taken: ASSAY
PROCEDURE
Result = (ri/rS) (CS/W) F 1000 [NOTEConduct this procedure without exposure to
daylight and with minimum exposure to artificial light.]
ri = peak response for each impurity of the Sample Solution A: 0.125 mg/mL ammonium carbonate
solution Mobile phase: Acetonitrile and Solution A (3:2)
rS = peak response for bromocriptine of the Standard Standard solution: 1.0 mg/mL of bromocriptine from USP
solution Bromocriptine Mesylate RS in dehydrated alcohol
CS = concentration of USP Bromocriptine Mesylate RS Sample solution: Remove, as completely as possible, the
in the Standard solution (mg/mL) contents of NLT 10 Capsules. Weigh and determine the
W = weight, in mg, of Bromocriptine Mesylate taken average weight/Capsule. Mix the combined contents, and
to prepare the Sample solution transfer a weighed quantity of the powder, nominally
F = relative response factor is equal to 0.7 for any equivalent to 50 mg of bromocriptine, to a 50-mL
peaks eluting at a relative retention time of volumetric flask. Add 30 mL of dehydrated alcohol, and
about 0.9 or less, and is equal to 1.0 for all shake for 15 min. Dilute with dehydrated alcohol to volume,
other peaks mix, and filter. [NOTEUse this solution without delay.]
Individual impurities: NMT 0.4% of bromcriptinin is (See Chromatography 621, System Suitability.)
found; NMT 0.1% of any individual impurity is found
SPECIFIC TESTS
COLOR OF SOLUTION 631
Matching solutions: Prepare three solutions, A, B, and C,
containing, respectively, the following parts of cobaltous
124 Bromocriptine / Official Monographs USP 32
Mode: LC AU = absorbance of the Sample solution

Detector: UV 300 nm AS = Absorbance of the Standard solution
Column: 4-mm 25-cm; packing L7 CS = concentration of USP Bromocriptine Mesylate RS
Flow rate: 2 mL/min in the Standard solution (mg/mL)
Injection size: 20 L CU = nominal concentration of bromocriptine in the
System suitability Sample solution (mg/mL)
Sample: Standard solution Mr1 = molecular weight of bromocriptine, 654.59
Suitability requirements Mr2 = molecular weight of bromocriptine mesylate,
Column efficiency: NLT 1000 theoretical plates 750.70
Relative standard deviation: NMT 2.0% IMPURITIES
Analysis Organic Impurities
Samples: Standard solution and Sample solution PROCEDURE
Calculate the percentage of C32H40BrN5O5 in the Capsules [NOTEConduct this test without exposure to daylight and
taken: with minimum exposure to artificial light. Perform the test
rapidly, preparing and spotting the Sample solution last.]
Bromocriptine Mesylate RS in methanol
rU = peak response from the Sample solution Standard solutions: Equivalent to 0.06, 0.04, 0.02, and
rS = peak response from the Standard solution 0.01 mg/mL of bromocriptine (equivalent to 3.0%, 2.0%,
CS = concentration of USP Bromocriptine Mesylate RS 1.0%, and 0.5%, respectively) from Standard stock solution
in the Standard solution (mg/mL) diluted with methanol
CU = nominal concentration of bromocriptine in the Sample solution: Equivalent to 20 mg of bromocriptine
Sample solution (mg/mL) from Capsule contents, into a conical flask. Add 10 mL of
Acceptance criteria: 90.0%110.0% methanol, and stir by mechanical means for 20 min.
Centrifuge the suspension for 10 min at about 3500 rpm.
PERFORMANCE TESTS The clear supernatant is the Sample solution.
DISSOLUTION 711 Chromatographic system
Medium: 0.1 N hydrochloric acid; 500 mL (See Chromatography 621, Thin-Layer Chromatography.)
Apparatus 2: 50 rpm Mode: TLC
Time: 60 min Adsorbent: 0.25-mm layer of chromatographic silica gel
Determine the amount of C32H40BrN5O5 CH4SO3 dissolved mixture
by the following: Application volume: As 1.5-cm bands, 50-L portions of
Sample solution: Sample per Dissolution 711, passed the Standard stock solution and of each of the four
through a glass-fiber filter. Standard solutions and 50 L of the Sample solution
Standard solution: USP Bromocriptine Mesylate RS at a Developing solvent system: Methylene chloride,
known concentration in Medium. dioxane, alcohol, and ammonium hydroxide (180:15:5:1)
[NOTEA volume of alcohol not to exceed 5% of the total Spray reagent: 2-in-1000 solution of o-phthalaldehyde in
volume of the Standard solution may be used to bring the sulfuric acid
standard into solution before dilution with Medium.] Analysis
Fluorometric conditions Samples: Standard solution and Sample solution
Excitation wavelength: 315 nm Develop under the exclusion of light in a tank lined with
Emmision wavelength: 445 nm filter paper, previously equilibrated for 30 min, using
Blank: Medium Developing solvent system until the solvent front has
Analysis moved a distance of 15 cm on the plate. Dry the plate
Sample solutions: Standard solution and Sample solution briefly in a current of cold air. Spray evenly with the
Tolerances: NLT 75% (Q) of the labeled amount of Spray reagent, and view the plate under long-wavelength
C32H40BrN5O5 CH4SO3 is dissolved. UV light.
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Acceptance criteria: Any major secondary spot, other
Procedure for content uniformity: [NOTEProtect all than the principal spot, obtained from the Sample solution
solutions from light.] is not greater in size and intensity than the spot obtained
Solution A: Dissolve 1.0 g of tartaric acid in 500 mL of from the Standard solution corresponding to 3.0%, and
water, add 500 mL of methanol, and mix. any remaining spots are not greater in size and intensity
Standard solution: 0.04 mg/mL of USP Bromocriptine than the spot obtained from the Standard solution
Mesylate RS in Solution A corresponding to 1.0%. The sum of the related substances
Sample solution: Transfer the contents of 1 Capsule into a is NMT 5.0%.
25-mL volumetric flask. Add 15 mL of Solution A, and shake
by mechanical means for 20 min. Dilute with Solution A to ADDITIONAL REQUIREMENTS
volume, and mix. Filter and dilute 10.0 mL of the clear PACKAGING AND STORAGE: Preserve in tight, light-resistant
filtrate with Solution A to 50.0 mL. containers.
Spectrometric conditions USP REFERENCE STANDARDS 11
Mode: UV USP Bromocriptine Mesylate RS
Cell: 1 cm
Blank: Solution A
Analysis
Samples: Blank, Sample solution, and Standard solution
Calculate the percentage of C32H40BrN5O5 in the Capsule
taken:
Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100
USP 32 Official Monographs / Bromocriptine 125
Bromocriptine Mesylate Tablets Sample solution: Sample per Dissolution711, passed

(Comment on this Monograph)id=m10260=Bromocriptine through a glass-fiber filter.
Mesylate Tablets=B-Monos.pdf) Standard solution: USP Bromocriptine Mesylate RS at a
known concentration in Medium.
DEFINITION [NOTEA volume of alcohol not to exceed 5% of the total
Bromocriptine Mesylate Tablets contain bromocriptine mesylate volume of the Standard solution may be used to bring the
(C32H40BrN5O5 CH4SO3) equivalent to NLT 90.0% and NMT standard into solution before dilution with Medium.]
110.0% of the labeled amount of bromocriptine Fluorometric conditions
(C32H40BrN5O5). Excitation wavelength: 315 nm
Emmision wavelength: 445 nm
The principal spot from the Sample solution corresponds, in RF Sample solutions: Standard solution and Sample solution
value and color, to that from the Standard solution, as Tolerances: NLT 80% (Q) of the labeled amount of
obtained under the Procedure for Organic Impurities. C32H40BrN5O5 is dissolved.
Test 2: If the product complies with this test, the labeling
ASSAY indicates that it meets USP Dissolution Test 2.
PROCEDURE Medium: 0.1 N hydrochloric acid; 500 mL
[NOTEConduct this procedure without exposure to Apparatus 2: 50 rpm
daylight and with minimum exposure to artificial light.] Time: 30 min
Mobile phase: Acetonitrile and 0.01 M ammonium Mobile phase: Acetonitrile and 0.01 M ammonium
carbonate (13:7) carbonate (13:7)
Standard solution: 0.22 mg/mL of USP Bromocriptine Sample solution: Sample per 711 Dissolution. Dilute with
Mesylate RS in methanol Medium to concentration that is similar to the Standard
Sample solution: Transfer an equivalent to 10 mg of solution.
bromocriptine, from powdered Tablets (NLT 20 Tablets), to Standard solution: USP Bromocriptine Mesylate RS in
an appropriate container. Add 40 mL of methanol, and stir methanol, and quantitatively dilute with Dissolution
for 20 min, protected from light. Quantitatively pass Medium to obtain a solution having a known
through a fine glass filtering funnel into a 50-mL volumetric concentration similar to the expected concentration of the
flask. Rinse the filter with methanol, adding the rinsing to Sample solution
the filtrate, and dilute with methanol to volume. Chromatographic system
(See Chromatography 621, System Suitability.) Mode: LC
Mode: LC Detector: UV 300 nm
Detector: UV 300 nm Column: 3.9-mm 30-cm; packing L1
Column: 250- 4-mm stainless steel; packing L1 Flow rate: 1.5 mL/min
Injection size: 50 L System suitability
System suitability Sample: Standard solution
Sample: Standard solution Suitability requirements
Suitability requirements Relative standard deviation: NMT 2.0%
Coefficient of variation: NMT 3.0% for 3 replicate Analysis
injections, Standard solution Samples: Sample solution and Standard solution
Analysis Calculate the percentage of C32H40BrN5O5 dissolved by
Samples: Standard solution and Sample solution comparison of the peak responses from the Standard
Calculate the percentage of C32H40BrN5O5 in the portion of solution and the solution under test.
Tablets taken: Tolerances: NLT 80% (Q) of the labeled amount of
C32H40BrN5O5 is dissolved.
Result = (rU/rS) (CS/CU) 100 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
rU = peak response from the Sample solution Procedure for content uniformity: [NOTEProtect all
rS = peak response from the Standard solution solutions from light.]
CS = concentration of USP Bromocriptine Mesylate RS Solution A: Dissolve 1.0 g of tartaric acid in 500 mL of
in the Standard solution (mg/mL) water, add 500 mL of methanol, and mix.
CU = nominal concentration of bromocriptine in the Standard solution: 0.04 mg/mL of USP Bromocriptine
Sample solution (mg/mL) Mesylate RS in Solution A
Acceptance criteria: 90.0%110.0% Sample solution: Transfer 1 Tablet into a 25-mL volumetric
flask. Add 15 mL of Solution A, and shake by mechanical
PERFORMANCE TESTS means for 30 min. Dilute with Solution A to volume, and
DISSOLUTION 711 mix. Filter and dilute 10.0 mL of the clear filtrate with
Test 1: If the product complies with this test, the labeling Solution A to 50.0 mL.
indicates that it meets USP Dissolution Test 1.
Time: 60 min
Determine the amount of C32H40BrN5O5 CH4SO3 dissolved
by:
126 Bromocriptine / Official Monographs USP 32
Spectrometric conditions and any remaining spots are not greater in size and
Mode: Spectrophotometry intensity than the spot obtained from the 1.0% Standard
Analytical wavelength: 306 nm solution. The sum of the related compounds is NMT 5.0%.
Cell: 1 cm
Blank: Solution A ADDITIONAL REQUIREMENTS
Analysis PACKAGING AND STORAGE: Preserve in tight, light-resistant
Samples: Blank, Sample solution, and Standard solution containers.
Calculate the percentage of C32H40BrN5O5 in the Tablet LABELING: The labeling indicates the Dissolution Test with
taken: which the product complies.
Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 USP Bromocriptine Mesylate RS
AU = absorbance of the solution from the Tablet

CS = concentration of USP Bromocriptine Mesylate RS Bromodiphenhydramine Hydrochloride
in the Standard solution (mg/mL) (Comment on this
CU = nominal concentration of bromocriptine in the Monograph)id=m10310=Bromodiphenhydramine
solution from the Tablet, based on the labeled Hydrochloride=B-Monos.pdf)
quantity per Tablet and the extent of dilution
(mg/mL)
Mr1 = molecular weight of bromocriptine, 654.59
Mr2 = molecular weight of bromocriptine mesylate,
750.70
IMPURITIES
Organic Impurities
PROCEDURE C17H20BrNO HCl 370.71
[NOTEConduct this test without exposure to daylight and Ethanamine, 2-(4-bromophenyl)phenylmethoxy-N,N-dimethyl-,
with minimum exposure to artificial light. Perform the test hydrochloride;
rapidly, preparing and spotting the Sample solution last.] 2-(p-Bromo--phenylbenzyl)oxy-N,N-dimethylethylamine
Standard stock solution: 1.15 mg/mL of USP hydrochloride [1808-12-4].
Bromocriptine Mesylate RS in methanol
Standard solution: Equivalent to 0.1, 0.3, and 0.5 mg/mL DEFINITION
of bromocriptine (equivalent to 1.0, 3.0, and 5.0%, Bromodiphenhydramine Hydrochloride contains NLT 98.0% and
respectively) from Standard stock solution in methanol NMT 101.0% of C17H20BrNO HCl, calculated on the dried
Sample solution: Transfer an equivalent to 20 mg of basis.
bromocriptine, from powdered Tablets, to a conical flask.
Add 10 mL of methanol, and stir by mechanical means for IDENTIFICATION
20 min. Centrifuge the suspension for 10 min at about A. INFRARED ABSORPTION 197K
4000 rpm. The clear supernatant is the Sample solution. B. ULTRAVIOLET ABSORPTION 197U
Chromatographic system Analytical wavelength: 228 nm
(See Chromatography 621, Thin-Layer Chromatography.) Solution: 15 g/mL in 0.1 N sulfuric acid
Mode: TLC Acceptance criteria: Absorptivities, calculated on the dried
Adsorbent: 0.25-mm layer of chromatographic silica gel basis, do not differ by more than 3.0%.
mixture ASSAY
Application volume: As 1.5-cm bands, 10-L portions of PROCEDURE
the Standard stock solution and of each of the three Sample solution: Dissolve 700 mg of
Standard solutions and 50 L of the Sample solution Bromodiphenhydramine Hydrochloride in 50 mL of glacial
Developing solvent system: Methylene chloride, acetic acid, and add 10 mL of benzene and 15 mL of
dioxane, alcohol, and ammonium hydroxide mercuric acetate TS. Add 2 drops of crystal violet TS.
(180:15:5:0.1) Analysis: Titrate with 0.1 N perchloric acid VS to a green
Spray reagent: 2-in-1000 solution of o-phthalaldehyde in endpoint. Perform a blank determination, and make any
sulfuric acid necessary correction (see Titrimetry 541). Each mL of 0.1 N
Analysis perchloric acid is equivalent to 37.07 mg of C17H20BrNO
Samples: Standard solution and Sample solution HCl.
Proceed as directed for Chromatography 621, Thin-Layer Acceptance criteria: 98.0%101.0%
Chromatography. Dry the plate for 5 min in a current of
cold air. Develop in a tank lined with filter paper, SPECIFIC TESTS
previously equilibrated for 20 min, using Developing MELTING RANGE OR TEMPERATURE 741: 148152
solvent system until the solvent front has moved a distance LOSS ON DRYING 731
of 10 cm on the plate. Dry the plate under vacuum at Sample: Dry it at 105 for 3 h.
room temperature for 15 min. Spray evenly with the Spray Acceptance criteria: It loses NMT 0.5% of its weight.
reagent, and view the plate under long-wavelength UV
light. ADDITIONAL REQUIREMENTS
Acceptance criteria: Any spot, other than the principal PACKAGING AND STORAGE: Preserve in tight containers.
spot, from the Sample solution is not greater in size and USP REFERENCE STANDARDS 11
intensity than the spot from the 3.0% Standard solution; USP Bromodiphenhydramine Hydrochloride RS
USP 32 Official Monographs / Bromodiphenhydramine 127
Bromodiphenhydramine Hydrochloride Bromodiphenhydramine Hydrochloride

Oral Solution and Codeine Phosphate Oral Solution
(Comment on this (Comment on this
Monograph)id=m10330=Bromodiphenhydramine Hydrochloride Monograph)id=m10340=Bromodiphenhydramine Hydrochloride
Oral Solution=B-Monos.pdf) and Codeine Phosphate Oral Solution=B-Monos.pdf)
Bromodiphenhydramine Hydrochloride Oral Solution contains Bromodiphenhydramine Hydrochloride and Codeine Phosphate
NLT 93.0% and NMT 107.0% of the labeled amount of Oral Solution contains NLT 90.0% and NMT 110.0% of the
bromodiphenhydramine hydrochloride (C17H20BrNO HCl). labeled amounts of bromodiphenhydramine hydrochloride
(C17H20 BrNO HCl) and codeine phosphate hemihydrate
IDENTIFICATION (C18H21NO3 H3PO4 1/2H2O).
Sample solution: Transfer the final solution obtained from IDENTIFICATION
the titration in the Assay to a separator, add about 1 mL of A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
0.1 N sulfuric acid, and shake with 25 mL of ether. (Methyl Standard solution: 10 mg/mL of each of USP
red enters the ether phase.) Drain the aqueous layer into Bromodiphenhydramine Hydrochloride RS and USP Codeine
another separator, add 5 mL of 1 N sodium hydroxide, and Phosphate RS in methanol
shake with 10 mL of chloroform. Drain the chloroform layer Sample solution: Transfer an equivalent to 10 mg of
into a small flask containing 2 g of anhydrous sodium codeine phosphate, from a volume of Oral Solution, into a
sulfate, and swirl. Pour the chloroform solution through a separator, and add 5 mL of water, 5 mL of methylene
small cotton pledget, pre-rinsed with chloroform, into a chloride, and 1 mL of ammonium hydroxide. Shake for 1
beaker, and evaporate to 5 mL. Apply a few drops of the min, allow the layers to separate, and use the clear, lower
solution directly to a potassium bromide plate, and layer.
completely remove the chloroform by warming for 23 min Developing solvent system: Alcohol and ammonium
under an IR lamp. hydroxide (49:1)
B. The retention times of the major peaks of the Sample
ASSAY solution correspond to those of the Standard solution, as
PROCEDURE obtained in the Assay.
Sample solution: Evaporate a volume of Oral Solution
nominally equivalent to 250 mg of bromodiphenhydramine ASSAY
hydrochloride to about half the original volume, using a PROCEDURE
suitable vacuum evaporator. Transfer the concentrated Diluent: Methanol and water (4:1)
solution to a 250-mL separator, with the aid of sufficient Mobile phase: Methanol, water, 0.1 N ammonium
warm water to bring the volume to the original volume. hydroxide solution, and 0.1 N ammonium nitrate solution
Add 20 g of sodium chloride, and shake until dissolved. Add (27:3:2:1)
5 mL of 1 N sodium hydroxide, shake with 100 mL of ether, Standard solution: 100 g/mL of USP
and drain the aqueous layer into a second separator Bromodiphenhydramine Hydrochloride RS and 80 g/mL of
containing 50 mL of ether. Shake, and discard the aqueous USP Codeine Phosphate RS in Diluent
layer. Wash the ether solutions with two 20-mL portions of Sample solution: Nominally 10 mg/mL of
water, shaking each aqueous portion successively in the two bromodiphenhydramine hydrochloride and 0.08 mg/mL of
separators, and then discard the aqueous solutions. Extract codeine phosphate from Oral Solution diluted with Diluent.
the ether solutions successively with 10.0 mL of 0.1 N Chromatographic system
sulfuric acid VS, followed by two 5-mL portions of water, (See Chromatography 621, System Suitability.)
and collect the aqueous extracts in a conical flask. Add Mode: LC
methyl red TS to the solution in the flask. Detector: UV 254 nm
Analysis: Titrate the excess acid with 0.02 N sodium Column: 3.9-mm 30.0-cm; packing L3
hydroxide VS. Perform a blank determination (see Titrimetry Flow rate: 1 mL/min
541, Residual Titrations). Each mL of 0.1 N sulfuric acid is Injection size: 20 L
equivalent to 37.07 mg of C17H20BrNO HCl. System suitability
Acceptance criteria: 93.0%107.0% Sample: Standard solution
[NOTEThe relative retention times for
OTHER COMPONENTS bromodiphenhydramine and codeine are about 1.0 and
ALCOHOL DETERMINATION, Method I 611: 12.0%15.0% of 1.4, respectively.]
C2H5OH Suitability requirements
Resolution: NLT 2.0 between bromodiphenhydramine
ADDITIONAL REQUIREMENTS and codeine
containers.
USP Bromodiphenhydramine Hydrochloride RS
128 Bromodiphenhydramine / Official Monographs USP 32
Relative standard deviation: NMT 2.0% Brompheniramine Maleate

Analysis (Comment on this Monograph)id=m10390=Brompheniramine
Samples: Standard solution and Sample solution Maleate=B-Monos.pdf)
Calculate the percentage of C17H20BrNO HCl in each mL of
Oral Solution taken:
rU = peak response for bromodiphenhydramine from
the Sample solution
rS = peak response for bromodiphenhydramine from
the Standard solution C16H19BrN2 C4H4O4 435.31
CS = concentration of USP Bromodiphenhydramine 2-Pyridinepropanamine, -(4-bromophenyl)-N,N-dimethyl-, ()-,
Hydrochloride RS in the Standard solution (Z)-2-butenedioate (1:1);
(mg/mL) ()-2-p-Bromo--2-(dimethylamino)ethylbenzylpyridine maleate
CU = nominal concentration of the Sample solution (1:1) [980-71-2].
(mg/mL)
Calculate the percentage of C18H21NO3 H3PO4 1/2H2O in DEFINITION
each mL of Oral Solution taken: Brompheniramine Maleate, dried at 105 for 3 h, contains NLT
98.0% and NMT 100.5% of C16H19BrN2 C4H4O4.
IDENTIFICATION
rU = peak response for codeine from the Sample A. INFRARED ABSORPTION 197K
solution B. ULTRAVIOLET ABSORPTION 197U
rS = peak response for codeine from the Standard Sample solution: 35 g/mL in methanol
solution
CS = concentration of USP Codeine Phosphate RS in ASSAY
the Standard solution (mg/mL) PROCEDURE
CU = concentration of the Sample solution (mg/mL) Sample solution: 425 mg of Brompheniramine Maleate,
Mr1 = molecular weight of codeine phosphate previously dried, in 50 mL of glacial acetic acid. Add 1 drop
hemihydrate, 406.37 of crystal violet TS.
Mr2 = molecular weight anhydrous codeine phosphate, Analysis: Titrate with 0.1 N perchloric acid VS to a green
397.36 endpoint. Perform a blank determination, and make any
Acceptance criteria: 90.0%110.0% necessary correction (see Titrimetry 541). Each mL of 0.1 N
perchloric acid is equivalent to 21.77 mg of C16H19BrN2
OTHER COMPONENTS C4H4O4.
ALCOHOL DETERMINATION, Method II 611: 4.0%6.0% is Acceptance criteria: 98.0%100.5%
found.
IMPURITIES
MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED RESIDUE ON IGNITION 281: NMT 0.2%
MICROORGANISMS 62: It meets the requirements of the Organic Impurities
tests for absence of Salmonella species, Escherichia coli, PROCEDURE
Staphylococcus aureus, and Pseudomonas aeruginosa. The Sample solution: 40 mg/mL of Brompheniramine Maleate
total aerobic microbial count does not exceed 100 cfu/mL, in methylene chloride
and the total combined molds and yeasts count does not Chromatographic system
exceed 50 cfu/mL. (See Chromatography 621, System Suitability.)
PH 791: 4.56.5 Mode: GC
ADDITIONAL REQUIREMENTS Column: 1.2-m 4-mm; glass column containing 3%
PACKAGING AND STORAGE: Preserve in tight, light-resistant phase G3 on support S1AB
containers. Temperature: Column, 190; injection port and detector,
LABELING: Label it to indicate the alcohol content. 250
USP Bromodiphenhydramine Hydrochloride RS
USP Codeine Phosphate RS
USP 32 Official Monographs / Brompheniramine 129
Carrier gas: Dry helium CU = nominal concentration of brompheniramine

Flow rate: Adjust to get a retention time of 67 min for maleate in the Sample solution (mg/mL)
the main peak. Acceptance criteria: 90.0%110.0%
Injection size: 1 L
System suitability SPECIFIC TESTS
Sample: Sample solution BACTERIAL ENDOTOXINS TEST 85: NMT 35.7 USP Endotoxin
Suitability requirements Units/mg of brompheniramine maleate
Tailing factor: NMT 1.8 for the brompheniramine PH 791: 6.37.3
maleate peak OTHER REQUIREMENTS: It meets the requirements under
Analysis Injections 1.
Record the chromatogram for a total time of NLT twice the ADDITIONAL REQUIREMENTS
retention time of the brompheniramine peak, and PACKAGING AND STORAGE: Preserve in single-dose or in
measure the areas of the peaks. multiple-dose containers, preferably of Type I glass,
Acceptance criteria: The total relative area of all protected from light.
extraneous peaks (except that of the solvent peak and USP REFERENCE STANDARDS 11
maleic acid, if observed) is NMT 2.0%. USP Brompheniramine Maleate RS
USP Endotoxin RS
SPECIFIC TESTS
PH 791: 4.05.0, in a solution (1 in 100) Brompheniramine Maleate Oral
LOSS ON DRYING 731: Drya sample at 105 for 3 h: it loses
NMT 0.5% of its weight. Solution
(Comment on this Monograph)id=m10400=Brompheniramine
ADDITIONAL REQUIREMENTS Maleate Oral Solution=B-Monos.pdf)
containers. DEFINITION
USP REFERENCE STANDARDS 11 Brompheniramine Maleate Oral Solution contains NLT 95.0%
USP Brompheniramine Maleate RS and NMT 105.0% of the labeled amount of brompheniramine
maleate (C16H19BrN2 C4H4O4).
IDENTIFICATION
Brompheniramine Maleate Injection PROCEDURE
(Comment on this Monograph)id=m10410=Brompheniramine Proceed as directed under IdentificationOrganic Nitrogenous
Maleate Injection=B-Monos.pdf) Bases 181.
Sample solution: Equivalent to 50 mg of brompheniramine
DEFINITION maleate from a volume of Oral Solution, to a separator.
Brompheniramine Maleate Injection is a sterile solution of Render distinctly alkaline with 1 N sodium hydroxide, and
Brompheniramine Maleate in Water for Injection. It contains extract with two 50-mL portions of chloroform, shaking
NLT 90.0% and NMT 110.0% of the labeled amount of gently to avoid emulsification. Wash the combined
C16H19BrN2 C4H4O4. chloroform extracts with 10 mL of water, and discard the
aqueous phase. Filter the combined chloroform extracts into
IDENTIFICATION a conical flask, and evaporate the solvent on a steam bath,
IDENTIFICATIONORGANIC NITROGENOUS BASES 181: Meets with the aid of a current of air. To the residue add 25 mL of
the requirements dilute hydrochloric acid (1 in 1200), and proceed as
Sample solution: Equivalent to 50 mg of brompheniramine directed under IdentificationOrganic Nitrogenous Bases
maleate from a volume of Injection, with dilute hydrochloric 181, beginning with Transfer the liquid to a separator.
acid (1 in 1200) to 25 mL, and proceed as directed under Acceptance criteria: The Oral Solution meets the
IdentificationOrganic Nitrogenous Bases 181, beginning requirements.
with Transfer the liquid to a separator.
ASSAY
ASSAY PROCEDURE
PROCEDURE Sample solution: Equivalent to 20 mg of brompheniramine
Analysis: Proceed with the Injection as directed under Salts maleate from a volume of Oral Solution, to a separator.
of Organic Nitrogenous Bases 501 to prepare the solution Render distinctly alkaline with 1 N sodium hydroxide, and
used for the determination of the absorbance, AU, at 262 extract with ten 10-mL portions of chloroform, shaking
nm. For the determination of AS, dissolve 25 mg of USP gently to avoid emulsification. Wash the combined
Brompheniramine Maleate RS in 20 mL of dilute sulfuric acid chloroform extracts with 10 mL of water, wash the latter
(1 in 350), and treat this solution the same as the portion of with 20 mL of chloroform, and discard the aqueous phase.
Injection being assayed. Quantitatively filter the combined chloroform extracts and
Calculate the quantity, as a percentage, of C16H19BrN2 washings into a conical flask, and evaporate the solvent on a
C4H4O4 in the Injection taken: steam bath, with the aid of a current of air. To the residue
add 25 mL of glacial acetic acid and 5 mL of acetic
Result = (AU/AS) (CS/CU) 100 anhydride, agitate, and allow to stand for about 15 min.
Add 1 drop of crystal violet TS.
AU = absorbance of Brompheniramine Maleate from Analysis: Titrate with 0.01 N perchloric acid VS to a blue-
the Sample solution green endpoint. Perform a blank determination, and make
AS = absorbance of USP Brompheniramine Maleate any necessary correction (see Titrimetry 541). Each mL of
from the Standard solution 0.01 N perchloric acid is equivalent to 2.177 mg of
CS = concentration of USP Brompheniramine Maleate brompheniramine maleate.
130 Brompheniramine / Official Monographs USP 32

OTHER COMPONENTS PERFORMANCE TESTS
ALCOHOL DETERMINATION, Method I 611: 2.7%3.3% of DISSOLUTION 711
C2H5OH Medium: Water; 500 mL
SPECIFIC TESTS Time: 45 min
PH 791: 2.53.5 Standard solution: USP Brompheniramine Maleate RS at a
known concentration in Medium
ADDITIONAL REQUIREMENTS Sample solution: Sample per 711 Dissolution, suitably
PACKAGING AND STORAGE: Preserve in well-closed, light- diluted with 3 N hydrochloric acid.
resistant containers. Spectrometric conditions
USP REFERENCE STANDARDS 11 Mode: UV
USP Brompheniramine Maleate RS Analytical wavelength: 264
Cuvette: 5 cm
Analysis
Brompheniramine Maleate Tablets Samples: Standard solution and Sample solution
(Comment on this Monograph)id=m10420=Brompheniramine C16H19BrN2 C4H4O4 is dissolved.
Maleate Tablets=B-Monos.pdf) UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Brompheniramine Maleate Tablets contain NLT 95.0% and PACKAGING AND STORAGE: Preserve in tight containers.
NMT 105.0% of the labeled amount of C16H19BrN2 C4H4O4. USP REFERENCE STANDARDS 11
IDENTIFICATION USP Brompheniramine Maleate RS
Tablets meet the requirements under IdentificationOrganic
Nitrogenous Bases 181.
ASSAY Brompheniramine Maleate and
PROCEDURE Pseudoephedrine Sulfate Oral Solution
Sample solution: Weigh and finely powder NLT 20 Tablets. (Comment on this Monograph)id=m10427=Brompheniramine
Weigh a portion of Tablets nominally equivalent to 4 mg of Maleate and Pseudoephedrine Sulfate Oral Solution=B-
brompheniramine maleate, mix with 50 mL of water for 10 Monos.pdf)
min, adjust with sodium hydroxide solution (1 in 10) to a
pH of 11, and cool to room temperature. Extract the DEFINITION
mixture with two 75-mL portions of solvent hexane, and Brompheniramine Maleate and Pseudoephedrine Sulfate Oral
combine the extracts in a second separator. Extract the Solution contains NLT 90.0% and NMT 110.0% of the labeled
solvent hexane solution with three 50-mL portions of dilute amounts of brompheniramine maleate (C16H19BrN2 C4H4O4)
hydrochloric acid (1 in 120), combining the acid extracts in and pseudoephedrine sulfate [(C10H15NO)2 H2SO4].
a 200-mL volumetric flask, and dilute with hydrochloric acid
(1 in 120) to volume. IDENTIFICATION
Standard stock solution: 160 g/mL of USP A. The retention times of the major peaks of the Sample
Brompheniramine Maleate RS solution correspond to those of the Standard solution, as
Standard solution: Transfer 25.0 mL of Standard stock obtained in the Assay.
solution to a separator containing 25 mL of water, mix, and B. IDENTIFICATION TESTSGENERAL, Sulfate 191
proceed as directed under Sample solution, beginning with C. THIN-LAYER CHROMATOGRAPHY
adjust with sodium hydroxide solution (1 in 10) to a pH of Standard solution: 1.2 mg/mL of USP Brompheniramine
11. The concentration of USP Brompheniramine Maleate RS Maleate RS and 9 mg/mL of USP Pseudoephedrine Sulfate
in the Standard solution is 20 g/mL. RS in methanol
Blank: Dilute hydrochloric acid (1 in 120) Sample solution: Equivalent to 6 mg of brompheniramine
Spectrometric conditions maleate in a separator. Add 0.5 mL of ammonium hydroxide
Mode: UV and 5 mL of methylene chloride, shake for 1 min, and allow
Analytical wavelength: 264 nm the layers to separate. Use the clear, lower layer as the
Cell: 1 cm Sample solution.
Samples: Blank, Sample solution, and Standard solution (See Chromatography 621, Thin-Layer Chromatography.)
Calculate the percentage of C16H19Br N2 C4H4O4 in the Mode: TLC
portion of Tablets taken: Adsorbent: 0.25-mm layer of chromatographic silica gel
mixture
Result = (AU/AS) (CS/CU) 100 Application volume: 5 L
Developing solvent system: Ethyl ether, methanol, and
AU = absorbance from the Sample solution ammonium hydroxide (16:3:1)
AS = absorbance from the Standard solution Analysis
CS = concentration of USP Brompheniramine Maleate Samples: Standard solution and Sample solution
RS in the Standard solution (g/mL) Allow the spots to dry, and develop the chromatogram
CU = nominal concentration of brompheniramine until the solvent front has moved about three-fourths of
maleate in the Sample solution (g/mL) the length of the plate. Remove the plate from the
USP 32 Official Monographs / Budesonide 131
developing chamber, mark the solvent front, and allow PERFORMANCE TESTS
the solvent to evaporate. Locate the spots on the plate by DELIVERABLE VOLUME 698: Meets the requirements for Oral
examination under short-wavelength UV light. Solution packaged in multiple-unit containers
Acceptance criteria: The RF values of the two principal UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
spots from the Sample solution correspond to those from the for Oral Solution packaged in multiple-unit containers
Standard solution.
PROCEDURE USP Brompheniramine Maleate RS
Mobile phase: Acetonitrile, methanol, tetrahydrofuran, and USP Pseudoephedrine Sulfate RS
water (320:80:50:550)
Transfer 1.0 mL of phosphoric acid, followed by 4.33 g of
dodecyl sulfate sodium to this mixture, and mix. Adjust
with ammonium hydroxide to a pH of 3.50 0.05 (see Budesonide
Chromatography 621, System Suitability). (Comment on this Monograph)id=m10458=Budesonide=B-
[NOTEThe pH of the Mobile phase is critical and may Monos.pdf)
cause a 14 min difference in the retention times of the
Internal standard solution and brompheniramine maleate.]
Internal standard solution: 0.5 mg/mL of naphazoline
hydrochloride in Mobile phase
Solution P: 6000J g/mL of USP Brompheniramine Maleate
RS in Mobile phase, J being the ratio of the labeled amount,
in mg/mL, of brompheniramine maleate to the labeled
amount, in mg/mL, of pseudoephedrine sulfate (Solution P)
Standard solution: Transfer 30 mg of USP Pseudoephedrine C25H34O6 430.53
Sulfate RS into a 25-mL volumetric flask. Add 5.0 mL each of Pregna-1,4-diene-3,20-dione, 16,17-butylidenebis(oxy)-11,21-
Solution P and Internal standard solution, and dilute with dihydroxy-, [11,16(R)], and 16,17-[(S)-
Mobile phase to volume. Butylidenebis(oxy)]-11,21-dihydroxypregna-1,4-diene-3,20-
[NOTEThe concentrations of the Standard solution are dione;
1200J g of USP Brompheniramine Maleate RS/mL and (RS)-11,16,17,21-Tetrahydroxypregna-1,4-diene-3,20-dione
about 1.2 mg of USP Pseudoephedrine Sulfate RS/mL.] cyclic 16,17-acetal with butyraldehyde [51372-29-3;
Sample solution: Transfer a volume of Oral Solution, 51372-28-2; 51333-22-3].
nominally equivalent to 30 mg of pseudoephedrine sulfate,
to a 25-mL volumetric flask. Add 5.0 mL of Internal standard DEFINITION
solution and dilute to volume with Mobile phase. Budesonide is a mixture of two epimeric forms, epimer
Chromatographic system A(C-22S) and epimer B(C-22R). It contains NLT 44.0% and
(See Chromatography 621, System Suitability.) NMT 51.0% of epimer A, and the sum of both epimers is NLT
Mode: LC 98.0% and NMT 102.0% of C25H34O6, calculated on the dried
Detector: UV 254 nm basis.
Column: 4-mm 30-cm; packing L11 [NOTEProtect all solutions containing budesonide from light.]
Injection size: 10 L A. INFRARED ABSORPTION 197K
System suitability B. ULTRAVIOLET ABSORPTION 197U
Sample: Standard solution Sample solution: 25 g/mL
[NOTEThe relative retention times for pseudoephedrine Medium: Methanol
sulfate, naphazoline hydrochloride, and brompheniramine
maleate are 1.0, 1.5, and 2.5, respectively.] ASSAY
Resolution: NLT 3 between the pseudoephedrine sulfate Solution A: 3.17 mg/mL of monobasic sodium phosphate
and naphazoline hydrochloride peaks; NLT 3 between the and 0.23 mg/mL of phosphoric acid. The pH is 3.2 0.1.
brompheniramine maleate and naphazoline hydrochloride Mobile phase: Acetonitrile and Solution A (32:68)
peaks Standard solution: Dissolve a quantity of USP Budesonide
Relative standard deviation: NMT 2.0% RS in acetonitrile and dilute quantitatively with Solution A to
Analysis obtain a solution having a concentration of 0.5 mg/mL,
Calculate the percentage of C16H19BrN2 C4H4O4 in the keeping the proportion of acetonitrile in this solution to
sample taken: NMT 30%.
Sample solution: Dissolve 25 mg of Budesonide in 15 mL
Result = (RU/RS) CS/CU) 100 of acetonitrile in a 50-mL volumetric flask, and dilute to
volume with Solution A.
RU = peak response ratio for brompheniramine Chromatographic system
maleate to naphazoline hydrochloride from the (See Chromatography 621, System Suitability.)
RS = peak response ratio for brompheniramine Detector: UV 254 nm
maleate to naphazoline hydrochloride from the Column: 4.6-mm 15-cm; 5-m packing L1
Standard solution Flow rate: 1.5 mL/min
CS = concentration of USP Brompheniramine Maleate Injection size: 20 L
RS in the Standard solution (mg/mL) System suitability
CU = nominal concentration of brompheniramine Sample: Standard solution
maleate in the Sample solution (mg/mL) [NOTEThe relative retention time for epimer A is 1.1 with
Acceptance criteria: 90.0%110.0% of the labeled respect to epimer B.]
amounts of C16H19BrN2 C4H4O4 and [(C10H15NO)2 H2SO4]
132 Budesonide / Official Monographs USP 32

Resolution: NMT 1.5 between the two budesonide Sample: Sample solution
epimer peaks Calculate the percentage of the 21-acetate of budesonide
Column efficiency: NLT 5500 theoretical plates, in the portion of C25H34O6 taken:
determined from the budesonide epimer B peak
Analysis Result = (rT1/rT2) 100
Calculate the percentage of epimer A (C25H34O6) in the rT1 = sum of the peak areas for the two epimers of the
portion taken: 21-acetate of budesonide
rT2 = sum of the areas of the two budesonide peaks
Result = [rUA/(rUA + rUB)] 100 Acceptance criteria: NMT 0.10% of the 21-acetate of
budesonide is found.
rUA = epimer A peak area response from the Sample PROCEDURE 2: LIMIT OF 11-KETOBUDESONIDE
solution Solution A: 3.17 mg/mL of monobasic sodium phosphate
rUB = epimer B peak area response from the Sample and 0.23 mg/mL of phosphoric acid. The pH is 3.2 0.1.
solution Mobile phase: Acetonitrile, isopropanol, and Solution A
Calculate the percentage of budesonide (C25H34O6) in the (26:9:65)
portion taken: Standard solution: Dissolve a quantity of USP Budesonide
RS in acetonitrile and dilute quantitatively with Solution A
Result = [(rUA + rUB)/(rSA + rSB)] (CS/CU) 100 to obtain a solution having a concentration of 0.5 mg/mL,
keeping the proportion of acetonitrile in this solution to
rUA = epimer A peak area response from the Sample NMT 30%.
solution Sample solution: Dissolve 25 mg of Budesonide in 15 mL
rUB = epimer B peak area response from the Sample of acetonitrile in a 50-mL volumetric flask, and dilute to
solution volume with Solution A.
rSA = epimer A peak area response from the Standard Chromatographic system
solution (See Chromatography 621, System Suitability.)
rSB = epimer B peak area response from the Standard Mode: LC
solution Detector: UV 254 nm
CS = concentration of USP Budesonide RS in the Column: 4.6-mm 15-cm; 3.5-m packing L1
Standard solution (mg/mL) Column temperature: 50
CU = concentration of Budesonide in the Sample Flow rate: 1.5 mL/min
solution (mg/mL) Injection size: 20 L
Epimer A: 44.0%51.0% Sample: Standard solution
Both epimers: 98.0%102.0% [NOTEThe relative retention times for the two epimers
of 11-ketobudesonide are 0.73 and 0.78, respectively;
IMPURITIES the relative retention times for 21-dehydrobudesonide,
Organic Impurities 14,15-dehydrobudesonide, and the first eluted epimer
PROCEDURE 1: LIMIT OF 21-ACETATE OF BUDESONIDE of budesonide (epimer B) are 0.68, 0.84, and 1.0,
Solution A: 3.17 mg/mL of monobasic sodium phosphate respectively.]
and 0.23 mg/mL of phosphoric acid. The pH is 3.2 0.1. Suitability requirements
Mobile phase: Acetonitrile and Solution A (45:55) Resolution: NLT 1.0 between the first epimer of 11-
Standard solution: Dissolve a quantity of USP Budesonide ketobudesonide and 21-dehydrobudesonide; NLT 1.2
RS in acetonitrile, and dilute quantitatively with Solution A between the second epimer of 11-ketobudesonide and
to obtain a solution having a concentration of 0.5 mg/mL, 14,15-dehydrobudesonide
keeping the proportion of acetonitrile in this solution to Column efficiency: NLT 5500 theoretical plates,
NMT 30%. determined from the budesonide epimer B peak
Sample solution: Dissolve 25 mg of Budesonide in 15 mL Analysis
of acetonitrile in a 50-mL volumetric flask, and dilute to Sample: Sample solution
volume with Solution A. Calculate the percentage of the 11-ketobudesonide in the
Chromatographic system portion of C25H34O6 taken:
Mode: LC Result = (rT1/rT2) 100
Detector: UV 254 nm
Column: 4.6-mm 15-cm; 5-m packing L1 rT1 = sum of the peak areas for the two
Flow rate: 1.5 mL/min ketobudesonide peaks
Injection size: 20 L rT2 = sum of the areas of the two budesonide peaks
System suitability Acceptance criteria: NMT 0.2% of the 11-ketobudesonide
Sample: Standard solution is found.
[NOTEThe relative retention times for the first eluted PROCEDURE 3
epimer of 21-acetate of budesonide, the second eluted Solution A: 3.17 mg/mL of monobasic sodium phosphate
epimer of 21-acetate of budesonide, the first eluted and 0.23 mg/mL of phosphoric acid. The pH is 3.2 0.1
epimer of budesonide (epimer B), and the second Mobile phase: Acetonitrile and Solution A (32:68)
eluted epimer of budesonide (epimer A) are 3.1, 3.2, Standard solution: Dissolve a quantity of USP Budesonide
1.0, and 1.1, respectively.] RS in acetonitrile, and dilute quantitatively with Solution A
Suitability requirements to obtain a solution having a concentration of 0.5 mg/mL,
Column efficiency: NLT 5500 theoretical plates,
determined from the budesonide epimer B peak
USP 32 Official Monographs / Bumetanide 133
keeping the proportion of acetonitrile in this solution to Bumetanide

NMT 30%. (Comment on this Monograph)id=m10463=Bumetanide=B-
Sample solution: Dissolve 25 mg of Budesonide in 15 mL Monos.pdf)
of acetonitrile in a 50-mL volumetric flask, and dilute to
volume with Solution A.
Mode: LC
Detector: UV 254 nm
Injection size: 20 L C17H20N2O5S 364.42
System suitability Benzoic acid, 3-(aminosulfonyl)-5-(butylamino)-4-phenoxy-;
Sample: Standard solution 3-(Butylamino)-4-phenoxy-5-sulfamoylbenzoic acid
Suitability requirements [28395-03-1].
determined from the budesonide epimer B peak DEFINITION
Analysis Bumetanide contains NLT 98.0% and NMT 102.0% of
Sample: Sample solution C17H20N2O5S, calculated on the dried basis.
Calculate the percentage of each impurity in the portion of
C25H34O6 taken: IDENTIFICATION
Result = (rU/rT) 100 B. ULTRAVIOLET ABSORPTION 197U
Sample solution: 50 g/mL in isopropyl alcohol
rU = peak area response for each impurity C. The principal spot from the Sample solution exhibits an RF
rT = sum of the responses of all of the peaks value corresponding to that of Standard solution A, as
Acceptance criteria: The impurities meet the requirements obtained in the Procedure for Organic Impurities.
listed in Impurity Table 1.
ASSAY
PROCEDURE
Impurity Table 1 Sample solution: Dissolve 1 g of Bumetanide in 150 mL of
Relative Relative Acceptance alcohol, and add phenol red TS.
Retention Response Criteria, Analysis: Titrate with 0.1 N sodium hydroxide VS. Perform a
Name Time Factor NMT (%) blank determination, and make any necessary correction
16-Hydroxylprednisolonea 0.11 0.2
(see Titrimetry 541). Each mL of 0.1 N sodium hydroxide is
equivalent to 36.44 mg of C17H20N2O5S.
D-Homobudesonideb 0.36 0.10 Acceptance criteria: 98.0%102.0%
21-Dehydrobudesonide 0.61; 0.07
(epimers)c 0.66 IMPURITIES
14,15-Dehydrobudesonide d 0.86 0.10 RESIDUE ON IGNITION 281: NMT 0.1%, a 1-g specimen
Total specified impurities 0.4 being used
Any other individual 0.10 HEAVY METALS, Method II 231: NMT 20 ppm
impurity Organic Impurities
PROCEDURE
Total unspecified impurities 0.4 Standard solution A: 25 mg/mL of USP Bumetanide RS in
a 11,16,17,21-Tetrahydroxypregna-1,4-diene-3,20-dione. methanol
b 16,17-[(1RS)-Ethylidenebis(oxo)]-11,21-dihydroxypregna-1,4- Standard solution B: 50 g/mL from Standard solution A
diene-3,20-dione. in methanol
c 16,17-[(1RS)-Butylidenebis(oxo)]-11-hydroxy-3,20-dioxopregna-1,4- Standard solution C: 50 g/mL of USP Bumetanide
dien-21-al. Related Compound B RS in methanol
d 16,17-[(1RS)-Butylidenebis(oxo)]-11,21-dihydroxypregna-1,4,14- Standard solution D: 25 g/mL of USP Bumetanide
triene-3,20-dione. Related Compound A RS in methanol
SPECIFIC TESTS Standard solution E: 25 g/mL of USP Butyl 3-
MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED (butylamino)-4-phenoxy-5-sulfamoylbenzoate RS in
MICROORGANISMS 62 methanol
Acceptance criteria Sample solution: 25 mg/mL of Bumetanide in methanol
Total aerobic microbial count: NMT 1000 cfu/g Chromatographic system
Total combined molds and yeast count: NMT 100 cfu/g (See Chromatography 621, Thin-Layer Chromatography.)
LOSS ON DRYING 731 Mode: TLC
Analysis: Dry a sample at 105 to constant weight; it loses Adsorbent: 0.25-mm layer of chromatographic silica gel
NMT 0.3% of its weight. mixture
ADDITIONAL REQUIREMENTS Developing solvent system: Chloroform, cyclohexane,
PACKAGING AND STORAGE: Preserve in tight, light-resistant glacial acetic acid, and methanol (160:20:20:5)
containers. Store at a controlled room temperature.
USP Budesonide RS
134 Bumetanide / Official Monographs USP 32
Visualization: Short-wavelength UV light Internal standard stock solution: 0.5 mg/mL of 4-

Analysis ethylbenzaldehyde in methanol
Samples: Standard solution A, Standard solution B, Internal standard solution: Add 10 mL of Internal standard
Standard solution C, Standard solution D, Standard solution stock solution, 10 mL of tetrahydrofuran, and 4.0 mL of
E, and Sample solution glacial acetic acid to a 100-mL volumetric flask, and dilute
Proceed as directed in Chromatography 621, except after to volume with methanol.
drying the application spots, place the plate in an Standard stock solution: 250 g/mL of USP Bumetanide
unlined and unsaturated chromatographic chamber. RS in Internal standard solution
Acceptance criteria: Any secondary spots from the Standard solution: 125 g/mL from Standard stock solution
Sample solution are not larger or more intense than the in water
corresponding principal spots from the corresponding Sample solution: Transfer a volume equivalent to 250 g of
standard solution identified in Impurity Table 1. bumetanide to a flask. Add an equal volume of Internal
Individual impurities: See Impurity Table 1. standard solution, and mix well.
Sum of other impurities: See Impurity Table 1. Chromatographic system
Impurity Table 1 Mode: LC
Detector: UV 254 nm
Acceptance Column: 3.9-mm 30-cm; packing L1
Corresponding Criteria, Flow rate: 1 mL/min
Name Standard Solution NMT (%) Injection size: 20 L
Bumetanide related Standard solution D 0.1 System suitability
compound A Sample: Standard solution
Bumetanide related Standard solution C 0.2
[NOTEThe relative retention times of ethylbenzaldehyde
compound B
and bumetanide are 0.7 and 1.0, respectively.]
Paroxetine butyl 3- Standard solution E 0.1 Resolution: NLT 1.5 between the analyte and internal
(butylamino)-4-phenoxy-5- standard peaks
sulfamoylbenzoate Tailing factor: NMT 1.4
Other individual impurities Standard solution B 0.2 Relative standard deviation: NMT 2.0%
Sum of other individual 0.4
Analysis
impurities
Calculate the percentage of C17H20N2O5S in each mL of the
Injection:
SPECIFIC TESTS
LOSS ON DRYING 731: Dry a sample at 105 for 4 h: it loses Result = (RU/RS) (CS/CU) 100
RU = peak response ratio from the Sample solution
ADDITIONAL REQUIREMENTS RS = peak response ratio from the Standard solution
PACKAGING AND STORAGE: Preserve in tight, light-resistant CS = concentration of USP Bumetanide RS in the
containers. Store at 25, excursions permitted between 15 Standard solution (mg/mL)
and 30. CU = nominal concentration of the Sample solution
USP Bumetanide Related Compound A RS Acceptance criteria: 90.0%110.0%
USP Bumetanide Related Compound B RS
USP Bumetanide RS IMPURITIES
USP Butyl 3-(butylamino)-4-phenoxy-5-sulfamoylbenzoate RS Organic Impurities
PROCEDURE
Identification solution: 10 mg/mL of USP Bumetanide RS
in methanol
Bumetanide Injection Standard solution A: 80 g/mL of USP Bumetanide RS
(Comment on this Monograph)id=m10465=Bumetanide from Identification solution, in methanol
Injection=B-Monos.pdf) Standard solution B: 60 g/mL of USP Bumetanide RS
from Standard solution A, in methanol
DEFINITION Standard solution C: 40 g/mL of USP Bumetanide RS
Bumetanide Injection is a sterile solution of Bumetanide in from Standard solution A, in methanol
Water for Injection, prepared with the aid of Sodium Standard solution D: 20 g/mL of USP Bumetanide RS
Hydroxide. It contains NLT 90.0% and NMT 110.0% of the from Standard solution A, in methanol
labeled amount of bumetanide (C17H20N2O5S). Standard solution E: 10 g/mL of USP Bumetanide RS
IDENTIFICATION Standard solution F: 20 g/mL of USP Bumetanide
A. The relative retention time of the major peak in the Related Compound A RS, in methanol
Sample solution corresponds to that in the Standard solution, Sample solution: Transfer a volume of Injection, nominally
both relative to the internal standard, as obtained in the equivalent to 5 mg of bumetanide, into a 125-mL
Assay. separator, and adjust with 0.1 N sodium hydroxide to a pH
B. The principal spot from the Sample solution exhibits an RF of 12. Extract with two 20-mL portions of ethyl ether,
value corresponding to that of the Identification solution, as discard the ethyl ether extracts, and adjust the aqueous
obtained in the Procedure for Organic Impurities. layer with 1 N acetic acid to a pH of 4. Extract with two
20-mL portions of ethyl ether, passing the extracts through
ASSAY anhydrous sodium sulfate. Wash the sodium sulfate with
PROCEDURE about 5 mL of ethyl ether. Evaporate the combined ethyl
Mobile phase: Methanol, tetrahydrofuran, glacial acetic
acid, and water (50:5:2:45)
USP 32 Official Monographs / Bumetanide 135
ether extracts with the aid of a stream of nitrogen to ASSAY

dryness, and dissolve the residue in 0.5 mL of methanol. PROCEDURE
Chromatographic system Mobile phase: Methanol, tetrahydrofuran, glacial acetic
(See Chromatography 621, Thin-Layer Chromatography.) acid, and water (50:5:2:45)
Mode: TLC Internal standard stock solution: 0.5 mg/mL of 4-
Adsorbent: 0.25-mm layer of chromatographic silica gel ethylbenzaldehyde in methanol
mixture Internal standard solution: Add 10 mL of Internal standard
Application volume: 50 L stock solution, 10 mL of tetrahydrofuran, and 4.0 mL of
Visualization: Short-wavelength UV light glacial acetic acid to a 100-mL volumetric flask, and dilute
Developing solvent system: Chloroform, cyclohexane, with methanol to volume.
glacial acetic acid, and methanol (160:20:20:5) Standard stock solution: 250 g/mL of USP Bumetanide
Analysis RS in Internal standard solution
Samples: Standard solution A, Standard solution B, Standard solution: 125 g/mL from Standard stock solution
Standard solution C, Standard solution D, Standard solution in water
E, Standard solution F, and Sample solution Sample solution: Weigh and finely powder NLT 20 Tablets.
Proceed as directed for Chromatography 621, Thin-Layer Transfer a portion of the powder, nominally equivalent to
Chromatography. 0.5 mg of bumetanide, to a 10-mL volumetric flask. Add 2.0
Acceptance criteria: Any secondary spots from the Sample mL of Internal standard solution and sonicate for 5 min. Add
solution are not larger or more intense than the 2.0 mL of water. Cool, and filter, discarding the first 1 mL.
corresponding principal spots from the corresponding Chromatographic system
standard solution identified in Impurity Table 1. (See Chromatography 621, System Suitability.)
Individual impurities: See Impurity Table 1. Mode: LC
Sum of all other impurities: See Impurity Table 1. Detector: UV 254 nm
Impurity Table 1 Flow rate: 1 mL/min
Acceptance System suitability
Corresponding Criteria, Sample: Standard solution
Name Standard Solution NMT (%) [NOTEThe relative retention times for 4-
Bumetanide related Standard solution F 0.2 ethylbenzaldehyde and bumetanide are 0.7 and 1.0,
compound A respectively.]
Any other individual Standard solutions AE 0.2
impurity
standard peaks
Sum of all other 0.8 Tailing factor: NMT 1.4

individual impurities Relative standard deviation: NMT 2.0%
Analysis
SPECIFIC TESTS Calculate the percentage of C17H20N2O5S in the portion of
BACTERIAL ENDOTOXINS TEST 85: NMT 350 USP Endotoxin Tablets taken:
Units/mg of bumetanide
PH 791: 6.87.8 Result = (RU/RS) (CS/CU) 100
Injections 1. RU = peak response ratio from the Sample solution
RS = peak response ratio from the Standard solution
ADDITIONAL REQUIREMENTS CS = concentration of USP Bumetanide RS in the
PACKAGING AND STORAGE: Preserve in single-dose or Standard solution (mg/mL)
multiple-dose containers, preferably of Type I glass, CU = nominal concentration of the bumetanide taken
protected from light. to prepare the Sample solution (mg/mL)
USP Bumetanide RS
USP Bumetanide Related Compound A RS IMPURITIES
USP Endotoxin RS Organic Impurities
PROCEDURE
Sample solution: Equivalent to 10 mg of bumetanide
Bumetanide Tablets from powdered Tablets in a 50-mL centrifuge tube. Add 20
mL of acetone (spectrophotometric or HPLC quality), and
(Comment on this Monograph)id=m10468=Bumetanide shake by mechanical means for 10 min. Centrifuge for 10
Tablets=B-Monos.pdf) min, decant the supernatant into a glass-stoppered, 25-mL
conical flask, and evaporate with the aid of a stream of
DEFINITION nitrogen to dryness. Dissolve the residue in 0.5 mL of
Bumetanide Tablets contain NLT 90.0% and NMT 110.0% of methanol.
the labeled amount of bumetanide (C17H20N2O5S). Identification solution: 20 mg/mL of USP Bumetanide RS
IDENTIFICATION in methanol
A. The relative retention time of the major peak in the Standard solution A: 160 g/mL of USP Bumetanide RS
Sample solution corresponds to that in the Standard solution, from Identification solution, in methanol
as obtained in the Assay. Standard solution B: 120 g/mL of USP Bumetanide RS
B. The principal spot of the Sample solution exhibits an RF from Standard solution A, in methanol
value corresponding to that of the Identification solution, as Standard solution C: 80 g/mL of USP Bumetanide RS
obtained in the Procedure for Organic impurities. from Standard solution A, in methanol
136 Bumetanide / Official Monographs USP 32
Standard solution D: 40 g/mL of USP Bumetanide RS Bupivacaine Hydrochloride

from Standard solution A, in methanol (Comment on this Monograph)id=m10490=Bupivacaine
Standard solution E: 20 g/mL of USP Bumetanide RS Hydrochloride=B-Monos.pdf)
Standard solution F: 0.04 mg/mL of USP Bumetanide
Related Compound A RS in methanol
(see Chromatography 621, Thin-Layer Chromatography.)
Mode: TLC
mixture
Application volume: 25 L C18H28N2O HCl H2O 342.90
Visualization: Short-wavelength UV light
Developing solvent system: Chloroform, cyclohexane,
glacial acetic acid, and methanol (160:20:20:5) Change to read:
Analysis
Samples: Sample solution, Standard solution A, Standard
solution B, Standard solution C, Standard solution D,
Standard solution E, and Standard solution F C18H28N2O HCl 324.90
Proceed as directed for Chromatography 621, Thin-Layer 2-Piperidinecarboxamide, 1-butyl-N-(2,6-dimethylphenyl)-,
Chromatography. monohydrochloride, monohydrate, ()-;
Acceptance criteria: Any secondary spots of the Sample ()-1-Butyl-2,6-pipecoloxylidide monohydrochloride,
solution are not larger or more intense than the monohydrate [73360-54-0USP32].
corresponding principal spots of the corresponding Standard Anhydrous [18010-40-7].
solution identified in the Impurity Table 1.
Individual Impurities: See Impurity Table 1. DEFINITION
Sum of all other impurities: See Impurity Table 1. Bupivacaine Hydrochloride contains NLT 98.5% and NMT
101.5% of C18H28N2O HCl, calculated on the anhydrous basis.
Impurity Table 1 IDENTIFICATION
Acceptance A. INFRARED ABSORPTION 197S
Corresponding Criteria, Sample solution: Dissolve 230 mg in 15 mL of water in a
Name Standard Solution NMT (%) separator, add 1 mL of 6 N ammonium hydroxide, and
Bumetanide related Standard solution F 0.2
extract with three 30-mL portions of chloroform. Evaporate
compound A
the chloroform at room temperature with the aid of a
stream of nitrogen, and dry the residue in a vacuum. Add 2
Any other individual Standard solutions A- 0.2 mL of chloroform to the residue, and dissolve.
impurity E B. ULTRAVIOLET ABSORPTION 197U
Sum of all other 0.8 Sample solution: 500 g/mL
individual impurities Medium: 0.1 N hydrochloric acid
Acceptance criteria: Absorptivities do not differ by more
PERFORMANCE TESTS than 3.0%, calculated on the anhydrous basis.
DISSOLUTION 711 C. IDENTIFICATION TESTSGENERAL, Chloride 191
Medium: Water; 900 mL Sample solution: Dissolve 50 mg in 10 mL of water in a
Apparatus 2: 50 rpm small separator, render alkaline with 6 N ammonium
Time: 30 min hydroxide, and extract with 10 mL of ether.
Stock solution A: 7.505 mg/mL of glycine and 5.85 Acceptance criteria: The aqueous layer of the Sample
mg/mL of sodium chloride in water solution meets the requirements.
Solution A: Stock solution A, 0.1 N hydrochloric acid, and
water (4:1:45). Adjust, if necessary, with 0.1 N hydrochloric ASSAY
acid or 0.1 N sodium hydroxide to a pH of 2.9. PROCEDURE
Sample solutions: Sample per Dissolution 711. Dilute with Sample: 600 mg
Solution A as needed. Analysis: Transfer the Sample to a 250-mL conical flask, and
Standard solution: USP Bumetanide RS at a known dissolve in 20 mL of glacial acetic acid. Add 10 mL of
concentration in Medium mercuric acetate TS and 3 drops of crystal violet TS, and
Fluorometric conditions titrate with 0.1 N perchloric acid VS to a green endpoint.
Excitation wavelength: 350 nm Perform a blank determination, and make any necessary
Emission wavelength: 450 nm correction (see Titrimetry 541). Each mL of 0.1 N perchloric
Analysis acid is equivalent to 32.49 mg of C18H28N2O HCl.
C17H20N2O5S is dissolved. IMPURITIES
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Inorganic Impurities
ADDITIONAL REQUIREMENTS HEAVY METALS, Method II 231: NMT 10 ppm
PACKAGING AND STORAGE: Preserve in tight, light-resistant Organic Impurities
containers. PROCEDURE 1: LIMIT OF RESIDUAL SOLVENTS
USP REFERENCE STANDARDS 11 Standard solution A: Pipet 2 mL of dehydrated alcohol
USP Bumetanide RS into a 100-mL volumetric flask, dilute with water to
USP Bumetanide Related Compound A RS volume, and mix. Transfer 2.0 mL of this solution to a 50-
USP 32 Official Monographs / Bupivacaine 137
mL volumetric flask, dilute with water to volume, and mix. the Sample solution does not exceed that of the principal
The resulting solution contains 0.08% alcohol. spot of the Standard solution B (0.5%). The total of the
Standard solution B: Pipet 2 mL of isopropyl alcohol into estimated sizes and intensities of all of the other spots of
a 1000-mL volumetric flask, dilute with water to volume, the Sample solution does not exceed four times that of the
and mix. Transfer 2.0 mL of this solution to a 100-mL principal spot of the Standard solution B (2.0%).
volumetric flask, dilute with water to volume, and mix. The
resulting solution contains 0.004% isopropyl alcohol. SPECIFIC TESTS
Sample solution: 40 mg/mL of Bupivacaine Hydrochloride PH 791
Chromatographic system Sample solution: 1 in 100
(See Chromatography 621, System Suitability.) Acceptance criteria: Between 4.5 and 6.0
Mode: GC WATER DETERMINATION, Method I 921: Between 4.0% and
Detector: Flame ionization 6.0%
Column: 4-mm 2-m; packing S3
Temperature ADDITIONAL REQUIREMENTS
Column: 175 PACKAGING AND STORAGE: Preserve in well-closed containers.
Injection port: 200 USP REFERENCE STANDARDS 11
Detector: 280 USP Bupivacaine Hydrochloride RS
Flow rate: 40 mL/min
Injection size: 5 L
Carrier gas: Nitrogen Bupivacaine Hydrochloride Injection
Analysis
Samples: Standard solution A, Standard solution B, and (Comment on this Monograph)id=m10498=Bupivacaine
Sample solution Hydrochloride Injection=B-Monos.pdf)
Determine the percentage of alcohol taken: DEFINITION
Bupivacaine Hydrochloride Injection is a sterile solution of
Result = 2(rU/rS) Bupivacaine Hydrochloride in Water for Injection. It contains
Determine the percentage of isopropyl alcohol taken: NLT 93.0% and NMT 107.0% of the labeled amount of
C18H28N2O HCl.
Result = 0.1(rU/rS) IDENTIFICATION
rU = peak response of the respective analytes in the A. IDENTIFICATIONORGANIC NITROGENOUS BASES 181
Sample solution Sample solution: Dilute a volume of Injection in 0.01 N
rS = peak response of the corresponding analytes in hydrochloric acid to give a nominal concentration of 2
Standard solution A and Standard solution B mg/mL of bupivacaine hydrochloride.
Acceptance criteria: The sum of the content of alcohol Analysis: Proceed as directed in the General Chapter
and the content of isopropyl alcohol is NMT 2%. beginning with Transfer the liquid to a separator.
PROCEDURE 2 Acceptance criteria: The Injection meets the requirements
Solution A: Chloroform and isopropylamine (99:1) of the test.
Sample solution: 20.0 mg/mL of Bupivacaine B. The retention time of the bupivacaine peak of the Sample
Hydrochloride in Solution A solution corresponds to that of the bupivacaine peak of the
Standard solution A: 20.0 mg/mL of USP Bupivacaine Standard solution, as obtained in the Assay.
Hydrochloride RS in Solution A ASSAY
Standard solution B: 100 g/mL of USP Bupivacaine PROCEDURE
Hydrochloride RS from Standard solution A diluted with Solution A: 1.94 mg/mL of monobasic potassium
Solution A phosphate and 2.48 mg/mL of dibasic potassium phosphate
Chromatographic system in water
(See Chromatography 621, Thin-Layer Chromatography.) Adjust, if necessary, with 1 N potassium hydroxide or 1 M
Mode: TLC phosphoric acid to a pH of 6.8.
Adsorbent: 0.25-mm layer of chromatographic silica gel Mobile phase: Acetonitrile and Solution A (65:35)
mixture Adjust, if necessary, with 1 M phosphoric acid to a pH of 7.7
Application volume: 10 L 0.2. Filter the solution through a membrane filter of 1-m
Developing solvent system: Hexanes and isopropylamine or finer porosity and degas.
(97:3) Internal standard solution: 1.3 mg/mL of dibutyl phthalate
Samples: Sample solution, Standard solution A, and in methanol
Standard solution B Standard solution: Dissolve 50 mg of USP Bupivacaine
Analysis: Proceed as directed under Chromatography 621. Hydrochloride RS in 10.0 mL of water, using sonication if
Develop the chromatogram in a suitable chamber until the necessary, in a 100-mL volumetric flask. Add 10 mL of
solvent has moved about three-fourths of the length of the Internal standard solution, and dilute with methanol to
plate. Remove the plate from the chamber, mark the volume.
solvent front, and dry it in warm air. Place the plate in a Sample solution: Transfer the amount of Injection
closed chamber with a dish containing 1 g of iodine in a equivalent to 50 mg of bupivacaine hydrochloride to a 100-
shallow layer, and allow to remain for about 5 min. mL volumetric flask, add 10.0 mL of Internal standard
Remove the plate from the chamber, spray it with 7 N solution, and dilute with methanol to volume.
sulfuric acid, and examine the chromatogram. Chromatographic system
Acceptance criteria: The RF value of the principal spot of (See Chromatography 621, System Suitability.)
the Sample solution corresponds to that of Standard solution
A, and the estimated size and intensity of any other spot of
138 Bupivacaine / Official Monographs USP 32
Mode: LC solution of USP Dextrose RS in water, and (C) a solution of

Detector: UV 263 nm USP Bupivacaine Hydrochloride RS in Standard solution B to
Column: 4-mm 30-cm; packing L1 obtain solutions having concentrations corresponding to the
Flow rate: 2 mL/min labeled concentrations of bupivacaine hydrochloride and
Injection size: 20 L dextrose in the Injection.
System suitability Sample solution: Bupivacaine Hydrochloride in Dextrose
Sample: Standard solution Injection
[NOTEThe relative retention times for bupivacaine Naphthalenediol reagent: Dissolve 20 mg of 1,3-
hydrochloride and dibutyl phthalate are about 1.0 and naphthalenediol in 10 mL of dehydrated alcohol containing
1.2, respectively.] 0.2 mL of sulfuric acid.
Suitability requirements Iodoplatinate reagent: Mix equal volumes of platinic
Resolution: NLT 2.0 between bupivacaine hydrochloride chloride solution (3 in 1000) and potassium iodide solution
and dibutyl phthalate (6 in 100).
Relative standard deviation: NMT 1.0% for the ratios of Application volume: See Analysis.
the bupivacaine hydrochloride peak to the dibutyl Developing solvent system: Butyl alcohol, dehydrated
phthalate peak (three replicate injections) alcohol, glacial acetic acid, and water (6:1:1:2)
Analysis Analysis: Separately apply 10 L each of the Sample
Samples: Standard solution and Sample solution solution, Standard solution A, and Standard solution C to a
Calculate the percentage of C18H28N2O HCl in the portion of the chromatographic plate, and separately apply
Injection: 1 L each of the Sample solution and Standard solution B to
the remaining portion of the plate. Dry the applications in
Result = (RU/RS) (CS/CU) 100 a current of warm air, develop the chromatograms in the
Developing solvent system, remove the plate from the
RU = peak response ratio of bupivacaine hydrochloride developing chamber, and mark the solvent front. Dry the
to the internal standard from the Sample plate in warm circulating air, and examine the plate under
solution short-wavelength UV light. [NOTEAcceptance criteria 1
RS = peak response ratio of bupivacaine hydrochloride should be met.]
to the internal standard from the Standard Spray the plate with Naphthalenediol reagent, heat at 90
solution for 5 min, and examine the plate. [NOTEAcceptance
CS = concentration of USP Bupivacaine Hydrochloride criteria 2 should be met.]
RS, calculated on the anhydrous basis, in the Cool the plate, spray it with Iodoplatinate reagent, and
Standard solution (mg/mL) examine the plate. [NOTEAcceptance criteria 3 should be
CU = nominal concentration of bupivacaine met. Also, bupivacaine appears as a blue-purple spot on a
hydrochloride taken to prepare the Sample salmon-colored background, and the dextrose spots fade
solution (mg/mL) slightly.]
Acceptance criteria: 93.0%107.0% Acceptance criteria 1: The RF value of the principal spot
from the Sample solution corresponds to those of Standard
SPECIFIC TESTS solutions A and C.
BACTERIAL ENDOTOXINS TEST 85: NMT 2.5 USP Endotoxin Acceptance criteria 2: The RF value of the principal blue-
Units/mg of bupivacaine hydrochloride purple spot from the Sample solution corresponds to that
PH 791: 4.06.5 from Standard solution B.
OTHER REQUIREMENTS: It meets the requirements under Acceptance criteria 3: The RF value of the bupivacaine
Injections 1. spot from the Sample solution corresponds to those from
ADDITIONAL REQUIREMENTS Standard solutions A and C.
PACKAGING AND STORAGE: Preserve in single-dose or in B. The retention time of the bupivacaine peak of the Sample
multiple-dose containers, preferably of Type I glass. Injection solution corresponds to that of the Standard solution, as
labeled to contain 0.5% or less of bupivacaine hydrochloride obtained in the Assay.
may be packaged in 50-mL, multiple-dose containers. ASSAY
USP REFERENCE STANDARDS 11 BUPIVACAINE HYDROCHLORIDE
USP Bupivacaine Hydrochloride RS Solution A: 1.94 mg/mL of monobasic potassium
USP Endotoxin RS phosphate and 2.48 mg/mL of dibasic potassium phosphate
Adjust, if necessary, with 1 N potassium hydroxide or 1 M
phosphoric acid to a pH of 6.8.
Bupivacaine Hydrochloride in Dextrose Mobile phase: Acetonitrile and Solution A (65:35)
Injection Adjust, if necessary, with 1 M phosphoric acid to a pH of 7.7
0.2. Pass the solution through a membrane filter of 1-m
(Comment on this Monograph)id=m10502=Bupivacaine or finer porosity, and degas.
Hydrochloride in Dextrose Injection=B-Monos.pdf) Internal standard solution: 1.3 mg/mL of dibutyl phthalate
in methanol
DEFINITION Standard solution: Dissolve 50 mg of USP Bupivacaine
Bupivacaine Hydrochloride in Dextrose Injection is a sterile Hydrochloride RS in 10.0 mL of water, using sonication if
solution of Bupivacaine Hydrochloride and Dextrose in Water necessary, in a 100-mL volumetric flask. Add 10 mL of
for Injection. It contains NLT 93.0% and NMT 107.0% of the Internal standard solution and dilute with methanol to
labeled amounts of bupivacaine hydrochloride (C18H28N2O volume.
HCl) and dextrose (C6H12O6). It contains no preservative. Sample solution: Transfer the amount of Injection
IDENTIFICATION nominally equivalent to 50 mg of bupivacaine hydrochloride
A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 to a 100-mL volumetric flask, add 10.0 mL of Internal
Adsorbent: Chromatographic silica gel mixture; 0.25 mm standard solution, and dilute with methanol to volume.
Standard solutions A, B, and C: Separately prepare (A) a Chromatographic system
solution of USP Bupivacaine Hydrochloride RS in water, (B) a (See Chromatography 621, System Suitability.)
USP 32 Official Monographs / Bupivacaine 139
Mode: LC Bupivacaine Hydrochloride and

Detector: UV 263 nm Epinephrine Injection
Flow rate: 2 mL/min (Comment on this Monograph)id=m10505=Bupivacaine
Injection size: 20 L Hydrochloride and Epinephrine Injection=B-Monos.pdf)
System suitability DEFINITION
Sample: Standard solution Bupivacaine Hydrochloride and Epinephrine Injection is a sterile
[NOTEThe relative retention times for dibutyl phthalate solution of Bupivacaine Hydrochloride and Epinephrine or
and bupivacaine hydrochloride are about 1.2 and 1.0, Epinephrine Bitartrate in Water for Injection. It contains NLT
respectively.] 93.0% and NMT 107.0% of the labeled amount of
Suitability requirements bupivacaine hydrochloride (C18H28N2O HCl). The content of
Resolution: NLT 2.0 between bupivacaine hydrochloride epinephrine (C9H13NO3) does not exceed 0.001% (1 in
and dibutyl phthalate 100,000). It contains the equivalent of NLT 90.0% and NMT
Relative standard deviation: NMT 1.0% for the ratios of 115.0% of the labeled amount of epinephrine (C9H13NO3).
the bupivacaine hydrochloride peak to the dibutyl
phthalate peak (three replicate injections) IDENTIFICATION
Analysis A. PROCEDURE
Samples: Standard solution and Sample solution Sample solution 1: Dilute a volume of Injection in 0.01 N
Calculate the percentage of C18H28N2O HCl in the portion hydrochloric acid to give a concentration of 2 mg/mL of
of Injection taken: bupivacaine hydrochloride.
Analysis 1: Proceed as directed under Identification
Result = (RU/RS) (CS/CU) 100/L Organic Nitrogenous Bases 181, beginning with Transfer
the liquid to a separator.
RU = peak response ratio of bupivacaine hydrochloride Acceptance criteria 1: The Injection meets the
to the internal standard from the Sample requirements of the test.
solution Sample 2: Sample solution and Standard solution, as
RS = peak response ratio of bupivacaine hydrochloride obtained from the Assay.
to the internal standard from the Standard Analysis 2: The retention time of the bupivacaine peak of
solution the Sample solution corresponds to that of the Standard
CS = concentration of USP Bupivacaine Hydrochloride solution, as obtained in the Assay.
RS, calculated on the anhydrous basis, in the B. PROCEDURE
Standard solution (mg/mL) Sample: Volume of Injection, nominally equivalent to 50 g
CU = nominal concentration of bupivacaine of epinephrine
hydrochloride in the Sample solution (mg/mL) Analysis: Pipet the Sample into a suitable container, add 0.1
L = label claim (mg/volume of Injection) mL of Ferro-citrate solution and 2.0 mL of Buffer solution
Acceptance criteria: 93.0%107.0% (prepared as directed under Epinephrine Assay 391), and
DEXTROSE allow the solution to stand for 10 min. Filter the solution.
Analysis: Determine the angular rotation of the Injection in Acceptance criteria: The filtrate is violet in color and may
a suitable polarimeter tube (see Optical Rotation 781). turn brownish.
Injection taken: ASSAY
BUPIVACAINE HYDROCHLORIDE
Result = {[A R)/F] 100} 100/L Solution A: 1.94 mg/mL of monobasic potassium
phosphate and 2.48 mg/mL of dibasic potassium phosphate
A = 100 mm divided by the length of the polarimeter in water. Adjust, if necessary, with 1 N potassium hydroxide
tube (mm) or 1 M phosphoric acid to a pH of 6.8.
R = observed rotation (degrees) Mobile phase: Acetonitrile and Solution A (65:35). Adjust, if
F = midpoint of the specific rotation range for necessary, with 1 M phosphoric acid to a pH of 7.7 0.2.
anhydrous dextrose, 52.9 Filter the solution through a membrane filter of 1-m or
L = label claim of dextrose (g/100 mL) finer porosity.
Acceptance criteria: 93.0%107.0% Internal standard solution: 1.3 mg/mL of dibutyl phthalate
in methanol
SPECIFIC TESTS Standard solution: Dissolve 50 mg of USP Bupivacaine
BACTERIAL ENDOTOXINS TEST 85: NMT 1.8 USP Endotoxin Hydrochloride RS in 10.0 mL of water, using sonication if
Units/mg of bupivacaine hydrochloride necessary, in a 100-mL volumetric flask. Add 10 mL of
PH 791: Between 4.0 and 6.5 Internal standard solution, and dilute with methanol to
OTHER REQUIREMENTS: It meets the requirements under volume.
Injections 1. Sample solution: Transfer a volume of Injection, nominally
equivalent to 50 mg of bupivacaine hydrochloride, to a 100-
ADDITIONAL REQUIREMENTS mL volumetric flask. Add 10.0 mL of Internal standard
PACKAGING AND STORAGE: Preserve in single-dose containers, solution, and dilute with methanol to volume.
preferably of Type I glass. Chromatographic system
USP Bupivacaine Hydrochloride RS
USP Dextrose RS
USP Endotoxin RS
140 Bupivacaine / Official Monographs USP 32

Detector: UV 263 nm Resolution: NLT 6.0 between the epinephrine and
Column: 4-mm 30-cm; packing L1 dopamine peaks, System suitability solution
Flow rate: 2 mL/min Relative standard deviation: NMT 2.0% for replicate
Injection size: 20 L injections, Standard solution
System suitability Analysis
Sample: Standard solution Samples: Standard solution and the Sample solution
[NOTEThe relative retention times for bupivacaine Calculate the percentage of C9H13NO3 in each mL of the
hydrochloride and dibutyl phthalate are about 1.0 and Injection:
1.2, respectively.]
Suitability requirements Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
Resolution: NLT 2.0 between bupivacaine hydrochloride
and dibutyl phthalate rU = peak response from the Sample solution
Relative standard deviation: NMT 1.0% for the ratios of rS = peak response from the Standard solution
the bupivacaine hydrochloride peak to the dibutyl CS = concentration of USP Epinephrine Bitartrate RS in
phthalate peak (three replicate injections) the Standard solution (g/mL)
Analysis CU = nominal concentration of epinephrine taken to
Samples: Standard solution and Sample solution prepare the Sample solution (g of
Calculate the percentage of C18H28N2O HCl in the epinephrine/mL)
Injection: Mr1 = molecular weight of epinephrine, 183.21
Mr2 = molecular weight of epinephrine bitartrate,
Result = (RU/RS) (CS/CU) 100 333.30
RU = peak response ratio of bupivacaine hydrochloride
to the internal standard peak from the Sample SPECIFIC TESTS
solution COLOR AND CLARITY
RS = peak response ratio of bupivacaine hydrochloride Standard solution: 0.1 N iodine VS and water (1:249)
to the internal standard peak from the Sample: Bupivacaine Hydrochloride and Epinephrine
Standard solution Injection
CS = concentration of USP Bupivacaine Hydrochloride Analysis: Visually examine a portion of the Sample in a
RS, calculated on the anhydrous basis, in the suitable clear glass test tube against a white background.
Standard solution (mg/mL) [NOTEAcceptance criteria 1 should be met.] If any yellow
CU = nominal concentration of bupivacaine color is observed in the Sample, concomitantly determine
hydrochloride taken to prepare the Sample the absorbances of the Sample and the Standard solution in
solution (mg/mL) 1-cm cells with a suitable spectrophotometer set at 460 nm.
Acceptance criteria: 93.0%107.0% [NOTEAcceptance criteria 2 should be met.]
EPINEPHRINE Acceptance criteria 1: The Sample is not pinkish, and it
Mobile phase: Prepare a mixture of water, methanol, and 2 contains no precipitate.
M monobasic sodium phosphate (900:50:50), containing 40 Acceptance criteria 2: The absorbance of the Sample does
mg/mL of edetate disodium, 0.4 mL/L of phosphoric acid, not exceed that of the Standard solution.
and 0.4 mg/mL of sodium 1-octanesulfonate. Make BACTERIAL ENDOTOXINS TEST 85: NMT 1.6 USP Endotoxin
adjustments, if necessary, to obtain a retention time of NLT Units/mg of bupivacaine hydrochloride
11 min for the epinephrine peak. PH 791: 3.35.5
System suitability solution: 2 g/mL of each of INJECTIONS 1: It meets the requirements.
epinephrine bitartrate and dopamine hydrochloride in
Mobile phase ADDITIONAL REQUIREMENTS
Standard solution: 2 g/mL of USP Epinephrine Bitartrate PACKAGING AND STORAGE: Preserve in single-dose or in
RS in Mobile phase multiple-dose containers, preferably of Type I glass,
Sample solution: Nominally equivalent to 1 g/mL of protected from light. Injection labeled to contain 0.5% or
epinephrine, from a volume of Injection, diluted with Mobile less of bupivacaine hydrochloride may be packaged in 50-mL
phase multiple-dose containers.
Chromatographic system LABELING: The label indicates that the Injection is not to be
(See Chromatography 621, System Suitability.) used if its color is pinkish or darker than slightly yellow or if
Mode: LC it contains a precipitate.
Detector: Electrochemical detector held at a potential of USP REFERENCE STANDARDS 11
+0.75 volt USP Bupivacaine Hydrochloride RS
Column: 4.6-mm 25-cm; packing L1 USP Endotoxin RS
Flow rate: 1.2 mL/min USP Epinephrine Bitartrate RS
System suitability
[NOTEThe relative retention times for epinephrine and
dopamine are about 1.0 and 2.0, respectively, in the
chromatograph of the System suitability solution. ]
USP 32 Official Monographs / Bupropion 141
Buprenorphine Hydrochloride Column efficiency: NLT 6500 theoretical plates

(Comment on this Monograph)id=m10510=Buprenorphine Relative standard deviation: NMT 2.0%
Hydrochloride=B-Monos.pdf) Analysis
Allow the Sample solution to elute for NLT two times the
retention time of buprenorphine hydrochloride.
of Buprenorphine Hydrochloride taken:
C29H41NO4 HCl 504.10 rU = peak response for each impurity from the
6,14-Ethenomorphinan-7-methanol, 17-(cyclopropyl-methyl)-- Sample solution
(1,1-dimethylethyl)-4,5-epoxy-18,19-dihydro-3-hydroxy-6- rS = peak response of buprenorphine hydrochloride
methoxy--methyl-, hydrochloride, [5,7 (S)]-; from the Standard solution
21-Cyclopropyl-7-[(S)-1-hydroxy-1,2,2-trimethylpropyl]-6,14- CS = concentration of USP Buprenorphine
endo-ethano-6,7,8,14-tetrahydrooripavine hydrochloride Hydrochloride RS in the Standard solution
[53152-21-9]. (mg/mL)
CU = concentration of Buprenorphine Hydrochloride
DEFINITION in the Sample solution (mg/mL)
Buprenorphine Hydrochloride contains NLT 98.5% and NMT Acceptance criteria
101.0% of C29H41NO4 HCl, calculated on the anhydrous basis. Individual impurity: NMT 0.25%
IDENTIFICATION
A. INFRARED ABSORPTION 197K SPECIFIC TESTS
B. PROCEDURE OPTICAL ROTATION, Specific Rotation 781S: 92 to 98
Sample solution: 50 mg/mL of Buprenorphine Sample solution: 20 mg/mL, in methanol
Hydrochloride in methanol PH 791: 4.06.0 in a solution containing 10 mg/mL
Analysis: To 0.5 mL of the Sample solution add 0.2 mL of a WATER DETERMINATION, Method I 921: NMT 1.0%
freshly prepared solution (1 in 10) of potassium ferricyanide
TS and 0.5 mL of ferric chloride TS. ADDITIONAL REQUIREMENTS
Acceptance criteria: A blue color appears immediately. PACKAGING AND STORAGE: Preserve in tight, light-resistant
C. IDENTIFICATION TESTSGENERAL, Chloride 191 containers.
Sample solution: 1 in 100 USP REFERENCE STANDARDS 11
USP Buprenorphine Hydrochloride RS
ASSAY USP Buprenorphine Related Compound A RS
PROCEDURE
Sample solution: 0.8 g of Buprenorphine Hydrochloride in
50 mL of glacial acetic acid. Add 10 mL of mercuric acetate.
Analysis: Titrate Sample solution with 0.1 N perchloric acid Bupropion Hydrochloride
VS, determining the green endpoint with 2 drops of crystal (Comment on this Monograph)id=m10520=Bupropion
violet TS. Perform a blank determination, and make any Hydrochloride=B-Monos.pdf)
necessary correction (see Titrimetry 541). Each mL of 0.1 N
perchloric acid is equivalent to 50.41 mg of C29H41NO4 HCl.
IMPURITIES
Organic Impurities
PROCEDURE C13H18ClNO HCl 276.21
Mobile phase: Methanol, 1% solution of ammonium 1-Propanone, 1-(3-chlorophenyl)-2-[(1,1-dimethylethyl)amino]-,
acetate, and glacial acetic acid (60:10:0.01) hydrochloride, ()-;
Standard solution: 12.5 g/mL each of USP ()-2-(tert-Butylamino)-3-chloropropiophenone hydrochloride
Buprenorphine Hydrochloride RS and USP Buprenorphine [31677-93-7].
Related Compound A RS in Mobile phase
Sample solution: 5 mg/mL of Buprenorphine DEFINITION
Hydrochloride in Mobile phase Bupropion Hydrochloride contains NLT 98.0% and NMT
Chromatographic system 102.0% of C13H18ClNO HCl, calculated on the anhydrous
(See Chromatography 621, System Suitability.) basis.
Detector: UV 288 nm A. INFRARED ABSORPTION 197K
Column: 4.6-mm 25-cm; packing L1 B. The retention time of the major peak of the Sample
Temperature: 40 solution corresponds to that of the Standard solution, as
Flow rate: 1 mL/min obtained in the Assay.
Injection size: 20 L C. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the
System suitability requirements for the silver nitrate precipate test
Resolution: NLT 3.0 between buprenorphine
hydrochloride and buprenorphine related compound A
142 Bupropion / Official Monographs USP 32
Sample solution: 1 mg/mL of Bupropion Hydrochloride Analysis

Samples: Standard solutions and Sample solution
ASSAY Proceed as directed under the General Chapter. Locate
PROCEDURE and quantitate the spots obtained by scanning the entire
Diluent: Methanol and water (1:1) plate with a suitable densitometer at 235 nm. Plot a
Solution A: Dissolve 6.8 g of monobasic potassium standard curve of area versus concentrations of the
phosphate in 1900 mL of water. Adjust with 1 N sodium Standard solutions. From the standard curve, determine
hydroxide to a pH of 7.0 and dilute with water to 2000 mL. the percentages of m-chlorobenzoic acid and any other
Mobile phase: Methanol, tetrahydrofuran, and Solution A impurity present.
(39:11:51) Acceptance criteria: NMT 0.2% of m-chlorobenzoic acid
Standard stock solution: 0.025 mg/mL of each of USP is found, and NMT 0.1% of any other individual impurity is
Bupropion Hydrochloride Related Compound A RS and USP found.
Bupropion Hydrochloride Related Compound B RS in Diluent PROCEDURE 2
Standard solution: Transfer 25 mg of USP Bupropion Diluent, Solution A, Mobile phase, Solution B, Standard
Hydrochloride RS into a 25-mL volumetric flask. Dissolve in a solution, Sample solution, Chromatographic system, and
portion of Diluent, pipet 2.0 mL of Solution B into the flask, System suitability: Proceed as directed in the Assay.
and dilute with Diluent to volume. Analysis
Sample solution: 1 mg/mL of Bupropion Hydrochloride in Samples: Standard solution and Sample solution
Diluent Calculate the percentage of each impurity in the portion
Chromatographic system of Bupropion Hydrochloride taken:
Mode: LC Result = (rU/rS) (CS/CU) 100
Detector: UV 250 nm
Column: 3.9-mm 15-cm; 5-m packing L7 rU = peak response for each impurity from the
Flow rate: 1.1 mL/min Sample solution
Injection size: 20 L rS = peak response for bupropion hydrochloride from
System suitability the Standard solution
Sample: Standard solution CS = concentration of USP Bupropion Hydrochloride
[NOTEThe relative retention times for bupropion RS in the Standard solution (mg/mL)
hydrochloride related compound A, bupropion CU = concentration of bupropion hydrochloride in the
hydrochloride, and bupropion hydrochloride related Sample solution (mg/mL)
compound B are about 0.92, 1.0, and 1.4, respectively.] Acceptance criteria
Suitability requirements Individual impurities: See Impurity Table 1.
Resolution: NLT 1.3 between bupropion hydrochloride Total impurities: The limits of impurities are specified in
related compound A and bupropion hydrochloride the accompanying table: NMT 0.3% of total unidentified
Relative standard deviation: NMT 2.0% determined impurities is found; and NMT 1.0% of total impurities is
from bupropion hydrochloride; NMT 5.0% determined found, the results of Procedure 1 and Procedure 2 being
from bupropion hydrochloride related compound B added.
Analysis
Calculate the percentage of C13H18ClNO HCl in the portion Impurity Table 1
of Bupropion Hydrochloride taken:
Result = (rU/rS) (CS/CU) 100 Retention Response Criteria,
Name Time Factor NMT (%)
rU = peak response from the Sample solution 2-(tert- 0.38 1.5 0.5
rS = peak response from the Standard solution Butylamino)propiophenone
CS = concentration of USP Bupropion Hydrochloride hydrochloride
CU = concentration of bupropion hydrochloride in the 1-(3- 0.58 1.0 0.2
Sample solution (mg/mL) Chlorophenyl)-1,2-
Acceptance criteria: 98.0%102.0% propanedione
2-(tert- 0.71 0.45 0.1
IMPURITIES Butylamino)-2-
Organic Impurities chloropropiophenone
PROCEDURE 1 hydrochloride
Standard solutions: 0.3, 0.2, and 0.1 mg/mL from
3- 0.78 1.2 0.1
Standard stock solution in methanol
Chloropropiophenone
Sample solution: 100 mg/mL of Bupropion Hydrochloride
in methanol Bupropion 0.92 1.4 0.2
Standard stock solution: 0.5 mg/mL of m-chlorobenzoic hydrochloride
acid in methanol related
Chromatographic system compound A
(See Chromatography 621, Thin-Layer Chromatography.) Bupropion 1.0
Mode: TLC
Adsorbent: A 0.25-mm layer of high-performance silica aThe percentage is determined by direct comparison to the area of the
gel, previously washed with methanol peak for bupropion hydrochloride related compound B obtained from the
Application volume: 4 L System suitability solution.
Developing solvent system: Toluene, cyclohexane, and
glacial acetic acid (47:47:6)
Impurity Table 1 (continued) concentration of nominally 3.0 mg/mL of bupropion

hydrochloride. Add a portion of Diluent, equivalent to about
one-half of the flask volume, and shake by mechanical
means until the Tablets have disintegrated (between 30 and
60 min). Sonicate for 5 min, dilute with Diluent to volume,
Bupropion 1.14 0.2a and mix. Allow to stand for at least 30 min, and pipet 10.0
hydrochloride mL of the supernatant into a 50-mL volumetric flask. Dilute
related with Diluent to volume, mix, and pass through a suitable
compound B filter.
2-Bromo-3- 1.63 0.88 0.1 Chromatographic system
chloropropiophenone (See Chromatography 621, System Suitability.)
2-(tert- 2.30 1.1 0.2
Mode: LC
Butylamino)-3,4-
Detector: UV 224 nm
chloropropiophenone
Column: 4.6-mm 15-cm; 5-m base-deactivated packing
hydrochloride
L1
2-(tert- 2.74 0.69 0.2 Injection size: 10 L
Butylamino)-3,5- System suitability
chloropropiophenone Sample: Standard solution
hydrochloride Suitability requirements
Unknown All other peaks 1.00 0.1, individual Tailing factor: NMT 2.5
aThe percentage is determined by direct comparison to the area of the
peak for bupropion hydrochloride related compound B obtained from the
Analysis
System suitability solution.
Calculate the pecentage of C13H18ClNO HCl in the portion
of Tablets taken:
SPECIFIC TESTS
WATER DETERMINATION, Method I 921: NMT 0.5% Result = (rU/rS) (CS/CU) 100
ADDITIONAL REQUIREMENTS rU = peak area response from the Sample solution
PACKAGING AND STORAGE: Preserve in well-closed, light- rS = peak area response from the Standard solution
resistant containers. CS = concentration of USP Bupropion Hydrochloride
USP REFERENCE STANDARDS 11 RS in the Standard solution (mg/mL)
USP Bupropion Hydrochloride RS CU = nominal concentration of bupropion
USP Bupropion Hydrochloride Related Compound A RS hydrochloride in the Sample solution (mg/mL)
USP Bupropion Hydrochloride Related Compound B RS Acceptance criteria: 90.0%110.0%
PERFORMANCE TESTS
DISSOLUTION 711
Bupropion Hydrochloride Tablets Medium: Water; 900 mL
(Comment on this Monograph)id=m10523=Bupropion Apparatus 2: 50 rpm
Hydrochloride Tablets=B-Monos.pdf) Time: 45 min
Standard solution: USP Bupropion Hydrochloride RS in
DEFINITION Medium at a known concentration
Bupropion Hydrochloride Tablets contain NLT 90.0% and NMT Sample solution: Sample per Dissolution 711, diluted if
110.0% of the labeled amount of bupropion hydrochloride necessary with 0.1 N hydrochloric acid.
(C13H18ClNO HCl). Spectrometric conditions
Mode: UV
IDENTIFICATION Analytical wavelength: 252 nm
A. INFRARED ABSORPTION 197K Analysis
Sample: Crush 1 Tablet using a mortar and pestle. Prepare Samples Standard solution and Sample solution
an approximate 1% (w/w) dispersion of the sample in Tolerances: NLT 80% (Q) of the labeled amount of
potassium bromide. C13H18ClNO HCl is dissolved.
Acceptance criteria: The Sample shows strong bands at UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
about 1690, 1560, and 1240 cm1 and a weaker band at
about 740 cm1 similar to the reference preparation. ADDITIONAL REQUIREMENTS
B. The retention time of the major peak of the Sample PACKAGING AND STORAGE: Preserve in tight containers.
solution corresponds to that of the Standard solution, as USP REFERENCE STANDARDS 11
obtained in the Assay. USP Bupropion Hydrochloride RS
ASSAY
PROCEDURE
Diluent: Methanol and water (13:7) Bupropion Hydrochloride Extended-
Buffer: 6.8 g of monobasic potassium phosphate and 1.164
g of sodium hydroxide in 1 L of water
Release Tablets
Mobile phase: Methanol and Buffer (13:7) (Comment on this Monograph)id=m10524=Bupropion
Standard solution: 0.6 mg/mL of USP Bupropion Hydrochloride Extended-Release Tablets=B-Monos.pdf)
Hydrochloride RS in Diluent DEFINITION
Sample solution: Transfer a number of Tablets to a Bupropion Hydrochloride Extended-Release Tablets contain NLT
volumetric flask suitable to obtain a solution having a final 90.0% and NMT 110.0% of the labeled amount of bupropion
hydrochloride (C13H18ClNO HCl).
IDENTIFICATION Suitability requirements

A. INFRARED ABSORPTION 197K Resolution: NLT 1.5, between bupropion hydrochloride
Sample: Crush 1 Tablet using a mortar and pestle. Prepare related compound C and bupropion hydrochloride related
an approximate 1% (w/w) dispersion of the sample in compound F, System Suitability solution A
potassium bromide. Tailing factor: NMT 1.9, Standard solution
Acceptance criteria: The sample shows strong bands at Relative standard deviation: NMT 1.5%, Standard
about 1690, 1560, and 1240 cm1 and a weaker band at solution
about 740 cm1 similar to the reference preparation. Relative response factor: Between 0.22 and 0.26 for m-
B. The retention time of the major peak of the Sample chlorobenzoic acid
solution corresponds to that of the Standard solution, as [NOTEUse the responses from System suitability solution B
obtained in the Assay. and Standard solution.]
Analysis: Calculate the percentage of C13H18ClNO HCl in
ASSAY the portion of Tablets taken:
PROCEDURE
Diluent: Methanol and 0.001 N hydrochloric acid (2:8) Result = (rU/rS) (CS/CU) 100
Solution A: Acetonitrile, trifluoroacetic acid, and water
(10:0.04:90) rU = peak response for bupropion hydrochloride from
Solution B: Acetonitrile, trifluoroacetic acid, and water the Sample solution
(95:0.03:5) rS = peak response for bupropion hydrochloride from
Mobile phase: See the gradient table below. the Standard solution
CS = concentration of USP Bupropion Hydrochloride
Time Solution A Solution B RS in the Standard solution (mg/mL)
(min) (%) (%) CU = nominal concentration of bupropion
0 90 10 Acceptance criteria: 90.0%110.0%
3.4 87 13
PERFORMANCE TESTS
10.0 15 85
DISSOLUTION 711
10.1 0 100 For products labeled for dosing every 12 h
13.0 0 100 Test 1
13.2 90 10
Sample solution: Sample per Dissolution 711, diluted
19.0 90 10 with Medium as necessary.
Apparatus 2: 50 rpm
System suitability solution A: Separate solutions of 0.20 Times: 1, 4, and 8 h
mg/mL each of USP Bupropion Hydrochloride Related Standard solution: USP Bupropion Hydrochloride RS at a
Compound C RS and USP Bupropion Hydrochloride Related known concentration in the same Medium
Compound F RS in methanol. Transfer suitable volumes of Spectrometric conditions
each solution to a single volumetric flask to obtain 0.0018 Mode: UV spectrometry
mg/mL of USP Bupropion Hydrochloride Related Compound Analytical wavelength: 298 nm
C RS and 0.018 mg/mL of USP Bupropion Hydrochloride Cell: 1.0 cm
Related Compound F RS. Dilute with Diluent to volume. Analysis
System suitability stock solution B: 0.09 mg/mL of m- Samples: Standard solution and Sample solution
chlorobenzoic acid in methanol Tolerances: The percentage of the labeled amount of
System suitability solution B: 0.0018 mg/mL of m- C13H18ClNO HCl dissolved at the times specified conforms
chlorobenzoic acid from System suitability stock solution B to Acceptance Table 1.
and Diluent
Standard solution: 0.6 mg/mL of USP Bupropion Time (h) Amount Dissolved
Hydrochloride RS in Diluent
Sample solution: Transfer a number of Tablets to a suitable 1 25%45%
homogenizer vessel containing sufficient methanol to obtain 4 60%85%
a concentration of 3.0 mg of bupropion hydrochloride/mL. 8 NLT 80%
Immediately homogenize the sample for 30 s at 20,000
rpm. Allow extraction for 3 min, and follow by two Test 2: If the product complies with this test, the labeling
additional 10-s pulses, each at 20,000 rpm, pausing 3 min indicates that it meets USP Dissolution Test 2.
between these pulses to ensure complete extraction. Pass a Medium: 0.1 N hydrochloric acid, pH 1.5 (prepared by
portion of the solution through a nylon filter having a 0.45- transferring 50 mL of concentrated hydrochloric acid to
m porosity, discarding the first 24 mL of the filtrate. Pipet 6000 mL of water, adding 18 g of sodium hydroxide,
10.0 mL of the filtrate into a 50-mL volumetric flask, and mixing, and adjusting with either diluted sodium
add about 25 mL of 0.001 N hydrochloric acid. Allow to hydroxide or hydrochloric acid to a pH of 1.5 0.05); 900
cool to room temperature, and dilute with 0.001 N mL
hydrochloric acid to volume. Apparatus 1: 50 rpm
Chromatographic system Times: 1, 2, 4, and 6 h
(See Chromatography 621, System Suitability.) Determine the percentages of the labeled amount of
Mode: LC C13H18ClNO HCl dissolved by using the following
Detector: UV 226 nm method.
Column: 4.6-mm 10-cm; 3.5-m packing L1 Buffer: 3.45 g of sodium phosphate monobasic
Flow rate: 1.5 mL/min monohydrate in 996 mL of water. Add 4.0 mL of
Injection size: 5 L triethylamine, and adjust with phosphoric acid to a pH of
System suitability 2.80 0.05.
Samples: System suitability solution A, System suitability
solution B, and Standard solution
Mobile phase: Methanol and Buffer (7:13) For products labeled for dosing every 24 h
Standard solution: USP Bupropion Hydrochloride RS in Medium: 0.1 N hydrochloric acid; 900 mL
Medium at a known concentration similar to the one Apparatus 1: 75 rpm
expected in the Sample solution Times: 2, 4, 8, and 16 h
Sample solution: Use portions of the solution under test, Standard solution: USP Bupropion Hydrochloride RS at a
and pass through a 0.45-m nylon filter. known concentration in the same Medium
Chromatographic system Sample solution: Sample per Dissolution 711.
(See Chromatography 621, System Suitability.) Spectrometric conditions
Mode: LC Mode: UV spectrometry
Detector: UV 298 nm Analytical wavelength: 252 nm
Column: 4.6-mm 15-cm; packing L1 Cell: 1.0 cm
System suitability Tolerances: The percentage of the labeled amount of
Sample: Standard solution C13H18ClNO HCl dissolved at the times specified conforms
Suitability requirements to Acceptance Table 4.
Tailing factor: NMT 2.0 Time (h) Amount Dissolved
Analysis 2 NMT 20%
Samples: Standard solution and Sample solution 4 20%45%
Tolerances: The percentage of the labeled amount of 8 65%90%
C13H18ClNO HCl dissolved at the times specified conforms
to Acceptance Table 2. 16 NLT 80%
Test 6: If the product complies with this test, the labeling

Time (h) Amount Dissolved indicates that it meets USP Dissolution Test 6.
1 25%50% Medium and Apparaus: Proceed as directed for Test 4.
2 40%65% Times: 1, 2, 4, 8, and 12 h
Standard solution: USP Bupropion Hydrochloride RS at a
4 65%90% known concentration in Medium
6 NLT 80% Spectrometric conditions
Mode: UV spectrometry
Test 3: If the product complies with this test, the labeling Analytical wavelength: 252 nm
indicates that it meets USP Dissolution Test 3. Cell: 1.0 cm
Medium, Apparatus, Standard solution, Sample solution, Analysis: Determine the amount of C13H18ClNO HCl
Spectometric conditions, and Analysis: Proceed as dissolved in filtered portions of the solution under test,
directed for Test 1, except the wavelength is about 250 suitably diluted with Medium, if necessary, in comparison
nm. with the Standard solution.
Times: 1, 2, 4, and 6 h Tolerances: The percentage of the labeled amount of
Tolerances: The percentage of the labeled amount of C13H18ClNO HCl dissolved at the times specified conforms
C13H18ClNO HCl dissolved at the times specified conforms to Acceptance Table 6.
to Acceptance Table 3.
Time (h) Amount Dissolved
Time (h) Amount Dissolved 1 Between 15% and 35%
1 30%55% 2 Between 25% and 50%
2 50%75% 4 Between 40% and 65%
4 70%90% 8 Between 65% and 90%
6 NLT 80% 12 NLT 80%
Test 5: If the product complies with this test, the labeling UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
indicates that it meets USP Dissolution Test 5. Procedure for content uniformity
Medium and Analysis: Proceed as directed for Test 1. Standard solution: 0.33 mg/mL of USP Bupropion
Spectrometric conditions: Proceed as directed for Test 1, Hydrochloride RS in water
except to use a 0.5-cm cell. Sample solution: Transfer 1 Tablet to a suitable
Times: 1, 3, and 6 h homogenizer vessel containing a volume of water to obtain
Tolerances: The percentage of the labeled amount of a concentration of about 0.33 mg of bupropion
C13H18ClNO HCl dissolved at the times specified conforms hydrochloride/mL. Immediately homogenize the sample
to Acceptance Table 5. using single 30-s pulses each at 5,000, 10,000, and 15,000
rpm, and follow by two pulses each at 20,000 rpm. After
Time (h) Amount Dissolved the homogenate has settled, mix at 5000 rpm for an
1 35%55% additional 30 s. Pass a portion of the solution through a
nylon filter having a 0.45-m porosity, discarding the first 4
3 65%85% mL of the filtrate.
6 NLT 80%
Analysis Buspirone Hydrochloride

Sample solution: Sample per Dissolution 711, filter (Comment on this Monograph)id=m10540=Buspirone
IMPURITIES Hydrochloride=B-Monos.pdf)
Organic Impurities
PROCEDURE
Solution A, Solution B, Mobile phase, System suitability
solution A, System suitability solution B, Standard
solution, Sample solution, and Chromatographic
system: Proceed as directed in the Assay.
Analysis: Calculate the percentage of each impurity in the
portion of Tablets taken: C21H31N5O2 HCl 421.96
8-Azaspiro[4,5]decane-7,9-dione, 8-[4-[4-(2-pyrimidinyl)-1-
Result = (rU/rS) (CS/CU) F 100 piperazinyl]butyl]-, monohydrochloride;
rU = peak response for each impurity from the Sample N-[4-[4-(2-Pyrimidinyl)-1-piperazinyl]butyl]-1,1-
solution cyclopentanediacetamide monohydrochloride [33386-08-2;
rS = peak response for bupropion hydrochloride from 36505-84-7].
the Standard solution DEFINITION
CS = concentration of USP Bupropion Hydrochloride Buspirone Hydrochloride contains NLT 97.5% and NMT
RS in the Standard solution (mg/mL) 102.5% of C21H31N5O2 HCl, calculated on the as-is basis.
CU = nominal concentration of bupropion
hydrochloride in the Sample solution (mg/mL) IDENTIFICATION
F = relative response factor for each impurity (see A. INFRARED ABSORPTION 197K
Impurity Table 1 for values) B. The relative retention time of the major peak of the
Acceptance criteria: See Impurity Table 1. Sample solution corresponds to that of the Standard solution,
PACKAGING AND STORAGE: Preserve in well-closed containers. ASSAY
LABELING: When more than one Dissolution Test is given, the PROCEDURE
labeling states the Dissolution Test used only if Test 1 is not Solution A: 1.36 mg/mL of monobasic potassium
used. phosphate in water, adjust the solution with 10% sodium
USP REFERENCE STANDARDS 11 hydroxide (w/v) to a pH of 7.5, and filter
USP Bupropion Hydrochloride RS
USP Bupropion Hydrochloride Related Compound C RS
USP Bupropion Hydrochloride Related Compound F RS
Impurity Table 1
Relative Retention Relative Response Fac- Acceptance Criteria, NMT (%)

Name Time tor 100 mg or less 150 mg or greater
2-Amino-1-(3- 0.38 0.80 0.3 0.3
chlorophenyl)-1-propa-
none
(3S,5S,6S)-6-(3- 0.56 0.86 1.0 1.5
Chlorophenyl)-6-hydroxy-
5-methyl-3-thi-
omorpholine carboxylic
acid
(3S,5R,6R)-6-(3- 0.78 0.88 0.5 0.4
Chlorophenyl)-6-hydroxy-
5-methyl-3-thi-
omorpholine carboxylic
acid
Bupropion 1.0
Bupropion related com- 1.71 0.55 1.2 2.3
pound F
pound C
m-Chlorobenzoic acid 1.80 0.24 0.3 0.3
pound E
Any unspecified impurity 1.00 0.2 0.2
Total impurities 3.2 3.3
USP 32 Official Monographs / Buspirone 147
Mobile phase: Acetonitrile and Solution A (2:3) Acceptance criteria: NLT 8.0% and NMT 8.8%
Internal standard stock solution: 2.5 mg/mL of
propylparaben in methanol IMPURITIES
Internal standard solution: 0.125 mg/mL from Internal Inorganic Impurities
standard stock solution in water RESIDUE ON IGNITION 281: NMT 0.5%
Standard stock solution: Dissolve 50 mg of USP Buspirone HEAVY METALS, Method II 231: NMT 20 ppm
Hydrochloride RS in 25 mL of 1 N hydrochloric acid in a
100-mL volumetric flask; dilute with water to volume. SPECIFIC TESTS
Standard solution: 10.0 mL from Standard stock solution. WATER DETERMINATION, Method I 921: NMT 0.5%
Add 10.0 mL of Internal standard solution in a 50.0-mL ADDITIONAL REQUIREMENTS
volumetric flask, and add water to volume. PACKAGING AND STORAGE: Preserve in tight, light-resistant
Sample stock solution: Dissolve 50 mg of Buspirone containers, at controlled room temperature.
Hydrochloride in 25 mL of 1 N hydrochloric acid in a 100- USP REFERENCE STANDARDS 11
mL volumetric flask; dilute with water to volume. USP Buspirone Hydrochloride RS
Sample solution: 10.0 mL from Sample stock solution plus
10.0 mL of Internal standard solution in a 50.0-mL
volumetric flask. Add water to volume.
Chromatographic system Buspirone Hydrochloride Tablets
(See Chromatography 621, System Suitability.) (Comment on this Monograph)id=m10550=Buspirone
Mode: LC Hydrochloride Tablets=B-Monos.pdf)
Detector: UV 254 nm
Column: 3.9-mm 30-cm; packing L1 DEFINITION
Flow rate: 2 mL/min Buspirone Hydrochloride Tablets contain NLT 90.0% and NMT
Injection size: 25 L 110.0% of the labeled amount of buspirone hydrochloride
System suitability (C21H31N5O2 HCl).
[NOTEThe relative retention times for propylparaben and IDENTIFICATION
buspirone hydrochloride are about 0.55 and 1.0, A. INFRARED ABSORPTION 197K
respectively.] Sample solution: Grind 20 Tablets to a fine powder, add
Suitability requirements 50 mL of chloroform, stir for 35 min, and filter into a 250-
Resolution: NLT 4 between buspirone hydrochloride and mL evaporating flask. Evaporate the solution with the aid of
the internal standard a rotary evaporator to dryness at low heat. Use the residue.
Relative standard deviation: NMT 2.0% B. The relative retention time of the major peak of the
Analysis Sample solution corresponds to that of the Standard solution,
Calculate the percentage of C21H31N5O2 HCl in the portion as obtained in the Assay.
of Buspirone Hydrochloride taken:
ASSAY
Result = (RU/RS) (CS/CU) 100 PROCEDURE
RU = peak response ratio of buspirone hydrochloride phosphate in water. Adjust the solution with 10% sodium
to propylparaben from the Sample solution hydroxide (w/v) to a pH of 7.5, and filter.
RS = peak response ratio of buspirone hydrochloride Mobile phase: Acetonitrile and Solution A (2:3)
to propylparaben from the Standard solution Internal standard stock solution: 2.5 mg/mL of
CS = concentration of USP Buspirone Hydrochloride RS propylparaben in methanol
in the Standard solution (mg/mL) Internal standard solution: 0.125 mg/mL from Internal
CU = concentration of buspirone hydrochloride in the standard stock solution in water
Sample solution (mg/mL) Standard stock solution: Dissolve 50 mg of USP Buspirone
Acceptance criteria: 97.5%102.5% Hydrochloride RS in 25 mL of 1 N hydrochloric acid in a
100-mL volumetric flask; dilute with water to volume.
OTHER COMPONENTS Standard solution: 10.0 mL from Standard stock solution.
CONTENT OF CHLORIDE: Between 8.0% and 8.8% is found. Add 10.0 mL of Internal standard solution in a 50.0-mL
Sample solution: 400 mg in 20 mL of water. Add 3 mL of volumetric flask, and add water to volume.
nitric acid and 20.0 mL of 0.1 N silver nitrate VS. Gently boil Sample stock solution: Equivalent to 100 mg of buspirone
the mixture for about 5 min. Filter, rinse the flask with hydrochloride from powdered Tablets, in a 200-mL
about 80 mL of water divided into small portions, and filter volumetric flask. Add 50 mL of 1 N hydrochloric acid, and
each portion. Add 2 mL of 8% ferric ammonium sulfate. shake for 15 min. Add about 100 mL of water, and shake
Analysis: While stirring rapidly, titrate the excess silver for 30 min. Dilute with water to volume, mix, and filter,
nitrate with 0.1 N ammonium thiocyanate VS to a faint red- discarding the first 20 mL of the filtrate.
brown endpoint. Perform a blank determination (see
Titrimetry 541, Residual Titrations). Each mL of 0.1 N silver
nitrate consumed is equivalent to 3.545 mg of chloride.
148 Buspirone / Official Monographs USP 32
Sample solution: Transfer 10.0 mL of the Sample stock Busulfan

solution and 10.0 mL of Internal standard solution into a 50- (Comment on this Monograph)id=m10570=Busulfan=B-
mL volumetric flask. Dilute with water to volume, and mix. Monos.pdf)
Mode: LC
Detector: UV 254 nm
Flow rate: 2 mL/min
System suitability
Sample: Standard solution C6H14O6S2 246.30
[NOTEThe relative retention times for propylparaben and 1,4-Butanediol, dimethanesulfonate;
buspirone hydrochloride are 0.55 and 1.0, respectively.] Butane-1,4-diyl dimethanesulfonate [55-98-1].
Resolution: NLT 4 between buspirone hydrochloride and DEFINITION
the internal standard Busulfan contains NLT 98.0% and NMT 100.5% of C6H14O6S2,
Relative standard deviation: NMT 2.0% calculated on the dried basis.
Analysis
Samples: Standard solution and Sample solution IDENTIFICATION
Calculate the percentage of C21H31N5O2 HCl in the Tablets A. PROCEDURE
taken: Sample: 100 mg
Analysis: Fuse Sample with about 100 mg of potassium
Result = (RU/RS) (CS/CU) 100 nitrate and a pellet of potassium hydroxide weighing
approximately 250 mg. Cool, dissolve the residue in water,
RU = peak response ratio of buspirone hydrochloride acidify with 3 N hydrochloric acid, and add a few drops of
to propylparaben from the Sample solution barium chloride TS.
RS = peak response ratio of buspirone hydrochloride Acceptance criteria: A white precipitate is formed.
to propylparaben from the Standard solution B. PROCEDURE
CS = concentration of USP Buspirone Hydrochloride Sample: 100 mg
RS in the Standard solution (mg/mL) Analysis: Add 10 mL of water and 5 mL of 1 N sodium
CU = nominal concentration of buspirone hydroxide. Heat until a clear solution is obtained.
hydrochloride in the Sample solution (mg/mL) Acceptance criteria: An odor characteristic of
Acceptance criteria: 90.0%110.0% methanesulfonic acid is perceptible.
C. PROCEDURE
PERFORMANCE TESTS Sample solution: Use the solution from Identification B.
DISSOLUTION 711 Analysis: Cool the Sample solution obtained in Identification
Medium: 0.01 N hydrochloric acid; 500 mL test B, and divide it into two equal portions. To one portion
Apparatus 2: 50 rpm add 1 drop of potassium permanganate TS. Acidify the
Time: 30 min second portion of the solution with 2 N sulfuric acid, and
Sample solutions: Sample per Dissolution 711. Dilute with add 1 drop of potassium permanganate TS.
Medium as needed at a known concentration. Acceptance criteria: The first portion becomes purple
Standard solution: USP Buspirone Hydrochloride RS in which changes to violet, then to blue and finally to emerald
Medium green. In the second portion (acidified) the color of the
Spectometric conditions permanganate is not discharged.
Mode: UV
Analytical wavelength: 235 nm ASSAY
Analysis PROCEDURE
Samples: Standard solution and Sample solution Sample solution: 80 mg of Busulfan in a 250-mL conical
Tolerances: NLT 80% (Q) of the labeled amount of flask. Add about 30 mL of water, swirl, add phenolphthalein
C21H31N5O2.HCl is dissolved. TS, and neutralize with 0.05 N sodium hydroxide. Connect
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements the flask to a reflux air condenser, and boil the mixture
gently for NLT 30 min, adding water occasionally to
ADDITIONAL REQUIREMENTS maintain the volume. Cool to room temperature, and add
PACKAGING AND STORAGE: Preserve in tight, light-resistant phenolphthalein TS.
containers, at controlled room temperature.
USP Buspirone Hydrochloride RS
USP 32 Official Monographs / Butabarbital 149
Analysis: Titrate with 0.05 N sodium hydroxide VS. Each mL Transfer an equivalent to 80 mg of busulfan, from powdered
of 0.05 N sodium hydroxide is equivalent to 6.158 mg of Tablets (NLT 40), to a 100-mL beaker. Extract with four 20-
C6H14O6S2. mL portions of acetone, each time stirring the mixture well,
Acceptance criteria: 98.0%100.5% then allowing the insoluble matter to settle, and finally
decanting the supernatant through a sintered-glass filter
IMPURITIES into a 250-mL conical flask. Evaporate the combined
Inorganic Impurities acetone extracts to about 10 mL, add phenolphthalein TS,
RESIDUE ON IGNITION 281: NMT 0.1% and neutralize with 0.05 N sodium hydroxide. Evaporate to
dryness, and add about 30 mL of water. Connect the flask
SPECIFIC TESTS to a reflux air condenser, and boil the mixture gently for
MELTING RANGE OR TEMPERATURE 741: 115118 NLT 30 min, adding water occasionally to maintain the
LOSS ON DRYING 731: Dry it in a vacuum at 60 to volume. Cool to room temperature, and add
constant weight: it loses NMT 2.0% of its weight. phenolphthalein TS.
ADDITIONAL REQUIREMENTS Analysis: Titrate with 0.05 N sodium hydroxide VS. Each mL
PACKAGING AND STORAGE: Preserve in tight containers. of 0.05 N sodium hydroxide is equivalent to 6.158 mg of
LABELING: The label bears a warning that great care should C6H14O6S2.
be taken to prevent inhaling particles of Busulfan and Acceptance criteria: 93.0%107.0%
exposing the skin to it. PERFORMANCE TESTS
DISINTEGRATION 701: 30 min, the use of disks being
omitted
Busulfan Tablets UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
(Comment on this Monograph)id=m10600=Busulfan Tablets=B- ADDITIONAL REQUIREMENTS
Monos.pdf) PACKAGING AND STORAGE: Preserve in well-closed containers.
DEFINITION
Busulfan Tablets contain NLT 93.0% and NMT 107.0% of the
labeled amount of C6H14O6S2. Butabarbital
IDENTIFICATION (Comment on this Monograph)id=m10640=Butabarbital=B-
A. PROCEDURE Monos.pdf)
Sample: A suitable number of Tablets
Analysis: Pulverize the Sample and extract the powder with
several portions of acetone. Evaporate the combined
acetone extracts, with the aid of a current of air, on a steam
bath.
Acceptance criteria: The dry residue melts at about 115.
B. PROCEDURE
Sample: 100 mg of the powder obtained in Identification A C10H16N2O3 212.25
Analysis: Fuse the Sample with 100 mg of potassium nitrate 2,4,6(1H,3H,5H)-Pyrimidinetrione, 5-ethyl-5-(1-methylpropyl)-;
and a pellet of potassium hydroxide weighing 250 mg. 5-sec-Butyl-5-ethylbarbituric acid [125-40-6].
Cool, dissolve the residue in water, acidify with 3 N
hydrochloric acid, and add a few drops of barium chloride DEFINITION
TS. Butabarbital contains NLT 98.5% and NMT 101.0% of
Acceptance criteria: A white precipitate is formed. C10H16N2O3, calculated on the dried basis.
C. PROCEDURE
Sample: 100 mg of the powder obtained in Identification A IDENTIFICATION
Analysis: To the Sample add 10 mL of water and 5 mL of 1 INFRARED ABSORPTION 197M
N sodium hydroxide. Heat until a clear solution is obtained.
Acceptance criteria: An odor characteristic of ASSAY
methanesulfonic acid is perceptible. PROCEDURE
D. PROCEDURE Internal standard solution: 2 mg/mL of tetracosane in
Sample solution: Use the solution from Identification C. chloroform
Analysis: Cool the Sample solution, and divide it into two Standard stock solution: 2 mg/mL of USP Butabarbital RS
equal portions. To one portion add 1 drop of potassium in chloroform
permanganate TS. Acidify the second portion of the Sample Standard solution: 1 mg/mL from Standard stock solution
solution with 2 N sulfuric acid, and add 1 drop of potassium diluted with Internal standard solution
permanganate TS. Sample stock solution: 2 mg/mL of Butabarbital in
Acceptance criteria: The first portion becomes purple chloroform
which changes to violet, then to blue, and finally to Sample solution: 1 mg/mL from Sample stock solution
emerald-green. In the second portion (acidified), the color of diluted with Internal standard solution
the permanganate is not discharged. Chromatographic system
ASSAY
PROCEDURE
Sample solution: [NOTEGuard against accidental
inhalation of the fine powder.]
150 Butabarbital / Official Monographs USP 32
Mode: GC Standard solution B, corresponding to NMT a total of 1% of

Detector: Flame ionization impurities.
Column: 4-mm 1.8-m
Packing: 10% phase G37 on support S1AB SPECIFIC TESTS
Temperature MELTING RANGE OR TEMPERATURE, Class Ia 741: 164167
Column: 260 LOSS ON DRYING 731: Dry a sample at 105 for 2 h: it loses
Injection port: 260 NMT 1.0% of its weight.
Detector block: 300
Carrier gas: Dry nitrogen ADDITIONAL REQUIREMENTS
Flow rate: 50 mL/min PACKAGING AND STORAGE: Preserve in tight containers.
System suitability USP Butabarbital RS
[NOTEThe relative retention times for butabarbital and
tetracosane are 0.6 and 1.0, respectively.] Butabarbital Sodium
Resolution: NLT 3.0 between the Butabarbital and (Comment on this Monograph)id=m10650=Butabarbital
internal standard peaks Sodium=B-Monos.pdf)
Tailing factor: NMT 1.3 for Butabarbital and NMT 1.2 C10H15N2NaO3 234.23
internal standard peaks 2,4,6(1H,3H,5H)-Pyrimidinetrione, 5-ethyl-5-(1-methylpropyl)-,
Relative standard deviation: NMT 1.0% monosodium salt;
Analysis Sodium 5-sec-butyl-5-ethylbarbiturate [143-81-7].
Butabarbital taken: DEFINITION
Butabarbital Sodium contains NLT 98.2% and NMT 100.5% of
Result = (RU/RS) (CS/CU) 100 C10H15N2NaO3, calculated on the dried basis.
RU = peak response ratio from the Sample solution IDENTIFICATION

RS = peak response ratio from the Standard solution A. INFRARED ABSORPTION 197K: 150 mg to a suitable
CS = concentration of USP Butabarbital RS in the separator, dissolve in 10 mL of water, and add 15 mL of 3 N
Standard solution (mg/mL) hydrochloric acid. Extract with three 20-mL portions of
CU = concentration of Butabarbital in the Sample chloroform, filter the extracts through anhydrous sodium
solution (mg/mL) sulfate, and collect the extracts in a suitable beaker.
Acceptance criteria: 98.5%101.0% Evaporate the combined chloroform extracts on a steam
bath with the aid of a current of air to dryness, and dry the
IMPURITIES residue at 105 for 2 h.
Inorganic Impurities B. ULTRAVIOLET ABSORPTION 197U
RESIDUE ON IGNITION 281: NMT 0.1% Analytical wavelength: 240 nm
Organic Impurities Standard solution: 10 g/mL in pH 9.6 alkaline borate
PROCEDURE buffer (see Reagents, Indicators, and SolutionsBuffer
Standard solution A: 4.0 mg/mL of USP Butabarbital RS Solutions)
in a mixture of chloroform and methanol (1:1) Sample solution: 10 g/mL in pH 9.6 alkaline borate buffer
Standard solution B: 0.4 mg/mL from Standard solution A Acceptance criteria: Absorptivities, calculated on the dried
in a mixture of chloroform and methanol (1:1) basis, do not differ by more than 3.0%
Sample solution: 40 mg/mL of Butabarbital in a mixture C. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the
of chloroform and methanol (1:1) requirements
Chromatographic system Analysis: Ignite about 100 mg.
Mode: TLC ASSAY
Adsorbent: 0.25-mm layer of chromatographic silica gel PROCEDURE
mixture Standard stock solution: 0.125 mg/mL of USP Butabarbital
Application volume: 10 L RS in pH 9.6 Alkaline Borate Buffer (see Reagents, Indicators,
Developing solvent system: Acetone, methylene and SolutionsBuffer Solutions)
chloride, methanol, and ammonium hydroxide (5:3:1:1) Standard solution: 0.0125 mg/mL from Standard stock
Analysis solution in pH 9.6 Alkaline Borate Buffer
Samples: Standard solution A, Standard solution B, and Sample stock solution: 0.14 mg/mL of Butabarbital
Sample solution Sodium in pH 9.6 Alkaline Borate Buffer
Proceed as directed under the General Chapter. Develop Standard solution: 0.014 mg/mL from Sample stock
the chromatogram until the solvent front has moved solution in pH 9.6 Alkaline Borate Buffer
about three-fourths of the length of the plate. Remove Spectrometric conditions
the plate from the developing chamber, and dry in a Mode: UV
current of air. Spray the plate with a solution of Analytical wavelength: 240 nm
mercurous nitrate dihydrate in 0.15 N nitric acid (1 in Samples: Standard solution and Sample solution
100), and immediately estimate the intensities of any Analysis: Calculate the percentage of C10H15N2NaO3 in the
spots in the chromatogram of the Sample solution, other portion of Butabarbital Sodium taken:
than the principal spot, in comparison with Standard
solution B. Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100
the Sample solution corresponds to that of Standard solution AS = absorbance of the solution from the Standard
A; and the sum of the intensities of any secondary spots solution
observed in the chromatogram of the Sample solution is AU = absorbance of the solution from the Sample
NMT the intensity of the principal spot produced by solution
USP 32 Official Monographs / Butabarbital 151
CS = concentration of USP Butabarbital RS in the IDENTIFICATION

Standard solution (g/mL) INFRARED ABSORPTION 197K
CU = concentration of Butabarbital Sodium in the Sample: Equivalent to 150 mg of butabarbital sodium from
Sample solution (g/mL) a volume of Oral Solution, to a separator. Render it distinctly
Mr1 = molecular weight of butabarbital sodium, 234.23 alkaline by the addition of 1 N sodium hydroxide, and
Mr2 = molecular weight of butabarbital, 212.25 saturate it with sodium chloride. Extract the mixture with
Acceptance criteria: 98.2%100.5% two 15-mL portions of ether, and discard the ether. Acidify
the solution with hydrochloric acid, and render it just
IMPURITIES alkaline to litmus by adding small portions of sodium
Inorganic Impurities bicarbonate (carbonate-free). Extract the liberated acid
HEAVY METALS, Method II 231: NMT 30 ppm barbiturate using five 20-mL portions of chloroform. Wash
Organic Impurities the combined chloroform extracts with 10 mL of water
PROCEDURE acidified with 1 drop of hydrochloric acid, then extract the
Standard solution A: 4.0 mg/mL of USP Butabarbital RS water with 10 mL of chloroform, adding the latter to the
in a mixture of chloroform and methanol (1:1) main chloroform solution. Pass the chloroform solution
Standard solution B: 0.4 mg/mL from Standard solution A through a pledget of cotton or other suitable filter,
in a mixture of chloroform and methanol (1:1) previously washed with chloroform, into a tared beaker, and
Sample solution: 44 mg/mL of Butabarbital Sodium in a finally wash the separator and the filter with three 5-mL
mixture of chloroform and methanol (1:1) portions of chloroform. Evaporate the combined chloroform
Chromatographic system solution and washings on a steam bath with the aid of a
(See Chromatography 621, Thin-Layer Chromatography.) current of air to dryness, and dry the residue at 105 for 2
Mode: TLC h.
mixture ASSAY
Application volume: 10 L PROCEDURE
Developing solvent system: Acetone, methylene chloride, Internal standard solution: 0.7 mg/mL of secobarbital in
methanol, and ammonium hydroxide (5:3:1:1) chloroform
Analysis Standard solution: 1 mg/mL of USP Butabarbital RS and
Samples: Standard solution A, Standard solution B, and 1.4 mg/mL of secobarbital in chloroform
Sample solution Sample solution: Equivalent to 30 mg of butabarbital
Proceed as directed under the General Chapter. Develop sodium from a volume of Oral Solution, to a separator. Add
the chromatogram in a solvent system until the solvent 1 mL of bromine water (prepared by dissolving 2.0 mL of
front has moved about three-fourths of the length of the bromine and 10 g of potassium bromide in 60 mL of water),
plate. Remove the plate from the developing chamber, and swirl. Allow to stand for 5 min, add 1 mL of sodium
and in a current of air. Spray the plate with a solution of metabisulfite solution (1 in 10), and swirl. Add 300 mg of
mercurous nitrate dihydrate in 0.15 N nitric acid (1 in sodium bicarbonate in small portions, with mixing, and
100), and immediately estimate the intensities of any extract with four 10-mL portions of chloroform. Filter the
spots in the chromatogram of the Sample solution, other extracts through about 15 g of anhydrous sodium sulfate
than the principal spot, in comparison with Standard that is supported on a funnel by a small pledget of glass
solution B. wool. Collect the combined filtrates in a 50-mL volumetric
Acceptance criteria: The RF value of the principal spot of flask, wash the sodium sulfate with 5 mL of chloroform,
the Sample solution corresponds to that of Standard solution collecting the washing with the filtrate, dilute with
A; and the sum of the intensities of any secondary spots chloroform to volume, and mix. Combine 2.0 mL of this
observed in the chromatogram of the Sample solution is solution with 2.0 mL of Internal standard solution in a
NMT the intensity of the principal spot produced by suitable container, and reduce the volume to about 1 mL by
Standard solution B, corresponding to NMT a total of 1% of evaporation, with the aid of a stream of dry nitrogen, at
impurities. room temperature.
[NOTEThis solution includes a bromination step for
SPECIFIC TESTS elimination of parabens and a carbonatechloroform
COMPLETENESS OF SOLUTION: 1.0 g in 10 mL of carbon extraction for elimination of benzoic acid.]
dioxide-free water: after 1 min, the solution is clear and free Chromatographic system and System suitability: Proceed
from undissolved solid as directed for Chromatographic System and System Suitability
PH 791: 10.011.2, in the solution prepared for the test under Barbiturate Assay 361.
for Completeness of Solution Resolution: NLT 2.4 between butabarbital and secobarbital
LOSS ON DRYING 731: Dry a sample at 150 to constant [NOTEThe relative retention times for butabarbital and
weight: it loses NMT 5.0% of its weight. secobarbital are 0.6 and 1.0, respectively.]
Analysis
ADDITIONAL REQUIREMENTS Samples: Standard solution and Sample solution
PACKAGING AND STORAGE: Preserve in tight containers. Proceed as directed for Barbiturate Assay 361, Analysis.
USP REFERENCE STANDARDS 11 Calculate the percentage of C10H15N2NaO3 in each mL of the
USP Butabarbital RS Oral Solution taken:
Butabarbital Sodium Oral Solution rU = peak response from the Sample solution
(Comment on this Monograph)id=m10670=Butabarbital rS = peak response from the Standard solution
Sodium Oral Solution=B-Monos.pdf) CS = concentration of USP Butabarbital RS in the
DEFINITION CU = nominal concentration of butabarbital sodium in
Butabarbital Sodium Oral Solution contains NLT 90.0% and the Sample solution (g/mL)
NMT 110.0% of the labeled amount of butabarbital sodium Mr1 = molecular weight of butabarbital sodium, 234.23
(C10H15N2NaO3).
152 Butabarbital / Official Monographs USP 32
Mr2 = molecular weight of butabarbital, 212.25 Analysis

Acceptance criteria: 90.0%110.0% Samples: Standard solution and Sample solution
Proceed as directed for Barbiturate Assay 361, Procedure.
OTHER COMPONENTS Calculate the percentage of C10H15N2NaO3 in the portion of
Alcohol Determination, Method II 611: Between 95.0% Tablets taken:
and 115.0% of the labeled amount of C2H5OH
USP Butabarbital RS CS = concentration of USP Butabarbital RS in the
CU = nominal concentration of butabarbital sodium in
Butabarbital Sodium Tablets the Sample solution (mg/mL)
Mr1 = molecular weight of butabarbital sodium, 234.23
(Comment on this Monograph)id=m10680=Butabarbital Mr2 = molecular weight of butabarbital, 212.25
Sodium Tablets=B-Monos.pdf) Acceptance criteria: 90.0%110.0%
DEFINITION PERFORMANCE TESTS
Butabarbital Sodium Tablets contain NLT 90.0% and NMT DISSOLUTION 711
110.0% of the labeled amount of butabarbital sodium Medium: Water; 900 mL
(C10H15N2NaO3). Apparatus 1: 100 rpm
Time: 45 min
IDENTIFICATION Sample solutions: Sample per Dissolution 711. Mix with
INFRARED ABSORPTION 197K sufficient ammonium hydroxide to provide a concentration
Sample: Equivalent to 150 mg of butabarbital sodium from of 0.5 N ammonium hydroxide. Dilute with Medium as
powdered Tablets, in 1 mL of dimethyl sulfoxide and 1 mL necessary.
of water. Add hydrochloric acid dropwise until the solution Standard solution: USP Butabarbital RS in Medium to a
is just acid to litmus, and mix. Add 3 g of chromotographic known concentration
siliceous earth, and mix. Proceed as directed for Spectrometric conditions
Chromatography 621, Column Partition Chromatography, Mode: UV
packing the chromatographic tube as follows. The lower Analytical wavelength: 239 nm
layer consists of 4 g of chromatographic siliceous earth Analysis
mixed with 3 mL of sodium carbonate solution (1 in 10), Samples: Standard solution and Sample solution
and the upper layer is the Sample. Wash the column with 75 Tolerances: NLT 75% (Q) of the labeled amount of
mL of a water-saturated mixture of isooctane and ether C10H15N2NaO3 is dissolved.
(4:1), and discard the washing. Elute the Butabarbital with UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
200 mL of water-saturated ether, collecting the eluate in a Spectrometric conditions
suitable vessel. Evaporate the eluate to dryness on a steam Detector: UV
bath under a current of air, and dry the residue at 105 for Analytical wavelength: 240 nm
2 h. Solution A: Methanol and 1 N hydrochloric acid (9:1)
ASSAY Standard solution: 0.45 mg/mL of USP Butabarbital RS in
PROCEDURE Solution A
Internal standard solution: 1.2 mg/mL of secobarbital in Sample solution: Transfer 1 finely powdered Tablet to a 25-
chloroform mL volumetric flask. Add Solution A to volume, and mix.
Standard solution: 0.8 mg/mL of USP Butabarbital RS and Filter, discarding the first 5 mL of the filtrate, and dilute the
1 mg/mL of secobarbital in chloroform subsequent filtrate, if necessary, with Solution A to obtain a
Sample solution: Transfer an equivalent to 50 mg of solution containing 0.50.6 mg of butabarbital sodium/mL.
butabarbital sodium, from powdered Tablets (NLT 20), to a Analysis: Transfer 2.0 mL each of the Standard solution and
50-mL volumetric flask. Add 35 mL of dilute ammonium the Sample solution to separate 100-mL volumetric flasks,
hydroxide (1 in 25), dilute with water to volume, and mix. and transfer 2.0 mL of Solution A to a third volumetric flask
Filter, if necessary, discarding the first 15 mL of the filtrate, to provide a blank. Dilute each flask with pH 9.6 Alkaline
and transfer 25.0 mL of the clear solution to a separator. Borate Buffer (see Reagents, Indicators, and Solutions
Add 2 mL of hydrochloric acid, and extract with three 25- Solutions), and mix. Concomitantly determine the
mL portions of chloroform. Filter the extracts through about absorbances of the solutions in 1-cm cells at the wavelength
15 g of anhydrous sodium sulfate that is supported on a of maximum absorbance at about 240 nm, with a suitable
funnel by a small pledget of glass wool. Collect the spectrophotometer, using the blank to set the instrument.
combined filtrate in a 100-mL volumetric flask, wash the Calculate the percentage of C10H15N2NaO3 in the Tablet:
sodium sulfate with 15 mL of chloroform, collecting the
washing with the filtrate, dilute with chloroform to volume, Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100
and mix. Combine 4.0 mL of this solution with 1.0 mL of AU = absorbance of the solution from the Sample
Internal standard solution in a suitable container, and reduce solution
the volume to about 1 mL by evaporation, with the aid of a AS = absorbance of the solution from the Standard
stream of nitrogen, at room temperature. solution
Chromatographic system and System suitability: Proceed CS = concentration of USP Butabarbital RS in the
as directed for Barbiturate Assay 361, Chromatographic Standard solution (mg/mL)
System and System Suitability. CU = nominal concentration of butabarbital sodium in
Resolution: NLT 2.4 between butabarbital and the Sample solution, based upon the labeled
secobarbital quantity/Tablet and the extent of dilution
[NOTEThe relative retention times for butabarbital and (mg/mL)
secobarbital are 0.6 and 1.0, respectively.]
USP 32 Official Monographs / Butalbital 153
Mr1 = molecular weight of butabarbital sodium, 234.23 Application volume: 10 L

Mr2 = molecular weight of butabarbital, 212.25 Developing solvent system: Acetone, dichloromethane,
methanol, and ammonium hydroxide (5:3:1:1)
ADDITIONAL REQUIREMENTS Spray reagent: Dissolve 5 g of potassium hydroxide in a
PACKAGING AND STORAGE: Preserve in well-closed containers. mixture of 25 mL of water and 75 mL of alcohol.
USP Butabarbital RS Samples: Standard solutions A, B, C, and D and Sample
solution
In a suitable chromatographic chamber, lined with filter
Butalbital paper, place a volume of the Developing solvent system
sufficient to develop the chromatogram. Cover the
(Comment on this Monograph)id=m10790=Butalbital=B- chamber, and allow it to equilibrate for 30 min. Apply 10
Monos.pdf) L each of the Sample solution and Standard solutions A,
B, C, and D to a suitable thin-layer chromatographic
plate. Develop the chromatogram until the solvent front
has moved three-fourths of the length of the plate.
Remove the plate from the developing chamber, mark
the solvent front, and dry the plate in a current of air.
Spray the plate with Spray reagent. Allow the plate to dry
in warm air for 10 min, and examine the chromatograms
under UV light.
C11H16N2O3 224.26 Acceptance criteria: The chromatograms show principal
2,4,6(1H,3H,5H)-Pyrimidinetrione, 5-(2-methylpropyl)-5-(2- spots at the same RF value; and the sum of the intensities
propenyl)-; of any secondary spots, if present in the chromatogram
5-Allyl-5-isobutylbarbituric acid [77-26-9]. from the Sample solution, are NMT 1% of that of the
DEFINITION principal spot from Standard solution A. [NOTEThe
Butalbital contains NLT 98.0% and NMT 102.0% of relative intensities of the principal spots from Standard
C11H16N2O3, calculated on the dried basis. solution A, Standard solution B, Standard solution C, and
Standard solution D are 1, 0.01, 0.005, and 0.0025,
IDENTIFICATION respectively.]
B. ULTRAVIOLET ABSORPTION 197U SPECIFIC TESTS
Analytical wavelength: 246 nm MELTING RANGE OR TEMPERATURE 741: 138141
Sample solution: 15 g/mL in 0.1 N sodium hydroxide LOSS ON DRYING 731: Dry a sample in a vacuum at room
Acceptance criteria: Absorptivities, calculated on the dried temperature to constant weight: it loses NMT 0.2% of its
basis, do not differ by more than 2.5% weight.
ASSAY ADDITIONAL REQUIREMENTS

PROCEDURE PACKAGING AND STORAGE: Preserve in well-closed containers.
Sample: 180 mg of Butalbital USP REFERENCE STANDARDS 11
Analysis: Dissolve in a mixture of 25 mL of alcohol and 25 USP Butalbital RS
mL of sodium carbonate solution (3 in 100). Titrate with 0.1
N silver nitrate VS, using a silver electrode, either with a
suitable reference electrode containing a saturated aqueous
solution of potassium nitrate, or a combination electrode in
Butalbital, Acetaminophen, and
which the reference portion of the electrode contains a Caffeine Capsules
saturated aqueous solution of potassium nitrate. Each mL of (Comment on this Monograph)id=m10798=Butalbital,
0.1 N silver nitrate is equivalent to 22.43 mg of C11H16N2O3. Acetaminophen, and Caffeine Capsules=B-Monos.pdf)
DEFINITION
IMPURITIES Butalbital, Acetaminophen, and Caffeine Capsules contain NLT
Inorganic Impurities 90.0% and NMT 110.0% of the labeled amounts of butalbital
RESIDUE ON IGNITION 281: NMT 0.1% (C11H16N2O3), acetaminophen (C8H9NO2), and caffeine
HEAVY METALS, Method II 231: NMT 20 ppm (C8H10N4O2).
Organic Impurities
PROCEDURE IDENTIFICATION
Solution A: Chloroform and methanol (1:1) The retention times of the butalbital, acetaminophen, and
Sample solution: 40 mg/mL of Butalbital in Solution A caffeine peaks of the Sample solution correspond to those of
Standard solution A: 40 mg/mL of USP Butalbital RS in the butalbital, acetaminophen, and caffeine peaks of the
Solution A Standard solution, as obtained in the Assay.
Standard solution B: 0.4 mg/mL from Standard solution A ASSAY
in Solution A PROCEDURE
Standard solution C: Standard solution B and Solution A Mobile phase: Transfer 800 mg of monobasic potassium
(1:1) phosphate to a 2000-mL volumetric flask. Dissolve in 1100
Standard solution D: Standard solution C and Solution A mL of water, and dilute with methanol to volume.
(1:1) Internal standard solution: 0.65 mg/mL of phenacetin in
Chromatographic system methanol
(See Chromatography 621, Thin-Layer Chromatography.) Standard stock solution A: 0.01B mg/mL of USP Butalbital
Mode: TLC RS in Internal standard solution, B being the labeled amount
mixture
154 Butalbital / Official Monographs USP 32
of butalbital in mg/Capsule, sonicating and shaking the the labeled amounts of acetaminophen, butalbital, and
solution, if necessary, to dissolve caffeine, respectively, in mg/Capsule
Standard stock solution B: 0.01C mg/mL of USP Caffeine Standard solution: Standard stock solution and water (1:19)
RS in Internal standard solution, C being the labeled amount Sample solution: Sample per Dissolution 711. Dilute with
of caffeine in mg/Capsule, sonicating and shaking the Medium to concentration that is similar to the Standard
solution, if necessary, to dissolve solution.
Standard solution: Transfer to a 50-mL volumetric flask Analysis
0.1A mg of USP Acetaminophen RS, A being the labeled Samples: Standard solution and Sample solution
amount of acetaminophen in mg/Capsule, 10.0 mL of Pass a portion of the Sample solution through a filter of 10-
Standard stock solution A, and 10.0 mL of Standard stock m or finer porosity. Separately inject equal volumes (20
solution B. Sonicate for 5 min, and dilute with water to L) of the filtrate and the Standard solution.
volume. This solution contains 0.002B mg/mL of butalbital, Calculate the percentage of butalbital (C11H16N2O3),
0.002A mg/mL of acetaminophen, and 0.002C mg/mL of acetaminophen (C8H9NO2), and caffeine (C8H10N4O2)
caffeine. Pass a portion of this solution through a suitable dissolved by the same formula:
filter having a 0.5-m or finer porosity, and use the filtrate
as the Standard solution. Result = (rU/rS) (CS/CU) 100
Sample stock solution: Equivalent to 1 average Capsule
weight (powdered NLT 20 Capsules), in a 200-mL rU = peak response of the corresponding analyte from
volumetric flask. Add Internal standard solution to volume. the Sample solution
Sonicate for 15 min, mix, and allow to cool and settle. rS = peak response of the corresponding USP
Sample solution: Transfer 20.0 mL of the clear supernatant Reference Standard from the Standard solution
of Sample stock solution to a 50-mL volumetric flask, and CS = concentration of the appropriate USP Reference
dilute with water to volume. Pass a portion of this solution Standard in the Standard solution (mg/mL)
through a filter of 0.5 m or finer porosity, discarding the CU = nominal concentration of the corresponding
first 5 mL of the filtrate. Use the clear filtrate. analyte in the Sample solution (mg/mL)
Chromatographic system Tolerances: NLT 80% (Q) of the labeled amounts of
(See Chromatography 621, System Suitability.) C11H16N2O, C8H9NO2, and C8H10N4O2 is dissolved.
Mode: LC UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Detector: UV 216 nm
Column: 4-mm 25-cm; packing L1 ADDITIONAL REQUIREMENTS
Flow rate: 2 mL/min PACKAGING AND STORAGE: Preserve in tight containers.
System suitability USP Acetaminophen RS
Sample: Standard solution USP Butalbital RS
[NOTEThe relative retention times for acetaminophen, USP Caffeine RS
caffeine, phenacetin, and butalbital are about 0.16, 0.33,
Suitability requirements Butalbital, Acetaminophen, and
Resolution: NLT 1.2 between any two peaks Caffeine Tablets
calculated from the butalbital peak (Comment on this Monograph)id=m10800=Butalbital,
Relative standard deviation: NMT 2.0% of the Acetaminophen, and Caffeine Tablets=B-Monos.pdf)
acetaminophen, caffeine, and butalbital responses
Analysis DEFINITION
Samples: Standard solution and Sample solution Butalbital, Acetaminophen, and Caffeine Tablets contain NLT
Calculate the percentage of butalbital (C11H16N2O3), 90.0% and NMT 110.0% of the labeled amounts of butalbital
acetaminophen (C8H9NO2), and caffeine (C8H10N4O2) in (C11H16N2O3), acetaminophen (C8H9NO2), and caffeine
the portion of Capsules taken: (C8H10N4O2).
IDENTIFICATION
Result = (RU/RS) (CS/CU) 100 The retention times of the butalbital, acetaminophen, and
RU = peak response ratio of the corresponding analyte caffeine peaks of the Sample solution correspond to those of
to phenacetin from the Sample solution the butalbital, acetaminophen, and caffeine peaks of the
RS = peak response ratio of the corresponding analyte Standard solution, as obtained in the Assay.
to phenacetin from the Standard solution ASSAY
CS = concentration of appropriate USP Reference PROCEDURE
Standard in the Standard solution (mg/mL) Mobile phase: Transfer 800 mg of monobasic potassium
CU = nominal concentration of the corresponding phosphate to a 2000-mL volumetric flask. Dissolve in 1100
analyte in the Sample solution (mg/mL) mL of water, and dilute with methanol to volume.
Acceptance criteria: 90.0%110.0% of C11H16N2O3, Internal standard solution: 0.65 mg/mL of phenacetin in
C8H9NO2, and C8H10N4O2 methanol
PERFORMANCE TESTS Standard stock solution A: 0.01B mg/mL of USP Butalbital
DISSOLUTION 711 RS in Internal standard solution, B being the labeled amount
Medium: Water; 900 mL of butalbital, in mg/Tablet, sonicating and shaking the
Apparatus 1: 100 rpm solution, if necessary, to dissolve
Time: 60 min Standard stock solution B: 0.01C mg/mL of USP Caffeine
Mobile phase, Chromatographic system, and System RS in Internal standard solution, C being the labeled amount
suitability: Proceed as directed in the Assay. of caffeine, in mg/Tablet, sonicating and shaking the
Standard stock solution: 0.02A mg/mL of USP solution, if necessary, to dissolve
Acetaminophen RS, 0.02B mg/mL of USP Butalbital RS, and Standard solution: Transfer to a 50-mL volumetric flask
0.02C mg/mL of USP Caffeine RS, in which A, B, and C are 0.1A mg of USP Acetaminophen RS, A being the labeled
amount of acetaminophen, in mg/Tablet, 10.0 mL of Analysis

Standard stock solution A, and 10.0 mL of Standard stock Samples: Standard solution and Sample solution
solution B, sonicate for 5 min, and dilute with water to Calculate the percentage of butalbital (C11H16N2O3),
volume. This solution contains 0.002B mg/mL of butalbital, acetaminophen (C8H9NO2), and caffeine (C8H10N4O2)
0.002A mg/mL of acetaminophen, and 0.002C mg/mL of dissolved by the same formula:
caffeine. Pass a portion of this solution through a suitable
filter having a 0.5-m or finer porosity, and use the filtrate Result = (rU/rS) CS V 100/L
as the Standard solution.
Sample stock solution: Equivalent to 1 average Tablet rU = peak response of the corresponding analyte from
weight (powdered NLT 20 Tablets), into a 200-mL the Sample solution
volumetric flask. Add Internal standard solution to volume. rS = peak response of the corresponding USP
Sonicate for 15 min, mix, and allow to cool and settle. Reference Standard from the Standard solution
Sample solution: Transfer 20.0 mL of the clear supernatant CS = concentration of the appropriate USP Reference
of Sample stock solution to a 50-mL volumetric flask, and Standard in the Standard solution (mg/mL)
dilute with water to volume. Pass a portion of this solution V = volume of Medium, 900 mL
through a filter of 0.5-m or finer porosity, discarding the L = Tablet label claim of corresponding analyte (mg)
first 5 mL of the filtrate. Use the clear filtrate. Tolerances: NLT 80% (Q) of the labeled amounts of
Chromatographic system C11H16N2O, C8H9NO2, and C8H10N4O2 is dissolved.
(See Chromatography 621, System Suitability.) UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Mode: LC
Column: 4-mm 25-cm; packing L1 PACKAGING AND STORAGE: Preserve in tight containers.
Injection size: 10 L USP Acetaminophen RS
System suitability USP Butalbital RS
Sample: Standard solution USP Caffeine RS
[NOTEThe relative retention times for acetaminophen,
caffeine, phenacetin, and butalbital are about 0.16, 0.33,
0.77, and 1.0, respectively.] Butalbital and Aspirin Tablets
Resolution: NLT 1.2 between any two peaks (Comment on this Monograph)id=m10805=Butalbital and
Column efficiency: NLT 1000 theoretical plates, Aspirin Tablets=B-Monos.pdf)
calculated from the butalbital peak DEFINITION
Relative standard deviation: NMT 2.0% of the Butalbital and Aspirin Tablets contain NLT 90.0% and NMT
acetaminophen, caffeine, and butalbital responses 110.0% of the labeled amounts of butalbital (C11H16N2O3) and
Analysis aspirin (C9H8O4).
Calculate the percentage of butalbital (C11H16N2O3), IDENTIFICATION
acetaminophen (C8H9NO2), and caffeine (C8H10N4O2) in The retention times of the butalbital and aspirin peaks of the
the portion of Tablets taken: Sample solution correspond to those of the butalbital and
aspirin peaks of Standard solution A and Standard solution B,
Result = (RU/RS) (CS/CU) 100 as obtained in the Assay.
RU = peak response ratio of the corresponding analyte ASSAY
to phenacetin from the Sample solution BUTALBITAL AND ASPIRIN
RS = peak response ratio of the corresponding analyte Mobile phase: Acetonitrile, phosphoric acid, and water
to phenacetin from the Standard solution (725:4:3100). Adjust the ratio as necessary.
CS = concentration of appropriate USP Reference Solvent mixture: Formic acid and acetonitrile (1:100)
Standard in the Standard solution (mg/mL) Standard stock solution A: 3250J g/mL of USP Butalbital
CU = nominal concentration of the corresponding RS in Solvent mixture, J being the ratio of the labeled
analyte in the Sample solution (mg/mL) amount, in mg, of butalbital to the labeled amount of
Acceptance criteria: 90.0%110.0% of C11H16N2O3, aspirin, in mg/Tablet
C8H9NO2, and C8H10N4O2 Standard stock solution B: 200 g/mL of USP Salicylic Acid
RS in Solvent mixture
PERFORMANCE TESTS Standard solution A: 25.0 mL of Standard stock solution A
DISSOLUTION, ANALYSIS FOR A POOLED SAMPLE 711 and 3.0 mL of Standard stock solution B to a 250-mL
Medium: Water; 900 mL volumetric flask. Dilute with Solvent mixture to volume. This
Apparatus 2: 50 rpm solution contains 325J g of butalbital and 2.4 g of salicylic
Time: 30 min acid/mL.
Mobile phase, Chromatographic system, and System Standard solution B: 325 g/mL of USP Aspirin RS in
suitability: Proceed as directed in the Assay, except use an Solvent mixture
injection size of 20 L for the Analysis. System suitability solution: 4.0 mL of Standard stock
Standard stock solution: 0.02A mg/mL of USP solution A and 3.0 mL of Standard stock solution B to a 50-
Acetaminophen RS, 0.02B mg/mL of USP Butalbital RS, and mL volumetric flask. Dilute with Solvent mixture to volume.
0.02C mg/mL of USP Caffeine RS, in which A, B, and C are Sample solution: Equivalent to 320 g/mL of aspirin from
the labeled amounts of acetaminophen, butalbital, and powdered Tablets (NLT 20 Tablets), in Solvent mixture.
caffeine, respectively, in mg/Tablet Sonicate for 15 min, and mix. Pass a portion of this solution
Standard solution: Standard stock solution and water (1:19) through a fliter of 0.5-m porosity before use.
Sample solution: Sample per Dissolution 711. Dilute with Chromatographic system
Medium as needed and pass through a filter of 10 m or (See Chromatography 621, System Suitability.)
finer porosity.
Mode: LC Mode: LC
Column: 3.9-mm 30-cm; 10-m packing L1 Column: 3.9-mm 30-cm; 10-m packing L1
Flow rate: 1.5 mL/min Flow rate: 1.5 mL/min
Injection size: 10 L Injection size: 1025 L
System suitability System suitability
Samples: System suitability solution, Standard solution A, Sample: Standard solution
and Standard solution B [NOTEThe relative retention times for aspirin, salicylic
[NOTEThe relative retention times for aspirin, salicylic acid, and butalbital are about 0.6, 0.85, and 1.0,
acid, and butalbital are about 0.6, 0.85, and 1.0, respectively.]
respectively.] Suitability requirements
Suitability requirements Resolution: NLT 3.0 between the butalbital and salicylic
Resolution: NLT 3.0 between the butalbital and salicylic acid peaks
acid peaks, System suitability solution Relative standard deviation: NMT 3.0%
Relative standard deviation: NMT 3.0% for butalbital Analysis
and aspirin, and NMT 6.0% for salicylic acid, Standard Samples: Standard solution and Sample solution
solutions A, and B [NOTEFilter a portion of the Sample solution through a
Analysis 0.5-m filter. After use, the column may be regenerated
Samples: Standard solution A, Standard solution B, and by passing through it at least 50 mL of a mixture of
Sample solution acetonitrile, methanol, and water (1:1:1), followed by 50
[NOTEAfter use, the column may be regenerated by mL of a mixture of acetonitrile and water (1:1).]
passing through it at least 50 mL of a mixture of Calculate the percentage of C11H16N2O3 dissolved:
acetonitrile, methanol, and water (1:1:1), followed by a
mixture of acetonitrile and water (1:1).] Result = (rU/rS) (CS/CU) 100
Tablets taken: rU = peak response of butalbital from the Sample
solution
Result = (rU/rS) (CS/CU) 100 rS = peak response butalbital from the Standard
solution
rU = peak response of butalbital from the Sample CS = concentration of USP Butalbital RS in the
solution Standard solution (g/mL)
rS = peak response of butalbital from Standard CU = nominal concentration of butalbital in the
solution A Sample solution (g/mL)
CS = concentration of USP Butalbital RS in Standard Determination of dissolved aspirin
solution A (g/mL) Solution A: 5.98 g of sodium acetate trihydrate in 500 mL
CU = nominal concentration of butalbital in the of water. Add 2.5 mL of glacial acetic acid, dilute with
Sample solution (g/mL) water to 1000 mL, and mix. Adjust this solution with
Calculate the percentage of C9H8O4 in the portion of glacial acetic acid to a pH of 4.5 0.05.
Tablets taken: Sample solution: Solution under test diluted with 4
volumes of Solution A
Result = (rU /rS) (CS/CU) 100 Standard solution: A known concentration of USP Aspirin
RS diluted with 4 volumes of Solution A
rU = peak response of aspirin from the Sample solution [NOTEPrepare the Standard solution at the time of use. An
rS = peak response of aspirin from Standard solution B amount of alcohol not to exceed 1% of the total volume
CS = concentration of USP Aspirin RS in Standard of the Standard solution may be used to bring the
solution B (g/mL) Reference Standard into solution before dilution first with
CU = nominal concentration of aspirin in the Sample water and then with 4 volumes of Solution A.]
solution (g/mL) Analysis: Determine the amount of aspirin (C9H8O4)
Acceptance criteria: 90.0%110.0% of the labeled dissolved from UV absorbances at the wavelength of the
amounts of C11H16N2O3 and C9H8O4 isosbestic point of aspirin and salicylic acid at 265 2 nm of
filtered portions of the Sample solution in comparison with
PERFORMANCE TESTS the Standard solution.
DISSOLUTION 711 Tolerances: NLT 75% (Q) of the labeled amounts of
Medium: Water; 900 mL butalbital (C11H16N2O3) and aspirin (C9H8O4) is dissolved.
Apparatus 1: 100 rpm UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Time: 60 min for Content Uniformity with respect to butalbital and for
Determination of dissolved butalbital Weight Variation with respect to aspirin.
Mobile phase: Acetonitrile, phosphoric acid, and water
(725:4:3100). Adjust the ratio as necessary. IMPURITIES
Sample solution: Sample per Dissolution 711. Dilute with Organic Impurities
Medium to concentration that is similar to the Standard PROCEDURE: Limit of Free Salicylic Acid
solution. Mobile Phase, Solvent mixture, Standard stock solution A,
Standard solution: 1 g/mL of USP Butalbital RS for each Standard stock solution B, Standard solution A, System
mg of the labeled amount/Tablet and 30 g/mL of salicylic suitability solution, Sample solution, Chromatographic
acid in Mobile phase system, and System suitability: Proceed as directed under
Chromatographic system Assay
Analysis Mode: LC
Samples: Standard solution and Sample solution Detector: Set at the wavelength of the isosbestic point of
Calculate the percentage of free salicylic acid in the Tablets aspirin and salicylic acid at 277 and 210 nm.
taken: Column: 3.9-mm 30-cm; packing L1
Temperature: 35 1
Result = (rU/rS) (CS/CU) 100 Flow rate: 1 mL/min
rU = peak response of salicylic acid from the Sample System suitability
solution Samples: Standard solution A and Standard solution B
rS = peak response of salicylic acid from Standard [NOTEThe relative retention times for caffeine, aspirin,
solution A and butalbital are about 0.45, 0.6, and 1.0, respectively,
CS = concentration of USP Salicylic Acid RS in in Standard solution A.]
Standard solution A [NOTEThe salicylic acid peak has the same retention time
CU = nominal concentration of aspirin in the Sample as that of the aspirin peak obtained in the chromatogram
solution of Standard solution B. If the retention time of the salicylic
Acceptance criteria: NMT 3.0% of free salicylic acid acid peak differs from that of the aspirin peak, adjust the
pH of the Mobile phase with 0.2 N potassium hydroxide or
ADDITIONAL REQUIREMENTS 1 M phosphoric acid so that the salicylic acid peak has the
PACKAGING AND STORAGE: Preserve in tight containers. same retention time as that of the aspirin peak. The
USP REFERENCE STANDARDS 11 retention time of the salicylic acid peak decreases 0.3 min
USP Aspirin RS for each 0.1 pH increase. The retention time of the aspirin
USP Butalbital RS peak is essentially unaffected by such pH adjustments.]
USP Salicylic Acid RS Suitability requirements
Resolution: NLT 2.0 between the caffeine and aspirin
peaks, Standard solution A
Butalbital, Aspirin, and Caffeine Column efficiency: NLT 2000 theoretical plates from the
Capsules butalbital peak, Standard solution A
Relative standard deviation: NMT 2.0% of the caffeine,
(Comment on this Monograph)id=m10810=Butalbital, Aspirin, aspirin, and butalbital responses, Standard solution A
and Caffeine Capsules=B-Monos.pdf) Analysis
Samples: Standard solution A and Sample solution
DEFINITION Record the chromatograms, using the 277-nm detector to
Butalbital, Aspirin, and Caffeine Capsules contain NLT 90.0% record the caffeine and aspirin peak responses and the
and NMT 110.0% of the labeled amounts of butalbital (C11H16 210-nm detector to record the butalbital peak responses
N2O3), aspirin (C9H8O4), and caffeine (C8H10N4O2). Calculate the percentage of caffeine (C8H10N4O2), aspirin
IDENTIFICATION (C9H8O4), and butalbital (C11H16 N2O3) in the portion of
The retention times of the butalbital, aspirin, and caffeine Capsules taken:
peaks of the Sample solution correspond to those of the
butalbital, aspirin, and caffeine peaks of the Standard Result = (rU/rS) (CS/CU) 100
solution A, as obtained in the Assay. rU = peak response of the corresponding analyte from
ASSAY the Sample solution
PROCEDURE rS = peak response of the corresponding USP
Solution A: 1.361 mg/mL of monobasic potassium Reference Standard from the Standard solution
phosphate A
Solution B: Methanol and Solution A (9:11). Adjust with CS = concentration of appropriate USP Reference
phosphoric acid to a pH of 2.5 0.05 Standard in Standard solution A (mg/mL)
Mobile phase: Methanol and Solution A (9:11). Adjust with CU = nominal concentration of the corresponding
phosphoric acid to a pH of 3.9. analyte in the Sample solution (mg/mL)
Standard stock solution: 1.6 mg/mL of USP Aspirin RS in Correct the amount of aspirin obtained for the amount of
Solution B, sonicating and shaking the solution, if necessary, salicylic acid present by the formula:
to dissolve. Use this solution within 24 h.
Standard solution A: 1.6J mg/mL of USP Butalbital RS and Result = A (0.01 F A)
1.6J mg/mL of USP Caffeine RS in Standard stock solution, J A = quantity of aspirin in the portion of Capsules
and J being the ratios of the respective labeled amounts, in taken to prepare the Sample solution (mg)
mg, of butalbital and caffeine to the labeled amount, in F = percentage of salicylic acid obtained in the
mg/Capsule of aspirin, sonicating and shaking the solution, Procedure for Limit of free salicylic acid
if necessary, to dissolve. Use this solution within 24 h. Acceptance criteria: 90.0%110.0% of C11H16 N2O3,
Standard solution B: 0.1 mg/mL of salicylic acid in Solution C9H8O4, and C8H10N4O2
B. Pass this solution through a suitable filter of 0.5-m or
finer porosity. PERFORMANCE TESTS
Sample solution: Equivalent to 325 mg of aspirin (remove, DISSOLUTION 711
as completely as possible, the contents of NLT 20 Capsules), Medium: Water; 1000 mL
into a 200-mL volumetric flask. Dilute with Solution B to Apparatus 2: 50 rpm
volume, sonicate for 30 min, and mix. Pass a portion of this Time: 60 min
solution through a suitable filter of 0.5-m or finer porosity, Sample solution: Samples per Dissolution 711.
and use the filtrate. Use this solution within 24 h. [NOTE Solution A, Solution B, Mobile phase, Standard stock
Reserve the remaining portion of the powder for the solution, Standard solution A, Standard solution B,
Procedure for Limit of Free Salicylic Acid]
Chromatographic system, System suitability, and IDENTIFICATION

Analysis: Proceed as directed in the Assay. The retention times of the butalbital, aspirin, and caffeine
Tolerances: NLT 75% (Q) of the labeled amounts of peaks of the Sample solution correspond to those of the
butalbital (C11H16N2O3), caffeine (C8H10N4O2), and aspirin butalbital, aspirin, and caffeine peaks of the Standard
(C9H8O4) solution A, as obtained in the Assay.
ASSAY
IMPURITIES PROCEDURE
Organic Impurities Solution A: 1.361 mg/mL of monobasic potassium
PROCEDURE: LIMIT OF FREE SALICYLIC ACID phosphate
[NOTEUse glassware in this test.] Solution B: Methanol and Solution A (9:11). Adjust with
Solution A: Phosphoric acid and methanol (1:999) phosphoric acid to a pH of 2.5 0.05
Standard solution: 0.0012 mg/mL of USP Salicylic Acid RS Mobile phase: Methanol and Solution A (9:11). Adjust with
in Solution A. [NOTEUse this solution promptly.] phosphoric acid to a pH of 3.9.
Sample solution: Equivalent to 65 mg of aspirin from a Standard stock solution: 1.6 mg/mL of USP Aspirin RS in
portion of the powder remaining from the solution of the Solution B, sonicating and shaking the solution, if necessary,
Sample solution in the Assay, to a 200-mL flask, add 100.0 to dissolve. Use this solution within 24 h.
mL of Solution A, and shake for 1 min. Promptly filter a Standard solution A: 1.6J mg/mL of USP Butalbital RS and
portion of this solution, discarding the first 15 mL of the 1.6J mg/mL of USP Caffeine RS in Standard stock solution, J
filtrate, and use the clear filtrate. Use this solution within 20 and J being the ratios of the respective labeled amounts, in
min after the addition of the Solution A. [NOTEPerform mg, of butalbital and caffeine to the labeled amount, in
this test on the same day the powder is removed from the mg/Capsule of aspirin, sonicating and shaking the solution,
Capsules] if necessary, to dissolve. Use this solution within 24 h.
Fluorimetric conditions Standard solution B: 0.1 mg/mL of salicylic acid in Solution
Excitation wavelength: 305 nm B. Pass this solution through a suitable filter of 0.5-m or
Emmission wavelength: 444 nm finer porosity.
Analysis Sample solution: Equivalent to 325 mg of aspirin from
Samples: Standard solution and Sample solution powdered Tablets (NLT 20 Tablets), into a 200-mL
Allow the Samples to equilibrate for 2 min in the volumetric flask. Dilute with Solution B to volume, and
fluorimeter. sonicate for 30 min. Pass a portion of this solution through
Calculate the percentage of salicylic acid in the portion of a suitable filter of 0.5-m or finer porosity, and use the
Capsules taken: filtrate. Use this solution within 24 h.
Result = (IU/IS) (CS/CU) 100 (See Chromatography 621, System Suitability.)
Mode: LC
IU = fluorescence intensity readings from the Sample Detector: Set at the wavelength of the isosbestic point of
solution aspirin and salicylic acid at 277 and 210 nm.
IS = fluorescence intensity readings from the Column: 3.9-mm 30-cm; packing L1
Standard solution Temperature: 35 1
CS = concentration of USP Salicylic Acid RS in the Flow rate: 1 mL/min
Standard solution (mg/mL) Injection size: 10 L
CU = nominal concentration of aspirin in the Sample System suitability
solution (mg/mL) Sample: Standard solution A and Standard solution B
[NOTEIf the intensity of the Sample solution greatly exceeds [NOTEThe relative retention times for caffeine, aspirin,
that of the Standard solution, immediately transfer 5.0 mL of and butalbital are about 0.45, 0.6, and 1.0, respectively,
the Sample solution to a 50-mL volumetric flask, dilute with in Standard solution A.]
Solution A to volume, and mix. Immediately determine the [NOTEThe salicylic acid peak has the same retention time
intensity of this solution. Calculate the percentage of salicylic as that of the aspirin peak obtained in the chromatogram
acid in the portion of Capsules taken, by using the same of the Standard solution B. If the retention time of the
formula.] salicylic acid peak differs from that of the aspirin peak,
Acceptance criteria: NMT 2.5% adjust the pH of the Mobile phase with 0.2 N potassium
hydroxide or 1 M phosphoric acid so that the salicylic acid
ADDITIONAL REQUIREMENTS peak has the same retention time as that of the aspirin
PACKAGING AND STORAGE: Preserve in tight containers. peak. The retention time of the salicylic acid peak
USP REFERENCE STANDARDS 11 decreases 0.3 min for each 0.1 pH increase. The retention
USP Aspirin RS time of the aspirin peak is essentially unaffected by such
USP Butalbital RS pH adjustments.]
USP Caffeine RS Suitability requirements
USP Salicylic Acid RS Resolution: NLT 2.0 between the caffeine and aspirin
peaks, Standard solution A
Butalbital, Aspirin, and Caffeine butalbital peak, Standard solution A
Tablets Relative standard deviation: NMT 2.0% of the caffeine,
aspirin, and butalbital responses, Standard solution A
(Comment on this Monograph)id=m10820=Butalbital, Aspirin, Analysis
and Caffeine Tablets=B-Monos.pdf) Samples: Standard solution A and Sample solution
Record the chromatograms, using the 277-nm detector to
DEFINITION record the caffeine and aspirin peak areas and the 210-nm
Butalbital, Aspirin, and Caffeine Tablets contain NLT 90.0% and detector to record the butalbital peak areas.
NMT 110.0% of the labeled amounts of butalbital
(C11H16N2O3), aspirin (C9H8O4), and caffeine (C8H10N4O2).
Calculate the percentage of butalbital (C11H16N2O3), aspirin IS = fluorescence intensity readings from the
(C9H8O4), and caffeine (C8H10N4O2) in the portion of Standard solution
Tablets taken: CS = concentration of USP Salicylic Acid RS in the
Result = (rU/rS) (CS/CU) 100 CU = nominal concentration of aspirin in the Sample
solution (mg/mL)
rU = peak response of the corresponding analyte from [NOTEIf the intensity of the Sample solution greatly exceeds
the Sample solution that of the Standard solution, immediately transfer 5.0 mL of
rS = peak response of the corresponding USP the Sample solution to a 50-mL volumetric flask, dilute with
Reference Standard from the Standard solution Solution A to volume, and mix. Immediately determine the
A intensity of this solution, and calculate the percentage of
CS = concentration of appropriate USP Reference salicylic acid in the portion of Tablets taken, by using the
Standard in Standard solution A (mg/mL) same formula.]
CU = nominal concentration of the corresponding Acceptance criteria: NMT 3.0% is found
analyte in the Sample solution (mg/mL)
Correct the amount of aspirin obtained for the amount of ADDITIONAL REQUIREMENTS
salicylic acid present by the formula: PACKAGING AND STORAGE: Preserve in tight containers.
Result = A (0.01 F A) USP Aspirin RS
USP Butalbital RS
A = quantity of aspirin in the portion of Tablets taken USP Caffeine RS
to prepare the Sample solution (mg) USP Salicylic Acid RS
F = percentage of salicylic acid obtained in the
Procedure for Limit of Free Salicylic Acid
Acceptance criteria: 90.0%110.0% of C11H16 N2O3,
C9H8O4, and C8H10N4O2 Butalbital, Aspirin, Caffeine, and
PERFORMANCE TESTS
Codeine Phosphate Capsules
DISSOLUTION 711 (Comment on this Monograph)id=m10825=Butalbital, Aspirin,
Medium: Water; 900 mL Caffeine, and Codeine Phosphate Capsules=B-Monos.pdf)
Apparatus 1: 100 rpm DEFINITION
Time: 60 min Butalbital, Aspirin, Caffeine, and Codeine Phosphate Capsules
Sample solution Sample per Dissolution 711 contain NLT 90.0% and NMT 110.0% of the labeled amounts
Solution A, Solution B, Mobile phase, Standard stock of butalbital (C11H16N2O3), aspirin (C9H8O4), caffeine
solution, Standard solution A, Standard solution B, (C8H10N4O2), and codeine phosphate (C18H21 NO3 H3PO4
Chromatographic system, System suitability, and 1
/2H2O).
Analysis: Proceed as directed under Assay.
Tolerances: NLT 80% (Q) of the labeled amounts of IDENTIFICATION
butalbital (C11H16N2O3), caffeine (C8H10N4O2), and aspirin The retention times of the butalbital, aspirin, caffeine, and
(C9H8O4) is dissolved. codeine peaks of the Sample solution correspond to those of
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements the butalbital, aspirin, caffeine, and codeine peaks of the
Standard solution, as obtained in the Assay.
IMPURITIES
PROCEDURE: LIMIT OF FREE SALICYLIC ACID PROCEDURE
[NOTEUse glassware in this test.] Solution A: 1.361 mg/mL of monobasic potassium
Solution A: Phosphoric acid and methanol (1:999) phosphate
Standard solution: 0.0012 mg/mL of USP Salicylic Acid RS Solution B: Methanol and Solution A (9:11). Adjust with
in Solution A [NOTEUse this solution promptly.] phosphoric acid to a pH of 2.5 0.05.
Sample solution: Equivalent to 65 mg of aspirin from Mobile phase: Methanol and Solution A (9:11). Adjust with
powdered Tablets (NLT 20 Tablets), to a 200-mL flask. Add phosphoric acid to a pH of 3.9.
100.0 mL of Solution A, and shake by mechanical means for Standard stock solution A: 1.6 mg/mL of USP Aspirin RS in
15 min. Filter a portion of this solution, discarding the first Solution B, sonicating and shaking the solution, if necessary,
15 mL of the filtrate, and use the clear filtrate. Use this to dissolve. Use this solution within 24 h.
solution within 20 min after the addition of the Solution A. Standard solution A: 1.6J mg/mL of USP Butalbital RS,
[NOTEPerform this test on the same day the Tablets are 1.6J mg/mL of USP Caffeine RS, and 1.6J mg/mL of USP
powdered.] Codeine Phosphate RS in Standard stock solution A, J, J, and
Fluorimetric conditions J being the ratios of the respective labeled amounts, in mg,
Excitation wavelength: 305 nm of butalbital, caffeine, and codeine phosphate to the labeled
Emmission wavelength: 444 nm amount, in mg/Capsule of aspirin, sonicating and shaking
Analysis the solution, if necessary, to dissolve.
Samples: Standard solution and Sample solution Standard solution B: 0.1 mg/mL of salicylic acid in Solution
Allow the Samples to equilibrate for 2 min in the B. Pass this solution through a suitable filter of 0.5-m or
fluorimeter. finer porosity.
Calculate the percentage of salicylic acid in the portion of Sample solution: Equivalent to 325 mg of aspirin (remove,
Tablets taken: as completely as possible, the contents of NLT 20 Capsules),
into a 200-mL volumetric flask. Dilute with Solution B to
Result = (IU/IS) (CS/IU) 100 volume, sonicate for 30 min, and mix. Pass a portion of this
solution through a suitable filter of 0.5-m or finer porosity,
IU = fluorescence intensity readings from the Sample and use the filtrate. Use this solution within 24 h. [NOTE
solution
Reserve the remaining portion of the powder for the Correct the amount of aspirin obtained for the amount of
Procedure for Limit of Free Salicylic Acid] salicylic acid present taken:
(See Chromatography 621, System Suitability.) Result = A (0.01 F A)
Mode: LC
Detector: Set at the wavelength of the isosbestic point of A = quantity of aspirin in the portion of Capsules
aspirin and salicylic acid at 277 and 210 nm. taken to prepare the Sample solution (mg)
Column: 3.9-mm 30-cm; packing L1 F = percentage of salicylic acid obtained in the
Temperature: 35 1 Procedure for Limit of Free Salicylic Acid
Flow rate: 1 mL/min Acceptance criteria: 90.0%110.0% of C11H16N2O3,
Injection size: 10 L C9H8O4, C8H10N4O2, and C18H21 NO3 H3PO4 1/2H2O
System suitability
Sample: Standard solution A and Standard solution B PERFORMANCE TESTS
[NOTEThe relative retention times for codeine, caffeine, DISSOLUTION 711
aspirin, and butalbital are about 0.3, 0.45, 0.6, and 1.0, Medium: Water; 1000 mL
respectively.] Apparatus 2: 50 rpm
[NOTEThe salicylic acid peak has the same retention time Time: 60 min
as that of the aspirin peak obtained in the chromatogram Solution A, Solution B, Mobile phase, and Standard
of the Standard solution B. If the retention time of the solution B: Prepare as directed in the Assay.
salicylic acid peak differs from that of the aspirin peak, Standard solution C: Transfer 5.0 mL of Standard solution
adjust the pH of the Mobile phase with 0.2 N potassium B, prepared as directed in the Assay, to a 50-mL volumetric
hydroxide or 1 M phosphoric acid so that the salicylic acid flask, and dilute with Solution B to volume.
peak has the same retention time as that of the aspirin Standard stock solution A: 0.16 mg/mL of USP Aspirin RS
peak. The retention time of the salicylic acid peak in a mixture of Solution B and Medium (1:1). Use this
decreases 0.3 min for each 0.1 pH increase. The retention solution within 24 h.
time of the aspirin peak is essentially unaffected by such Standard solution A: 0.16J mg/mL of USP Butalbital RS,
pH adjustments.] 0.16J mg/mL of USP Caffeine RS, and 0.16J mg/mL of USP
Suitability requirements Codeine Phosphate RS in Standard stock solution A, J, J, and
Resolution: NLT 2.0 between the caffeine and aspirin J being the ratios of the respective labeled amounts, in mg,
peaks, Standard solution A of butalbital, caffeine, and codeine phosphate to the labeled
Column efficiency: NLT 2000 theoretical plates from the amount of aspirin, in mg/Capsule, sonicating and shaking
butalbital peak, Standard solution A the solution, if necessary to dissolve. Pass a portion of this
Relative standard deviation: NMT 2.0% of the codeine, solution through a suitable filter of 0.5-m or finer porosity.
caffeine, aspirin, and butalbital responses, Standard Use this solution within 24 h.
solution A Sample solution: Pass 20 mL of the solution under test
Analysis through a suitable filter of 0.5-m or finer porosity,
Samples: Standard solution A and Sample solution discarding the first 2 mL of the filtrate. Mix 5.0 mL of the
Record the chromatograms, using the 277-nm detector to filtrate and 5.0 mL of Solution B.
record the caffeine and aspirin peak responses and the Chromatographic system: Proceed as directed in the
210-nm detector to measure the codeine and butalbital Assay, except to inject 100 L, instead of 10 L, into the
responses. chromatograph, and use Standard solution C instead of
Calculate the percentage of caffeine (C8H10N4O2), aspirin Standard solution B.
(C9H8O4), and butalbital (C11H16N2O3) in the portion of System suitability: Proceed as directed under Assay.
Capsules taken: Analysis
Samples: Standard solution A and Sample solution
Result = (rU/rS) (CS/CU) 100 Use the 277-nm detector to record the caffeine and aspirin
peaks and the 210-nm detector to record the butalbital
rU = peak response of the corresponding analyte from and codeine responses.
the Sample solution Calculate the percentage of caffeine (C8H10N4O2), aspirin
rS = peak response of the corresponding USP (C9H8O4), and butalbital (C11H16N2O3) dissolved:
Reference Standard from Standard solution A
CS = concentration of appropriate USP Reference Result = (rU/rS) (CS/CU) 100
Standard in Standard solution A (mg/mL)
CU = nominal concentration of the corresponding rU = peak response of the corresponding analyte from
analyte in the Sample solution (mg/mL) the Sample solution
Calculate the percentage of codeine phosphate (C18H21NO3 rS = peak response of the corresponding analyte from
H3PO4 1/2H2O) in the portion of Capsules taken: Standard solution A
CS = concentration of the appropriate USP Reference
Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 Standard in Standard solution A (mg/mL)
CU = nominal concentration of the corresponding
rU = peak response of codeine from the Sample analyte in the Sample solution (mg/mL)
solution Calculate the percentage of codeine phosphate (C18H21NO3
rS = peak response of codeine from the Standard H3PO4 1/2H2O) dissolved:
solution
CS = concentration of USP Codeine Phosphate RS in Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
CU = nominal concentration of codeine phosphate in rU = peak response of codeine from the Sample
the Sample solution (mg/mL) solution
Mr1 = molecular weight of codeine phosphate rS = peak response of codeine from Standard solution
hemihydrate, 406.37 A
Mr2 = molecular weight of anhydrous codeine CS = concentration of USP Codeine Phosphate RS in
phosphate, 397.37 Standard solution A (mg/mL)
USP 32 Official Monographs / Butamben 161
CU = nominal concentration of codeine in the Sample Butamben

solution (mg/mL) (Comment on this Monograph)id=m10840=Butamben=B-
Mr1 = molecular weight of codeine phosphate Monos.pdf)
hemihydrate, 406.37
Mr2 = molecular weight of anhydrous codeine
phosphate, 397.37
Tolerances: NLT 75% (Q) of the labeled amounts of
butalbital (C11H16N2O3), aspirin (C9H8O4), caffeine
(C8H10N4O2), and codeine phosphate (C18H21NO3 H3PO4
1
/2H2O) is dissolved.
C11H15NO2 193.24
IMPURITIES Benzoic acid, 4-amino-, butyl ester;
Organic Impurities Butyl p-aminobenzoate [94-25-7].
PROCEDURE: LIMIT OF FREE SALICYLIC ACID
[NOTEUse glassware in this test.] DEFINITION
Solution A: Phosphoric acid and methanol (1:999) Butamben, dried over phosphorus pentoxide for 3 h, contains
Standard solution: 0.0012 mg/mL of USP Salicylic Acid RS NLT 98.0% and NMT 101.0% of C11H15NO2.
in Solution A [NOTEUse this solution promptly.]
Sample solution: Equivalent to 65 mg of aspirin from a IDENTIFICATION
portion of the powder remaining from the solution of the INFRARED ABSORPTION 197K
Sample solution in the Assay, to a 200-mL flask. Add 100.0
mL of Solution A, and shake for 1 min. Promptly filter a ASSAY
portion of this solution, discarding the first 15 mL of the PROCEDURE
filtrate, and use the clear filtrate. Use this solution within 20 Solution A: Dissolve, without warming, 0.5 g of
min after the addition of the Solution A. [NOTEPerform ferrocyphen in 50 mL of sulfuric acid.
this test on the same day the powder is removed from the Sample solution: Dissolve 400 mg of Butamben, previously
Capsules.] dried, in a mixture of 100 mL of water and 20 mL of
Fluorimetric conditions hydrochloric acid. Add 1 mL of Solution A, and cool the
Excitation wavelength: 305 nm solution in an ice bath to 10.
Emmission wavelength: 444 nm Analysis: Titrate the Sample solution with 0.1 M sodium
Analysis nitrite VS to a violet endpoint that is stable for NLT 3 min.
Samples: Standard solution and Sample solution Perform a blank determination, and make any necessary
Allow to equilibrate for 2 min in the fluorimeter. correction (see Titrimetry 541). Each mL of 0.1 M sodium
Calculate the percentage of salicylic acid in the portion of nitrite is equivalent to 19.32 mg of C11H15NO2.
Capsules taken: Acceptance criteria: 98.0%101.0%
Result = (IU/IS) (CS/CU) 100 IMPURITIES
IU = fluorescence intensity readings from the Sample RESIDUE ON IGNITION 281: NMT 0.2%
solution CHLORIDE AND SULFATE, Chloride 221: To a solution of 200
IS = fluorescence intensity readings from the mg in 10 mL of alcohol, add 1 mL of 2 N nitric acid and a
Standard solution few drops of silver nitrate TS: no opalescence is produced.
CS = concentration of USP Salicylic Acid RS in the HEAVY METALS, Method I 231: Dissolve 2 g in 2 mL of 1 N
Standard solution (mg/mL) acetic acid and sufficient alcohol to make 25 mL: NMT 10
CU = nominal concentration of aspirin in the Sample ppm
solution (mg/mL)
[NOTEIf the intensity of the Sample solution greatly SPECIFIC TESTS
exceeds that of the Standard solution, immediately COMPLETENESS AND COLOR OF SOLUTION: One g dissolves
transfer 5.0 mL of the Sample solution to a 50-mL completely in 30 mL of alcohol and in 30 mL of ether, and
volumetric flask, dilute with Solution A to volume, and the solutions are colorless.
mix. Immediately determine the intensity of this MELTING RANGE OR TEMPERATURE, Class I 741: 5759
solution. Calculate the percentage of salicylic acid in the REACTION: Dissolve 1 g in 10 mL of neutralized alcohol: a
portion of Capsules taken.] clear solution results. Dilute with 10 mL of water this
Acceptance criteria: NMT 3.0% solution, and add 2 drops of phenolphthalein TS and 1 drop
of 0.1 N sodium hydroxide: a red color is produced.
ADDITIONAL REQUIREMENTS LOSS ON DRYING 731: Dry it over phosphorus pentoxide for
PACKAGING AND STORAGE: Preserve in tight, light-resistant 3 h: it loses NMT 1.0% of its weight.
containers.
USP REFERENCE STANDARDS 11 ADDITIONAL REQUIREMENTS
USP Aspirin RS PACKAGING AND STORAGE: Preserve in well-closed containers.
USP Butalbital RS USP REFERENCE STANDARDS 11
USP Caffeine RS USP Butamben RS
USP Codeine Phosphate RS
USP Salicylic Acid RS
162 Butoconazole / Official Monographs USP 32
Butoconazole Nitrate Developing solvent system: Chloroform, tetrahydrofuran,

(Comment on this Monograph)id=m10900=Butoconazole cyclohexane, and ammonium hydroxide (18:18:13:1)
Nitrate=B-Monos.pdf) Visualization: 22
Acceptance criteria: Any spot of the Sample solution,
except for the prinicpal spot, is not more intense than the
principal spot of the Standard solution, 0.01 mg/mL. NMT
2.0% of total impurities is found.
SPECIFIC TESTS
LOSS ON DRYING 731: Dry a sample in a vacuum at 60 for
3 h: it loses NMT 1.0% of its weight.
C19H17Cl3N2S HNO3 474.79 ADDITIONAL REQUIREMENTS
1H-Imidazole, 1-[4-(4-chlorophenyl)-2-[(2,6- PACKAGING AND STORAGE: Preserve in well-closed, light-
dichlorophenyl)thio]butyl-, mononitrate, ()-; resistant containers.
()-1-[4-(p-Chlorophenyl)-2-[(2,6-dichlorophenyl)- USP REFERENCE STANDARDS 11
thio]butyl]imidazole mononitrate [64872-77-1]. USP Butoconazole Nitrate RS
DEFINITION
Butoconazole Nitrate contains NLT 98.0% and NMT 102.0% of
C19H17Cl3N2S HNO3, calculated on the dried basis. Butoconazole Nitrate Vaginal Cream
IDENTIFICATION (Comment on this Monograph)id=m10904=Butoconazole
INFRARED ABSORPTION 197K Nitrate Vaginal Cream=B-Monos.pdf)
ASSAY DEFINITION
PROCEDURE Butoconazole Nitrate Vaginal Cream contains Butoconazole
Buffer: Monobasic potassium phosphate 2.18 g/L and Nitrate in a suitable cream base. It contains NLT 90.0% and
dibasic potassium phosphate 4.18 g/L in water NMT 110.0% of the labeled amount of butoconazole nitrate
Mobile phase: Methanol and Buffer (3:1) (C19H17Cl3N2S HNO3).
Standard solution: USP Butoconazole Nitrate RS 0.2 IDENTIFICATION
mg/mL in Mobile phase PROCEDURE
Sample solution: Butoconazole Nitrate 0.2 mg/mL in Analysis: Prepare a mixture of the Standard solution and the
Mobile phase Sample solution (1:1), prepared as directed in the Assay, and
Chromatographic system chromatograph as directed in the Assay.
(See Chromatography 621, System Suitability.) Acceptance criteria: The chromatogram so obtained
Mode: LC exhibits two main peaks, corresponding to butoconazole
Detector: UV 229 nm nitrate and the internal standard.
Temperature: 40 ASSAY
Injection size: 10 L Buffer: Potassium acetate 1.4 g in 980 mL of water. Adjust
System suitability with about 2 mL of glacial acetic acid to a pH of 4.3 0.1,
Sample: Standard solution dilute with water to 1000 mL, and mix. Adjust the buffer
Suitability requirements molarity (0.0180.072 M) as necessary to obtain suitable
Column efficiency: NLT 2800 theoretical plates chromatographic performance. Increased retention time may
Tailing factor: NMT 1.5 be achieved by a decrease in the buffer molarity.
Relative standard deviation: NMT 1.5% Diluent: Methanol and Buffer (3:2)
Analysis Mobile phase: Methanol and Buffer (13:7)
Samples: Standard solution and Sample solution Internal standard solution: 1-Benzylimidazole 1.6 mg/mL
Calculate the percentage of C19H17Cl3N2S HNO3 in the in methanol
portion of Butoconazole Nitrate taken: Standard stock solution: USP Butoconazole Nitrate RS 0.4
mg/mL in methanol
Result = (rU/rS) (CS/CU) 100 Standard solution: Transfer 2.0 mL of Standard stock
solution and 3.0 mL of Internal standard solution to a 50-mL
rU = peak response from the Sample solution flask, and add 35.0 mL of Diluent.
rS = peak response from the Standard solution Sample stock solution: Sonicate a quantity of
CS = concentration of USP Butoconazole Nitrate RS in Butoconazole.
the Standard solution (mg/mL) Sample solution: Transfer 2.0 mL of Sample stock solution
CU = concentration of Butoconazole Nitrate in the and 3.0 mL of Internal standard solution to a 50-mL flask,
Sample solution (mg/mL) and add 35.0 mL of Diluent. Allow the precipitated
Acceptance criteria: 98.0%102.0% excipients that form to rise to the top of the solution,
IMPURITIES remove them by aspiration, and discard. Centrifuge or filter
Inorganic Impurities the remaining solution.
RESIDUE ON IGNITION 281: NMT 0.1% Chromatographic system
Organic Impurities (See Chromatography 621, System Suitability.)
PROCEDURE: ORDINARY IMPURITIES 466 Mode: LC
Standard solution: 0.01, 0.05, 0.1, and 0.2 mg/mL in Detector: UV 225 nm
methylene chloride and methanol (2:1) Column: 4.6-mm 25-cm; packing L9 that has been
Sample solution: 10 mg/mL in methylene chloride and converted to the potassium form by the use of 0.555 M
methanol (2:1) potassium acetate solution
USP 32 Official Monographs / Butorphanol 163
Flow rate: 1 mL/min Analysis: Titrate with 0.1 N perchloric acid VS. Perform a
Injection size: 20 L blank determination, and make any necessary correction
System suitability (see Titrimetry 541). Each mL of 0.1 N perchloric acid is
Sample: Standard solution equivalent to 47.76 mg of C21H29NO2 C4H6O6.
[NOTEThe relative retention times for butoconazole nitrate Acceptance criteria: 98.0%102.0%
and 1-benzylimidazole are 0.6 and 1.0, respectively.]
Suitability requirements IMPURITIES
Resolution: NLT 4.0 between the analyte and internal Inorganic Impurities
standard peaks RESIDUE ON IGNITION 281: NMT 0.1%
Column efficiency: NLT 1100 theoretical plates HEAVY METALS, Method III 231: NMT 30 ppm
Tailing factor: NMT 2.1 Organic Impurities
Relative standard deviation: NMT 1.5% PROCEDURE 1
Analysis Standard solution: 1 mg/mL of USP Butorphanol Tartrate
Samples: Standard solution and Sample solution RS in methanol
Calculate the percentage of C19H17Cl3N2S HNO3 in the Sample solution: 10 mg/mL of Butorphanol Tartrate in
portion of Vaginal Cream taken: methanol
Result = (RU/RS) (CS/CU) 100 (See Chromatography 621, Thin-Layer Chromatography.)
Mode: TLC
RU = peak response ratio of butoconazole nitrate to Adsorbent: 0.25-mm layer of chromatographic silica gel
the internal standard from the Sample solution mixture
RS = peak response ratio of butoconazole nitrate to Application volume: 50 L of Sample solution, 5 L and
the internal standard from the Standard 10 L of Standard solution
solution Developing solvent system: Chloroform, methanol,
CS = concentration of USP Butoconazole Nitrate RS in benzene, and ammonium hydroxide (17:5:4:1)
the Standard solution (g/mL) Spray reagent: Prepare a 1in10 solution of
CU = nominal concentration of butoconazole nitrate in chloroplatinic acid in water. To 0.5 mL of this solution,
the Sample solution (g/mL) add 33 mL of water and 1 g of potassium iodide. Prepare
Acceptance criteria: 90.0%110.0% fresh daily.
Analysis
PERFORMANCE TESTS Samples: Standard solution and Sample solution
MINIMUM FILL 755: Meets the requirements Proceed as directed under General Chapter. Place the
plate in a developing chamber containing, and
ADDITIONAL REQUIREMENTS equilibrated with the Developing solvent system. Develop
PACKAGING AND STORAGE: Preserve in collapsible tubes or the chromatogram until the solvent front has moved 10
tight containers. Avoid excessive heat and avoid freezing. cm above the line of application. Remove the plate, mark
USP REFERENCE STANDARDS 11 the solvent front, and allow the solvent to evaporate.
USP Butoconazole Nitrate RS Spray the plate with Spray reagent. Estimate the
percentage of the impurities present in the Sample
solution by comparing the intensities of secondary spots,
Butorphanol Tartrate if present, with the intensities of the principal spots of
the Standard solutions.
(Comment on this Monograph)id=m11000=Butorphanol Acceptance criteria: The sum of the impurities observed
Tartrate=B-Monos.pdf) is NMT 2.0%.
PROCEDURE 2
Sample solution: 10 mg/mL of Butorphanol Tartrate in
methanol
Mode: GC
Column: 1.8-m 4-mm; glass column containing 3%
C21H29NO2 C4H6O6 477.55 liquid phase G3 on support S1AB
Morphinan-3,14-diol, 17-(cyclobutylmethyl)-, ()-, [S- Temperature
(R*,R*)]-2,3-dihydroxybutanedioate (1:1) (salt); Injection port: 280
()-17-(Cyclobutylmethyl)morphinan-3,14-diol D-()-tartrate Column: 250
(1:1) (salt) [58786-99-5]. Detector: 290
Carrier gas: Nitrogen
DEFINITION Injection size: 1 L
Butorphanol Tartrate contains NLT 98.0% and NMT 102.0% of Analysis
C21H29NO2 C4H6O6, calculated on the anhydrous basis. Sample: Sample solution
[NOTEThe retention time for the alpha isomer of
IDENTIFICATION butorphanol tartrate, for butorphanol tartrate, and for
A. INFRARED ABSORPTION 197K butorphanol tartrate is 1.2, 1.0, and NLT 15 min,
B. The RF value of the principal spot of the Sample solution respectively.]
corresponds to that of the Standard solution, as obtained in Record a 30-min chromatogram. Preferably using an
Procedure 1. electronic integrator, determine the areas of all peaks in
the chromatogram excluding the area of the solvent.
ASSAY
PROCEDURE
Sample solution: 500 mg of Butorphanol Tartrate in 75 mL
of glacial acetic acid, and add crystal violet TS
164 Butorphanol / Official Monographs USP 32
Calculate the percentage of synthesis precursors in the Standard solution: 5 mL of the Standard stock solution into
Sample solution taken: a 50-mL volumetric flask containing 10.0 mL of Internal
standard solution. Add water to volume, mix, and pass
Result = (AV/AS) 100 through a microporous filter, discarding the first 5 mL of the
filtrate and collecting the remainder in a suitable container.
AV = sum of the areas of all minor peaks Sample solution: Equivalent to 10 mg of butorphanol
AS = sum of the areas of the major and minor peaks tartrate from Injection, into a 50-mL volumetric flask. Add
Acceptance criteria: NMT 2.0% 10.0 mL of Internal standard solution, mix, and add water to
volume. Pass through a microporous filter, discarding the
SPECIFIC TESTS first 5 mL of the filtrate and collecting the remainder in a
OPTICAL ROTATION, Specific Rotation 781S: 60 to 66 suitable container.
Sample solution: 4 mg/mL, in methanol Chromatographic system
PACKAGING AND STORAGE: Preserve in tight containers. Store Detector: UV 280 nm
at 25, excursions permitted between 15 and 30. Column: 4-mm 30-cm; packing L11
USP REFERENCE STANDARDS 11 Flow rate: 2 mL/min
USP Butorphanol Tartrate RS Injection size: 20 L
System suitability
Sample: Standard solution (five replicate injections)
[NOTEThe relative retention times for propylparaben and
Butorphanol Tartrate Injection butorphanol tartrate are about 1.7 and 1.0, respectively.]
(Comment on this Monograph)id=m11010=Butorphanol Suitability requirements
Tartrate Injection=B-Monos.pdf) Relative standard deviation: NMT 1.5%
Capacity factor: NLT 2.0 for butorphanol tartrate
DEFINITION Analysis
Butorphanol Tartrate Injection is a sterile solution of Samples: Standard solution and Sample solution
Butorphanol Tartrate in Water for Injection. It contains NLT Adjust the flow rate and other operating parameters, if
90.0% and NMT 110.0% of the labeled amount of C21H29NO2 necessary, until satisfactory chromatography and peak
C4H6O6. It may contain a suitable preservative and a buffer. responses are obtained. Record the chromatograms, and
measure the responses for the major peaks.
IDENTIFICATION Calculate the percentage of C21H29NO2 C4H6O6 in each mL
THIN-LAYER CHROMATOGRAPHY of the Injection taken:
Standard solution: USP Butorphanol Tartrate RS at the
same concentration as the Injection Result = (RU/RS) (CS/CU) 100
Sample solution: Injection
Chromatographic system RU = peak response ratio of the butorphanol tartrate
(See Chromatography 621.) peak to the internal standard peak from the
Mode: TLC Sample solution
Adsorbent: 0.25-mm layer of chromatographic silica gel RS = peak response ratio of the butorphanol tartrate
mixture peak to the internal standard peak from the
Application volume: 10 L Standard solution
Developing solvent system: Chloroform, ethyl acetate, CS = concentration of USP Butorphanol Tartrate RS in
and methanol (40:10:9) the Standard solution (mg/mL)
Spray reagent: bromocresol purpl in dehydrated alcohol CU = nominal concentration of butorphanol tartrate in
(1 in 250) the Sample solution (mg/mL)
Develop the chromatogram until the solvent front has SPECIFIC TESTS
moved 10 cm above the line of application. Remove the PH 791: 3.05.5
plate, mark the solvent front, and allow the solvent to BACTERIAL ENDOTOXINS TEST 85: NMT 88.0 USP Endotoxin
evaporate. Examine the plate under short-wavelength UV Units/mg of butorphanol tartrate
light. Visualize the butorphanol spots by lightly spraying OTHER REQUIREMENTS: It meets the requirements under
the plate with Spray reagent: butorphanol appears as a Injections 1.
blue spot against a light yellow background. ADDITIONAL REQUIREMENTS
Benzethonium chloride, if present, is observed as a PACKAGING AND STORAGE: Preserve in single-dose or in
streaked zone near the point of application. multiple-dose containers, preferably of Type I glass,
Acceptance criteria: The RF value of the principal spot of protected from light.
the Sample solution corresponds to that of the Standard USP REFERENCE STANDARDS 11
solution. USP Butorphanol Tartrate RS
ASSAY USP Endotoxin RS
PROCEDURE
Mobile phase: Acetonitrile and 0.05 M ammonium acetate
(1:3), adjusted by the addition of glacial acetic acid to a pH
of 4.1
Butorphanol Tartrate Nasal Solution
Internal standard solution: 0.2 mg/mL of propylparaben (Comment on this Monograph)id=m11015=Butorphanol
dissolved in methanol and diluted with water (1:49) Tartrate Nasal Solution=B-Monos.pdf)
Standard stock solution: 50 mg of USP Butorphanol DEFINITION
Tartrate RS in a 25-mL volumetric flask containing 1.0 mL of Butorphanol Tartrate Nasal Solution is an aqueous solution of
1 N sulfuric acid. Swirl the flask to dissolve the powder butorphanol tartrate for administration as a metered spray to
completely, add water to volume, and mix.
USP 32 Official Monographs / Butorphanol 165
the nasal mucosa. It contains the equivalent of NLT 90.0% and rS = peak response of butorphanol tartrate from the
NMT 110.0% of the labeled amount of butorphanol tartrate Standard solution
(C21H29NO2 C4H6O6). CS = concentration of USP Butorphanol Tartrate RS in
IDENTIFICATION CU = nominal concentration of butorphanol tatrtrate
A. The retention time of the major peak of the Sample in the Sample solution (mg/mL)
solution corresponds to that of the Standard solution, as Acceptance criteria: 90.0%110.0%
B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 IMPURITIES
Standard solution: 1.0 mg/mL of USP Butorphanol Tartrate Organic Impurities
RS in methanol PROCEDURE
Sample solution: Prepare a composite solution by pooling Buffer: Prepare as directed in the Assay.
the contents of three containers of Nasal Solution into a Mobile phase: Acetonitrile, triethylamine, and Buffer
suitable vessel. Transfer 1.0 mL of pooled sample to a 10-mL (15:5.1:85)
volumetric flask, and dilute with methanol to volume. Mix thoroughly, and adjust with 85.0% phosphoric acid to
Developing solvent system: Chloroform, methanol, a pH of 3.0 0.1.
benzene, and ammonium hydroxide (17:5:4:1) Standard solution: 0.005 mg/mL of USP Butorphanol
[CautionPrepare in a hood while wearing appropriate Tartrate RS
safety gloves, lab coat, and protective eyewear.] Sensitivity solution: Transfer 2.5 mL of the Standard
Spray reagent: Prepare a 1-in-10 solution of chloroplatinic solution to a 50-mL volumetric flask, dilute with water to
acid in water. To 0.5 mL of this solution, add 33 mL of volume, and mix. Do not filter.
water and 1 g of potassium iodide. Prepare fresh daily. Sample solution: Prepare a composite solution by pooling
Analysis a minimum of four containers of Nasal Solution into a
Samples: Standard solution and Sample solution suitable glass vessel. Transfer the equivalent of 50 mg of
Proceed as directed in the chapter, except to spray the butorphanol tartrate to a 50-mL volumetric flask. Dilute
plate with Spray reagent. with water to volume, and mix. Do not filter.
Acceptance criteria: The typical RF value is 0.7 for [NOTEThe Sample solution has a nominal concentration
butorphanol tartrate. of 1 mg/mL of butorphanol tartrate.]
ASSAY (See Chromatography 621, System Suitability.)
PROCEDURE Mode: LC
Buffer: 3.4 g/L of monobasic potassium phosphate Detector: UV 280 nm
Mobile phase: Acetonitrile, triethylamine, and Buffer Column: 4.6-mm 25-cm; 5-m packing L11
(15:2:85) Guard column: 4.6-mm 1-cm; 5-m packing L11
Mix thoroughly, and adjust with 85.0% phosphoric acid to a Temperature: 40
pH of 3.0 0.1. Flow rate: 2.0 mL/min
Standard solution: 0.2 mg/mL of USP Butorphanol Tartrate Injection size: 60 L
RS in Mobile phase System suitability
Mix, and filter, discarding the first 2 mL of the filtrate. Samples: Standard solution and Sensitivity solution
[NOTEThe Standard solution is stable for at least 108 h.] Suitability requirements
Sample solution: Prepare a composite solution by pooling Relative standard deviation: NMT 10.0% in the
a minimum of four containers of Nasal Solution into a Standard solution
suitable glass vessel. Transfer the equivalent of 20 mg of [NOTEThe peak height for butorphanol tartrate is
butorphanol tartrate to a 100-mL volumetric flask. Dilute greater than or equal to three times the baseline noise
with Mobile phase to volume, mix, and filter, discarding the in the Sensitivity solution.]
first 2 mL of the filtrate. Analysis
[NOTEThe Sample solution has a nominal concentration of Samples: Standard solution and Sample solution
0.2 mg/mL of butorphanol tartrate.] Record the chromatograms, and measure the responses
Chromatographic system for the butorphanol tartrate peak in the Standard
(See Chromatography 621, System Suitability.) solution, and for all known and unknown related
Mode: LC compounds in the Sample solution. The chromatographic
Detector: UV 280 nm run time is 40 min.
Column: 4.6-mm 15-cm; 5-m packing L11 Calculate the percentage of each related compound (see
Guard column: 4.6-mm 1-cm; 5-m packing L11 Impurity Table 1) and each unknown impurity in the
Temperature: 30 portion of Nasal Solution taken:
Flow rate: 2 mL/min
Injection size: 20 L Result = (rU/rS) (CS/CU) 100
System suitability
Sample: Standard solution rU = peak response of each known or unknown
Suitability requirements related compound from the Sample solution
Tailing factor: NMT 2.0 rS = peak response of butorphanol tartrate from the
Relative standard deviation: NMT 2.0% Standard solution
Analysis CS = concentration of USP Butorphanol Tartrate RS in
Calculate the percentage of C21H29NO2 C4H6O6 in the CU = nominal concentration of butorphanol tartrate
portion of Nasal Solution taken: in the Sample solution (mg/mL)
rU = peak response of butorphanol tartrate from the
Sample solution
166 Butorphanol / Official Monographs USP 32
Acceptance criteria: See Impurity Table 1 SPECIFIC TESTS

MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED
Impurity Table 1 MICROORGANISMS 62: The total aerobic microbial count
does not exceed 1000 cfu/g or mL, and the total combined
Relative Relative Acceptance molds and yeasts count does not exceed 100 cfu/g or mL. It
Retention Response Criteria, meets the requirements for absence of Staphylococcus aureus
Name Time Factor NMT (%) and Pseudomonas aeruginosa.
3,14- PH 791: 4.06.0
0.3 0.3 OSMOLALITY AND OSMOLARITY 785: 252292 mOsmol/kg
Dihydroxymorphinan
6-Butorphanol 0.7 0.5 ADDITIONAL REQUIREMENTS
Butorphanol tartrate 1.0 PACKAGING AND STORAGE: Preserve in tight containers at
Unknown impurity 0.3 controlled room temperature. Store at 25, excursions
permitted between 15 and 30.
Total 1.0 USP REFERENCE STANDARDS 11
USP Butorphanol Tartrate RS
Cabergoline Calcium Levulinate

Caffeine Calcium Levulinate Injection
Caffeine Citrate Injection Calcium Pantothenate
Caffeine Citrate Oral Solution Calcium Pantothenate Tablets
Caffeine and Sodium Benzoate Injection Racemic Calcium Pantothenate
Calamine Dibasic Calcium Phosphate Dihydrate
Calamine Topical Suspension Anhydrous Dibasic Calcium Phosphate
Phenolated Calamine Topical Suspension Dibasic Calcium Phosphate Tablets
Calcifediol Calcium Polycarbophil
Calcifediol Capsules Calcium Saccharate
Calcitriol Calcium Undecylenate
Calcitriol Injection Camphor
Calcitonin Salmon Camphor Spirit
Calcitonin Salmon Injection Capecitabine
Calcitonin Salmon Nasal Solution Capecitabine Tablets
Calcium Acetate Capreomycin Sulfate
Calcium Acetate Tablets Capreomycin for Injection
Calcium Ascorbate Capsaicin
Calcium Carbonate Capsicum
Calcium Carbonate Lozenges Capsicum Oleoresin
Calcium Carbonate Oral Suspension Captopril
Calcium Carbonate Tablets Captopril Oral Solution
Calcium Carbonate and Magnesia Tablets Captopril Oral Suspension
Calcium Carbonate and Magnesia Chewable Tablets Captopril Tablets
Calcium Carbonate, Magnesia, and Simethicone Tablets Captopril and Hydrochlorothiazide Tablets
Calcium Carbonate, Magnesia, and Simethicone Chewable Carbachol
Tablets
Carbachol Intraocular Solution
Calcium and Magnesium Carbonates Oral Suspension
Carbachol Ophthalmic Solution
Calcium and Magnesium Carbonates Tablets
Carbamazepine
Calcium Chloride
Carbamazepine Oral Suspension
Calcium Chloride Injection
Carbamazepine Tablets
Calcium Citrate
Carbamazepine Extended-Release Tablets
Calcium Glubionate Syrup
Carbamide Peroxide
Calcium Gluceptate
Carbamide Peroxide Topical Solution
Calcium Gluceptate Injection
Carbenicillin Disodium
Calcium Gluconate
Carbenicillin for Injection
Calcium Gluconate Injection
Carbenicillin Indanyl Sodium
Calcium Gluconate Tablets
Carbenicillin Indanyl Sodium Tablets
Calcium Hydroxide
Carbidopa
Calcium Hydroxide Topical Solution
Carbidopa and Levodopa Tablets
Calcium Lactate
Carbinoxamine Maleate
Calcium Lactate Tablets
Carbinoxamine Maleate Tablets
Calcium Lactobionate
Carbol-Fuchsin Topical Solution
Carbon Dioxide Cefadroxil for Oral Suspension

Carbon Monoxide C 11 Cefadroxil Tablets
Flumazenil C 11 Injection Cefamandole Nafate
Mespiperone C 11 Injection Cefamandole Nafate for Injection
Methionine C 11 Injection Cefazolin
Raclopride C 11 Injection Cefazolin Sodium
Sodium Acetate C 11 Injection Cefazolin Injection
Urea C 13 Cefazolin for Injection
Urea C 13 for Oral Solution Cefazolin Ophthalmic Solution
Urea C 14 Capsules Cefdinir
Carboplatin Cefdinir Capsules
Carboplatin for Injection Cefdinir for Oral Suspension
Carboprost Tromethamine Cefepime Hydrochloride
Carboprost Tromethamine Injection Cefepime for Injection
Carboxymethylcellulose Sodium Cefixime
Carboxymethylcellulose Sodium Paste Cefixime for Oral Suspension
Carboxymethylcellulose Sodium Tablets Cefixime Tablets
Carisoprodol Cefmenoxime Hydrochloride
Carisoprodol Tablets Cefmenoxime for Injection
Carisoprodol and Aspirin Tablets Cefmetazole
Carisoprodol, Aspirin, and Codeine Phosphate Tablets Cefmetazole Injection
Carprofen Cefmetazole Sodium
Carprofen Tablets Cefmetazole for Injection
Carteolol Hydrochloride Cefonicid Sodium
Carteolol Hydrochloride Ophthalmic Solution Cefonicid for Injection
Carteolol Hydrochloride Tablets Cefoperazone Sodium
Casanthranol Cefoperazone Injection
Cascara Sagrada Cefoperazone for Injection
Cascara Sagrada Extract Ceforanide
Cascara Tablets Ceforanide for Injection
Cascara Sagrada Fluidextract Cefotaxime Sodium
Aromatic Cascara Fluidextract Cefotaxime Injection
Castor Oil Cefotaxime for Injection
Castor Oil Capsules Cefotetan
Castor Oil Emulsion Cefotetan Injection
Aromatic Castor Oil Cefotetan for Injection
Cefaclor Cefotetan Disodium
Cefaclor Capsules Cefotiam Hydrochloride
Cefaclor for Oral Suspension Cefotiam for Injection
Cefaclor Chewable Tablets Cefoxitin Sodium
Cefaclor Extended-Release Tablets Cefoxitin Injection
Cefadroxil Cefoxitin for Injection
Cefadroxil Capsules Cefpiramide
Cefpiramide for Injection Cephapirin Benzathine

Cefpodoxime Proxetil Cephapirin Benzathine Intramammary Infusion
Cefpodoxime Proxetil for Oral Suspension Cephapirin Sodium
Cefpodoxime Proxetil Tablets Cephapirin for Injection
Cefprozil Cephapirin Sodium Intramammary Infusion
Cefprozil for Oral Suspension Cephradine
Cefprozil Tablets Cephradine Capsules
Ceftazidime Cephradine for Injection
Ceftazidime Injection Cephradine for Oral Suspension
Ceftazidime for Injection Cephradine Tablets
Ceftizoxime Sodium Cetylpyridinium Chloride
Ceftizoxime Injection Cetylpyridinium Chloride Lozenges
Ceftizoxime for Injection Cetylpyridinium Chloride Topical Solution
Ceftriaxone Sodium Activated Charcoal
Ceftriaxone Injection Chloral Hydrate
Ceftriaxone for Injection Chloral Hydrate Capsules
Cefuroxime Axetil Chloral Hydrate Oral Solution
Cefuroxime Axetil for Oral Suspension Chlorambucil
Cefuroxime Axetil Tablets Chlorambucil Tablets
Cefuroxime Sodium Chloramphenicol
Cefuroxime Injection Chloramphenicol Capsules
Cefuroxime for Injection Chloramphenicol Cream
Oxidized Cellulose Chloramphenicol Injection
Oxidized Regenerated Cellulose Chloramphenicol Ophthalmic Ointment
Cellulose Sodium Phosphate Chloramphenicol Ophthalmic Solution
Cellulose Sodium Phosphate for Oral Suspension Chloramphenicol for Ophthalmic Solution
Cephalexin Chloramphenicol Oral Solution
Cephalexin Hydrochloride Chloramphenicol Otic Solution
Cephalexin Capsules Chloramphenicol Tablets
Cephalexin for Oral Suspension Chloramphenicol and Hydrocortisone Acetate for
Ophthalmic Suspension
Cephalexin Tablets
Chloramphenicol and Polymyxin B Sulfate Ophthalmic
Cephalexin Tablets for Oral Suspension Ointment
Cephalothin Sodium Chloramphenicol, Polymyxin B Sulfate, and Hydrocortisone
Cephalothin Injection Acetate Ophthalmic Ointment
Cephalothin for Injection
USP 32 Official Monographs / Cabergoline 1
132270

Add the following: IMPURITIES
Organic Impurities
PROCEDURE
Cabergoline [NOTEPrepare solutions immediately before use, and
(Comment on this Monograph)id=m11140=Cabergoline=Ca- protect from light.]
Chl-Monos.pdf) Buffer: Proceed as directed in the Assay.
Mobile phase: Proceed as directed in the Assay.
System suitability solution: To 10 mL of 0.1 M sodium
hydroxide, add 50 mg of Cabergoline. Stir for about 15
min. To 1 mL of the suspension, add 1 mL of 0.1 M
hydrochloric acid, and dilute with Mobile phase to 10.0 mL.
Sonicate until dissolution is complete. [NOTEThe main
degradation product obtained is cabergoline related
compound A.]
C26H37N5O2 451.60 Sample solution: 0.25 mg/mL of Cabergoline in Mobile
Ergoline-8-carboxamide, N-[3-(dimethylamino)propyl]-N- phase [NOTESonicate if needed.]
[(ethy- quinolamino)carbonyl]-6-(2-propenyl)-; Chromatographic system
1-[(6-Allylergolin-8-yl)carbonyl]-1-[3-(dimethylamino)propyl]-3- (See Chromatography 621, System Suitability.)
ethylurea [81409-90-7]. Mode: LC
Cabergoline contains NLT 98.0% and NMT 102.0% of the Column: 4.0-mm 25-cm; 10 m packing L1
labeled amount of C26H37N5O2, calculated on the anhydrous Flow rate: 1.3 mL/min
basis. Injection size: 100 L
System suitability
IDENTIFICATION Sample: System suitability solution
A. INFRARED ABSORPTION 197K Suitability requirements
B. The retention time of the major peak of the Sample Resolution: NLT 3.0 between cabergoline and
solution corresponds to that of the Standard solution, as cabergoline related compound A
Analysis
obtained in the Assay. Sample: Sample solution
ASSAY Calculate the percentage of each impurity in the portion
PROCEDURE of Cabergoline taken:
[NOTE Prepare solutions immediately before use and
protect from light.] Result = (rU/rS) 100
Buffer: Dissolve 6.8 g of monobasic potassium phosphate in rU = peak response of each impurity from the Sample
900 mL water, adjust with phosphoric acid to a pH of 2.0, solution
and dilute to 1 L. Add 0.2 mL of triethylamine to the rS = sum of the peak responses for all peaks from the
resulting solution, and mix. Sample solution
Mobile phase: Acetonitrile and Buffer (4:21) Acceptance criteria
Standard solution: 0.25 mg/mL of USP Cabergoline RS in Individual impurities: See Impurity Table 1.
Mobile phase [NOTESonicate if needed.] Total impurities: NMT 0.8%
Sample solution: 0.25 mg/mL of Cabergoline in Mobile
phase [NOTESonicate if needed.]
Chromatographic system Impurity Table 1
(See Chromatography 621, System Suitability.) Relative Acceptance
Mode: LC Retention Criteria,
Detector: UV 280 nm Name Time NMT (%)
Column: 4.0-mm 25-cm; 10 m packing L1
Flow rate: 1.3 mL/min Cabergoline related 0.3 0.1
Injection size: 100 L compound Da
System suitability Cabergoline related 0.6 0.1
Samples: Standard solution compound Bb
Suitability requirements Cabergoline related 0.8 0.3
Column efficiency: NLT 1000 theoretical plates compound Ac
Analysis Cabergoline 1.0
Samples: Standard solution and Sample solution a (6aR,9R,10aR)-N-[3-(Dimethylamino)propyl]-7-(prop-2-
Calculate the percentage of C26H37N5O2 in the portion of enyl)-4,6,6a,7,8,9,10,10aoctahydroindolo[4,3-fg]quinoline-9-carboxamide.
Cabergoline taken: b (6aR,9R,10aR)-N9-[3-(Dimethylamino)propyl]-N4-ethyl-7-(prop-2-
enyl)6a,7,8,9,10,10a-hexahydroindolo[4,3-fg]quinoline-4,9(6H)-
dicarboxamide.
c (6aR,9R,10aR)-7-(Prop-2-enyl)-4,6,6a,7,8,9,10,10a-octahydroindolo[4,3-
rS = peak response of the Standard solution fg]quinoline-9-carboxylic acid.
d (6aR,9R,10aR)-N9-[3-(Dimethylamino)propyl]-N4-ethyl-N9-
CS = concentration of USP Cabergoline RS in the
Standard solution (mg/mL) (ethylcarbamoyl)-7-(prop2-enyl)-6a,7,8,9,10,10a-hexahydroindolo[4,3-
CU = concentration of caberrgoline the Sample solution fg]quinoline-4,9(6H)-dicarboxamide.
(mg/mL)
2 Cabergoline / Official Monographs USP 32
Impurity Table 1 (continued) ASSAY

PROCEDURE
Relative Acceptance Solution A: 0.82 mg/mL of anhydrous sodium acetate
Retention Criteria, Mobile phase: Acetonitrile, tetrahydrofuran, and Solution A
Name Time NMT (%) (5:4:191). Adjust with glacial acetic acid to a pH of 4.5.
Cabergoline related 2.9 0.3 System suitability solution: 0.02 mg/mL of theophylline in
compound Cd Mobile phase [NOTEShake, and sonicate, if necessary, to
Any other individual, 0.10 dissolve.]
unidentified Standard solution: 0.2 mg/mL of USP Caffeine RS in
impurity System suitability solution and Mobile phase (1:4) [NOTE
Shake, and sonicate, if necessary, to dissolve.]
a (6aR,9R,10aR)-N-[3-(Dimethylamino)propyl]-7-(prop-2- Sample solution: 0.2 mg/mL of Caffeine in Mobile phase
enyl)-4,6,6a,7,8,9,10,10aoctahydroindolo[4,3-fg]quinoline-9-carboxamide. [NOTEShake, and sonicate, if necessary, to dissolve.]
b (6aR,9R,10aR)-N9-[3-(Dimethylamino)propyl]-N4-ethyl-7-(prop-2-
enyl)6a,7,8,9,10,10a-hexahydroindolo[4,3-fg]quinoline-4,9(6H)- (See Chromatography 621, System Suitability.)
dicarboxamide. Mode: LC
c (6aR,9R,10aR)-7-(Prop-2-enyl)-4,6,6a,7,8,9,10,10a-octahydroindolo[4,3-
Detector: UV 275 nm
fg]quinoline-9-carboxylic acid. Column: 4.6-mm 15-cm; packing L1
d (6aR,9R,10aR)-N9-[3-(Dimethylamino)propyl]-N4-ethyl-N9-
Flow rate: 1 mL/min
(ethylcarbamoyl)-7-(prop2-enyl)-6a,7,8,9,10,10a-hexahydroindolo[4,3- Injection size: 10 L
fg]quinoline-4,9(6H)-dicarboxamide. System suitability
SPECIFIC TESTS Sample: Standard solution
OPTICAL ROTATION, Specific Rotation 781S: 77 to 83 [NOTEThe relative retention times for theophylline and
Sample solution: 1 mg/mL in alcohol, on the anhydrous caffeine are about 0.69 and 1.0, respectively. ]
basis Suitability requirements
WATER DETERMINATION, Method I 921: NMT 5.0% Resolution: NLT 6.0 between theophylline and caffeine
Tailing factor: NMT 2.0 for each of the peaks
ADDITIONAL REQUIREMENTS Relative standard deviation: NMT 2.0%
PACKAGING AND STORAGE: Preserve in tight containers, Analysis
protected from light. Samples: Standard solution and Sample solution
USP REFERENCE STANDARDS 11 Calculate the percentage of C8H10N4O2 in the portion of
USP Cabergoline RSUSP32 Caffeine taken:
Caffeine rU = peak response for Caffeine from the Sample

solution
(Comment on this Monograph)id=m11170=Caffeine=Ca-Chl- rS = peak response for Caffeine from the Standard
Monos.pdf) solution
CS = concentration of USP Caffeine RS in the Standard
solution (mg/mL)
CU = nominal concentration of caffeine in the Sample
solution (mg/mL)
IMPURITIES
C8H10N4O2 (anhydrous) 194.19 RESIDUE ON IGNITION 281: NMT 0.1%
1H-Purine-2,6-dione, 3,7-dihydro-1,3,7-trimethyl-; HEAVY METALS, Method II 231: NMT 10 ppm
1,3,7-Trimethylxanthine. [58-08-2] Organic Impurities
Monohydrate 212.21 PROCEDURE 1: READILY CARBONIZABLE SUBSTANCES 271:
[5743-12-4]. Dissolve 0.5 g in 5 mL of sulfuric acid TS: the solution has
DEFINITION no more color than Matching Fluid D.
Caffeine is anhydrous or contains one molecule of water of PROCEDURE 2
hydration. It contains NLT 98.5% and NMT 101.0% of Mobile phase, System suitability solution, Standard
C8H10N4O2, calculated on the anhydrous basis. solution, Sample solution, and Chromatographic
system: Proceed as directed in the Assay.
A. INFRARED ABSORPTION 197M Samples: Standard solution and Sample solution
B. PROCEDURE Calculate the percentage of each impurity in the portion
Sample solution: 5 mg in 1 mL of hydrochloric acid, in a of Caffeine taken:
porcelain dish
Analysis: Add 50 mg of potassium chlorate, and evaporate Result = (ri/rT) 100
on a steam bath to dryness. Invert the dish over a vessel
containing a few drops of 6 N ammonium hydroxide. ri = peak response for each impurity, Sample solution
Acceptance criteria: The residue acquires a purple color, rT = sum of the responses of all the peaks, Sample
which disappears upon the addition of a solution of 1 N solution
sodium hydroxide.
USP 32 Official Monographs / Caffeine 3

Individual impurities: NMT 0.1% (See Chromatography 621, System Suitability.)
Total impurities: NMT 0.1% Mode: LC
Detector: UV 275 nm
SPECIFIC TESTS Column: 4.6-mm 150-cm; 5-m packing L1
MELTING RANGE OR TEMPERATURE 741: 235239, Flow rate: 1 mL/min
determined after drying at 80 for 4 h Injection size: 10 L
WATER DETERMINATION, Method III 921: Dry it at 80 for 4 System suitability
h: the anhydrous form loses NMT 0.5% and the hydrous Sample: Standard solution
form NMT 8.5% of its weight. [NOTEThe relative retention times for theophylline and
OTHER ALKALOIDS: To 5 mL of a solution (1 in 50), add caffeine are 0.7 and 1.0, respectively. ]
mercuricpotassium iodide TS: no precipitate is formed. Suitability requirements
Resolution: NLT 6.0 between theophylline and caffeine
ADDITIONAL REQUIREMENTS Tailing factor: NMT 2.0, theophylline and caffeine
PACKAGING AND STORAGE: Preserve hydrous Caffeine in tight Relative standard deviation: NMT 2.0% for caffeine
containers. Preserve anhydrous Caffeine in well-closed Analysis
containers. Samples: Standard solution and Sample solution
LABELING: Label it to indicate whether it is anhydrous or Calculate the percentage of C14H18N4O9 in the volume of
hydrous. Injection taken:
USP Caffeine RS Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
Caffeine Citrate Injection rS = peak response of the Standard solution
CS = concentration of USP Caffeine RS in the Standard
(Comment on this Monograph)id=m11175=Caffeine Citrate solution (mg/mL)
Injection=Ca-Chl-Monos.pdf) CU = nominal concentration of caffeine citrate in the
DEFINITION Mr1 = molecular weight of caffeine citrate, 386.31
Caffeine Citrate Injection is a sterile solution containing Caffeine Mr2 = molecular weight of caffeine, 194.19
and citric acid in Water for Injection. It contains NLT 90.0% Acceptance criteria: 90.0%110.0%
and NMT 110.0% of the labeled amount of caffeine citrate
(C14H18N4O9). It contains no bacteriostat or other preservative. IMPURITIES
Organic Impurities
A. The retention time of the major peak of the Sample Mobile phase, Identification solution, Standard solution,
solution corresponds to that of the Standard solution, as and Sample solution: Proceed as directed in the Assay.
obtained in the Assay. System suitability solution: Standard solution and water
B. IDENTIFICATION TESTSGENERAL, Citrate 191 (1:39)
C. PROCEDURE Chromatographic system: Proceed as directed in the
Sample solution: 4 g of potassium iodide in a 100-mL Assay.
volumetric flask. Add 10 mL of water, and shake until the Suitability requirements
potassium iodide is dissolved. Transfer 2 g of iodine to the Sensitivity: A peak for theophylline is discernable.
volumetric flask, and shake until dissolved. Dilute with water Analysis
to volume. Transfer 5 drops to a 25-mL centrifuge tube Samples: Standard solution and Sample solution
containing 5.0 mL of Injection, and mix. Calculate the percentage of any impurity in the portion of
Analysis: Add 0.5 mL of 2.0 M hydrochloric acid solution, Injection taken:
and mix.
Acceptance criteria: A brown precipitate that dissolves on Result = (rU/rS) (CS/CU) (Mr1/Mr2) F 100
neutralization with 0.5 mL of sodium hydroxide TS is
produced. rU = individual peak response for each impurity in
the Sample solution
ASSAY rS = peak response for caffeine from the Standard
PROCEDURE solution
Mobile phase: Acetonitrile, tetrahydrofuran, and 0.01 M CS = concentration of USP Caffeine RS in the
sodium acetate (5:4:191). Adjust with glacial acetic acid to a Standard solution (mg/mL)
pH of 4.5. CU = nominal concentration of caffeine citrate in the
Identification solution: 0.02 mg/mL of theophylline Sample solution (mg/mL)
Standard solution: 0.2 mg/mL of USP Caffeine RS in Mr1 = molecular weight of caffeine citrate, 386.31
Identification solution and water (1:4) Mr2 = molecular weight of caffeine, 194.19
Sample solution: Equivalent to 0.2 mg/mL of caffeine from F = relative response factor as given in Impurity
a volume of Injection in water. [NOTEPass through a Table 1
polyvinylidene difluoride or equivalent membrane having a
porosity of 0.45 m.]
4 Caffeine / Official Monographs USP 32
Acceptance criteria ASSAY

Individual impurities: See Impurity Table 1. PROCEDURE
Total impurities: NMT 0.1% Mobile phase: Acetonitrile, tetrahydrofuran, and 0.01 M
sodium acetate (5:4:191). Adjust with glacial acetic acid to a
Impurity Table 1 pH of 4.5.
Identification solution: 0.02 mg/mL of theophylline
Relative Relative Acceptance Standard solution: 0.2 mg/mL of USP Caffeine RS in
Retention Response Criteria, Identification solution and water (1:4)
Name Time Factor NMT (%) Sample solution: Equivalent to 0.2 mg/mL of caffeine from
Theobromine 0.4 0.878 0.1 a volume of Oral Solution in water. [NOTEPass through a
Paraxanthine 0.6 1.10 0.1
polyvinylidene difluoride or equivalent membrane having a
0.45-m porosity.]
Theophylline 0.7 0.905 0.1 Chromatographic system
Any other 1.0 0.1 (See Chromatography 621, System Suitability.)
impurity Mode: LC
Detector: UV 275 nm
COLOR AND CLARITY: Transfer a suitable portion of the Injection size: 10 L
Injection to a clear glass test tube, and visually examine the System suitability
solution in a well-lighted area: the solution is colorless and Sample: Standard solution
free of haze, obvious turbidity, and precipitate. [NOTEThe relative retention times for theophylline and
BACTERIAL ENDOTOXINS TEST 85: NMT 0.25 USP Endotoxin caffeine are 0.7 and 1.0, respectively.]
Unit/mg of caffeine Suitability requirements
STERILITY TESTS 71: It meets the requirements for Test for Resolution: NLT 6.0 between theophylline and caffeine
Sterility of the Product to Be Examined, Membrane Filtration. Tailing factor: NMT 2.0 determined from the
PH 791: 4.25.2 theophylline and caffeine peaks
PARTICULATE MATTER IN INJECTIONS 788: NMT 150 particles Relative standard deviation: NMT 2.0% for caffeine
are equal to or greater than 10 m, and NMT 25 particles peaks
are equal to or greater than 25 m Analysis
OTHER REQUIREMENTS: It meets the requirements under Samples: Standard solution and Sample solution
Injections 1. Calculate the percentage of C14H18N4O9 in the volume of
Oral Solution taken:
PACKAGING AND STORAGE: Preserve in single-dose, tight Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
containers of Type I glass, and store between 15 and 30.
USP REFERENCE STANDARDS 11 rU = peak response of the Sample solution
USP Caffeine RS rS = peak response of the Standard solution
USP Endotoxin RS CS = concentration of USP Caffeine RS in the Standard
solution (mg/mL)
CU = nominal concentration of caffeine citrate in the
Caffeine Citrate Oral Solution Mr1 = molecular weight of caffeine citrate, 386.31
(Comment on this Monograph)id=m11180=Caffeine Citrate Mr2 = molecular weight of caffeine, 194.19
Oral Solution=Ca-Chl-Monos.pdf) Acceptance criteria: 90.0%110.0%
DEFINITION IMPURITIES
Caffeine Citrate Oral Solution is a sterile aqueous solution Organic Impurities
containing Caffeine and citric acid. It contains NLT 90.0% and PROCEDURE
NMT 110.0% of the labeled amount of caffeine citrate Mobile phase, Identification solution, Standard solution,
(C14H18N4O9). It contains no bacteriostat or other preservative. and Sample solution: Proceed as directed in the Assay.
System sensitivity solution: Standard solution and water
IDENTIFICATION (1:39)
A. The retention time of the major peak of the Sample Chromatographic system: Proceed as directed in the
solution corresponds to that of the Standard solution, as Assay. C
obtained in the Assay. Suitability requirements
B. IDENTIFCATION TESTSGENERAL, Citrate 191 Sensitivity: A peak for theophylline is discernable.
C. PROCEDURE Analysis
Sample solution: 10 mg/mL of potassium iodide and 20 Samples: Standard solution and Sample solution
mg/ml of iodine in water [NOTEInitially combine potassium Calculate the percentage of any impurity in the portion of
iodide and water, and shake until dissolved, then add iodine Oral Solution taken:
to the solution]
Analysis: Transfer 5 drops of the Sample solution to a 25- Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
mL centrifuge tube containing 5 mL of the Oral Solution,
and mix. Add 0.5 mL of 2.0 M hydrochloric acid solution, rU = individual peak response for each impurity for
and mix. the Sample solution
Acceptance criteria: A brown precipitate is produced that rS = peak response for caffeine from the Standard
dissolves on neutralization with 0.5 mL of sodium hydroxide solution
TS.
USP 32 Official Monographs / Calamine 5
CS = concentration of USP Caffeine RS in the caffeine, passing each chloroform extract through a small
Standard solution (mg/mL) filter previously moistened with chloroform into a tared dish.
CU = nominal concentration of caffeine citrate in the [NOTERetain the water layer for the Assay for Sodium
Sample solution (mg/mL) Benzoate.] Wash the stem of the separator, the filter, and the
Mr1 = molecular weight of caffeine citrate, 396.31 funnel with 10 mL of hot chloroform, adding the washings
Mr2 = molecular weight of caffeine, 194.19 to the dish. Evaporate the combined chloroform solutions
Acceptance criteria on a steam bath, adding 2 mL of alcohol just before the last
Individual impurities: See Impurity Table I. trace of chloroform is expelled. Complete the evaporation of
Total impurities: NMT 0.1% the solvent; dry the residue, consisting of C8H10N4O2, at 80
for 4 h; cool; and weigh.
Impurity Table 1 Acceptance criteria: 90.0%110.0%
SODIUM BENZOATE
Relative Relative Acceptance Sample solution: To the water layer obtained in the Assay
Retention Response Criteria, for Caffeine add 75 mL of ether and 5 drops of methyl
Name Time Factor NMT (%) orange TS.
Theobromine 0.4 0.878 0.1 Analysis: Titrate with 0.1 N hydrochloric acid VS, mixing
Paraxanthine 0.6 1.10 0.1
the liquids by vigorous shaking, until a permanent pink color
is produced in the water layer. Each mL of 0.1 N
Theophylline 0.7 0.905 0.1 hydrochloric acid is equivalent to 14.41 mg of C7H5NaO2.
Any other Acceptance criteria: 90.0%110.0%
individual,
1.0 0.1 SPECIFIC TESTS
unidentified
impurity BACTERIAL ENDOTOXINS TEST 85: NMT 0.7 USP Endotoxin
Unit/mg of caffeine and sodium benzoate, based on the
total, in mg, of the labeled amounts
SPECIFIC TESTS PH 791: 6.58.5
STERILITY TESTS 71: It meets the requirements when tested OTHER REQUIREMENTS: It meets the requirements under
as directed for Test for Sterility of the Product to Be Examined, Injections 1.
PH 791: 4.25.2 ADDITIONAL REQUIREMENTS
ADDITIONAL REQUIREMENTS preferably of Type I glass.
PACKAGING AND STORAGE: Preserve in single-dose, tight USP REFERENCE STANDARDS 11
containers, and store at a temperature between 15 and 30. USP Caffeine RS
USP REFERENCE STANDARDS 11 USP Endotoxin RS
USP Caffeine RS
Calamine
Caffeine and Sodium Benzoate (Comment on this Monograph)id=m11250=Calamine=Ca-Chl-
Injection Monos.pdf)
(Comment on this Monograph)id=m11200=Caffeine and Iron oxide (Fe2O3), mixture with zinc oxide;
Sodium Benzoate Injection=Ca-Chl-Monos.pdf) Calamine (pharmaceutical solution) [8011-96-9].
Caffeine and Sodium Benzoate Injection is a sterile solution Calamine is Zinc Oxide with a small proportion of ferric oxide,
containing equal amounts of Caffeine and Sodium Benzoate in and contains, after ignition, NLT 98.0% and NMT 100.5% of
Water for Injection. It contains NLT 90.0% and NMT 110.0% ZnO.
of the labeled amounts of anhydrous caffeine (C8H10N4O2) and
sodium benzoate (C7H5NaO2). IDENTIFICATION
A. IDENTIFICATION TESTSGENERAL, Zinc 191: Combine 1 g
IDENTIFICATION of Calamine with 10 mL of 3 N hydrochloric acid, and filter:
A. INFRARED ABSORPTION 197K: The caffeine obtained in the filtrate meets the requirements of tests.
the Assay for Caffeine meets the requirements of the test. B. PROCEDURE
B. Dip the end of a platinum wire into a portion of Analysis: Combine 1g of Calamine with 100 mL of 3 N
Injection, and introduce it into a nonluminous flame: the hydrochloric acid, heat to boiling, and filter.
flame is colored intensely yellow. Acceptance criteria: The filtrate assumes a reddish color
C. To 0.5 mL of Injection, add a few drops of ferric chloride upon the addition of ammonium thiocyanate TS.
TS: a salmon-colored precipitate is formed. To another
portion of Injection, add 3 N hydrochloric acid: a white ASSAY
precipitate is formed. PROCEDURE
Sample solution: Digest 1.5 g of freshly ignited Calamine
ASSAY in 50.0 mL of 1 N sulfuric acid VS, applying gentle heat until
CAFFEINE no further solution occurs. Filter the mixture, and wash the
Sample solution: Equivalent to 250 mg each of caffeine residue on the filter with hot water until the last washing is
and sodium benzoate from a volume of Injection. Transfer it neutral to litmus paper. To the combined filtrate and
completely with the aid of 5 mL of water to a small washings, add 2.5 g of ammonium chloride, cool, add
separator, add 1 drop of phenolphthalein TS, and add 0.1 N methyl orange TS.
sodium hydroxide, dropwise, until a permanent pink color is Analysis: Titrate with 1 N sodium hydroxide VS. Each mL of
just produced. 1 N sulfuric acid is equivalent to 40.69 mg of ZnO.
Analysis: Shake the mixture with three or more 20-mL
portions of chloroform to effect complete extraction of the
6 Calamine / Official Monographs USP 32

IMPURITIES MICROORGANISMS 62: It meets the requirements of the
Inorganic Impurities tests for absence of Staphylococcus aureus and Pseudomonas
LOSS ON IGNITION 733: Weigh 2 g, and ignite at 500 to aeruginosa.
constant weight: it loses NMT 2.0% of its weight.
CALCIUM: Digest 1 g in 25 mL of 3 N hydrochloric acid for ADDITIONAL REQUIREMENTS
30 min, filter to remove the insoluble ferric oxide, and add PACKAGING AND STORAGE: Preserve in tight containers.
6 N ammonium hydroxide to the filtrate until the precipitate
first formed is redissolved, then add 5 mL more of 6 N
ammonium hydroxide. To 10 mL of this solution, add 2 mL
of ammonium oxalate TS: NMT a slight turbidity is Phenolated Calamine Topical
produced. Suspension
CALCIUM OR MAGNESIUM: To another 10-mL portion of the (Comment on this Monograph)id=m11270=Phenolated
solution prepared in the Calcium Procedure, add 2 mL of Calamine Topical Suspension=Ca-Chl-Monos.pdf)
dibasic sodium phosphate TS: NMT a slight turbidity is Prepare Phenolated Calamine Topical Suspension as follows:
produced.
ARSENIC, Method I 211: NMT 8 ppm
LEAD: To 1 g add 15 mL of water, stir, then add 3 mL of Liquefied Phenol 10 mL
glacial acetic acid, and warm on a steam bath until Calamine Topical Suspension 990 mL
dissolved. Filter, and add 5 drops of potassium chromate TS: To make 1000 mL
no turbidity is produced.
Mix the ingredients.
SPECIFIC TESTS
[NOTEShake Phenolated Calamine Topical Suspension before
dispensing.]
MICROORGANISMS 62: It meets the requirements of the
tests for absence of Staphylococcus aureus and Pseudomonas ADDITIONAL REQUIREMENTS
aeruginosa. PACKAGING AND STORAGE: Preserve in tight containers.
ACID-INSOLUBLE SUBSTANCES: Dissolve 2.0 g in 50 mL of 3 N
hydrochloric acid. If an insoluble residue remains, collect it
on a tared filter, wash with water, dry at 105 for 1 h, cool,
and weigh: the weight of the residue does not exceed 40 Calcifediol
mg (2.0%). (Comment on this Monograph)id=m11290=Calcifediol=Ca-Chl-
ALKALINE SUBSTANCES: Digest 1.0 g with 20 mL of water on Monos.pdf)
a steam bath for 15 min, filter, and add 2 drops of
phenolphthalein TS: if a red color is produced, NMT 0.20
mL of 0.10 N sulfuric acid is required to discharge it.
Calamine Topical Suspension C27H44O2 H2O 418.65

9,10-Secocholesta-5,7,10(19)-triene-3,25-diol monohydrate,
(Comment on this Monograph)id=m11260=Calamine Topical (3,5Z,7E)-;
Suspension=Ca-Chl-Monos.pdf) 25-Hydroxycholecalciferol;
(Former monograph title is Calamine Lotion) Monohydrate [63283-36-3].
Prepare Calamine Topical Suspension as follows.
DEFINITION
Calamine 80 g Calcifediol contains NLT 97.0% and NMT 103.0% of C27H44O2
H2O.
Zinc Oxide 80 g
Glycerin 20 mL IDENTIFICATION
Bentonite Magma 250 mL A. INFRARED ABSORPTION 197M
Calcium Hydroxide Topical A sufficient quantity ASSAY
Solution PROCEDURE
To make 1000 mL Internal standard solution: 0.10 mg/mL of testosterone in
ethyl acetate
Dilute the Bentonite Magma with an equal volume of Calcium Mobile phase: Heptane, methylene chloride, ethyl acetate,
Hydroxide Topical Solution. Mix the powders intimately with and water-saturated heptane (6:3:5:6)
the Glycerin and about 100 mL of the diluted magma, Standard solution: 20 g/mL of USP Calcifediol RS in
triturating until a smooth, uniform paste is formed. Gradually Internal standard solution
incorporate the remainder of the diluted magma. Finally, add Sample solution: 20 g/mL of Calcifediol in Internal
enough Calcium Hydroxide Topical Solution to make 1000 mL, standard solution
and shake. Chromatographic system
If a more viscous consistency in the Calamine Topical (See Chromatography 621, System Suitability.)
Suspension is desired, the quantity of Bentonite Magma may Mode: LC
be increased to NMT 400 mL. Detector: UV 254 nm, and a pump capable of providing
[NOTEShake the Calamine Topical Suspension before constant flow at over 2000 psi
dispensing.]
USP 32 Official Monographs / Calcifediol 7
Column: 4-mm 30-cm; packing L3 Mobile phase: Heptane, methylene chloride, ethyl acetate,
Injection size: Equal volumes and water-saturated heptane (6:3:5:6)
System suitability Standard solution: 7 g/mL of USP Calcifediol RS in
Sample: Standard solution Internal standard solution
Suitability requirements Sample solution: 7 g/mL of calcifediol from Capsules in
Resolution factor: NLT 3.0 Internal standard solution
Relative standard deviation: NMT 3.0% (four replicate [NOTEUsing a suitable implement, shear open a number
injections) of Capsules inside the container. Wash the implement
Analysis with a volume of Internal standard solution that will yield
Samples: Standard solution and Sample solution the final concentration. Collect the rinsings in the
Calculate the percent of C27H44O2 H2O taken: container, and mix to obtain a homogeneous solution of
the Capsule contents.]
Result = (RU/RS) (CS/CU) 100 Chromatographic system
RU = peak response ratio of calcifediol to testosterone Mode: LC
from the Sample solution Detector: UV 254 nm and a pump capable of providing
RS = peak response ratio of calcifediol to testosterone constant flow of greater than 2000 psi
from the Standard solution Column: 4-mm 30-cm; packing L3
CS = concentration of USP Calcifediol RS in the Injection size: Equal volumes
Standard solution (g/mL) System suitability
CU = concentration of Calcifediol in the Sample Sample: Standard solution
solution (g/mL) Suitability requirements
Acceptance criteria: 97.0%103.0% Resolution factor: NLT 3.0
Relative standard deviation: NMT 3.0% (four replicate
SPECIFIC TESTS injections)
WATER DETERMINATION, Method Ia 921: 3.8%5.0%, Analysis
determined on a 0.2 g specimen Samples: Standard solution and Sample solution
ADDITIONAL REQUIREMENTS Calculate the percent label claim of C27H44O2 H2O taken:
containers at controlled room temperature. Result = (RU/RS) (CS/CU) 100
USP REFERENCE STANDARDS 11 RU = peak response ratio of calcifediol to testosterone
USP Calcifediol RS of the Sample solution
RS = peak response ratio of calcifediol to testosterone
of the Standard solution
Calcifediol Capsules CS = concentration of USP Calcifediol RS in the
(Comment on this Monograph)id=m11293=Calcifediol CU = concentration of calcifediol in the Sample
Capsules=Ca-Chl-Monos.pdf) solution (g/mL)
DEFINITION Acceptance criteria: 90.0%120.0%
Calcifediol Capsules contain NLT 90.0% and NMT 120.0% of PERFORMANCE TESTS
the labeled amount of C27H44O2 H2O. DISSOLUTION 711
IDENTIFICATION Medium: Water; 500 mL
Standard solution: 150 g/mL of USP Calcifediol RS in Time: 15 min
chloroform Analysis: Place 1 Capsule in each vessel, and allow the
Sample solution: Equivalent to 150 g of calcifediol from Capsule to sink to the bottom of the vessel before starting
Capsules, in 1 mL of methanol, and shake vigorously for 1 rotation of the blade. Observe the Capsules, and record the
min. Separate the layers by centrifugation, and transfer as time taken for each capsule shell to rupture.
much of the top, methanol layer as possible to a second Tolerances: Meets the requirements if all of the Capsules
container. Evaporate this extract to dryness, and dissolve the tested rupture in NMT 15 min. If 1 or 2 of the Capsules
residue in 1 mL of chloroform. rupture in more than 15 but NMT 30 min, repeat the test
Analysis: Proceed as directed under Thin-Layer on 12 additional Capsules. NMT 2 of the total of 18
Chromatographic Identification Test 201. Capsules tested rupture in more than 15 but NMT 30 min.
Application volume: 20 L UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Developing solvent system: Cyclohexane and ethyl acetate ADDITIONAL REQUIREMENTS
(3:2) PACKAGING AND STORAGE: Preserve in tight, light-resistant
ASSAY containers.
Internal standard solution: 35 g/mL of testosterone in USP Calcifediol RS
ethyl acetate
8 Calcitriol / Official Monographs USP 32
Calcitriol Column efficiency: NLT 10,000 theoretical plates

(Comment on this Monograph)id=m11360=Calcitriol=Ca-Chl- Relative standard deviation: NMT 1.0%
Monos.pdf) Analysis
Calcitriol taken:
rU = sum of the peak responses of calcitriol and pre-
calcitriol from the Sample solution
C27 H44 O3 416.64 rS = sum of the peak responses of calcitriol and pre-
9,10-Secocholesta-5,7,10(19)-triene-1,3,25-triol, (1,3,5Z,7E)-; calcitriol from the Standard solution
(5Z,7E)-9,10-Secocholesta-5,7,10(19)-triene-1,3,25-triol CS = concentration of USP Calcitriol RS in the
[32222-06-3]. Standard solution (g/mL)
Monohydrate 434.65 CU = concentration of Calcitriol in the Sample solution
[77326-95-5]. (g/mL)
Acceptance criteria: 97.0%103.0% (anhydrous form,
DEFINITION calculated on the solvent-free basis); 97.0%103.0%
Calcitriol is anhydrous or contains one molecule of hydration. (monohydrate form, calculated on the anhydrous basis)
The anhydrous form contains NLT 97.0% and NMT 103.0% of
C27 H44 O3, calculated on the solvent-free basis. The IMPURITIES
monohydrate form contains NLT 97.0% and NMT 103.0% of Organic Impurities
C27H44O3, calculated on the anhydrous basis. PROCEDURE
[CAUTIONCare should be taken to prevent inhaling particles of [NOTECarry out the procedure as rapidly as possible,
calcitriol, and exposing the skin to it.] avoiding unnecessary exposure of solutions to light and
air.]
IDENTIFICATION Solution A, Mobile phase, System suitability solution,
A. INFRARED ABSORPTION 197K Sample solution, and Chromatographic system:
B. The retention time of the major peak of the Sample Proceed as directed in the Assay.
solution corresponds to that of the Standard solution, as Analysis
obtained in the Assay. Sample: Sample solution [NOTERecord chromatograms
for at least two times the retention time of the calcitriol
ASSAY peak.]
PROCEDURE Calculate the percentage of any individual impurity in the
[NOTECarry out the procedure as rapidly as possible, portion of Calcitriol taken:
avoiding unnecessary exposure of solutions to light and
air.] Result = (ri/rs) 100
Solution A: 1.0 mg/mL of
tris(hydroxymethyl)aminomethane in water, adjusted with ri = peak response of any individual peak other than
phosphoric acid to a pH of 7.07.5 the main calcitriol peak and the pre-calcitriol
Mobile phase: Acetonitrile and Solution A (11:9) peak
Standard solution: 100 g/mL of calcitriol from USP rs = sum of the responses of all the peaks
Calcitriol RS, dissolve first in acetonitrile (without heating), Acceptance criteria
using 55% of the final volume, then dilute with Solution A to Individual impurities: See Impurity Table 1.
volume Total impurities: NMT 1.0%
[NOTEAllow the solution to warm up to room [NOTEDisregard any peak less than 0.1%.]
temperature before diluting with Solution A to final
volume.] Impurity Table 1
System suitability solution: Heat 2.0 mL of the Standard
solution at 80 for 30 min. Relative Relative Acceptance
Sample solution: 100 g/mL of Calcitriol, dissolve first in Retention Response Criteria,
acetonitrile (without heating), using 55% of the final Name Time Factor NMT (%)
volume, then dilute with Solution A to volume Triazoline adduct of
0.43 0.1
[NOTEAllow the solution to warm up to room pre-calcitriol
temperature before diluting with Solution A to final trans-Calcitriola 0.96 0.25
volume.]
Chromatographic system 1-Calcitriolb 1.15 0.1
(See Chromatography 621, System Suitability.) Methylene calcitriolc 1.5 0.25
Mode: LC Any other individual
Detector: UV 230 nm 0.1
unidentified impurity
a(5E,7E)-9,10-secocholesta-5,7,10(19)-triene-1,3,25-triol
Flow rate: 1 mL/min
b(5Z,7E)-9,10-secocholesta-5,7,10(19)-triene-1,3,25-triol
c(5Z,7E)-1,3-dihydroxy-17-((R)-7-hydroxy-7-methyloctan-2-yl)-9,10-
System suitability secoandrosta-5,7,10(19)-triene
Samples: Standard solution and System suitability solution. SPECIFIC TESTS
[NOTEThe relative retention times for pre-calcitriol and WATER DETERMINATION, Method Ic 921: Where it is labeled
calcitriol are 0.9 and 1.0, respectively.] as a monohydrate, 3.5%5.5%
Resolution: NLT 3.5 between the pre-calcitriol and
calcitriol peaks
USP 32 Official Monographs / Calcitonin 9
ADDITIONAL REQUIREMENTS PH 791: 5.98.0, determined on a portion to which, if

PACKAGING AND STORAGE: Preserve in tight, light-resistant necessary, 0.30 mL of saturated potassium chloride solution
containers. Store as per labeling instructions. has been added for each 100 mL of Injection
LABELING: Where it is a monohydrate form, the label so OTHER REQUIREMENTS: It meets the requirements under
indicates. Injections 1.
USP Calcitriol RS ADDITIONAL REQUIREMENTS
preferably of Type I glass, protected from light. Store at
controlled room temperature.
Calcitriol Injection USP REFERENCE STANDARDS 11
(Comment on this Monograph)id=m11380=Calcitriol USP Calcitriol Solution RS
Injection=Ca-Chl-Monos.pdf) USP Endotoxin RS
DEFINITION
Calcitriol Injection is a sterile solution of Calcitriol. It contains an
amount of Calcitriol equivalent to NLT 90.0% and NMT Calcitonin Salmon
115.0% of the labeled amount of calcitriol (C27H44O3). It (Comment on this Monograph)id=m11340=Calcitonin
contains no antimicrobial agents. Salmon=Ca-Chl-Monos.pdf)
IDENTIFICATION
corresponds to that of the Standard solution, as obtained in
the Assay.
ASSAY
PROCEDURE
[NOTEAvoid unnecessary exposure of solutions to light or C145H240N44O48S2 3432 daltons
air.] [47931-85-1].
Mobile phase: Methanol and water (37:13). Make
adjustments if necessary so that the retention time of DEFINITION
calcitriol is NLT 20 min. Calcitonin Salmon is a polypeptide that has the same sequence
Standard solution: 3.0 mL of USP Calcitriol Solution RS, as that of the hormone that regulates calcium metabolism and
equilibrated to room temperature, to a container, and add is secreted by the ultimobranchial gland of salmon. It is
3.0 mL of water produced from either synthetic processes or microbial
Sample solution: Equivalent to 3 g of calcitriol from a processes using recombinant DNA (rDNA) technology. The
volume of Injection, in a sufficient amount of water to dilute host cell-derived protein content and the host cell- or vector-
to a total volume of 3.0 mL, and add 3.0 mL of methanol derived DNA content of Calcitonin Salmon produced from an
Chromatographic system rDNA process are determined by validated methods. It
(See Chromatography 621, System Suitability.) contains NLT 90.0% and NMT 105.0% of calcitonin salmon,
Mode: LC calculated on an acetic acid-free and dried basis.
Detector: UV 264 nm [NOTEOne mg of acetic acid-free, anhydrous Calcitonin
Column: 4.6-mm 7.5-cm; 3-m packing L1 Salmon is equivalent to 6000 USP Calcitonin Salmon Units.]
Guard column: 4.6-mm 4.5-cm; 5-m packing L1
Injection size: 100 L The retention time of the major peak of the Sample solution
System suitability corresponds to that of the Standard solution, as obtained in
Sample: Standard solution the Assay
Analysis PROCEDURE
Samples: Standard solution and Sample solution Solution A: Dissolve 3.26 g of tetramethylammonium
Calculate the percentage of C27H44O3 in the portion of hydroxide pentahydrate in 900 mL of water, add 100 mL of
Injection taken: acetonitrile, and mix. Adjust with phosphoric acid to a pH of
2.5.
Result = (rU/rS) (CS/CU) 100 Solution B: Dissolve 1.45 g of tetramethylammonium
hydroxide pentahydrate in 400 mL of water, add 600 mL of
rU = peak response of the Sample solution acetonitrile, and mix. Adjust with phosphoric acid to a pH of
rS = peak response of the Standard solution 2.5.
CS = concentration of calcitriol in the USP Calcitriol Mobile phase: See gradient table below.
Solution RS (g/mL)
CU = nominal concentration of calcitriol in the Sample Time Solution A Solution B
solution (g/mL) (min) (%) (%)
Acceptance criteria: 90.0%115.0% 0 72 28
SPECIFIC TESTS 30 48 52
BACTERIAL ENDOTOXINS TEST 85: NMT 100 USP Endotoxin 32 72 28
Units/g of calcitriol
PARTICULATE MATTER IN INJECTIONS 788: Meets the 55 72 28
10 Calcitonin / Official Monographs USP 32
Standard solution: 1.0 mg of USP Calcitonin Salmon RS in rU = peak area response of each impurity in the
Solution A Sample solution
System suitability solution: Dissolve the contents of a vial rS = sum of all areas in the Sample solution
of USP Calcitonin Salmon Related Compound A RS in 0.4 Acceptance criteria:
mL of Solution A, add 0.1 mL of the Standard solution, and Individual impurities: NMT 3.0%
mix. Total impurities: NMT 5.0%
Sample solution: 1.0 mg/mL of Calcitonin Salmon in PROCEDURE 2
Solution A [NOTEThis test needs to be performed only on material
Chromatographic system produced using rDNA technology.]
(See Chromatography 621, System Suitability.) Buffer A: 2.72 mg/mL of monobasic potassium phosphate
Mode: LC in water
Detector: UV 220 nm Buffer B: 2.72 mg/mL of monobasic potassium phosphate
Column: 4.6-mm 25-cm; packing L1 and 29.2 mg/mL of sodium chloride in water
Column temperature: 65 Buffer C: 4.8 mg/mL of citric acid in water. Adjust with 1
Flow rate: 1 mL/min M sodium hydroxide to a pH of 3.0, prior to final dilution
Injection size: 20 L Solution A: Acetonitrile and Buffer A (3:17). Adjust with
System suitability 45% w/w potassium hydroxide to a pH of 5.0
Sample: System suitability solution Solution B: Acetonitrile and Buffer B (3:17). Adjust with
[NOTEThe relative retention times for calcitonin salmon 45% w/w potassium hydroxide to a pH of 4.6
and calcitonin salmon related compound A are 1.0 and Mobile phase: See gradient table below.
1.15, respectively.]
Suitability requirements Time Solution A Solution B
Resolution: NLT 3 between calcitonin salmon and (min) (%) (%)
calcitonin salmon related compound A
Tailing factor: NMT 2.5 for calcitonin salmon 0 100 0
Relative standard deviation: NMT 3% 10 0 100
Analysis 15 0 100
Calculate the percentage of C145H240N44O48S2 in the portion 15.1 100 0
of Calcitonin Salmon taken: 22.1 100 0
Result = (rU/rS) (CS/CU) 100 System suitability solution: Prepare a solution containing 1
mg/mL of USP Calcitonin Salmon RS. Combine equal
rU = peak response of Calcitonin Salmon from the volumes of this solution with USP Calcitonin Salmon Related
Sample solution Compound B RS. To 1 mL of this mixture, add 100 L of
rS = peak response of calcitonin salmon from the Buffer C.
Standard solution Sample solution: 0.5 mg/mL of Calcitonin Salmon. To 1
CS = concentration of USP Calcitonin Salmon RS in mL of this solution, add 100 mL of Buffer C.
the Standard solution (mg/mL) Chromatographic system
CU = concentration of the Sample solution (mg/mL) (See Chromatography 621, System Suitability.)
Detector: UV 214 nm
OTHER COMPONENTS Column: 4.6-mm 20-cm; packing L9
ACETIC ACID CONTENT 503 Flow rate: 1.2 mL/min
Sample solution: 1 mg/mL of Calcitonin Salmon in Diluent Injection size: 50 L
(see Acetic acid in peptides, Diluent) System suitability
Acceptance criteria: 4%20% Sample: System suitability solution
IMPURITIES [NOTEThe relative retention times for [1,7-bis(3-sulpho-l-
Inorganic Impurities alanine)]calcitonin salmon-glycine, [1,7-bis(3-sulpho-l-
HEAVY METALS, Method II 231: NMT 50 ppm alanine)]calcitonin salmon, and calcitonin salmon related
Organic Impurities compound B (calcitonin salmon-glycine) are 0.4, 0.6, and
PROCEDURE 1 0.9, respectively; and the retention time for calcitonin
[NOTEThis test is performed on material produced by both salmon is about 9 min.]
chemical processes and recombinant DNA processes.] Suitability requirements
Solution A, Solution B, Mobile phase, System suitability Resolution: NLT 3.0 between calcitonin salmon and
solution, Chromatographic system, and System calcitonin salmon related compound B
suitability: Proceed as directed in the Assay. Analysis
Sample solution: Prepare as directed for the Sample Sample: Sample solution
solution under Assay. Calculate the percentage of each impurity in the portion of
Analysis Calcitonin Salmon taken:
Calculate the percentage of each impurity in the portion Result = (rU/rS) 100
of Calcitonin Salmon taken: rU = peak response for each impurity
Result = (rU/rS) 100 rS = sum of the responses of all peaks
Acceptance criteria 0.91.1; lysine, 1.82.2; serine, 3.24.2; threonine, 4.25.2;

Individual impurities: See Impurity Table. tyrosine, 0.71.1; half cystine, 1.42.1.
PEPTIDE MAPPING
Relative Retention (See Biotechnology-Derived ArticlesPeptide Mapping 1055.)
Name Time Limit NMT (%) [NOTEThis test needs to be performed only on material
produced using rDNA technology.]
Calcitonin Salmon 0.9 0.6 Solution A: Trifluoroacetic acid and water (1:1000)
Related Solution B: Acetonitrile, trifluoroacetic acid, and water
Compound B (800:0.85:200)
[1, 7 bis(3-sulpho- 0.4 0.2 Mobile phase: See gradient table below.
L-alanine)]
Calcitonin Time Solution A Solution B
Salmon-glycine (min) (%) (%)
[1, 7 bis(3-sulpho- 0.6 0.2 0 100 0
L-alanine)]
Calcitonin 50 65 35
Salmon 60 40 60
Any other impurity 0.1 60.1 0 100
65.1 0 100
SPECIFIC TESTS 65.2 100 0
AMINO ACID PROFILE 80.2 100 0
(See Biotechnology-Derived ArticlesAmino Acid Analysis
1052.) Trypsin solution: Freshly prepare a solution containing 0.1
[NOTEThis test is performed only on material of synthetic mg/mL of trypsin (previously treated with L- 1-tosylamido-2-
origin. The concentration of amino acids in the Internal phenylethyl chloromethyl ketone (TPCK) to remove
standard solution, the Standard stock solution, and the chymotrypsin activity).
Standard solution and the amount of material used to Tris buffer: 1 M tris(hydroxymethyl) aminomethane, 10
prepare the Sample solution can be adjusted depending on mM calcium chloride, adjust with hydrochloric acid to a pH
the method used for amino acid analysis. The of 8.0
concentrations given are based on analysis using Method Stopping solution: Water and trifluoroacetic acid (1:1)
1.] Standard solution: 1.0 mg/mL of USP Calcitonin Salmon
Internal standard solution: 1 mM solution of - RS. Transfer 1 mL of this solution to a clean vial. Add 100
aminobutyric acid mL of Tris buffer and 50 mL of Trypsin solution, mix, and
Standard stock solution: 2.5 mM ammonia and the L form incubate at 2 to 8 for 1620 h. Quench the digestion by
of lysine, histidine, arginine, aspartic acid, threonine, serine, adding 10 mL of Stopping solution.
proline, valine, glutamic acid, glycine, leucine, tyrosine, and Sample solution: 1.0 mg/mL of Calcitonin Salmon. Transfer
L-cystine, in 0.1 M hydrochloric acid 1 mL of this solution to a clean vial. Add 100 mL of Tris
Standard solution: Transfer 5 mL of the Internal standard buffer and 50 mL of Trypsin solution, mix, and incubate at 2
solution and 2 mL of the Standard stock solution into a 50- to 8 for 1620 h. Quench the digestion by adding 10 mL
mL volumetric flask, and dilute with 0.1 M hydrochloric acid of Stopping solution.
to volume. Chromatographic system
Sample solution: Place 1.5 mg of Calcitonin Salmon into a (See Chromatography, 621 System Suitability.)
heavy-wall ignition tube, add 1.0 mL of 6 N hydrochloric Mode: LC
acid, allow to cool, immerse the lower half of the tube in a Detector: UV 214 nm
freezing mixture until the contents are frozen, evacuate to Column: 4.6-mm 25-cm; packing L1
approximately 10 mm of Hg, purge with nitrogen (repeat Flow rate: 1.2 mL/min
the evacuation and nitrogen purge three times), and seal Injection size: 20 L
the tube while it is under a 10 mm of Hg vacuum. Heat for Analysis
16 h at 110 to 115 in an air oven. Cool, open the tube, Samples: Standard solution and Sample solution
dry in a vacuum desiccator, remove the contents, and allow [NOTECondition the chromatographic system by running
to cool to room temperature. Dissolve in 0.1 M hydrochloric a blank gradient program prior to injecting the digests.]
acid, transfer to a 10-mL volumetric flask, add 1 mL of Acceptance criteria: The chromatographic profile of the
Internal standard solution, and dilute with 0.1 M hydrochloric Sample solution is similar to that of the Standard solution
acid to volume. BIOIDENTITY
Analysis RPMI 1640 with L-glutamine: Prepare a mixture of the
Samples: Standard solution and Sample solution ingredients, in the quantities shown below, in sufficient
Standardize the amino acid analyzer, using the Standard water to obtain 1 L of RPMI 1640 with L-glutamine solution,
solution. Inject the Sample solution into the amino acid and sterilize by filtration.
analyzer, and determine the relative proportion of amino
acids.
Calcium Nitrate 100.00 mg
Express the content of each amino acid in moles, using an
internal standard calibration technique. Calculate the Potassium Chloride 400.00 mg
relative proportions of the amino acids by taking as Magnesium Sulfate, Anhydrous 48.84 mg
equivalent to 1 the sum of the number of moles of
Potassium Chloride 400 mg
aspartic acid, glutamic acid, proline, glycine, valine,
leucine, histidine, arginine, and lysine divided by 20. Sodium Chloride 6000 mg
Acceptance criteria: The requirements are met if the values Sodium Phosphate, Dibasic, Anhydrous 800 mg
fall within the following limits: aspartic acid, 1.82.2;
Sodium Bicarbonate 2000 mg
glutamic acid, 2.73.3; proline, 1.72.3; glycine, 2.73.3;
valine, 0.91.1; leucine, 4.55.3; histidine, 0.91.1; arginine,
Glycine 10 mg Positive control solution: 1 ng/mL of USP Calcitonin

L-Arginine 200 mg Salmon RS, from Standard stock solution in Medium B
Negative control solution: Medium B
L-Asparagine 50 mg Standard solution A: 0.1 ng of USP Calcitonin Salmon RS,
L-Aspartic Acid 20 mg from Standard stock solution in Medium B
L-Cystine Dihydrochloride 65 mg Standard solution B: 33 pg/mL USP Calcitonin Salmon RS,
from Standard solution A in Medium B
L-Glutamic Acid 20 mg Standard solution C: 11 pg/mL of USP Calcitonin Salmon
L-Glutamine 300 mg RS, from Standard solution B in Medium B
L-Histidine 15 mg Standard solution D: 3.7 pg/mL USP Calcitonin Salmon RS,
from Standard solution C in Medium B
L-Hydroxyproline 20 mg Sample stock solution: 20 g/mL of Calcitonin Salmon in
L-Isoleucine 50 mg Solution B
L-Leucine 50 mg Sample solution A: 0.1 ng/mL of Calcitonin Salmon, from
Sample stock solution in Medium B
L-Lysine Hydrochloride 40 mg Sample solution B: 33 pg/mL of Calcitonin Salmon, from
L-Methionine 15 mg Sample solution A in Medium B
L-Phenylalanine 15 mg Sample solution C: 11 pg/mL of Calcitonin Salmon, from
Sample solution B in Medium B
L-Proline 20 mg Sample solution D: 3.7 pg/mL of Calcitonin Salmon, from
L-Serine 30 mg Sample solution C in Medium B
L-Threonine 20 mg Cell culture preparation: Prepare a cell culture of the
human mammary tumor cell line T-47D. Cells are
L-Tryptophan 5 mg propagated using Medium A at 37 and 5% carbon dioxide.
L-Tyrosine Disodium Salt Dihydrate 29 mg The medium is changed every 2 days, and cells are
L-Valine 20 mg passaged every 5 to 9 days, using TrypsinEDTA solution with
a 1:4 subculture.
Biotin 0.2 mg Cell suspension: For the test, use a cell culture that is 59
Choline Chloride 3 mg days old. Remove the cell culture medium from the flask by
D-Calcium Pantothenate 0.25 mg aspiration, add 10 mL of Solution D, and rock the culture
flask to rinse the entire monolayer. Remove the liquid by
Folic Acid 1 mg aspiration, add 2 mL of Solution C, spread over the entire
i-Inositol 35 mg monolayer, allow to stand for 35 min, and add 10 mL of
Niacinamide 1 mg Medium A. Homogenize the cell suspension using a pipet,
transfer to a 15-mL polypropylene tube, centrifuge at about
para-Aminobenzoic Acid 1 mg 220 g for 5 min, pour off the supernatant, and resuspend
Pyridoxine Hydrochloride 1 mg the cell pellet in 10 mL of Medium A. Count the cells, and
Riboflavin 0.2 mg adjust the cell density through dilution, using Medium A, to
2.5 104 cells/mL.
Thiamine Hydrochloride 1 mg Analysis: Place 200 L of the Cell suspension into each well
Vitamin B12 0.005 mg of a 96-well culture plate (the tissue culture plate), and
incubate for 1824 h at 37 and 5% carbon dioxide. Fill
Medium A (growth medium): Using aseptic technique, each well of an empty round-bottomed 96-well culture plate
prepare the following tissue culture medium. (the prepared plate) with 150 L of one of the following
solutions: Positive control solution, Negative control solution,
RPMI 1640 with L-glutamine 500 mL Standard solutions AD, and Sample solutions AD, so that
each solution fills at least five wells on the prepared plate.
Fetal bovine serum 50 mL After incubation, remove the culture medium from the tissue
1 M HEPES 5 mL culture plate. Using an 8-channel or 12-channel pipet,
Penicillin/streptomycin solution (10,000 IU per 5 mL rapidly transfer 100 L of solution from each well of the
mL/10 mg per mL) prepared plate to each well of the tissue culture plate.
Incubate for 15 min at ambient temperature, remove the
Human insulin 10 IU solution from each well, stop stimulation by immediately
Hydrocortisone 0.5 mg adding an appropriate cell-lysis buffer, and quantitate cAMP
produced within the cells, using a validated kit. Perform the
Medium B (stimulation medium): Dissolve 5 g of albumin test three times, using three different 96-well culture plates.
bovine serum (BSA) in 500 mL of 2 mM RPMI 1640 with L- [NOTESome kits include a cell-lysis reagent and a
glutamine. sequestering agent for the cell-lysis reagent. The range of
Solution A: Dissolve 500 mg of albumin bovine serum in the test kit is between 0.05 ng and 10 ng/mL of cAMP. The
25 mL of water. [NOTEUse within 1 day.] number of cells used in the assay may vary depending on
Solution B: Add 25 mL of 0.1 M formic acid and 5 mL of the validated kit used to quantitate cAMP.] Potency is
Solution A to a 50-mL volumetric flask, dilute with water to determined by a 3-dose, 6-point parallel-line assay, using
volume, and mix. [NOTEUse within 2 days.] standard statistical methods. The calculation is carried out
Solution C: Prepare a sterile filtered solution containing using both the lower three concentrations and the upper
0.25% trypsin and 0.53 mM EDTA. three concentrations. For the assay to be valid, the
Solution D: Dissolve 8 g of sodium chloride, 1.15 g of requirements for regression and parallelism must be met. If
dibasic sodium phosphate, 0.2 g of monobasic potassium the requirements for validity are met to the same extent in
phosphate, 0.2 g of potassium chloride, 0.1 g of calcium both assessments (the lower and the higher assessments),
chloride, and 0.1 g of magnesium chloride in 1 L of water. the final result is determined from the concentration range
Standard stock solution: 20 g of USP Calcitonin Salmon that shows the higher value when the common slope is
RS in Solution B. divided by the mean square error.
Acceptance criteria: The potency levels determined from 0.4 mL of Solution A, and add 0.1 mL of the Standard
all three performances of the test are homogeneous, and solution.
the confidence limits for all three determinations are System suitability solution: System suitability stock solution
between 64% and 156% of the calculated potency. and Solution A (1:9)
Sample solution: Use the Injection.
Change to read: (See Chromatography 621, System Suitability.)
Mode: LC
MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED Detector: UV 220 nm
MICROORGANISMS 62 Column: 4.6-mm 25-cm; packing L1
Sample: 0.2 g Temperature: 65
Acceptance criteria: The total aerobic microbial count Flow rate: 1 mL/min
NMT 100 cfu/g and the total combined molds and yeasts Injection size: 200 L
is NMT 100 cfu/gUSP32 System suitability
Samples: Standard solution and System suitability solution
WATER DETERMINATION, Method Ic 921: NMT 10% [NOTEThe relative retention times for calcitonin salmon
ADDITIONAL REQUIREMENTS and calcitonin salmon related compound A are 1.0 and
PACKAGING AND STORAGE: Preserve in tight containers. Store 1.15, respectively.]
in a refrigerator or maintain in a frozen state, protected from Suitability requirements
light. Resolution: NLT 3 between calcitonin salmon and
LABELING: The labeling states that the material is synthetic calcitonin salmon related compound A
or of recombinant DNA origin. Tailing factor: NMT 2.5
USP REFERENCE STANDARD 11 Relative standard deviation: NMT 3.0%
USP Calcitonin Salmon RS Analysis
USP Calcitonin Salmon Related Compound A RS (N-acetyl- Samples: Standard solution and Sample solution
cys1 -calcitonin salmon) Calculate the potency, in USP Calcitonin Salmon Units/mL,
USP Calcitonin Salmon Related Compound B RS (calcitonin in the portion of Injection taken:
salmon-glycine)
rU = peak area response of the Sample solution
Calcitonin Salmon Injection rS = peak area response of the Standard solution
CS = concentration of USP Calcitonin Salmon RS in
(Comment on this Monograph)id=m11350=Calcitonin Salmon the Standard solution (mg/mL)
Injection=Ca-Chl-Monos.pdf) CU = nominal concentration of calcitonin salmon in
DEFINITION the Sample solution (mg/mL)
Calcitonin Salmon Injection is a sterile solution of Calcitonin Acceptance criteria: 80.0%110.0%
Salmon in a suitable diluent. Each mL of Calcitonin Salmon SPECIFIC TESTS
Injection possesses an activity of NLT 80% and NMT 110% of BACTERIAL ENDOTOXINS TEST 85: NMT 0.625 USP
that stated on the label. Endotoxin Unit/USP Calcitonin Salmon Unit
IDENTIFICATION STERILITY TESTS 71: Meets the requirements when tested as
The retention time of the major peak of the Sample solution directed under Test for Sterility of the Product to Be Examined,
corresponds to that of the Standard solution, as obtained in Membrane Filtration
the Assay. PARTICULATE MATTER IN INJECTIONS 788: Meets the
ASSAY PH 791: 3.94.5
PROCEDURE INJECTIONS 1: Meets the requirements
Solution A: 3.26 mg/mL of tetramethylammonium
hydroxide pentahydrate in water and acetonitrile (9:1). ADDITIONAL REQUIREMENTS
Adjust with phosphoric acid to a pH of 2.5. PACKAGING AND STORAGE: Preserve in single-dose or
Solution B: 1.45 mg/mL of tetramethylammonium multiple-dose containers, preferably of Type I glass. Avoid
hydroxide pentahydrate in water and acetonitrile (2:3). freezing. Store in a refrigerator.
Adjust with phosphoric acid to a pH of 2.5. LABELING: Label it to indicate the activity in USP Calcitonin
Mobile phase: See the gradient table below. Salmon Units per mL. The labeling states that the material is
synthetic. Label it to state that it is to be stored in a
refrigerator, and that freezing is to be avoided.
Time Solution A Solution B USP REFERENCE STANDARDS 11
(min) (%) (%) USP Calcitonin Salmon RS
0 72 28 USP Calcitonin Salmon Related Compound A RS
30 48 52 USP Endotoxin RS
32 72 28
55 72 28
Calcitonin Salmon Nasal Solution
Standard stock solution: 1.0 mg/mL of USP Calcitonin (Comment on this Monograph)id=m11342=Calcitonin Salmon
Salmon RS in Solution A Nasal Solution=Ca-Chl-Monos.pdf)
Standard solution: 0.1 mg/mL of USP Calcitonin Salmon
RS from Standard stock solution in Solution A DEFINITION
System suitability stock solution: Dissolve the contents of Calcitonin Salmon Nasal Solution is a solution of Calcitonin
a vial of USP Calcitonin Salmon Related Compound A RS in Salmon in a suitable diluent. It contains suitable preservatives,
and is packaged in a form suitable for nasal administration so CU = nominal concentration of calcitonin salmon in
that the required dosage can be controlled as required. Each the Sample solution (mg/mL)
mL of Calcitonin Salmon Nasal Solution possesses an activity of Acceptance criteria: 80.0%110.0%
NLT 80% and NMT 110% of that stated on the label.
SPECIFIC TESTS
IDENTIFICATION MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED
The retention time of the major peak of the Sample solution MICROORGANISMS 62: Total aerobic microbial count: NMT
corresponds to that of the Standard solution, as obtained in 100 cfu/g Total combined molds and yeast count: NMT 50
the Assay. cfu/g.
It meets the requirements for absence of Staphylococcus
ASSAY aureus and Pseudomonas aeruginosa.
PROCEDURE PH 791: 3.54.5
Solution A: 3.26 mg/mL of tetramethylammonium
hydroxide pentahydrate in water and acetonitrile (9:1). ADDITIONAL REQUIREMENTS
Adjust with phosphoric acid to a pH of 2.5. PACKAGING AND STORAGE: Preserve in containers suitable for
Solution B: 1.45 mg/mL of tetramethylammonium spraying the contents into the nasal cavities in a controlled
hydroxide pentahydrate in acetonitrile and water (3:2). individualized dosage. Store unopened containers in a
Adjust with phosphoric acid to a pH of 2.5. refrigerator, and opened containers at room temperature.
Mobile phase: See the gradient table below. LABELING: Label it to indicate that it is for intranasal
administration only. The labeling also states that it has been
Time Solution A Solution B prepared either with Calcitonin Salmon of synthetic origin or
(min) (%) (%) Calcitonin Salmon of rDNA origin. Label it to state that it is
to be stored in a refrigerator, and that freezing is to be
0 72 28 avoided. Label it to indicate the activity in USP Calcitonin
30 48 52 Salmon Units/mL.
32 72 28 USP REFERENCE STANDARDS 11
USP Calcitonin Salmon RS
55 72 28 USP Calcitonin Salmon Related Compound A RS
Standard stock solution: 1.0 mg/mL of USP Calcitonin
Salmon RS in Solution A
Standard solution: 0.1 mg/mL of USP Calcitonin Salmon Calcium Acetate
RS from Standard stock solution in Solution A (Comment on this Monograph)id=m11400=Calcium
System suitability stock solution: Dissolve the contents of Acetate=Ca-Chl-Monos.pdf)
a vial of USP Calcitonin Salmon Related Compound A RS in
0.4 mL of Solution A, and add 0.1 mL of the Standard
solution.
System suitability solution: System suitability stock solution
and Solution A (1:9)
Diluent: 7.5 mg/mL of sodium chloride, 2 mg/mL of
sodium acetate, and 2 mg/mL of glacial acetic acid in water
Sample solution: Nasal Solution in Diluent (1:9)
Chromatographic system C4H6CaO4 158.17
(See Chromatography 621, System Suitability.) Acetic acid, calcium salt;
Mode: LC Calcium acetate [62-54-4].
Detector: UV 220 nm
Column: 4.6-mm 25-cm; packing L1 DEFINITION
Temperature: 65 Calcium Acetate contains NLT 99.0% and NMT 100.5% of
Flow rate: 1 mL/min C4H6CaO4, calculated on the anhydrous basis.
Samples: Standard solution and System suitability solution IDENTIFICATION TESTSGENERAL, Calcium, 191 AND Acetate,
[NOTEThe relative retention times for calcitonin salmon 191
and calcitonin salmon related compound A are 1.0 and Sample solution: 50 mg/mL
1.15, respectively.] Acceptance criteria: Meets the requirements
Resolution: NLT 3 between calcitonin salmon and PROCEDURE
calcitonin salmon related compound A Sample solution: 50 mg/mL
Tailing factor: NMT 2.5 Acceptance criteria: Meets the requirements
Relative standard deviation: NMT 3.0% Sample: 300 mg
Analysis Analysis: Dissolve the Sample in 150 mL of water
Samples: Standard solution and Sample solution containing 2 mL of 3 N hydrochloric acid. While stirring,
Calculate the potency, in USP Calcitonin Salmon Units/mL, add about 30 mL of 0.05 M edetate disodium VS from a 50-
in the portion of Nasal Solution taken: mL buret. Add 15 mL of 1 N sodium hydroxide and 300 mg
Result = (rU/rS) (CS/CU) 100 of hydroxy naphthol blue, and continue the titration to a
blue endpoint. Each mL of 0.05 M edetate disodium is
rU = peak area response of the Sample solution equivalent to 7.909 mg of C4H6CaO4.
rS = peak area response of the Standard solution Acceptance criteria: 99.0%100.5%, calculated on the
CS = concentration of USP Calcitonin Salmon RS in anhydrous basis
USP 32 Official Monographs / Calcium 15
IMPURITIES Dissolve about 100 mg of the treated wire in a mixture of

Inorganic Impurities 10 mL of hydrochloric acid and 2 mL of nitric acid by
LIMIT OF FLUORIDE heating at about 80 for approximately 30 min. Continue
[NOTEPrepare and store all solutions in plastic containers.] heating until the volume is reduced to about 4 mL. Cool to
Solution A: 294 mg/mL of sodium citrate dihydrate in room temperature, and add 4 mL of water. Evaporate to
water about 2 mL by heating. Cool, and transfer this solution,
Standard stock solution: 1.1052 mg/mL of USP Sodium with the aid of water, to a 100-mL volumetric flask, dilute
Fluoride RS with water to volume, and mix.
Standard solution: Combine 20.0 mL of Standard stock [NOTEEquivalent to 1 mg/mL of Aluminum.]
solution with 50.0 mL of Solution A and dilute to 100.0 mL Aluminum standard solution: 1.0 g/mL of Aluminum,
with water. from Aluminum standard stock solution in water.
Electrode system: Use a fluoride-specific, ion-indicating Standard solution: Prepare a solution containing 2.0 mL
electrode and a silversilver chloride reference electrode of Aluminum standard solution, 5 mL of Solution A, and 48
connected to a pH meter capable of measuring potentials mL of water, and extract this solution with successive
with a minimum reproducibility of 0.2 mV (see pH portions of 10, 10, and 5 mL of a 0.5% 8-hydroxyquinoline
791). in chloroform, combining the chloroform extracts in a 50-
Standard response line: Transfer 50.0 mL of Solution A mL volumetric flask. Dilute the combined extracts with
and 2.0 mL of hydrochloric acid to a beaker, and add chloroform to volume.
water to make 100 mL. Add a plastic-coated stirring bar, Sample solution: Dissolve 1.0 g of Calcium Acetate in 50
insert the electrodes into the solution, stir for 15 min, and mL of water and add 5 mL of Solution A. Extract this
read the potential, in mV. Continue stirring, and at 5-min solution with successive portions of 10, 10, and 5 mL of a
intervals add 100, 100, 300, and 500 L of Standard 0.5% 8-hydroxyquinoline in chloroform, combining the
solution, reading the potential 5 min after each addition. chloroform extracts in a 50-mL volumetric flask. Dilute the
Plot the logarithms of the cumulative fluoride ion combined extracts with chloroform to volume.
concentrations (0.1, 0.2, 0.5, and 1.0 g/mL) versus Blank: Prepare a solution containing 50 mL of water and 5
potential, in mV. mL of Solution A, and extract this solution with successive
Sample solution: Transfer 2.0 g of the Calcium Acetate to portions of 10, 10, and 5 mL of a 0.5% 8-hydroxyquinoline
a beaker containing a plastic-coated stirring bar, add 20.0 in chloroform, combining the chloroform extracts in a 50-
mL of water and 2.0 mL of hydrochloric acid, and stir until mL volumetric flask. Dilute the combined extracts with
dissolved. Add 50.0 mL of Solution A and sufficient water to chloroform to volume.
make 100 mL. Spectrometric conditions
Analysis: Rinse and dry the electrodes, insert them into Mode: fluorescence
the Sample solution, stir for 5 min, and read the potential, Excitation wavelength: 392 nm
in mV. Emission wavelength: 518 nm
From the measured potential and the Standard response line Analysis
determine the concentration, C, in g/mL, of fluoride ion Samples: Sample solution and Standard solution.
in the Sample solution. Acceptance criteria: The fluorescence of the Sample
Calculate the amount of fluoride (ppm) in the specimen solution is NMT that of the Standard solution (2 ppm).
taken by multiplying C by 0.005. LIMIT OF BARIUM: [NOTEUse where it is labeled as intended
Acceptance criteria: 50 ppm for use in hemodialysis or peritoneal dialysis.]
CHLORIDE AND SULFATE, Chloride 221: A 1.0-g portion Barium chloride solution: 0.758 mg/mL of anhydrous
shows no more chloride than corresponds to 0.70 mL of barium chloride [NOTEThis solution contains 500 g/mL
0.020 N hydrochloric acid (0.05%). of barium.]
CHLORIDE AND SULFATE, Sulfate 221: A 0.25-g portion Standard solution: To a fifth tube, add 1 g of ammonium
shows no more sulfate than corresponds to 0.15 mL of acetate, 2 mL of 1 N hydrochloric acid, 3.0 mL of Barium
0.020 N sulfuric acid (0.06%). chloride solution, and sufficient water to bring the volume
ARSENIC, Method I 211: NMT 3 ppm to 40 mL.
LEAD 251: NMT 10 ppm Sample stock solution: 250 mg/mL of Calcium Acetate
HEAVY METALS, Method I 231 and 25 mg/mL of ammonium acetate in 1 N hydrochloric
Sample solution: Dissolve 0.8 g of Calcium Acetate in acid. The pH of this solution is 4.55.5. [NOTEFilter and
20.0 mL of water, add 3.0 mL of glacial acetic acid, dilute cover the solution.]
with water to 25 mL, and adjust with glacial acetic acid to Sample solution: To four separate tubes, add 1.0, 1.5,
a pH of 3.84.0, measured with a pH meter. 2.0, and 2.5 mL of Barium chloride solution. To each tube,
Monitor preparation: Prepare as directed for Sample add a sufficient volume of the Sample stock solution to
solution, 2.0 mL of Standard lead solution being added. bring the volume to 40 mL.
Acceptance criteria: NMT 25 ppm Solution A: Ammonium sulfate solution (1 in 10)
LIMIT OF NITRATE Analysis: To the Sample solutions and the Standard solution
Sample solution: 100 mg/mL of Calcium Acetate in water add, with brisk stirring, 3.0 mL of Solution A, and allow to
Analysis: To 10 mL of the Sample solution, add 5 mg of stand for 20 min.
sodium chloride, 0.05 mL of indigo carmine TS, and, with Acceptance criteria: The Sample solutions containing 1.0
stirring, 10 mL of nitrogen-free sulfuric acid. and 1.5 mL of Barium chloride solution remain clear or are
Acceptance criteria: The blue color persists for NLT 10 only faintly turbid. The Sample solution containing 2.0 mL
min. of Barium chloride solution is not more turbid than the
LIMIT OF ALUMINUM [NOTEUse where it is labeled as Standard solution.
intended for parenteral use or for use in hemodialysis or LIMIT OF MAGNESIUM: [NOTEUse where it is labeled as
peritoneal dialysis.] intended for use in hemodialysis or peritoneal dialysis.]
Solution A: Dissolve 200 mg/mL of ammonium acetate in [NOTEThe Standard solution and the Sample solutions may
water, and adjust with glacial acetic acid to a pH of 6.0 be modified, if necessary, to obtain solutions of suitable
prior to final dilution. concentrations, adaptable to the linear or working range of
Aluminum standard stock solution: Treat some aluminum the instrument.]
wire with 6 N hydrochloric acid at 80 for a few min.
16 Calcium / Official Monographs USP 32
Standard stock solution A: 1.516 mg/mL of magnesium Acceptance criteria: NMT 0.05%
oxide in 1 N nitric acid [NOTEThis solution contains 1000 LIMIT OF SODIUM: [NOTEUse where it is labeled as intended
g/mL of magnesium.] for use in hemodialysis or peritoneal dialysis.]
Standard stock solution B: 5.0 g/mL of magnesium, [NOTEThe Standard solution and the Sample solutions may
from Standard stock solution be modified, if necessary, to obtain solutions of suitable
Standard solution A: Dilute 20.0 mL of Sample solution to concentrations, adaptable to the linear or working range of
25.0 mL with water the instrument.]
Standard solution B: Dilute 2.0 mL of Standard stock Standard stock solution A: 25.42 mg/mL of sodium
solution B and 20.0 mL of Sample solution to 25.0 mL with chloride, using sodium chloride previously dried at 105 for
water 2 h [NOTEThis solution contains 10.0 mg/mL of sodium.]
Standard solution C: Dilute 4.0 mL of Standard stock Standard stock solution B: 250 g/mL of sodium: from
solution B and 20.0 mL of Sample solution to 25.0 mL with Standard stock solution
water Standard solution A: Dilute 20.0 mL of Sample solution to
Sample solution: 2 mg/mL of Calcium Acetate 25.0 mL with water
Blank: water Standard solution B: Dilute 2.0 mL of Standard stock
Spectometric conditions solution B and 20.0 mL of Sample solution to 25.0 mL with
Mode: Atomic absorption spectroscopy water
Source: Air-acetylene flame Standard solution C: Dilute 4.0 mL of Standard stock
Detector magnesium hollow-cathode lamp solution B and 20.0 mL of Sample solution to 25.0 mL with
Analytical wavelength: 285.2 water
Analysis Sample solution: 10 mg/mL
Samples: Standard solution A, Standard solution B, and Blank: water
Standard solution C Spectometric conditions
Plot the absorbances of the Standard solutions versus their (see Spectrophotometry and Light-Scattering 851)
contents of magnesium, in g/mL, draw the straight line Mode: Atomic absorption spectroscopy
best fitting the three points, and extrapolate the line Source: air-acetylene flame
until it intercepts the concentration axis. From the Detector sodium hollow-cathode lamp
intercept determine the amount, in g, of magnesium in Analytical wavelength: 589.0 nm
each mL of the Sample solution. Analysis: Plot the absorbances of the treated Sample
Calculate the percentage of magnesium in the specimen solutions versus their contents of sodium, in g/mL, draw
by multiplying this value by 0.0625. the straight line best fitting the three points, and
Acceptance criteria: NMT 0.05% extrapolate the line until it intercepts the concentration
LIMIT OF POTASSIUM: [NOTEUse where it is labeled as axis. From the intercept determine the amount, in g, of
intended for use in hemodialysis or peritoneal dialysis.] sodium in each mL of the Sample solution.
[NOTEThe Standard solution and the Sample solutions may Calculate the percentage of sodium in the specimen by
be modified, if necessary, to obtain solutions of suitable multiplying this value by 0.0125.
concentrations, adaptable to the linear or working range of Acceptance criteria: NMT 0.5%
the instrument.] LIMIT OF STRONTIUM: [NOTEUse where it is labeled as
Standard stock solution A: 23.836 mg/mL of potassium intended for use in hemodialysis or peritoneal dialysis.]
chloride, using potassium chloride previously dried at 105 [NOTEThe Standard solution and the Sample solutions may
for 2 h [NOTEThis solution contains 12.5 mg/mL of be modified, if necessary, to obtain solutions of suitable
potassium.] concentrations, adaptable to the linear or working range of
Standard stock solution B: 31.25 g/mL of potassium, the instrument.]
from Standard stock solution Standard stock solution A: 2.45 mg/mL of strontium
Standard solution A: Dilute 20.0 mL of Sample solution to acetate in water. [NOTEThis solution contains 1000
25.0 mL with water g/mL of strontium.]
Standard solution B: Dilute 2.0 mL of Standard stock Standard stock solution B: 50.0 g/mL of strontium, from
solution B and 20.0 mL of Sample solution to 25.0 mL with Standard stock solution in water
water Standard solution A: Dilute 20.0 mL of Sample solution to
Standard solution C: Dilute 4.0 mL of Standard stock 25.0 mL with water
solution B and 20.0 mL of Sample solution to 25.0 mL with Standard solution B: Dilute 2.0 mL of Standard stock
water solution B and 20.0 mL of Sample solution to 25.0 mL with
Sample solution: 12.5 mg/mL water
Blank: water Standard solution C: Dilute 4.0 mL of Standard stock
Spectometric conditions solution B and 20.0 mL of Sample solution to 25.0 mL with
(see Spectrophotometry and Light-Scattering 851) water
Mode: Atomic absorption spectroscopy Sample solution: 20 mg/mL
Source: Air-acetylene flame Blank: water
Detector potassium hollow-cathode lamp Spectometric conditions
Analytical wavelength: 766.7 nm (see Spectrophotometry and Light-Scattering 851)
Analysis Mode: Atomic absorption spectroscopy
Samples: Standard solution A, Standard solution B, and Source: Nitrous oxide-acetylene
Standard Solution C. Detector strontium hollow-cathode lamp
Plot the absorbances of the Standard solutions versus their Analytical wavelength: 460.7 nm
contents of potassium, in g/mL, draw the straight line Analysis: Plot the absorbances of the treated Sample
best fitting the three points, and extrapolate the line solutions versus their contents of strontium, in g/mL, draw
until it intercepts the concentration axis. From the the straight line best fitting the three points, and
intercept determine the amount, in g, of potassium in extrapolate the line until it intercepts the concentration
each mL of the Sample solution. axis. From the intercept determine the amount, in g, of
Calculate the percentage of potassium in the specimen by strontium in each mL of the Sample solution.
multiplying this value by 0.01.
Calculate the percentage of strontium in the specimen by Sample solution: Sample per Dissolution 711. Dilute with
multiplying this value by 0.00625. Medium to a concentration that is similar to Standard
Acceptance criteria: NMT 0.05% solution.
Organic Impurities Acceptance criteria: NLT 80% (Q) of the labeled amount
PROCEDURE: READILY OXIDIZABLE SUBSTANCES of C4H6CaO4 is dissolved.
Sample solution: 20 mg/mL of Calcium acetate in boiling UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
water
Analysis: Add a few glass beads to 100 mL of the Sample IMPURITIES
solution, 6 mL of 10 N sulfuric acid, and 0.3 mL of 1 N Inorganic Impurities
potassium permanganate, mix, boil gently for 5 min, and LIMIT OF ALUMINUM
allow the precipitate to settle. [NOTEUse where it is labeled as intended for parenteral use
Acceptance criteria: The pink color in the supernatant is or for use in hemodialysis or peritoneal dialysis.]
not completely discharged. Solution A: Dissolve 200 mg/mL of ammonium acetate in
water, and adjust with glacial acetic acid to a pH of 6.0
SPECIFIC TESTS prior to final dilution.
PH 791 Aluminum standard stock solution: Treat some aluminum
Sample solution: 50 mg/mL wire with 6 N hydrochloric acid at 80 for a few min.
Acceptance criteria: 6.39.6 Dissolve about 100 mg of the treated wire in a mixture of
WATER DETERMINATION, Method I 921 10 mL of hydrochloric acid and 2 mL of nitric acid by
Sample: 0.7 g heating at about 80 for approximately 30 min. Continue
Analysis: Proceed as directed, adding 20.0 mL of glacial heating until the volume is reduced to about 4 mL. Cool to
acetic acid to the titration vessel in addition to the room temperature, and add 4 mL of water. Evaporate to
methanol. about 2 mL by heating. Cool, and transfer this solution,
Acceptance criteria: NMT 7.0% with the aid of water, to a 100-mL volumetric flask, and
ADDITIONAL REQUIREMENTS [NOTEEquivalent to 1 mg/mL of aluminum]
PACKAGING AND STORAGE: Preserve in tight containers. Aluminum standard solution 1.0 g/mL of aluminum
LABELING: Where Calcium Acetate is intended for use in from Aluminum standard stock solution in water
hemodialysis or peritoneal dialysis, it is so labeled. Standard solution: Prepare a solution containing 2.0 mL
of Aluminum standard stock solution, 5 mL of Solution A, and
48 mL of water, and extract this solution with successive
Calcium Acetate Tablets portions of 10, 10, and 5 mL of a 0.5% 8-hydroxyquinoline
in chloroform, combining the chloroform extracts in a 50-
(Comment on this Monograph)id=m11405=Calcium Acetate mL volumetric flask. Dilute the combined extracts with
Tablets=Ca-Chl-Monos.pdf) chloroform to volume.
Sample solution: Prepare a solution containing an amount
DEFINITION of powdered Tablets equivalent to 1.0 g of calcium acetate
Calcium Acetate Tablets contain NLT 90.0% and NMT 110.0% (powder NLT 20 Tablets), 50 mL of water, and 5 mL of
of the labeled amount of calcium acetate (C4H6CaO4). Solution A. Extract this solution with successive portions of
IDENTIFICATION 10, 10, and 5 mL of a 0.5% 8-hydroxyquinoline in
IDENTIFICATION TESTSGENERAL, Calcium and Acetate 191 chloroform, combining the chloroform extracts in a 50-mL
Sample solution: 100 mg/mL of calcium acetate, from volumetric flask. Dilute the combined extracts with
powdered Tablets, in water chloroform to volume.
Acceptance criteria: Meet the requirements of the tests Blank: Prepare a solution containing 50 mL of water and 5
mL of Solution A, and extract as described for the Aluminum
ASSAY standard solution. Extract this solution with successive
PROCEDURE portions of 10, 10, and 5 mL of a 0.5% 8-hydroxyquinoline
Sample: Amount equivalent to 300 mg of calcium acetate in chloroform, combining the chloroform extracts in a 50-
from NLT 20 powdered Tablets mL volumetric flask. Dilute the combined extracts with
Analysis: Dissolve the Sample in 150 mL of water chloroform to volume.
containing 2 mL of 3 N hydrochloric acid. While stirring, Spectrometric conditions
add 30 mL of 0.05 M edetate disodium VS from a 50-mL Mode: fluorescence
buret, and add 15 mL of 1 N sodium hydroxide and 300 Excitation wavelength: 392 nm
mg of hydroxy naphthol blue. Continue the titration with Emission wavelength: 618 nm
the 0.05 M edetate disodium VS to a blue endpoint. Each Analysis
mL of 0.05 M edetate disodium is equivalent to 7.909 mg of Samples: Sample solution and Standard solution
C4H6CaO4. Acceptance criteria: The fluorescence of the Sample
Acceptance criteria: 90.0%110.0% solution does not exceed that of the Standard solution NMT
2 g/g of aluminum calcium acetate
PERFORMANCE TESTS
DISSOLUTION 711
Apparatus 2: 50 rpm
Time: 30 min
Mode: Atomic absorption spectrometry
Detector: calcium hollow-cathode tube
Analytical wavelength: 422.8 nm
Standard solution: Calcium at a known concentration in
Medium
ADDITIONAL REQUIREMENTS Calculate the concentration, in ppm, of fluoride in the

PACKAGING AND STORAGE: Preserve in well-closed containers. Calcium Ascorbate taken:
Result = (C/CU) x F1
Calcium Ascorbate C = measured concentration of fluoride ion in the
(Comment on this Monograph)id=m11410=Calcium Sample solution (g/mL)
Ascorbate=Ca-Chl-Monos.pdf) CU = concentration of Calcium Ascorbate in the
C12H14CaO12 2H2O 426.34 F1 = unit conversion factor, 1 ppm-mg/g
DEFINITION Acceptance criteria: NMT 10 ppm
Calcium Ascorbate contains NLT 98.0% and NMT 101.0% of SPECIFIC TESTS
C12H14CaO12 2H2O, calculated on the as-is basis. OPTICAL ROTATION, Specific Rotation 781S: +95 to +97,
IDENTIFICATION from optical rotation measurements made immediately
A. INFRARED ABSORPTION 197M following the preparation of the Sample solution
B. A 100 mg/mL solution of Calcium Ascorbate decolorizes Sample solution: 50 mg/mL, in carbon dioxide-free water
a 100 mg/mL solution of dichlorophenol-indophenol PH 791: 6.87.4, in a 100 mg/mL solution
C. IDENTIFICATION TESTSGENERAL, Calcium 191 LOSS ON DRYING 731: Dry 3 g of it at 105 for 2 h: it loses
Sample: 100 mg/mL NMT 0.1% of its weight.

PROCEDURE PACKAGING AND STORAGE: Preserve in tight, light-resistant
Sample solution: 60 mg/mL of Calcium Ascorbate in 50 containers.
mL of water USP REFERENCE STANDARDS 11
Analysis: Immediately titrate 50 mL of Sample solution with USP Calcium Ascorbate RS
0.1 N iodine VS, adding 3 mL of starch TS as the endpoint is USP Sodium Fluoride RS
approached. Each mL of 0.1 N iodine is equivalent to 10.66
mg of C12H14CaO12 2H2O.
Acceptance criteria: 98.0%101.0% Calcium Carbonate
IMPURITIES (Comment on this Monograph)id=m11430=Calcium
Inorganic Impurities Carbonate=Ca-Chl-Monos.pdf)
ARSENIC, Method I 211: 3 ppm CaCO3 100.09
HEAVY METALS, Method II 231: NMT 10 ppm Carbonic acid, calcium salt (1:1);
LIMIT OF FLUORIDE Calcium carbonate (1:1) [471-34-1].
[NOTEPrepare and store all solutions in plastic containers.]
Solution A: 294 mg/mL of sodium citrate dihydrate DEFINITION
Standard stock solution: 1.1052 mg/mL of USP Sodium Calcium Carbonate, dried at 200 for 4 h, contains calcium
Fluoride RS equivalent to NLT 98.0% and NMT 100.5%
Standard solution: Combine 20.0 mL of Standard stock
solution with 50.0 mL of Solution A, and dilute with water IDENTIFICATION
to 100 mL IDENTIFICATION TESTSGENERAL, Calcium 191: The addition
Sample solution: Transfer 2.0 g of Calcium Ascorbate to a of acetic acid to it produces effervescence (presence of
beaker containing a plastic-coated stirring bar, add 20 mL carbonate), and the resulting solution, after boiling, meets
of water and 2.0 mL of hydrochloric acid, and stir until the requirements of the tests.
dissolved. Add 50.0 mL of Solution A and sufficient water to
make 100 mL. ASSAY
Electrode system: Use a fluoride-specific, ion-indicating PROCEDURE
electrode and a silversilver chloride reference electrode Sample: 200 mg of Calcium Carbonate, previously dried at
connected to a pH meter capable of measuring potentials 200 for 4 h
with a minimum reproducibility of 0.2 mV (see pH Analysis: Transfer the Sample to a 250-mL beaker. Moisten
791). thoroughly with a few mL of water, and add, dropwise,
Analysis sufficient 3 N hydrochloric acid to dissolve. Add 100 mL of
Samples: Standard solution and Sample solution water, 15 mL of 1 N sodium hydroxide, and 300 mg of
Standard response line: Transfer 50.0 mL of Solution A hydroxy naphthol blue. Titrate with 0.05 M edetate
and 2.0 mL of hydrochloric acid to a beaker, and add disodium VS until the solution is a distinct blue in color.
water to make 100 mL. Add a plastic-coated stirring bar, Each mL of 0.05 M edetate disodium is equivalent to 5.004
insert the electrodes into the solution, stir for 15 min, and mg of CaCO3.
read the potential, in mV. Continue stirring, and at 5-min Acceptance criteria: 98.0%100.5%
intervals add 100, 100, 300, and 500 L of Standard IMPURITIES
solution, reading the potential 5 min after each addition. Inorganic Impurities
Plot the logarithms of the cumulative fluoride ion LIMIT OF FLUORIDE
concentrations (0.1, 0.2, 0.5, and 1.0 g/mL) versus [NOTEPrepare and store all solutions in plastic containers.]
potential, in mV. Solution A: 294 mg/mL of sodium citrate dihydrate in
Rinse and dry the electrodes, insert them into the Sample water
solution, stir for 5 min, and read the potential, in mV. Sample: 2.0 g
From the measured potential and the Standard response Standard stock solution: 1.1052 mg/mL of USP Sodium
line determine the concentration, C (in g/mL) of Fluoride RS in water
fluoride ion in the Sample solution.
Standard solution: Combine 20.0 mL of Standard stock Acceptance criteria: The absorbance of the solution from
solution with 50.0 mL of Solution A and dilute to 100.0 mL the specimen under test does not exceed that of the
with water Standard solution (0.1%).
Electrode system: Use a fluoride-specific, ion-indicating MERCURY, Method IIa 261
electrode and a silversilver chloride reference electrode Mercury stock solution and Standard mercury solution:
connected to a pH meter capable of measuring potentials Proceed as directed under Mercury 261.
with a minimum reproducibility of 0.2 mV (see pH Standard solution: Pipet 2.0 mL of Standard mercury
791). solution into a 100-mL beaker, and add 35 mL of water, 3
Standard response line: Transfer 50.0 mL of Buffer solution mL of hydrochloric acid, and 1 mL of Potassium
and 2.0 mL of hydrochloric acid to a beaker, and add permanganate solution. Cover the beaker with a watch
water to make 100 mL. Add a plastic-coated stirring bar, glass, boil for a few s, and cool.
insert the electrodes into the solution, stir for 15 min, and Sample stock solution: 4.0 g in a 100-mL beaker, and
read the potential, in mV. Continue stirring, and at 5-min cautiously dissolve in 14 mL of 6 N hydrochloric acid
intervals add 100, 100, 300, and 500 L of Standard Sample solution: Transfer the Sample stock solution to a
solution, reading the potential 5 min after each addition. 100-mL beaker, and add 35 mL of water. Stir, and warm to
Plot the logarithms of the cumulative fluoride ion assist solution, if necessary. Add 2 drops of phenolphthalein
concentrations (0.1, 0.2, 0.5, and 1.0 g/mL) versus TS, and, as necessary, slowly neutralize with constant
potential, in mV. stirring, using 1 N sodium hydroxide or 1 N sulfuric acid.
Analysis: Transfer the Sample to a beaker containing a Add 3 mL of hydrochloric acid and 1 mL of Potassium
plastic-coated stirring bar, add 20 mL of water and 4.0 mL permanganate solution. Cover the beaker with a watch
of hydrochloric acid, and stir until dissolved. Add 50.0 mL glass, boil for a few s, and cool.
of Buffer solution and sufficient water to make 100 mL of Analysis
test solution. Rinse and dry the electrodes, insert them into Samples: Standard solution and Sample solution
the Sample solution, stir for 5 min, and read the potential, Proceed as directed in Mercury 261.
in mV. From the measured potential and the Standard Acceptance criteria: 0.5 ppm
response line, determine the concentration, C, in g/mL, of LIMIT OF MAGNESIUM and ALKALI SALTS
fluoride ion in the Sample solution. Calculate the Sample solution: 1.0 g
percentage of fluoride in the specimen taken by Analysis: Mix the Sample with 35 mL of water. Carefully
multiplying C by 0.005. add 3 mL of hydrochloric acid, heat the solution, and boil
Acceptance criteria: 50 ppm for 1 min. Rapidly add 40 mL of oxalic acid TS, and stir
ACID-INSOLUBLE SUBSTANCES: Mix 5.0 g with 10 mL of water, vigorously until precipitation is well-established. Add
and add hydrochloric acid, dropwise, with agitation, until it immediately to the warm mixture 2 drops of methyl red TS
ceases to cause effervescence, then add water to make the and then 6 N ammonium hydroxide, dropwise, until the
mixture measure 200 mL, and filter. Wash the insoluble mixture is just alkaline. Cool to room temperature, transfer
residue with water until the last washing shows no chloride, to a 100-mL graduated cylinder, dilute with water to 100
and ignite: the weight of the residue does not exceed 10 mL, mix, and allow to stand for 4 h or overnight. Filter,
mg (0.2%). and to 50 mL of the clear filtrate in a platinum dish add
BARIUM: A platinum wire, dipped in the filtrate obtained in 0.5 mL of sulfuric acid, and evaporate the mixture on a
the test for Acid-Insoluble Substances and held in a steam bath to a small volume. Carefully heat over a free
nonluminous flame, does not impart a green color. flame to dryness, and continue heating to complete
ARSENIC, Method I 211 decomposition and volatilization of ammonium salts.
Sample solution: Slowly dissolve 1.0 g in 15 mL of Finally, ignite the residue to constant weight.
hydrochloric acid, and dilute with water to 55 mL. Acceptance criteria: The weight of the residue is NMT 5
Acceptance criteria: NMT 3 ppm. The resulting solution mg (NMT 1.0%).
meets the requirements of the test, the addition of 20 mL HEAVY METALS 231
of 7 N sulfuric acid specified under Procedure being Sample solution: Mix 1.0 g with 5 mL of water, slowly
omitted. add 8 mL of 3 N hydrochloric acid, and evaporate on a
LEAD 251 steam bath to dryness. Dissolve the residue in 20 mL of
Sample solution: 1.0 g in 5 mL of water water, filter, and add water to the filtrate to make 25 mL.
Analysis: To the Sample solution slowly add 8 mL of 3 N Acceptance criteria: NMT 20 ppm
hydrochloric acid, evaporate on a steam bath to dryness,
and dissolve the residue in 5 mL of water. SPECIFIC TESTS
Acceptance criteria: NMT 3 ppm LOSS ON DRYING 731: Dry it at 200 for 4 h: it loses NMT
IRON 241 2.0% of its weight.
Sample: 40 mg
Spectrometric conditions ADDITIONAL REQUIREMENTS
Analytical wavelength: 530 nm PACKAGING AND STORAGE: Preserve in well-closed containers.
Blank: Water
Analysis: Dissolve the Sample in 5 mL of 2 N hydrochloric
acid. Transfer to a beaker with the aid of water, and dilute Calcium Carbonate Lozenges
with water to 10 mL. Prepare a Standard solution by
transferring 4.0 mL of Standard iron solution, prepared as (Comment on this Monograph)id=m11439=Calcium Carbonate
directed under Iron 241, to a beaker and by diluting with Lozenges=Ca-Chl-Monos.pdf)
water to 10 mL. To each beaker, add 2 mL of citric acid DEFINITION
solution (1 in 5) and 2 drops of thioglycolic acid, adjust to Calcium Carbonate Lozenges contain NLT 90.0% and NMT
a pH of 9.5 0.1 with ammonia TS, dilute with water to 20 110.0% of the labeled amount of calcium carbonate (CaCO3).
mL, mix, and allow to stand for 5 min. Dilute with water to
50 mL. Concomitantly determine the absorbances of the
solutions from the Sample and the Standard solution.
IDENTIFICATION Standard stock solution: 2.542 mg/mL of sodium chloride,

IDENTIFICATION TESTSGENERAL, Calcium 191: The addition previously dried at 105 for 2 h (equivalent to 1.00 mg/mL
of 6 N hydrochloric acid to a Lozenge produces sodium)
effervescence, and the resulting solution, after being boiled Standard solution A: 1.0 g/mL of sodium from Standard
to expel carbon dioxide and then neutralized with 6 N stock solution
ammonium hydroxide, meets the requirements of the tests. Standard solution B: 3.0 g/mL of sodium from Standard
stock solution
ASSAY Standard solution C: 5.0 g/mL of sodium from Standard
PROCEDURE stock solution
[NOTEThe Standard solutions and the Sample solution may Sample solution: Sample stock solution in the Assay. Pass, if
be modified, if necessary, to obtain solutions, of suitable necessary, through a filter having a 0.5-m or finer porosity
concentrations, adaptable to the linear or working range of to obtain a clear solution. Transfer 10.0 mL of the clear
the instrument.] solution to a 25-mL volumetric flask, and dilute with water
Solution A: Transfer 10 g of potassium chloride and 20 g of to volume.
lanthanum chloride to a 2000-mL volumetric flask. Add Blank: Water
about 1000 mL of water and 40 mL of hydrochloric acid, Spectrometric conditions
mix, and allow to cool. Dilute with water to volume, and (See Spectrometry and Light-Scattering 851.)
mix. Mode: Atomic absorption spectrometer
Solution B: 1 N hydrochloric acid Flame: Airacetylene
Standard stock solution: 250 mg of chelometric standard Lamp: Sodium hollow-cathode
calcium carbonate, previously dried at 110 for 2 h and then Analytical wavelength: 589.6 nm
cooled in a desiccator, in a 500-mL volumetric flask. Add Analysis
about 100 mL of water and 12 mL of Solution B, swirl to Samples: Standard solution A, Standard solution B, Standard
dissolve the calcium carbonate, and allow to cool. Dilute solution C, Sample solution, and Blank
with water to volume. [NOTEThis stock solution contains Plot the absorbances of the Standard solutions versus their
about 500 g/mL of calcium carbonate.] contents of sodium, in g/mL, by drawing a straight line
Standard solution A: 10, 15, and 20 g/mL of calcium best fitting the three plotted points. From the graph
carbonate from Standard stock solution in Solution A determine the quantity, C, in g, of sodium in each mL of
Standard solution B: 15 g/mL of calcium carbonate from the Sample solution.
Standard stock solution in Solution A Calculate the quantity of sodium, in mg, per Lozenge:
Standard solution C: 20 g/mL calcium carbonate from
Standard stock solution in Solution A Result = 2.5 C/N
Sample stock solution: Equivalent to 3000 mg of calcium
carbonate, from powdered lozenges, to a 1000-mL C = measured quantity of sodium in the Sample
volumetric flask, add 100 mL of Solution B and 300 mL of solution (g/mL)
water, and sonicate to dissolve the powder. Dilute with N = number of Lozenges taken to prepare the
water to volume. Sample solution prepared as directed in the
Sample solution: Dilute 5.0 mL of Sample stock solution Assay
with Solution A to 1000 mL. Acceptance criteria: NMT 115.0% of the declared amount
Blank: Solution A
Spectrometric conditions SPECIFIC TESTS
(See Spectrometry and Light-Scattering 851.) ACID-NEUTRALIZING CAPACITY 301
Mode: Atomic absorption spectrometer Analysis: The acid consumed by the minimum single dose
Source: Nitrous oxideacetylene recommended in the labeling is NLT 5 mEq of acid and NLT
Lamp: Calcium hollow-cathode the number of mEq calculated:
Analysis Result = F (Ta C)
Standard solution C F = acceptable lower limits of the required acid-
Plot the absorbances of the Standard solutions versus their neutralizing capacity, 0.9
concentrations of calcium carbonate, in g/mL, by Ta = theoretical acid-neutralizing capacity, in mEq of
drawing a straight line best fitting the three plotted CaCO3 (0.02)
points. From the graph, determine the concentration, C, C = quantity of CaCO3 in the sample tested (mg),
in g/mL, of calcium carbonate in the Sample solution. based on the labeled quantity
Calculate the percentage of label claim of CaCO3 per PERFORMANCE TESTS
Lozenge: UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Result = (C/CU) 100 ADDITIONAL REQUIREMENTS
C = measured concentration of calcium carbonate in
the Sample solution (g/mL)
(g/mL) Calcium Carbonate Oral Suspension
Acceptance criteria: 90.0%110.0% (Comment on this Monograph)id=m11440=Calcium Carbonate
OTHER COMPONENTS Oral Suspension=Ca-Chl-Monos.pdf)
SODIUM CONTENT (if so labeled) DEFINITION
[NOTEThe Standard solutions and the Sample solution may Calcium Carbonate Oral Suspension contains NLT 90.0% and
be modified, if necessary, to obtain solutions of suitable NMT 110.0% of the labeled amount of calcium carbonate
concentrations adaptable to the linear or working range of (CaCO3).
the instrument.]
IDENTIFICATION container, in 15 mL of hydrochloric acid, and dilute with

IDENTIFICATION TESTSGENERAL, Calcium 191: The addition water to 55 mL: the resulting solution meets the
of acetic acid to it produces effervescence, and the resulting requirements of the test, the addition of 20 mL of 7 N
solution, after being boiled, meets the requirements. sulfuric acid specified under Analysis being omitted: NMT 3
ppm.
ASSAY LEAD 251: Mix equivalent to 1.0 g from Oral Suspension,
PROCEDURE previously shaken in its original container, with 5 mL of
Sample solution: Equivalent to 1 g of calcium carbonate water. Slowly add 8 mL of 3 N hydrochloric acid, evaporate
from Oral Suspension, previously shaken in its original on a steam bath to dryness, and dissolve the residue in 5
container, in a beaker with the aid of 25 mL of water, and mL of water: NMT 3 ppm.
add 20 mL of 1 N hydrochloric acid. Heat on a steam bath HEAVY METALS 231: Mix equivalent to 1.0 g from Oral
for 30 min, allow to cool, transfer with the aid of water to a Suspension, previously shaken in its original container, with
100-mL volumetric flask, dilute with water to volume, mix, 5 mL of water. Slowly add 8 mL of 3 N hydrochloric acid,
and filter. Transfer 20.0 mL of the filtrate to a suitable and evaporate on a steam bath to dryness. Dissolve the
container, dilute with water to 100 mL, and add 15 mL of 1 residue in 20 mL of water, filter, and add water to the
N sodium hydroxide, 5 mL of triethanolamine, and 100 mg filtrate to make 25 mL: NMT 20 ppm.
of hydroxy naphthol blue.
Analysis: Titrate with 0.05 M edetate disodium VS until the SPECIFIC TESTS
solution is deep blue. Each mL of 0.05 M edetate disodium MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED
is equivalent to 5.004 mg of CaCO3. MICROORGANISMS 62 and TESTS FOR SPECIFIED
Acceptance criteria: 90.0%110.0% MICROORGANISMS 62: Its total aerobic microbial count:
NMT 100 cfu/mL, and it meets the requirements of the test
IMPURITIES for absence of Escherichia coli and Pseudomonas aeruginosa.
Inorganic Impurities PH 791: 7.58.7
LIMIT OF FLUORIDE
[NOTEPrepare and store all solutions in plastic containers.] ADDITIONAL REQUIREMENTS
Solution A: 294 mg/mL of sodium citrate dihydrate PACKAGING AND STORAGE: Preserve in tight containers, and
Standard stock solution: 1.1052 mg/mL of USP Sodium avoid freezing.
Fluoride RS USP REFERENCE STANDARDS 11
Standard solution: Combine 20.0 mL of Standard stock USP Sodium Fluoride RS
solution with 50.0 mL of Solution A, and dilute to 100 mL
with water.
Sample solution Transfer an equivalent to 2.0 g of calcium
carbonate, to a beaker containing a plastic-coated stirring Calcium Carbonate Tablets
bar, add 20 mL of water and 2.0 mL of hydrochloric acid, (Comment on this Monograph)id=m11450=Calcium Carbonate
and stir until dissolved. Add 50.0 mL of Solution A and Tablets=Ca-Chl-Monos.pdf)
sufficient water to make 100.0 mL.
Electrode system: Use a fluoride-specific, ion-indicating DEFINITION
electrode and a silversilver chloride reference electrode Calcium Carbonate Tablets contain NLT 90.0% and NMT
connected to a pH meter capable of measuring potentials 110.0% of the labeled amount of calcium carbonate (CaCO3).
with a minimum reproducibility of 0.2 mV (see pH For Tablets labeled for any indication other than, or in addition
791). to, antacid use the Tablets contain NLT 90.0% and NMT
Analysis 115.0% of the labeled amount of calcium carbonate.
Standard response line: Transfer 50.0 mL of Solution A IDENTIFICATION
and 2.0 mL of hydrochloric acid to a beaker, and add A. IDENTIFICATION TESTSGENERAL, Calcium 191: The
water to make 100 mL. Add a plastic-coated stirring bar, addition of 6 N acetic acid to the Tablets produces
insert the electrodes into the solution, stir for 15 min, and effervescence, and the resulting solution, after being boiled
read the potential, in mV. Continue stirring, and at 5-min to expel carbon dioxide and neutralized with 6 N
intervals add 100, 100, 300, and 500 L of Standard ammonium hydroxide, meets the requirements of the tests.
solution, reading the potential 5 min after each addition. ASSAY
Plot the logarithms of the cumulative fluoride ion PROCEDURE
concentrations (0.1, 0.2, 0.5, and 1.0 g/mL) versus Sample: Equivalent to 200 mg of calcium carbonate, from
potential, in mV. powdered Tablets (NLT 20), in a suitable crucible
Rinse and dry the electrodes, insert them into the Sample Analysis: Ignite to constant weight. Cool the crucible, add
solution, stir for 5 min, and read the potential, in mV. 10 mL of water, and dissolve the residue by adding
From the measured potential and the Standard response sufficient 3 N hydrochloric acid, dropwise, to achieve
line determine the concentration, C (in g/mL) of fluoride complete solution. Transfer the solution completely to a
ion in the Sample solution. suitable container, dilute with water to 150 mL, and add 15
Calculate the concentration, in ppm, of fluoride in the mL of 1 N sodium hydroxide and 300 mg of hydroxy
Suspension taken: naphthol blue. Titrate with 0.05 M edetate disodium VS
until the solution is deep blue. Each mL of 0.05 M edetate
Result = (C/CU) x F1 disodium is equivalent to 5.004 mg of CaCO3.
C = measured concentration of fluoride ion in the Acceptance criteria: 90.0%110.0% of the labeled amount
Sample solution (g/mL) of CaCO3; for Tablets labeled for any indication other than,
CU = concentration of Calcium Carbonate in the or in addition to, antacid use, 90.0%115.0% of the labeled
Sample solution (mg/mL) amount of CaCO3
F1 = unit conversion factor, 1 ppm-mg/g
ARSENIC, Method I 211: Slowly dissolve equivalent to 1.0 g
from Oral Suspension, previously shaken in its original
SPECIFIC TESTS UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements

Analysis: Where Tablets are labeled for antacid use, NLT 5 ADDITIONAL REQUIREMENTS
mEq of acid is consumed by the minimum single dose PACKAGING AND STORAGE: Preserve in well-closed containers.
recommended in the labeling, and NLT the number of mEq LABELING: Label it to indicate whether it is for use as an
calculated: antacid, or as a dietary supplement, or both.
Result = (Ta C)
Ta = theoretical acid-neutralizing capacity of CaCO3 Calcium Carbonate and Magnesia
(mEq), 0.02 Tablets
C = quantity of CaCO3 in the sample tested (mg), (Comment on this Monograph)id=m11474=Calcium Carbonate
based on the labeled quantity and Magnesia Tablets=Ca-Chl-Monos.pdf)
(Current titlenot to change until February 1, 2010)
PERFORMANCE TESTS Monograph title changeto become official February 1, 2010
DISSOLUTION 711 See Calcium Carbonate and Magnesia Chewable Tablets
[NOTEFor Tablets labeled for any indication other than, or
in addition to, antacid use.] DEFINITION
Medium: 0.1 N hydrochloric acid; 900 mL Calcium Carbonate and Magnesia Tablets contain NLT 90.0%
Apparatus 2: 75 rpm and NMT 110.0% of the labeled amount of calcium carbonate
Time: 30 min (CaCO3) and NLT 90.0% and NMT 115.0% of the labeled
Analysis: Determine the amount of CaCO3 dissolved by amount of magnesium hydroxide [Mg(OH)2].
employing the following method.
Solution A: 50 mg/mL of lanthanum chloride in 0.1 N IDENTIFICATION
hydrochloric acid A. IDENTIFICATION TESTSGENERAL, Calcium 191: The
Blank: Solution A and 0.1 N hydrochloric acid (1:9) addition of 3 N hydrochloric acid to the Tablets produces
Standard stock solution: 100 g/mL of calcium carbonate effervescence, and the resulting solution, after being boiled
in 0.1 N hydrochloric acid to expel carbon dioxide and neutralized with 6 N
Standard solution A: Dilute 3 mL of Standard stock solution ammonium hydroxide, meets the requirements of the tests.
and 10 mL of Solution A with 0.1 N hydrochloric acid to 100 B. IDENTIFICATION TESTSGENERAL, Magnesium 191
mL. Sample solution: Heat 2 Tablets in 20 mL of 1 N sulfuric
Standard solution B: Dilute 4 mL of Standard stock solution acid.
and 10 mL of Solution A with 0.1 N hydrochloric acid to 100 Analysis: Cool, add 20 mL of alcohol, mix, and allow to
mL. stand for 30 min. Filter this solution, and add 2 mL of 1 N
Standard solution C: Dilute 5 mL of Standard stock solution hydrochloric acid to the filtrate.
and 10 mL of Solution A with 0.1 N hydrochloric acid to 100 Acceptance criteria: Meet the requirements
mL.
Standard solution D: Dilute 6 mL of Standard stock solution ASSAY
and 10 mL of Solution A with 0.1 N hydrochloric acid to 100 PROCEDURE 1: CALCIUM CARBONATE
mL. Sample solution: Transfer an equivalent to 400 mg of
Sample solution: Sample per Dissolution 711. Filter a calcium carbonate, from powdered Tablets (NLT 20), in 25
portion of the solution under test. Pipet a volume of the mL of water, and add 40 mL of 1 N hydrochloric acid. Heat
filtrate, estimated to contain 1 mg of calcium, into a 250-mL on a steam bath for 30 min, allow to cool, transfer to a 100-
volumetric flask, add 25.0 mL of Solution A, and dilute with mL volumetric flask with the aid of water, dilute with water
0.1 N hydrochloric acid to volume. to volume, mix, and filter. Transfer 20.0 mL of the filtrate to
Spectrometric conditions a suitable container, dilute with water to 100 mL, and add
(See Spectrometry and Light-Scattering 851.) 30 mL of 1 N sodium hydroxide, 5 mL of triethanolamine,
Mode: Atomic absorption spectrometer and 100 mg of hydroxy naphthol blue.
Lamp: Calcium hollow-cathode Analysis: Titrate with 0.05 M edetate disodium VS until the
Flame: Airacetylene solution is deep blue in color. Each mL of 0.05 M edetate
Analytical wavelength: 422.8 nm disodium is equivalent to 5.004 mg of CaCO3.
Analysis PROCEDURE 2: MAGNESIUM HYDROXIDE
Samples: Standard solution A, Standard solution B, Standard Sample solution: Equivalent to 120 mg of calcium
solution C, Standard solution D, Sample solution, and Blank carbonate and magnesium hydroxide combined, from the
Concomitantly determine the absorbances of the Standard portion of the filtrate remaining from the Assay for Calcium
solutions and the Sample solution, against the Blank. Carbonate, to a suitable container, dilute with water to 100
Construct a standard curve by plotting absorbances versus mL, and add 10 mL of ammoniaammonium chloride buffer
calcium concentrations of the Standard solutions, then from TS, 5 mL of triethanolamine, and 0.3 mL of eriochrome
it obtain the concentration, Cs, in g/mL of calcium, of the black TS.
Sample solution. Analysis: Titrate with 0.05 M edetate disodium VS to a blue
Calculate the percentage of CaCO3 dissolved: endpoint. The volume, in mL, of 0.05 M edetate disodium
consumed, less the volume of 0.05 M edetate disodium
Result = (Mr/Ar) (CS/CU) 100 corresponding to the content of calcium carbonate in the
volume, in mL, of the filtrate taken, represents the volume,
Mr = molecular weight of calcium carbonate, 100.09 in mL, of 0.05 M edetate disodium equivalent to the
Ar = atomic weight of calcium, 40.08 quantity of magnesium hydroxide present. Each mL of 0.05
CS = measured concentration of calcium in the Sample M edetate disodium is equivalent to 2.916 mg of Mg(OH)2.
solution (g/mL) Acceptance criteria: 90.0%110.0% of the labeled amount
CU = nominal concentration of the Sample solution of CaCO3; 90.0%115.0% of the labeled amount of
(g/mL) Mg(OH)2
Tolerances: NLT 75% (Q) of the labeled amount of CaCO3
is dissolved.
SPECIFIC TESTS Analysis: Titrate with 0.05 M edetate disodium VS until the
ACID-NEUTRALIZING CAPACITY 301 solution is deep blue in color. Each mL of 0.05 M edetate
Analysis: NLT 5 mEq of acid is consumed by the minimum disodium is equivalent to 5.004 mg of CaCO3.
single dose recommended in the labeling, and NLT the PROCEDURE 2: MAGNESIUM HYDROXIDE
number of mEq calculated: Sample solution: Equivalent to 120 mg of calcium
carbonate and magnesium hydroxide combined, from the
Result = Fa (Ta M) + Fb (Tb C) portion of the filtrate remaining from the Assay for Calcium
Carbonate, to a suitable container, dilute with water to 100
Fa = monograph correction factor for Mg(OH)2, 0.8 mL, and add 10 mL of ammoniaammonium chloride buffer
Ta = theoretical acid-neutralizing capacity of Mg(OH)2 TS, 5 mL of triethanolamine, and 0.3 mL of eriochrome
(mEq), 0.0343 black TS.
M = quantity of Mg(OH)2 in the specimen tested Analysis: Titrate with 0.05 M edetate disodium VS to a blue
(mg), based on the labeled quantities endpoint. The volume, in mL, of 0.05 M edetate disodium
Fb = monograph correction factor for CaCO3, 0.9 consumed, less the volume of 0.05 M edetate disodium
Tb = theoretical acid-neutralizing capacities of CaCO3 corresponding to the content of calcium carbonate in the
(mEq), 0.02 volume, in mL, of the filtrate taken, represents the volume,
C = quantity of CaCO3 in the specimen tested (mg), in mL, of 0.05 M edetate disodium equivalent to the
based on the labeled quantities quantity of magnesium hydroxide present. Each mL of 0.05
M edetate disodium is equivalent to 2.916 mg of Mg(OH)2.
PERFORMANCE TESTS Acceptance criteria: 90.0%110.0% of the labeled amount
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements of CaCO3; 90.0%115.0% of the labeled amount of
for Weight Variation with respect to calcium carbonate and Mg(OH)2
to magnesia
SPECIFIC TESTS
ADDITIONAL REQUIREMENTS ACID-NEUTRALIZING CAPACITY 301
PACKAGING AND STORAGE: Preserve in well-closed containers. Analysis: NLT 5 mEq of acid is consumed by the minimum
LABELING: Label the Tablets to indicate that they must be single dose recommended in the labeling, and NLT the
chewed before being swallowed. number of mEq calculated:
Result = Fa (Ta M) + Fb (Tb C)
Calcium Carbonate and Magnesia Fa = monograph correction factor for Mg(OH)2, 0.8
Chewable Tablets Ta = theoretical acid-neutralizing capacity of Mg(OH)2
(Comment on this Monograph)id=m2520=Calcium Carbonate (mEq), 0.0343
and Magnesia Chewable Tablets=Ca-Chl-Monos.pdf) M = quantity of Mg(OH)2 in the specimen tested
(Monograph under this new titleto become official February (mg), based on the labeled quantities
1, 2010) Fb = monograph correction factor for CaCO3, 0.9
(Current monograph title is Calcium Carbonate and Magnesia Tb = theoretical acid-neutralizing capacities of CaCO3
Tablets) (mEq), 0.02
C = quantity of CaCO3 in the specimen tested (mg),
DEFINITION based on the labeled quantities
Calcium Carbonate and Magnesia Chewable Tablets contain
NLT 90.0% and NMT 110.0% of the labeled amount of PERFORMANCE TESTS
calcium carbonate (CaCO3) and NLT 90.0% and NMT 115.0% UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
of the labeled amount of magnesium hydroxide [Mg(OH)2]. for Weight Variation with respect to calcium carbonate and
to magnesia
IDENTIFICATION
A. IDENTIFICATION TESTSGENERAL, Calcium 191: The ADDITIONAL REQUIREMENTS
addition of 3 N hydrochloric acid to the Chewable Tablets PACKAGING AND STORAGE: Preserve in well-closed containers.
produces effervescence, and the resulting solution, after LABELING: Label the Chewable Tablets to indicate that they
being boiled to expel carbon dioxide and neutralized with 6 must be chewed before being swallowed.
N ammonium hydroxide, meets the requirements of the
tests.
B. IDENTIFICATION TESTSGENERAL, Magnesium 191
Sample solution: Heat 2 Chewable Tablets in 20 mL of 1 N Calcium Carbonate, Magnesia, and
sulfuric acid. Simethicone Tablets
Analysis: Cool, add 20 mL of alcohol, mix, and allow to (Comment on this Monograph)id=m11476=Calcium Carbonate,
stand for 30 min. Filter this solution, and add 2 mL of 1 N Magnesia, and Simethicone Tablets=Ca-Chl-Monos.pdf)
hydrochloric acid to the filtrate. (Current titlenot to change until February 1, 2010)
Acceptance criteria: Meet the requirements Monograph title changeto become official February 1, 2010
See Calcium Carbonate, Magnesia, and Simethicone Chewable
ASSAY Tablets
PROCEDURE 1: CALCIUM CARBONATE
Sample solution: Transfer an equivalent to 400 mg of DEFINITION
calcium carbonate, from powdered Chewable Tablets (NLT Calcium Carbonate, Magnesia, and Simethicone Tablets contain
20), in 25 mL of water, and add 40 mL of 1 N hydrochloric NLT 90.0% and NMT 110.0% of the labeled amounts of
acid. Heat on a steam bath for 30 min, allow to cool, calcium carbonate (CaCO3) and magnesium hydroxide
transfer to a 100-mL volumetric flask with the aid of water, [Mg(OH)2], and an amount of polydimethylsiloxane
dilute with water to volume, mix, and filter. Transfer 20.0 [-(CH3)2SiO-]n that is NLT 85.0% and NMT 115.0% of the
mL of the filtrate to a suitable container, dilute with water to labeled amount of simethicone.
100 mL, and add 30 mL of 1 N sodium hydroxide, 5 mL of
triethanolamine, and 100 mg of hydroxy naphthol blue.
IDENTIFICATION volumetric flask, add 1.0 mL of Solution A, and dilute with 4-

A. INFRARED ABSORPTION 197S methyl-2-pentanone to volume.
Standard solution: Transfer 33 mg of USP Blank solution: 1.0 mL Solution A diluted with 4-methyl-2-
Polydimethylsiloxane RS, to a suitable round, narrow-mouth, pentanone to 50 mL
screw-capped, 120-mL bottle, add 40 mL of 0.1 N sodium Spectrometric conditions
hydroxide, and swirl to disperse. Add 20.0 mL of toluene, (See Spectrometry and Light-Scattering 851.)
close the bottle securely with a cap having an inert liner, Mode: Atomic absorption spectrometer
and shake for 30 min on a reciprocating shaker (e.g., about Lamp: Silicon hollow-cathode lamp
200 oscillations/min and a stroke of 38 2 mm). Transfer Flame: Nitrous oxcideacetylene
the mixture to a 125-mL separator, and allow to separate. Analytical wavelength: 251.6 nm
Remove the upper, organic layer to a screw-capped, Analysis
centrifuge tube containing about 2 g of anhydrous sodium Samples: Standard solution, Sample solution, and Blank
sulfate. Close the tube with a screw-cap having an inert solution
liner, agitate vigorously, and centrifuge the mixture until a Calculate the percent label claim of polydimethylsiloxane:
clear supernatant is obtained.
Sample solution: Transfer an equivalent to 33 mg of Result = (AU/AS) (CS/CU) 100
simethicone from powdered Tablets (NLT 20 Tablets), to a
suitable round, narrow-mouth, screw-capped, 120-mL AU = absorbance of the Sample solution
bottle. Add 40 mL of 0.1 N sodium hydroxide, and swirl to AS = absorbance of the the Standard solution
disperse. Add 20.0 mL of toluene, close the bottle securely CS = concentration of USP Polydimethylsiloxane RS in
with a cap having an inert liner, and shake for 30 min on a the Standard solution (mg/mL)
reciprocating shaker (e.g., about 200 oscillations/min and a CU = concentration of the Sample solution (mg/mL)
stroke of 38 2 mm). Transfer the mixture to a 125-mL CALCIUM CARBONATE
separator, and allow to separate. Remove the upper, organic Solution A: Transfer 26.8 g of lanthanum chloride to a
layer to a screw-capped, centrifuge tube containing about 2 200-mL volumetric flask, add 100 mL of water, and
g of anhydrous sodium sulfate. Close the tube with a screw- carefully add 50 mL of hydrochloric acid, mix and allow to
cap having an inert liner, agitate vigorously, and centrifuge cool. Dilute with water to volume.
the mixture until a clear supernatant is obtained. Solution B: Prepare as directed in the Assay for
Blank: 10 mL of toluene with about 1 g of anhydrous Polydimethylsiloxane.
sodium sulfate, centrifuged to obtain a clear supernatant Standard stock solution A: Transfer 499.5 mg of primary
Cell: 0.5 mm standard calcium carbonate to a 200-mL volumetric flask,
Analysis: Concomitantly determine the absorbances of the and add 10 mL of water. Carefully add 5 mL of Solution B,
Sample solution and Standard solution at a wavelength of and swirl to dissolve the calcium carbonate. Dilute with
maximum absorbance at about 7.9 m (1265.8 cm1). water to volume.
B. IDENTIFICATION TESTSGENERAL, Calcium 191: The [NOTEThis solution contains 1000 g/mL of calcium
addition of 1 N hydrochloric acid to a Tablet produces (Ca).]
effervescence, and the resulting solution, after having been Standard stock solution B: 1.000 g of magnesium metal
filtered, meets the requirements. to a 1000-mL volumetric flask containing 10 mL of water,
C. IDENTIFICATION TESTSGENERAL, Magnesium 191 slowly add 10 mL of hydrochloric acid, and swirl to dissolve
Sample solution: Heat 2 Tablets in 20 mL of 1 N sulfuric the metal. Dilute with water to volume.
acid. [NOTEThis solution contains 1000 g/mL of magnesium
Analysis: Cool, add 20 mL of alcohol, mix, and allow to (Mg).]
stand for 30 min. Filter this solution, and to the filtrate, add Standard solution A: To a 250-mL volumetric flask, add
2 mL of 1 N hydrochloric acid: this solution meets the 10.0 mL of Standard stock solution A and 5.0 mL of
requirements. Standard stock solution B, and dilute with water to volume.
[NOTEThis solution contains 40 g/mL of calcium (Ca)
ASSAY and 20 g/mL of magnesium (Mg).]
POLYDIMETHYLSILOXANE Standard solution: On the day of use, transfer 4.0 mL of
Solution A: 12.5 mg/mL of saccharin in 4-methyl-2- Standard solution A to a 100-mL volumetric flask containing
pentanone 2.0 mL of Solution A, and dilute with water to volume.
Solution B: 2.4 M hydrochloric acid [NOTEThis solution contains 1.6 g/mL of calcium (Ca)
Standard stock solution: 1 mg/mL of USP and 0.8 g/mL of magnesium (Mg).]
Polydimethylsiloxane RS in 4-methyl-2-pentanone Sample stock solution: Transfer a volume of the aqueous
Standard solution: Transfer 20.0 mL of Standard stock layer retained from the Sample solution in the Assay for
solution and 5.0 mL of Solution A into a 250-mL volumetric Polydimethylsiloxane, equivalent to about 28 mg of calcium
flask. Dilute to volume with 4-methyl-2-pentanone. carbonate, to a 200-mL volumetric flask, and dilute with
[NOTEPrepare the Standard solution on the day of use water to volume.
only. This solution contains about 0.08 mg/mL of USP Sample solution: Transfer 3.0 mL of Sample stock solution
Polydimethylsiloxane RS.] to a 100-mL volumetric flask containing 2.0 mL of Solution
Sample solution: Transfer an equivalent to 20 mg of A to dilute with water to volume.
polydimethylsiloxane from powdered Tablets (NLT 20 Blank solution: Transfer 5.0 mL of Solution A to a 250-mL
Tablets), to a 125-mL separator. Cautiously add 50.0 mL of volumetric flask, and dilute with water to volume.
Solution B, and swirl until the reaction subsides. Insert the Spectrometric conditions
stopper, and mix. Carefully release the pressure, add 50.0 (See Spectrometry and Light-Scattering 851.)
mL of 4-methyl-2-pentanone, and mix for 10 min. Allow the Mode: Atomic absorption spectrometer
layers to separate, and drain the aqueous layer into a Lamp: Calcium hollow-cathode
suitable stoppered container. [NOTERetain this aqueous Flame: Nitrous oxideacetylene
layer for use in preparing the Sample solution in the Assay for Analytical wavelength: 422.7 nm
Calcium carbonate and Magnesium hydroxide and for the Analysis
Sample solution in the test for Sodium content.] Pass the Samples: Standard solution, Sample solution, and Blank
organic layer through a filter containing 50 g of anhydrous solution
sodium sulfate. Transfer 10.0 mL of the filtrate to a 50-mL
Calculate the percent label claim of CaCO3: Mode: Atomic absorption spectrometer
Lamp: Sodium hollow-cathode
Result = (Mr/Ar) (CS/CU) (AU/AS) 100 Flame: Airacetylene
Mr = molecular weight of calcium carbonate, 100.09 Analysis
Ar = atomic weight of calcium, 40.08 Samples: Standard solution, Sample solution, and Blank
CS = concentration of calcium in the Standard solution
solution (g/mL) Calculate the mg of sodium in each Tablet:
CU = nominal concentration in the Sample solution
(g/mL) (5C/6) (A/W) (AU/AS)
AS = absorbance of the the Standard solution C = concentration of sodium from the Standard
MAGNESIUM HYDROXIDE solution (g/mL)
Solution A, Solution B, Standard stock solution A, A = average weight of each Tablet (mg)
Standard stock solution B, Standard solution A, W = weight of the portion of Tablets taken to prepare
Standard solution B, Sample stock solution, Sample the Sample solution in the Assay for
solution, and Blank solution: Proceed as directed in Polydimethylsiloxane (mg)
Assay for Calcium Carbonate. AU = absorbance of the Sample solution
Spectrometric conditions AS = absorbance of the Standard solution
(See Spectrometry and Light-Scattering 851.)
Mode: Atomic absorption spectrometer PERFORMANCE TESTS
Lamp: Magnesium hollow-cathode UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Flame: Nitrous oxideacetylene for Weight Variation with respect to calcium carbonate and
Analytical wavelength: 285.2 nm to magnesium hydroxide
Analysis
Samples: Standard solution, Sample solution, and Blank SPECIFIC TESTS
solution ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is
Calculate the percent label claim of Mg(OH)2: consumed by the minimum single dose recommended in
the labeling
Result = (AU/AS) (CS/CU) (Mr/Ar) 100 ADDITIONAL REQUIREMENTS
AU = absorbance of the Sample solution PACKAGING AND STORAGE: Preserve in well-closed containers.
AS = absorbance of the the Standard solution LABELING: Label it to indicate that the Tablets are to be
CS = concentration of calcium of the Standard chewed before swallowing. Label the Tablets to state the
solution (g/mL) sodium content, in mg/Tablet, if it is greater than 5
CU = concentration of the Sample solution (g/mL) mg/Tablet.
Mr = molecular weight of magnesium hydroxide, USP REFERENCE STANDARDS 11
58.34 USP Polydimethylsiloxane RS
Ar = atomic weight of magnesium, 24.305
amounts of CaCO3 and Mg(OH)2; and an amount of Calcium Carbonate, Magnesia, and
polydimethylsiloxane [(CH3)2SiO]n, 85.0%115.0% of the Simethicone Chewable Tablets
labeled amount of simethicone
(Comment on this Monograph)id=m11476=Calcium Carbonate,
OTHER COMPONENTS Magnesia, and Simethicone Chewable Tablets=Ca-Chl-
SODIUM CONTENT (if so labeled): Each Tablet contains NMT Monos.pdf)
the number of mg of sodium stated on the label. Monograph title changeto become official February 1, 2010
Solution A: Prepare as directed in the Assay for Calcium (Current titlenot to change until February 1, 2010)
Carbonate. See Calcium Carbonate, Magnesia, and Simethicone Tablets
Solution B: Prepare as directed in the Assay for
Polydimethylsiloxane. DEFINITION
Standard stock solution: 2.542 mg/mL of sodium chloride, Calcium Carbonate, Magnesia, and Simethicone Chewable
previously dried at 105 for 2 h, in water. Transfer 5.0 mL of Tablets contain NLT 90.0% and NMT 110.0% of the labeled
this solution to a 100-mL volumetric flask, and dilute with amounts of calcium carbonate (CaCO3) and magnesium
water to volume. Transfer 4.0 mL of this solution to a hydroxide [Mg(OH)2], and an amount of polydimethylsiloxane
second 100-mL volumetric flask containing 6.0 mL of [-(CH3)2SiO-]n that is NLT 85.0% and NMT 115.0% of the
Solution B and 2.0 mL of Solution A, and dilute with water to labeled amount of simethicone.
volume.
This solution contains 2.0 g of sodium/mL. IDENTIFICATION
Sample solution: Transfer 3.0 mL of the aqueous layer A. INFRARED ABSORPTION 197S
retained from the solution of the Sample solution in the Standard solution: Transfer 33 mg of USP
Assay for Polydimethylsiloxane to a 50-mL volumetric flask Polydimethylsiloxane RS, to a suitable round, narrow-mouth,
containing 1.0 mL of Solution A, and dilute with water to screw-capped, 120-mL bottle, add 40 mL of 0.1 N sodium
volume. hydroxide, and swirl to disperse. Add 20.0 mL of toluene,
Blank solution: 15.0 mL Solution B and 5.0 mL Solution A close the bottle securely with a cap having an inert liner,
diluted with water to 250 mL and shake for 30 min, on a reciprocating shaker (e.g., about
Spectrometric conditions 200 oscillations/min and a stroke of 38 2 mm). Transfer
(See Spectrometry and Light-Scattering 851.) the mixture to a 125-mL separator, and allow to separate.
Remove the upper, organic layer to a screw-capped, Mode: Atomic absorption spectrometer
centrifuge tube containing about 2 g of anhydrous sodium Lamp: Silicon hollow-cathode lamp
sulfate. Close the tube with a screw-cap having an inert Flame: Nitrous oxcideacetylene
liner, agitate vigorously, and centrifuge the mixture until a Analytical wavelength: 251.6 nm
clear supernatant is obtained. Analysis
Sample solution: Transfer an equivalent to 33 mg of Samples: Standard solution, Sample solution, and Blank
simethicone from powdered Chewable Tablets (NLT 20 solution
Chewable Tablets), to a suitable round, narrow-mouth, Calculate the percent label claim of polydimethylsiloxane:
screw-capped, 120-mL bottle. Add 40 mL of 0.1 N sodium
hydroxide, and swirl to disperse. Add 20.0 mL of toluene, Result = (AU/AS) (CS/CU) 100
close the bottle securely with a cap having an inert liner,
and shake for 30 min on a reciprocating shaker (e.g., about AU = absorbance of the Sample solution
200 oscillations/min and a stroke of 38 2 mm). Transfer AS = absorbance of the the Standard solution
the mixture to a 125-mL separator, and allow to separate. CS = concentration of USP Polydimethylsiloxane RS in
Remove the upper, organic layer to a screw-capped, the Standard solution (mg/mL)
centrifuge tube containing about 2 g of anhydrous sodium CU = concentration of the Sample solution (mg/mL)
sulfate. Close the tube with a screw-cap having an inert CALCIUM CARBONATE AND MAGNESIUM HYDROXIDE
liner, agitate vigorously, and centrifuge the mixture until a Solution A: Transfer 26.8 g of lanthanum chloride to a
clear supernatant is obtained. 200-mL volumetric flask, add 100 mL of water, and
Blank: 10 mL of toluene with about 1 g of anhydrous carefully add 50 mL of hydrochloric acid, mix and allow to
sodium sulfate, centrifuged to obtain a clear supernatant cool. Dilute with water to volume.
Cell: 0.5 mm Solution B: Prepare as directed in the Assay for
Analysis: Concomitantly determine the absorbances of the Polydimethylsiloxane.
Sample solution and Standard solution at a wavelength of Standard stock solution A: Transfer 499.5 mg of primary
maximum absorbance at about 7.9 m (1265.8 cm1). standard calcium carbonate to a 200-mL volumetric flask,
B. IDENTIFICATION TESTSGENERAL, Calcium 191: The and add 10 mL of water. Carefully add 5 mL of Solution B,
addition of 1 N hydrochloric acid to a Chewable Tablet and swirl to dissolve the calcium carbonate. Dilute with
produces effervescence, and the resulting solution, after water to volume.
having been filtered, meets the requirements. [NOTEThis solution contains 1000 g/mL of calcium
C. IDENTIFICATION TESTSGENERAL, Magnesium 191 (Ca).]
Sample solution: Heat 2 Chewable Tablets in 20 mL of 1 N Standard stock solution B: 1.000 g of magnesium metal
sulfuric acid. to a 1000-mL volumetric flask containing 10 mL of water,
Analysis: Cool, add 20 mL of alcohol, mix, and allow to slowly add 10 mL of hydrochloric acid, and swirl to dissolve
stand for 30 min. Filter this solution, and to the filtrate, add the metal. Dilute with water to volume.
2 mL of 1 N hydrochloric acid: this solution meets the [NOTEThis solution contains 1000 g/mL of magnesium
requirements. (Mg).]
Standard solution A: To a 250-mL volumetric flask, add
ASSAY 10.0 mL of Standard stock solution A and 5.0 mL of
POLYDIMETHYLSILOXANE Standard stock solution B, and dilute with water to volume.
Solution A: 12.5 mg/mL of saccharin in 4-methyl-2- [NOTEThis solution contains 40 g/mL of calcium (Ca)
pentanone and 20 g/mL of magnesium (Mg).]
Solution B: 2.4 M hydrochloric acid Standard solution: On the day of use, transfer 4.0 mL of
Standard stock solution: 1 mg/mL of USP Standard solution A to a 100-mL volumetric flask containing
Polydimethylsiloxane RS in 4-methyl-2-pentanone 2.0 mL of Solution A, and dilute with water to volume.
Standard solution: Transfer 20.0 mL of Standard stock [NOTEThis solution contains 1.6 g/mL of calcium (Ca)
solution and 5.0 mL of Solution A into a 250-mL volumetric and 0.8 g/mL of magnesium (Mg).]
flask. Dilute with 4-methyl-2-pentanone to volume. Sample stock solution: Transfer volume of the aqueous
[NOTEPrepare the Standard solution on the day of use layer retained from the Sample solution in the Assay for
only. This solution contains about 0.08 mg/mL of USP Polydimethylsiloxane, equivalent to about 28 mg of calcium
Polydimethylsiloxane RS.] carbonate, to a 200-mL volumetric flask, and dilute with
Sample solution: Transfer an equivalent to 20 mg of water to volume.
polydimethylsiloxane from powdered Chewable Tablets (NLT Sample solution: Transfer 3.0 mL of Sample stock solution
20 Chewable Tablets), to a 125-mL separator. Cautiously to a 100-mL volumetric flask containing 2.0 mL of Solution
add 50.0 mL of Solution B, and swirl until the reaction A to dilute with water to volume.
subsides. Insert the stopper, and mix. Carefully release the Blank solution: Transfer 5.0 mL of Solution A to a 250-mL
pressure, add 50.0 mL of 4-methyl-2-pentanone, and mix volumetric flask, and dilute with water to volume.
for 10 min. Allow the layers to separate, and drain the CALCIUM CARBONATE
aqueous layer into a suitable stoppered container. [NOTE Spectrometric conditions
Retain this aqueous layer for use in preparing the Sample (See Spectrometry and Light-Scattering 851.)
solution in the Assay for Calcium carbonate and Magnesium Mode: Atomic absorption spectrometer
hydroxide and for the Sample solution in the test for Sodium Lamp alcium hollow-cathode lamp
content.] Pass the organic layer through a filter containing Flame: nitrous oxide-acetylene flame
50 g of anhydrous sodium sulfate. Transfer 10.0 mL of the Analytical wavelength: 422.7 nm
filtrate to a 50-mL volumetric flask, add 1.0 mL of Solution Analysis
A, and dilute with 4-methyl-2-pentanone to volume. Samples: Standard solution, Sample solution, and Blank
Blank solution: 1.0 mL Solution A diluted with 4-methyl-2- solution
pentanone to 50 mL Calculate the percent label claim, of CaCO3:
(See Spectrometry and Light-Scattering 851.) Result = (AU/AS) (CS/CU) (Mr/Ar) 100
AU = absorbance of the Sample solution Calculate the mg of sodium in each Chewable Tablet:
AS = absorbance of the the Standard solution
CS = concentration of calcium in the Standard Result = (AU/AS) (A/W) (5C/6)
solution (g/mL)
CU = nominal concentration of the Sample solution AU = absorbance of the Sample solution
(g/mL) AS = absorbance of the Standard solution
Mr = molecular weight of calcium carbonate, 100.09 A = average weight of each Chewable Tablet (mg)
Ar = atomic weight of calcium, 40.08 W = weight of the portion of Chewable Tablets taken
MAGNESIUM HYDROXIDE to prepare the Sample solution in the Assay for
Solution A, Solution B, Standard stock solution A, Polydimethylsiloxane (mg)
Standard stock solution B, Standard solution A, C = concentration of sodium in the Standard solution
Standard solution B, Sample stock solution, Sample (g/mL)
solution, and Blank solution: Proceed as directed in
Assay for Calcium carbonate. PERFORMANCE TESTS
Spectrometric conditions UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
(See Spectrometry and Light-Scattering 851.) for Weight Variation with respect to calcium carbonate and
Mode: Atomic absorption spectrometer to magnesium hydroxide
Lamp: Magnesium hollow-cathode SPECIFIC TESTS
Flame: Nitrous oxideacetylene ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is
Analytical wavelength: 285.2 nm consumed by the minimum single dose recommended in
Analysis the labeling
Samples: Standard solution, Sample solution, and Blank
solution ADDITIONAL REQUIREMENTS
Calculate the percent label claim of [Mg(OH)2]: PACKAGING AND STORAGE: Preserve in well-closed containers.
LABELING: Label it to indicate that the Chewable Tablets are
Result = (Mr/Ar) (CS/CU) (AU/AS) 100 to be chewed before swallowing. Label the Chewable Tablets
to state the sodium content, in mg/Chewable Tablet, if it is
Mr = molecular weight of magnesium hydroxide, greater than 5 mg/Chewable Tablet.
58.34 USP REFERENCE STANDARDS 11
Ar = atomic weight of magnesium, 24.305 USP Polydimethylsiloxane RS
CS = concentration of calcium in the Standard
solution (g/mL)
AU = absorbance of the Sample solution Calcium and Magnesium Carbonates
AS = absorbance of the the Standard solution Oral Suspension
amounts of CaCO3 and Mg(OH)2; and an amount of (Comment on this Monograph)id=m11478=Calcium and
polydimethylsiloxane [-(CH3)2SiO-]n, 85.0%115.0% of the Magnesium Carbonates Oral Suspension=Ca-Chl-Monos.pdf)
labeled amount of simethicone DEFINITION
OTHER COMPONENTS Calcium and Magnesium Carbonates Oral Suspension contains
SODIUM CONTENT (if so labeled): Each Chewable Tablet NLT 90.0% and NMT 110.0% of the labeled amount of
contains NMT the number of mg of sodium stated on the calcium carbonate (CaCO3) and NLT 85.0% and NMT 115.0%
label. of the labeled amount of MgCO3.
Solution A: Prepare as directed in the Assay for Calcium IDENTIFICATION
carbonate and Magnesium hydroxide. A. IDENTIFICATION TESTSGENERAL, Calcium 191: The
Solution B: Prepare as directed in the Assay for addition of 3 N hydrochloric acid to a quantity of Oral
Polydimethylsiloxane. Suspension, equivalent to 500 mg of calcium carbonate,
Standard stock solution: 2.542 mg/mL of sodium chloride, produces effervescence, and the resulting solution, after
previously dried at 105 for 2 h, in water. Transfer 5.0 mL of having been filtered, meets the requirements.
this solution to a 100-mL volumetric flask, and dilute with B. IDENTIFICATION TESTSGENERAL, Magnesium 191
water to volume. Transfer 4.0 mL of this solution to a Sample solution: Heat a quantity of Oral Suspension,
second 100-mL volumetric flask containing 6.0 mL of equivalent to 800 mg of magnesium carbonate, with 20 mL
Solution B and 2.0 mL of Solution A, and dilute with water to of 1 N sulfuric acid.
volume. Analysis: Cool, add 20 mL of alcohol, mix, and allow to
This solution contains 2.0 g of sodium/mL. stand for 30 min. Filter this solution, and add 2 mL of 1 N
Sample solution: Transfer 3.0 mL of the aqueous layer hydrochloric acid to the filtrate.
retained from the solution of the Sample solution in the Acceptance criteria: Meets the requirements
Assay for Polydimethylsiloxane to a 50-mL volumetric flask
containing 1.0 mL of Solution A, and dilute with water to ASSAY
volume. CALCIUM CARBONATE
Blank solution: 15.0 mL Solution B and 5.0 mL Solution A Sample solution: Transfer the equivalent to 400 mg of
diluted with water to 250 mL calcium carbonate from Oral Suspension, previously well
Spectrometric conditions shaken in its original container and free of air bubbles, to a
(See Spectrometry and Light-Scattering 851.) beaker, with the aid of 20 mL of water, and add 10 mL of 1
Mode: Atomic absorption spectrometer N hydrochloric acid. Heat on a steam bath for 30 min, allow
Lamp: Sodium hollow-cathode to cool, transfer with the aid of water to a 100-mL
Flame: AirAcetylene volumetric flask, dilute with water to volume, and filter.
Analytical wavelength: 589.0 nm Analysis: Transfer 20.0 mL of Sample solution to a suitable
Analysis container, dilute with water to 100.0 mL, add 15 mL of 1 N
Samples: Standard solution, Sample solution, and Blank sodium hydroxide, 5 mL of triethanolamine, and 100 mg of
solution
hydroxynaphthol blue trituration. Titrate with 0.05 M B. IDENTIFICATION TESTSGENERAL, Magnesium 191
edetate disodium VS until the solution is deep blue. Each mL Sample solution: Heat 2 Tablets in 20 mL of 1 N sulfuric
of 0.05 M edetate disodium is equivalent to 5.004 mg of acid.
CaCO3. Analysis: Cool, add 20 mL of alcohol, mix, and allow to
MAGNESIUM CARBONATE stand for 30 min. Filter this solution, and add 2 mL of 1 N
Sample solution: Equivalent to 120 mg of calcium hydrochloric acid to the filtrate.
carbonate and magnesium carbonate from the Sample Acceptance criteria: The filtrate meets the requirements.
solution in the Assay for Calcium carbonate, dilute with water
to 100 mL, and add 10 mL of ammonia-ammonium chloride ASSAY
buffer TS, 5 mL of triethanolamine, and 0.3 mL of CALCIUM CARBONATE
eriochrome black TS. Sample solution: Transfer the equivalent of 400 mg of
Analysis: Titrate with 0.05 M edetate disodium VS to a blue calcium carbonate, from NLT 20 powdered Tablets, to a
endpoint. From the volume of 0.05 M edetate disodium beaker with the aid of 25 mL of water, and add 10 mL of 1
consumed, subtract the volume of 0.05 M edetate disodium N hydrochloric acid. Heat on a steam bath for 30 min, allow
used in Assay. The difference is the volume of 0.05 M to cool, transfer with the aid of water to a 100-mL
edetate disodium equivalent to the amount of magnesium volumetric flask, dilute with water to volume, mix, and filter.
carbonate present. Each mL of 0.05 M edetate disodium is Analysis: Transfer 20.0 mL of Sample solution to a suitable
equivalent to 4.216 mg of MgCO3. container, dilute with water to 100 mL, add 15 mL of 1 N
Acceptance criteria: 90.0%110.0% of the labeled amount sodium hydroxide, 5 mL of triethanolamine, and 100 mg of
of CaCO3; 85.0%115.0% of the labeled amount of MgCO3 hydroxy naphthol blue. Titrate with 0.05 M edetate
disodium VS until the solution is deep blue. Each mL of 0.05
PERFORMANCE TESTS M edetate disodium is equivalent to 5.004 mg of CaCO3.
DELIVERABLE VOLUME 698: Meets the requirements Acceptance criteria: 90.0%110.0% for CaCO3
MAGNESIUM CARBONATE
SPECIFIC TESTS Sample solution: Equivalent of 120 mg of calcium
MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED carbonate and magnesium carbonate from the Sample
MICROORGANISMS 62: Total aerobic microbial count: NMT solution in the Assay for Calcium carbonate. Dilute with water
100 cfu/mL, and it meets the requirements of the tests for to 100 mL, and add 10 mL of ammoniaammonium
absence of Escherichia coli and Pseudomonas aeruginosa chloride buffer TS, 5 mL of triethanolamine, and 0.3 mL of
PH 791: 7.08.6 eriochrome black TS.
ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is Analysis: Titrate with 0.05 M edetate disodium VS to a blue
consumed by the minimum single dose recommended in endpoint. From the volume of 0.05 M edetate disodium
the labeling, and NLT the number of mEq calculated: consumed, deduct the volume of 0.05 M edetate disodium
used in Assay. The difference is the volume of 0.05 M
Result = 0.8 (0.024 M) + 0.9 (0.02 C) edetate disodium equivalent to the amount of magnesium
carbonate present. Each mL of 0.05 M edetate disodium is
0.024 = theoretical acid-neutralizing capacity MgCO3 equivalent to 4.216 mg of MgCO3.
(mEq) Acceptance criteria: 85.0%115.0% for MgCO3
0.02 = theoretical acid-neutralizing capacities CaCO3
(mEq) PERFORMANCE TESTS
M = quantity of MgCO3 in the specimen tested, based DISINTEGRATION 701
on the labeled quantities (mg) Immersion fluid: Substitute simulated gastric fluid TS for
C = quantity of CaCO3 in the specimen tested, based water.
on the labeled quantities (mg) Time: 10 min; except that where Tablets are labeled as
gelatin-coated, the time is 30 min
ADDITIONAL REQUIREMENTS UNIFORMITY OF DOSAGE UNITS, Weight Variation 905: Meet
PACKAGING AND STORAGE: Preserve in tight containers, and the requirements for CaCO3 and MgCO3
avoid freezing.
SPECIFIC TESTS
Calcium and Magnesium Carbonates Analysis: NLT 5 mEq of acid is consumed by the minimum
single dose recommended in the labeling, and not less than
Tablets the number of mEq calculated.
(Comment on this Monograph)id=m11480=Calcium and Calculate mEq:
Magnesium Carbonates Tablets=Ca-Chl-Monos.pdf)
Result = [0.8 (ANC1M)] + [0.9 (ANC2C)]
DEFINITION
Calcium and Magnesium Carbonates Tablets contain NLT ANC1 = theoretical acid-neutralizing capacity of MgCO3,
90.0% and NMT 110.0% of the labeled amount of calcium 0.024 mEq
carbonate (CaCO3) and NLT 85.0% and NMT 115.0% of the M = quantity of MgCO3 in the specimen tested, based
labeled amount of magnesium carbonate (MgCO3). on the labeled quantities (mg)
ANC2 = theoretical acid-neutralizing capacity of CaCO3,
IDENTIFICATION 0.02 mEq
A. IDENTIFICATION TESTSGENERAL, Calcium 191: The C = quantity of CaCO3 in the specimen tested, based
addition of 1 N hydrochloric acid to 1 Tablet produces on the labeled quantities (mg)
effervescence, and the resulting solution, after having been
filtered, meets the requirements of the tests.
ADDITIONAL REQUIREMENTS volume. Carefully heat over a free flame to dryness, and
PACKAGING AND STORAGE: Preserve in well-closed containers. continue heating to complete decomposition and
LABELING: Tablets that are gelatin-coated are so labeled. volatilization of ammonium salts. Finally ignite the residue
to constant weight.
Acceptance criteria: The weight of the residue is NMT 5
mg (1.0%).
Calcium Chloride
(Comment on this Monograph)id=m11500=Calcium SPECIFIC TESTS
Chloride=Ca-Chl-Monos.pdf) PH 791: 4.59.2, in 50 mg/mL solution
CaCl2 2H2O 147.01 ADDITIONAL REQUIREMENTS
Calcium chloride dihydrate [10035-04-8]. PACKAGING AND STORAGE: Preserve in tight containers.
Anhydrous 110.98 LABELING: Where Calcium Chloride is intended for use in
[10043-52-4]. hemodialysis, it is so labeled.
DEFINITION
Calcium Chloride contains an amount of CaCl2 equivalent to
NLT 99.0% and NMT 107.0% of CaCl2 2H2O. Calcium Chloride Injection
IDENTIFICATION (Comment on this Monograph)id=m11510=Calcium Chloride
A. IDENTIFICATION TESTSGENERAL, Calcium 191: 100 Injection=Ca-Chl-Monos.pdf)
mg/mL DEFINITION
B. IDENTIFICATION TESTSGENERAL, Chloride 191: 100 Calcium Chloride Injection is a sterile solution of Calcium
mg/mL Chloride in Water for Injection. It contains NLT 95.0% and
NMT 105.0% of the labeled amount of CaCl2 2H2O.
ASSAY
PROCEDURE IDENTIFICATION
Sample solution: 1 g of Calcium Chloride in 0.15 M A. IDENTIFICATION TESTSGENERAL, Calcium 191
hydrochloric acid (20:1). Transfer the solution to a 250-mL B. IDENTIFICATION TESTSGENERAL, Chloride 191
volumetric flask, and dilute with water to volume. Pipet 50
mL of the solution into a suitable container, add 100 mL of ASSAY
water, 15 mL of 1 N sodium hydroxide, and 300 mg of PROCEDURE
hydroxy naphthol blue. Sample solution: 1 g of calcium chloride equivalent
Analysis: Titrate with 0.05 M edetate disodium VS until the Injection in a 250-mL volumetric flask, add 5 mL of 3 N
solution is deep blue. Each mL of 0.05 M edetate disodium hydrochloric acid, and dilute with water to volume. Pipet 50
is equivalent to 7.351 mg of CaCl2 2H2O. mL of the resulting solution into a suitable container, add
Acceptance criteria: 99.0%107.0% 100 mL of water, 15 mL of 1 N sodium hydroxide, and 300
mg of hydroxy naphthol blue.
IMPURITIES Analysis: Titrate with 0.05 M edetate disodium VS until the
Inorganic Impurities solution is deep blue. Each mL of 0.05 M edetate disodium
HEAVY METALS 231: NMT 10 ppm, 2.0 g in 25 mL is equivalent to 7.351 mg of CaCl2 2H2O.
ALUMINUM 206 (where it is labeled as intended for use in Acceptance criteria: 95.0%105.0%
hemodialysis): NMT 1 ppm
Sample: 2.0 g of Calcium Chloride SPECIFIC TESTS
IRON, ALUMINUM, AND PHOSPHATE BACTERIAL ENDOTOXINS TEST 85: NMT 0.2 USP Endotoxin
Sample solution: 50 mg/mL Unit/mg of calcium chloride
Analysis: Add 2 drops of 3 N hydrochloric acid and 1 drop PARTICULATE MATTER IN INJECTIONS 788: Meets the
of phenolphthalein TS. Then add ammonium requirements for small-volume injections
chlorideammonium hydroxide TS, dropwise, until the PH 791: 5.57.5 in the undiluted Injection, except where
solution is faintly pink, add 2 drops in excess, and heat the the concentration is greater than 50 mg/mL, in which case
liquid to boiling. this range applies to the Injection diluted with water to yield
Acceptance criteria: No turbidity or precipitate is a concentration of 50 mg/mL
produced. OTHER REQUIREMENTS: It meets the requirements under
LIMIT OF MAGNESIUM AND ALKALI SALTS Injections 1.
Sample: 1 g
Analysis: Dissolve in 50 mL of water, add 500 mg of ADDITIONAL REQUIREMENTS
ammonium chloride, heat the solution, and boil for 1 min. PACKAGING AND STORAGE: Preserve in single-dose containers,
Rapidly add 40 mL of oxalic acid TS, and stir vigorously preferably of Type I glass.
until precipitation is well established. Add immediately to LABELING: The label states the total osmolar concentration in
the warm mixture 2 drops of methyl red TS and then 6 N mOsmol/L. Where the contents are less than 100 mL, or
ammonium hydroxide, dropwise, until the mixture is just where the label states that the Injection is not for direct
alkaline. Cool to room temperature, transfer to a 100-mL injection but is to be diluted before use. The label
graduated cylinder, dilute with water to 100 mL, mix, and alternatively may state the total osmolar concentration in
allow to stand for 4 h or overnight. Filter, and to 50 mL of mOsmol/mL.
the clear filtrate in a platinum dish, add 0.5 mL of sulfuric USP REFERENCE STANDARDS 11
acid, and evaporate the mixture on a steam bath to a small USP Endotoxin RS
Calcium Citrate Standard solution: 5 g/mL of fluoride ion from Standard

(Comment on this Monograph)id=m11520=Calcium stock solution [NOTEPrepare on the day of use.]
Citrate=Ca-Chl-Monos.pdf) Electrode system: Use a fluoride-specific, ion-indicating
electrode and a silversilver chloride reference electrode
connected to a pH meter capable of measuring potentials
with a minimum reproducibility of 0.2 mV (see pH 791).
Standard response line: Transfer 1.0, 5.0, and 10.0 mL of
the Standard solution to separate 250-mL plastic beakers, to
each add 50 mL of water, 5 mL of 1 N hydrochloric acid,
10 mL of 1.0 M sodium citrate, and 10 mL of 0.2 M
edetate disodium. If necessary, adjust with 1 N sodium
C12H10Ca3O14 4H2O 570.49 hydroxide or 1 N hydrochloric acid to a pH of 5.5. Transfer
1,2,3-Propanetricarboxylic acid, 2-hydroxy-, calcium salt (2:3), each solution to a separate 100-mL volumetric flask, and
tetrahydrate; dilute with water to volume. Transfer 50 mL of each
Calcium citrate (3:2), tetrahydrate [5785-44-4]. solution to separate 250-mL plastic beakers, and measure
the potential of each solution with the Electrode system.
DEFINITION Between each reading wash the electrodes with water, and
Calcium Citrate contains four molecules of water of hydration. absorb any residual water by blotting the electrodes dry.
It contains NLT 97.5% and NMT 100.5% of Ca3(C6H5O7)2, on Plot the logarithms of the fluoride concentrations (0.05,
the previously dried basis. 0.25, and 0.50 g/mL, respectively) versus potential, in
mV.
IDENTIFICATION Sample solution: 1.0 g of Calcium Citrate in a 100-mL
A. PROCEDURE beaker, add 10 mL of water and, while stirring, 10 mL of 1
Sample solution: 40 mg/mL of Calcium citrate in 0.4 M N hydrochloric acid. When dissolved, boil rapidly for 1 min,
nitric acid transfer the solution to a 250-mL plastic beaker, and cool
Analysis: To 12.5 mL of Sample solution add 1 mL of in ice water. Add 15 mL of 1.0 M sodium citrate and 10
mercuric sulfate TS, heat to boiling, and add 1 mL of mL of 0.2 M edetate disodium, and adjust with 1 N sodium
potassium permanganate TS. hydroxide or 1 N hydrochloric acid to a pH of 5.5. Transfer
Acceptance criteria: A white precipitate is formed. this solution to a 100-mL volumetric flask, and dilute with
B. PROCEDURE water to volume.
Sample: 0.5 g Analysis: Transfer 50 mL of Sample solution to a 250-mL
Analysis: Ignite completely the Sample at as low a plastic beaker, and measure the potential with the Electrode
temperature as possible, cool, and dissolve the residue in a system. From the measured potential and the Standard
mixture of 10 mL of water and 1 mL of glacial acetic acid. response line determine the concentration, C, in g/mL, of
Filter, and add 10 mL of ammonium oxalate TS to the fluoride ion in the Sample solution.
filtrate. Calculate the percentage of fluoride in the specimen taken
Acceptance criteria: A voluminous white precipitate is by multiplying C by 0.01.
formed, and it is soluble in hydrochloric acid. Acceptance criteria: NMT 30 ppm
PROCEDURE PROCEDURE: LIMIT OF ACID-INSOLUBLE SUBSTANCES: NMT 10
Sample solution: 350 mg of Calcium Citrate in 12 mL of mg (0.2%)
0.5 M hydrochloric acid, and dilute with water to 100 mL Sample solution: 5 g dissolved by heating with a mixture
Analysis: While stirring, add 30 mL of 0.05 M edetate of 10 mL of hydrochloric acid and 50 mL of water for 30
disodium VS from a 50-mL buret. Add 15 mL of 1 N sodium min
hydroxide and 300 mg of hydroxy naphthol blue, and Analysis: Weigh the residue so obtained, filtered, washed,
continue the titration to a blue endpoint. Each mL of 0.05 and dried at 105 for 2 h.
M edetate disodium is equivalent to 8.307 mg of SPECIFIC TESTS
Ca3(C6H5O7)2. LOSS ON DRYING 731: Dry it at 150 for 4 h: it loses
Acceptance criteria: 97.5%100.5% between 10.0%13.3% of its weight.
IMPURITIES ADDITIONAL REQUIREMENTS
Inorganic Impurities PACKAGING AND STORAGE: Preserve in well-closed containers.
ARSENIC, Method I 211: NMT 3 ppm USP REFERENCE STANDARDS 11
Sample solution: 1 g in 5 mL of 3 N hydrochloric acid, USP Sodium Fluoride RS
and dilute with water to 35 mL
LEAD 251: NMT 10 ppm
Sample solution: 0.5 g in 20 mL of 3 N hydrochloric acid,
evaporating on a steam bath to 10 mL, diluting with water Calcium Glubionate Syrup
to 20 mL, and cooling (Comment on this Monograph)id=m11570=Calcium Glubionate
Analysis: Use 5 mL of Diluted Standard Lead Solution (5 g Syrup=Ca-Chl-Monos.pdf)
of Pb) for the test.
HEAVY METALS, Method I 231: NMT 20 ppm DEFINITION
Sample solution: 1 g in 22 mL of 11 M hydrochloric acid, Calcium Glubionate Syrup is a solution containing equimolar
add 1.5 mL of ammonium hydroxide, and dilute with amounts of Calcium Gluconate and Calcium Lactobionate or
water to 25 mL with Calcium Lactobionate predominating. It contains NLT
LIMIT OF FLUORIDE 95.0% and NMT 105.0% of the labeled amount of calcium
[NOTEPrepare and store all solutions in plastic containers.] (Ca).
Standard stock solution: 2.210 mg/mL of USP Sodium
Fluoride RS
IDENTIFICATION alpha epimer of glucoheptonic acid or of a mixture of the

A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 alpha and beta epimers of glucoheptonic acid. It contains NLT
Adsorbent: 0.25-mm layer of chromatographic silica gel 95.0% and NMT 102.0% of C14H26CaO16, calculated on the
Standard solution: 2 mg/mL of calcium gluconate and 4 dried basis.
mg/mL of calcium lactobionate
Sample solution: equivalent to 0.4 mg/mL of calcium, from IDENTIFICATION
Syrup A. INFRARED ABSORPTION 197K
Application volume: 5 L B. A solution of 20 mg/mL meets the requirements of the
Developing solvent system: Alcohol, ethyl acetate, test for Identification TestsGeneral, Calcium 191.
ammonium hydroxide, and water (5:1:1:3)
Spray reagent: 2.5 g of ammonium molybdate in 50 mL of ASSAY
2 N sulfuric acid in a 100-mL volumetric flask, add 1.0 g of PROCEDURE
ceric sulfate, swirl to dissolve, and dilute with 2 N sulfuric Sample solution: 800 mg of Calcium Gluceptate in 150 mL
acid to volume of water containing 2 mL of 3 N hydrochloric acid
Analysis Analysis: While stirring, preferably with a magnetic stirrer,
Samples: Standard solution and Sample solution add 25 mL of 0.05 M edetate disodium VS from a 50-mL
Proceed as directed in Thin-Layer Chromatographic buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of
Identification Test 201 and dry the plate at 100 for 20 hydroxy naphthol blue, and continue the titration to a blue
min. Allow to cool, and spray with Spray reagent. Heat the endpoint. Each mL of 0.05 M edetate disodium is equivalent
plate at 110 for 10 min. to 24.52 mg of C14H26CaO16.
Acceptance criteria: The two principal spots of the Sample Acceptance criteria: 95.0%102.0% of C14H26CaO16
solution correspond in color, size, and RF value to those of IMPURITIES
the Standard solution. [NOTESucrose, if present in the Inorganic Impurities
Sample solution, appears in the chromatogram as a spot CHLORIDE AND SULFATE, Chloride 221: A 1.0-g portion
between two principal spots.] shows no more chloride than corresponds to 1.0 mL of 20.0
B. IDENTIFICATION TESTSGENERAL, Calcium 191: Syrup in mM hydrochloric acid (0.07%).
water (1 in 10) CHLORIDE AND SULFATE, Sulfate 221: A 2.0-g portion shows
ASSAY no more sulfate than corresponds to 1.0 mL of 20 mN
PROCEDURE sulfuric acid (0.05%).
Sample solution: 70 mg of Ca equivalent Syrup in a HEAVY METALS 231: NMT 20 ppm
suitable beaker, add 2 mL of 3 N hydrochloric acid, and Sample solution: 40 mg/mL, use 25 mL
dilute with water to 150 mL SPECIFIC TESTS
Analysis: While stirring with a magnetic stirrer, add 20 mL PH 791: 6.08.0, in 100 mg/mL solution
of 0.05 M edetate disodium VS from a 50-mL buret. Add 15 LOSS ON DRYING 731:
mL of 1 N sodium hydroxide and 300 mg of hydroxy (see Thermal Analysis 891)
naphthol blue, and continue the titration to a blue [NOTEThe quantity taken for the determination may be
endpoint. Each mL of 0.05 M edetate disodium is equivalent adjusted, if necessary, for instrument sensitivity. Weight loss
to 2.004 mg of Ca. occurring at temperatures above about 160, indicative of
Acceptance criteria: 95.0%105.0% decomposition, is not to be interpreted as Loss on Drying.]
SPECIFIC TESTS Sample: 1025 mg
PH 791: 3.44.5 Analysis: Determine the percentage of volatile substances
by thermogravimetric analysis on an appropriately calibrated
ADDITIONAL REQUIREMENTS instrument. Heat the specimen under test at a rate of
PACKAGING AND STORAGE: Preserve in tight containers, at a 5/min in an atmosphere of nitrogen, at a flow rate of 40
temperature not exceeding 30, and avoid freezing. mL/min. Record the thermogram to 150.
Acceptance criteria: The anhydrous form loses NMT 1.0%,
the 2H2O form NMT 6.9%, and the 31/2H2O form NMT
11.4%, of its weight.
Calcium Gluceptate REDUCING SUGARS
(Comment on this Monograph)id=m11600=Calcium Sample: 0.50 g
Gluceptate=Ca-Chl-Monos.pdf) Analysis: Dissolve the Sample in 10 mL of hot water, add 2
mL of 3 N hydrochloric acid, boil for about 2 min, and cool.
Add 5 mL of sodium carbonate TS, allow to stand for 5 min,
dilute with water to 20 mL, and filter. Add 5 mL of the clear
filtrate to 2 mL of alkaline cupric tartrate TS, and boil for 1
min.
Acceptance criteria: No red precipitate is formed
immediately.
C14H26CaO16 (anhydrous) 490.42 ADDITIONAL REQUIREMENTS

Glucoheptonic acid, calcium salt (2:1); PACKAGING AND STORAGE: Preserve in well-closed containers.
Calcium glucoheptonate (1:2) [29039-00-7]. LABELING: Label to indicate whether hydrous or anhydrous;
if hydrous, label to indicate also the degree of hydration.
Calcium Gluceptate is anhydrous or contains varying amounts USP Calcium Gluceptate RS
of water of hydration. It consists of the calcium salt of the
Calcium Gluceptate Injection DEFINITION

(Comment on this Monograph)id=m11610=Calcium Gluceptate Calcium Gluconate is anhydrous or contains one molecule of
Injection=Ca-Chl-Monos.pdf) water of hydration. The anhydrous form contains NLT 98.0%
and NMT 102.0% of C12H22CaO14, calculated on the dried
DEFINITION basis. The monohydrate form contains NLT 99.0% and NMT
Calcium Gluceptate Injection is a sterile solution of Calcium 101.0% of C12H22CaO14 H2O where labeled as intended for
Gluceptate in Water for Injection. It contains NLT 95.0% and use in preparing injectable dosage forms, and NLT 98.5% and
NMT 105.0% of the labeled amount of calcium (Ca). NMT 102.0% of C12H22CaO14 H2O where labeled as not
intended for use in preparing injectable dosage forms.
IDENTIFICATION
A. INFRARED ABSORPTION 197K IDENTIFICATION
Sample: Transfer 5 mL of the Injection to a separator, add A. IDENTIFICATION TESTSGENERAL, Calcium 191: It meets
10 mL of chloroform, shake, and allow the layers to the requirements.
separate. Draw off and discard the chloroform layer, and Sample solution: 20 mg/mL
repeat the extraction with a second 10-mL portion of B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
chloroform. Drain the water layer into a small beaker, Adsorbent: 0.25-mm layer of chromatographic silica gel
evaporate to dryness, and dry in a vacuum at 60 for 16 h. Standard solution: 10 mg/mL of USP Potassium Gluconate
B. IDENTIFICATION TESTSGENERAL, Calcium 191: A dilution RS, heating in a water bath at 60, if necessary, to dissolve
of the Injection (1 in 5) meets the requirements. Sample solution: 10 mg/mL, heating in a water bath at
60, if necessary, to dissolve
ASSAY Application volume: 5 L
PROCEDURE Developing solvent system: Alcohol, ethyl acetate,
Sample solution: To a volume of the Injection, equivalent ammonium hydroxide, and water (5:1:1:3)
to 45 mg of calcium, add 2 mL of 3 N hydrochloric acid Spray reagent: Dissolve 2.5 g of ammonium molybdate in
and 148 mL of water. 50 mL of 2 N sulfuric acid in a 100-mL volumetric flask, add
Analysis: While stirring, preferably with a magnetic stirrer, 1.0 g of ceric sulfate, swirl to dissolve, dilute with 2 N
add 15 mL of 0.05 M edetate disodium VS from a 50-mL sulfuric acid to volume, and mix.
buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of Analysis: Develop the chromatogram until the solvent front
hydroxy naphthol blue, and continue the titration to a blue has moved about three-fourths of the length of the plate.
endpoint. Each mL of 0.05 M edetate disodium is equivalent Remove the plate from the chamber, and dry at 110 for 20
to 2.004 mg of Ca. min. Allow to cool, and spray with the Spray reagent. Heat
Acceptance criteria: 95.0%105.0% the plate at 110 for about 10 min.
Acceptance criteria: The principal spot of the Sample
SPECIFIC TESTS solution corresponds in color, size, and RF value to that of
BACTERIAL ENDOTOXINS TEST 85: Contains NMT 0.32 USP the Standard solution.
Endotoxin Unit/mg of calcium gluceptate
PARTICULATE MATTER IN INJECTIONS 788: Meets the ASSAY
requirements for small-volume injections PROCEDURE
PH 791: 5.67.0 Sample solution: 800 mg of Calcium Gluconate in 150 mL
OTHER REQUIREMENTS: It meets the requirements under of water containing 2 mL of 3 N hydrochloric acid
Injections 1. Analysis: While stirring, preferably with a magnetic stirrer,
add 30 mL of 0.05 M edetate disodium VS from a 50-mL
ADDITIONAL REQUIREMENTS buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of
PACKAGING AND STORAGE: Preserve in tight, single-dose hydroxy naphthol blue, and continue the titration to a blue
containers, preferably of Type I or Type II glass. endpoint. Each mL of 0.05 M edetate disodium is equivalent
LABELING: The label states the total osmolar concentration in to 21.52 mg of C12H22CaO14.
mOsmol/L. Where the contents are less than 100 mL, or Acceptance criteria: Anhydrous form, 98.0%102.0% of
where the label states that the Injection is not for direct C12H22CaO14; monohydrate form, 99.0%101.0% of
injection but is to be diluted before use, the label C12H22CaO14 H2O where labeled as intended for use in
alternatively may state the total osmolar concentration in preparing injectable dosage forms, and 98.5%102.0% of
mOsmol/mL. C12H22CaO14 H2O where labeled as not intended for use in
USP REFERENCE STANDARDS 11 preparing injectable dosage forms
USP Calcium Gluceptate RS
USP Endotoxin RS IMPURITIES
CHLORIDE AND SULFATE, Chloride 221: A 1.0-g portion
shows no more chloride than corresponds to 0.07 mL of
Calcium Gluconate 0.020 N hydrochloric acid (0.005%). Where it is labeled as
(Comment on this Monograph)id=m11660=Calcium not intended for use in the preparation of injectable dosage
Gluconate=Ca-Chl-Monos.pdf) forms, a 1.0-g portion shows no more chloride than
corresponds to 1 mL of 0.020 N hydrochloric acid (0.07%).
CHLORIDE AND SULFATE, Sulfate 221: A 2.0-g portion
dissolved in boiling water shows no more sulfate than
corresponds to 0.1 mL of 0.020 N sulfuric acid (0.005%).
Where it is labeled as not intended for use in the
preparation of injectable dosage forms, a 2.0-g portion
dissolved in boiling water shows no more sulfate than
corresponds to 1 mL of 0.020 N sulfuric acid (0.05%).
C12H22CaO14 (anhydrous) 430.37 ARSENIC, Method I 211
D-Gluconic acid, calcium salt (2:1); Sample solution: Dissolve 1.0 g in a mixture of 10 mL of
Calcium D-gluconate (1:2) [299-28-5]. hydrochloric acid and 20 mL of water, and dilute with
Monohydrate 448.39
water to 55 mL. [NOTEThe addition of 20 mL of 7 N Acceptance criteria: NMT 5 ppm

sulfuric acid specified under Analysis is omitted.] LIMIT OF PHOSPHATE
Acceptance criteria: Meets the requirements of the test: Standard stock solution: 0.716 mg/mL of monobasic
NMT 3 ppm potassium phosphate.
HEAVY METALS, Method II 231: NMT 10 ppm Standard solution: Dilute 1.0 mL of Standard stock
[NOTEWhere Calcium Gluconate is labeled as not intended solution with water to obtain 100 mL. To 2.0 mL of this
for use in the preparation of injectable dosage forms, the solution add 98 mL of water.
limit is 20 ppm.] Sample stock solution: To 10.0 g add 90 mL of hot water
LIMIT OF MAGNESIUM AND ALKALI METALS (7080), and heat to boiling, with swirling, for 10 s to
Sample: 1.0 g obtain a clear solution.
Analysis: Dissolve completely the Sample in 100 mL of Sample solution: Dilute 1 mL of Sample stock solution with
boiling water. Add 10 mL of ammonium chloride TS, 1 mL water to obtain 100 mL.
of ammonium hydroxide, and 50 mL of hot (maintained at Analysis: To the Standard solution and Sample solution add
7080) ammonium oxalate TS. Allow to stand for 4 h, the 4 mL of sulfomolybdic acid TS, and mix. To both
dilute with water to 200 mL, and filter. Evaporate 100 mL solutions add 0.1 mL of a freshly prepared mixture of 3 N
of the filtrate to dryness, and ignite to constant weight. hydrochloric acid and stronger acid stannous chloride TS
Acceptance criteria: The weight of the residue does not (10:1), and mix.
exceed 2 mg (0.4%). Acceptance criteria: After 10 min any color in the Sample
[NOTECalcium Gluconate labeled as not intended for use solution is not more intense than that in the Standard
in preparing injectable dosage forms is exempt from this solution (0.01%).
requirement.] [NOTECalcium Gluconate labeled as not intended for use
LIMIT OF IRON in the preparation of injectable dosage forms is exempt
Standard solutions: 0.2, 0.4, and 1.0 g/mL of iron, from this requirement.]
prepared as follows. Separately transfer 2.0, 4.0, and 10.0 Organic Impurities
mL of Standard Iron Solution, prepared as directed under PROCEDURE: LIMIT OF OXALATE
Iron 241, to 100-mL volumetric flasks, each containing [NOTEUse deionized water where water is indicated.]
1.37 g of calcium chloride, previously tested and shown to Mobile phase: Prepare a solution containing 0.0017 M
contain less than 5 ppm of iron, and dilute with 2 N sodium bicarbonate and 0.0018 M sodium carbonate in
hydrochloric acid to volume. water.
Sample solution: Transfer 1.0 g of Calcium Gluconate to a Solution A: 0.0125 M sulfuric acid
100-mL quartz glass flask, add 20 mL of 12 N nitric acid, Solution B: Dilute 1 mL of hydrochloric acid with water to
and heat to boiling until fumes are evolved. Add 0.5 mL of 1200 mL.
30% hydrogen peroxide, and heat again until fumes are Standard solution: 1.5 g/mL of sodium oxalate in Solution
evolved. Repeat this process until the volume is reduced to B
about 5 mL. Cool, add 1.0 mL of perchloric acid, and heat Sample solution: 20 mg/mL of Calcium Gluconate in
to boiling. [CautionDo not heat above 190 or evaporate Solution B. Sonicate if necessary.
to dryness because of danger of explosion.] Transfer this Chromatographic system
solution to a 25-mL volumetric flask, and dilute with 2 N (See Chromatography 621, System Suitability.)
hydrochloric acid to volume. Mode: Ion chromatography
Blank solution: Using 0.34 g of calcium chloride, Detector: Conductance
previously tested and shown to contain less than 5 ppm of Analytical column: 4-mm 25-cm; 15-m packing L12
iron, instead of Calcium Gluconate, prepare as directed Guard column: 4-mm 5-cm; 15-m packing L12
under Sample solution. Anion suppressor column: Micromembrane anion
Spectrometric conditions suppressor column connected in series with the guard and
(See Spectrophotometry and Light-Scattering 851.) analytical columns. The anion suppressor column is
Mode: Atomic absorption spectrophotometer equipped with a micromembrane that separates the Mobile
Lamp: Iron hollow-cathode phase from the Solution A flowing countercurrent to the
Flame: Airacetylene Mobile phase at a rate of about 7 mL/min.
Analytical wavelength: Iron emission line of 248.3 nm [NOTECondition the system for about 15 min with Mobile
Analysis phase at a flow rate of 2 mL/min.]
Samples: Standard solutions, Sample solution, and Blank Flow rate: 2 mL/min
solution Injection size: 50 L
Determine the absorbances of the Standard solutions and System suitability
the Sample solution, using the Blank solution as the blank Sample: Standard solution
and making deuterium background corrections. Plot the Suitability requirements
absorbances of the Standard solutions versus Column efficiency: NLT 2500 theoretical plates from the
concentration, in g/mL, of iron, and draw the straight analyte peak
line best fitting the three plotted points. From the graph Tailing factor: NMT 1.2
so obtained, determine the concentration, in g/mL, of Relative standard deviation: NMT 2.0%
iron in the Sample solution. Analysis
Calculate the concentration of iron, in ppm, in the Samples: Standard solution and Sample solution
specimen taken: Calculate the percentage of oxalate in the sample taken:
Result = F C Result: (rU/rS) (CS/CU) (Mr1/Mr2) F 100
F = monograph correction factor, 25 rU = peak response for oxalate from the Sample
C = concentration of iron in the Sample solution solution
(g/mL) rS = peak response for oxalate from the Standard
[NOTECalcium Gluconate labeled as not intended for solution
use in the preparation of injectable dosage forms is CS = concentration of sodium oxalate in the Standard
exempt from this requirement.] solution (g/mL)
CU = concentration of calcium gluconate in the IDENTIFICATION

Sample solution (mg/mL) A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 621
Mr1 = molecular weight of oxalate, 88.03 Adsorbent: 0.25-mm layer of chromatographic silica gel
Mr2 = molecular weight of sodium oxalate, 134.00 Standard solution: 10 mg/mL of USP Potassium Gluconate
F = unit conversion factor (g to mg), 0.001 RS, heating in a water bath at 60, if necessary, to dissolve
Acceptance criteria: NMT 0.01% Sample solution: 10 mg/mL from Injection in water
[NOTECalcium Gluconate labeled as not intended for use in Application volume: 5 L
the preparation of injectable dosage forms is exempt from Developing solvent system: Alcohol, ethyl acetate,
this requirement.] ammonium hydroxide, and water (5:1:1:3)
Spray reagent: Dissolve 2.5 g of ammonium molybdate in
SPECIFIC TESTS 50 mL of 2 N sulfuric acid in a 100-mL volumetric flask, add
LOSS ON DRYING 731: Dry it at 105 for 16 h: the 1.0 g of ceric sulfate, swirl to dissolve, dilute with 2 N
anhydrous form loses NMT 3.0% of its weight; the sulfuric acid to volume, and mix.
monohydrate form, where labeled as intended for use in Analysis: Develop the chromatogram until the solvent front
preparing injectable dosage forms, loses NMT 1.0% of its has moved about three-fourths of the length of the plate.
weight, and where labeled as not intended for use in Remove the plate from the chamber, and dry at 110 for 20
preparing injectable dosage forms, loses NMT 2.0% of its min. Allow to cool, and spray with the Spray reagent. Heat
weight. the plate at 110 for about 10 min.
REDUCING SUBSTANCES Acceptance criteria: The principal spot of the Sample
Sample solution: Transfer 1.0 g to a 250-mL conical flask, solution corresponds in color, size, and RF value to that of
dissolve in 20 mL of hot water, cool, and add 25 mL of the Standard solution.
alkaline cupric citrate TS. Cover the flask, boil gently for 5 B. IDENTIFICATION TESTSGENERAL, Calcium 191: It meets
min, accurately timed, and cool rapidly to room the requirements.
temperature. Sample solution: 200 mg/mL from Injection in water
Analysis: Add 25 mL of 0.6 N acetic acid, 10.0 mL of 0.1 N
iodine VS, and 10 mL of 3 N hydrochloric acid, and titrate ASSAY
with 0.1 N sodium thiosulfate VS, adding 3 mL of starch TS PROCEDURE
as the endpoint is approached. Perform a blank Sample solution: To a volume of Injection, equivalent to
determination, omitting the specimen, and note the 500 mg of calcium gluconate, add 2 mL of 3 N hydrochloric
difference in volumes required. Each mL of the difference in acid, and dilute with water to 150 mL.
volume of 0.1 N sodium thiosulfate consumed is equivalent Analysis: While stirring, preferably with a magnetic stirrer,
to 2.7 mg of reducing substances (as dextrose). add 20 mL of 0.05 M edetate disodium VS from a 50-mL
Acceptance criteria: NMT 1.0% buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of
hydroxy naphthol blue, and continue the titration to a blue
ADDITIONAL REQUIREMENTS endpoint. Each mL of 0.05 M edetate disodium is equivalent
PACKAGING AND STORAGE: Preserve in well-closed containers. to 2.004 mg of Ca.
LABELING: Label it to indicate whether it is anhydrous or Acceptance criteria: 95.0%105.0%
monohydrate. Where the quantity of calcium gluconate is
indicated in the labeling of any solution containing Calcium SPECIFIC TESTS
Gluconate, this shall be understood to be in terms of BACTERIAL ENDOTOXINS TEST 85: NMT 0.17 USP Endotoxin
anhydrous calcium gluconate (C12H22CaO14). Calcium Unit/mg of calcium gluconate
Gluconate intended for use in preparing injectable dosage PARTICULATE MATTER IN INJECTIONS 788: Meets the
forms is so labeled. Calcium Gluconate not intended for use requirements for small-volume injections
in preparing injectable dosage forms is so labeled; in PH 791: 6.08.2, except that in the case where it is
addition, it may be labeled also as intended for use in labeled as intended for veterinary use only and as containing
preparing oral dosage forms. boric acid, it is 2.54.5
USP REFERENCE STANDARDS 11 OTHER REQUIREMENTS: It meets the requirements under
USP Potassium Gluconate RS Injections 1.
Calcium Gluconate Injection preferably of Type I glass.
(Comment on this Monograph)id=m11670=Calcium Gluconate LABELING: Label the Injection to indicate its content, if any,
Injection=Ca-Chl-Monos.pdf) of added calcium salts, calculated as percentage of calcium
in the Injection. The label states the total osmolar
DEFINITION concentration in mOsmol/L. Where the contents are less
Calcium Gluconate Injection is a sterile solution of Calcium than 100 mL, or where the label states that the Injection is
Gluconate in Water for Injection. It contains NLT 95.0% and not for direct injection but is to be diluted before use, the
NMT 105.0% of the labeled amount of total Ca. The calcium label alternatively may state the total osmolar concentration
is in the form of calcium gluconate, except that a small in mOsmol/mL. The labeling indicates that the Injection
amount may be replaced with an equal amount of calcium in must be clear at the time of use, and that if crystallization
the form of Calcium Saccharate, or other suitable calcium salts, has occurred, warming may redissolve the precipitate.
for the purpose of stabilization. It may contain sodium Injection intended for veterinary use only is so labeled. If
hydroxide added for adjustment of the pH. Injection contains Boric Acid, it is so labeled.
Injection intended for veterinary use only may be prepared USP REFERENCE STANDARDS 11
from Calcium Gluconate solubilized with Boric Acid, or from USP Endotoxin RS
Glyconolactone, Boric Acid, and Calcium Carbonate. USP Potassium Gluconate RS
Calcium Gluconate Tablets UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
(Comment on this Monograph)id=m11680=Calcium Gluconate ADDITIONAL REQUIREMENTS
Tablets=Ca-Chl-Monos.pdf) PACKAGING AND STORAGE: Preserve in well-closed containers.
Calcium Gluconate Tablets contain NLT 95.0% and NMT USP Potassium Gluconate RS
105.0% of the labeled amount of calcium gluconate
(C12H22CaO14).
IDENTIFICATION Calcium Hydroxide
A. IDENTIFICATION TESTSGENERAL, Calcium 191: It meets (Comment on this Monograph)id=m11710=Calcium
the requirements. Hydroxide=Ca-Chl-Monos.pdf)
Sample solution: 20 mg/mL, made from a warm, filtered Ca(OH)2 74.09
solution of Tablets equivalent to 100 mg/mL of calcium Calcium hydroxide [1305-62-0].
gluconate in water
B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 DEFINITION
Adsorbent: 0.25-mm layer of chromatographic silica gel Calcium Hydroxide contains NLT 95.0% and NMT 100.5% of
Standard solution: 10 mg/mL of USP Potassium Gluconate Ca(OH)2.
RS, heating in a water bath at 60, if necessary, to dissolve
Sample solution: 10 mg/mL, made from a warm, filtered IDENTIFICATION
solution of Tablets equivalent to 100 mg/mL of calcium A. PROCEDURE
gluconate When mixed with three to four times its weight of water, it
Application volume: 5 L forms a smooth magma. The clear supernatant from the
Developing solvent system: Alcohol, ethyl acetate, magma is alkaline to litmus.
ammonium hydroxide, and water (5:1:1:3) B. IDENTIFICATION TESTSGENERAL, Calcium 191: The
Spray reagent: Dissolve 2.5 g of ammonium molybdate in Sample solution meets the requirements.
50 mL of 2 N sulfuric acid in a 100-mL volumetric flask, add Sample solution: Mix 1 g with 20 mL of water, and add
1.0 g of ceric sulfate, swirl to dissolve, dilute with 2 N sufficient 6 N acetic acid to dissolve the solution.
sulfuric acid to volume, and mix.
Analysis: Develop the chromatogram until the solvent front ASSAY
has moved about three-fourths of the length of the plate. PROCEDURE
Remove the plate from the chamber, and dry at 110 for 20 Sample solution: To 1.5 g of Calcium Hydroxide in a
min. Allow to cool, and spray with the Spray reagent. Heat beaker, gradually add 30 mL of 3 N hydrochloric acid. When
the plate at 110 for about 10 min. dissolved, transfer the solution to a 500-mL volumetric flask,
Acceptance criteria: The principal spot of the Sample and rinse the beaker thoroughly, adding the rinsings to the
solution corresponds in color, size, and RF value to that of flask. Dilute with water to volume, and mix. Pipet 50 mL of
the Standard solution. the solution into a suitable container, and add 100 mL of
water, 15 mL of 1 N sodium hydroxide, and 300 mg of
ASSAY hydroxy naphthol blue.
PROCEDURE Analysis: Titrate with 0.05 M edetate disodium VS to a blue
Sample solution: Transfer an equivalent to 500 mg of endpoint. Each mL of 0.05 M edetate disodium is equivalent
calcium gluconate, from finely powdered Tablets (NLT 20), to 3.705 mg of Ca(OH)2.
to a suitable crucible, and ignite, gently at first, until free Acceptance criteria: 95.0%100.5%
from carbon. Cool the crucible, add 10 mL of water, and
dissolve the residue by adding sufficient 3 N hydrochloric IMPURITIES
acid, dropwise, to achieve complete solution. Transfer the Inorganic Impurities
solution to a suitable container, and dilute with water to HEAVY METALS 231
150 mL. Sample solution: Dissolve 2.0 g in 20 mL of 3 N
Analysis: While stirring, preferably with a magnetic stirrer, hydrochloric acid, and evaporate on a steam bath to
add 20 mL of 0.05 M edetate disodium VS from a 50-mL dryness. Dissolve the residue in 20 mL of water, and filter.
buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of Dilute the filtrate with water to 40 mL. To 20 mL of the
hydroxy naphthol blue, and continue the titration to a blue resulting solution, add 1 mL of 0.1 N hydrochloric acid,
endpoint. Each mL of 0.05 M edetate disodium is equivalent then add water to make 25 mL.
to 21.52 mg of C12H22CaO14. Acceptance criteria: NMT 20 ppm
Acceptance criteria: 95.0%105.0% LIMIT OF MAGNESIUM AND ALKALI SALTS
Sample solution: 0.50 g in a mixture of 30 mL of water
PERFORMANCE TESTS and 10 mL of 3 N hydrochloric acid
DISSOLUTION 711 Analysis: Heat the solution, and boil for 1 min. Rapidly add
Medium: Water; 900 mL 40 mL of oxalic acid TS, and stir vigorously until
Apparatus 2: 50 rpm precipitation is well established. Add immediately to the
Time: 45 min warm mixture 2 drops of methyl red TS and then 6 N
Analysis: Determine the amount of C12H22CaO14 dissolved, ammonium hydroxide, dropwise, until the mixture is just
employing atomic absorption spectrophotometry at a alkaline. Cool to room temperature, transfer to a 100-mL
wavelength of about 422.8 nm on filtered portions of the graduated cylinder, dilute with water to 100 mL, mix, and
solution under test, suitably diluted with water, in allow to stand for 4 h or overnight. Filter, and to 50 mL of
comparison with a Standard solution having a known the clear filtrate in a platinum dish add 0.5 mL of sulfuric
concentration of calcium in the same Medium. acid, and evaporate the mixture on a steam bath to a small
Tolerances: NLT 75% (Q) of the labeled amount of volume. Carefully heat over a free flame to dryness, and
C12H22CaO14 continue heating to complete decomposition and
volatilization of ammonium salts. Finally, ignite the residue IMPURITIES

to constant weight. Inorganic Impurities
Acceptance criteria: The weight of the residue does not ALKALIES AND THEIR CARBONATES: A portion of it, saturated
exceed 12 mg (4.8%). with carbon dioxide and subsequently boiled, is neutral in
LIMIT OF ACID-INSOLUBLE SUBSTANCES reaction.
Sample solution: Dissolve 2.0 g in 30 mL of 4 N
hydrochloric acid, and heat to boiling. ADDITIONAL REQUIREMENTS
Analysis: Filter the mixture, wash the residue with hot PACKAGING AND STORAGE: Preserve in well-filled, tight
water, and ignite. containers, at a temperature not exceeding 25.
Acceptance criteria: The weight of the residue is NMT 10
mg (0.5%).
CARBONATE Calcium Lactate
Sample: 2 g
Analysis: Mix the Sample with 50 mL of water, and add an (Comment on this Monograph)id=m11790=Calcium
excess of 3 N hydrochloric acid to the mixture. Lactate=Ca-Chl-Monos.pdf)
Acceptance criteria: The addition of an excess of 3 N
hydrochloric acid to the mixture does not cause more than
a slight effervescence.
C6H10CaO6 xH2O (anhydrous) 218.22

Calcium Hydroxide Topical Solution Propanoic acid, 2-hydroxy-, calcium salt (2:1), hydrate;
(Comment on this Monograph)id=m11730=Calcium Hydroxide Calcium lactate (1:2) hydrate [41372-22-9].
Topical Solution=Ca-Chl-Monos.pdf) Calcium lactate (1:2) pentahydrate 308.30
[5743-47-5].
DEFINITION Anhydrous [814-80-2].
Calcium Hydroxide Topical Solution is a solution containing, in
each 100 mL, NLT 140 mg of calcium hydroxide [Ca(OH)2]. DEFINITION
Prepare Calcium Hydroxide Topical Solution as follows (see Calcium Lactate contains NLT 98.0% and NMT 101.0% of
Pharmaceutical CompoundingNonsterile Preparations 795): C6H10CaO6, calculated on the dried basis.
IDENTIFICATION
Calcium Hydroxide 3g A. IDENTIFICATION TESTSGENERAL, Calcium 191: Meets the
Purified Water 1000 mL requirements
Add the Calcium Hydroxide to 1000 mL of cool Purified Water, B. INFRARED ABSORPTION 197K
and agitate the mixture vigorously and repeatedly during 1 h.
Allow the excess calcium hydroxide to settle. Dispense only ASSAY
the clear supernatant. PROCEDURE
[NOTEThe solubility of calcium hydroxide, which varies with Sample solution: Transfer an amount of Calcium Lactate,
the temperature at which the solution is stored, is about 170 equivalent to 350 mg of C6H10CaO6, to a suitable container,
mg/100 mL at 15, and less at a higher temperature. The and dissolve in a mixture of water and 3 N hydrochloric acid
official concentration is based upon a temperature of 25.] (150:2).
The undissolved portion of the mixture is not suitable for Analysis: While stirring, preferably with a magnetic stirrer,
preparing additional quantities of Calcium Hydroxide Topical add 30 mL of 0.05 M edetate disodium VS from a 50-mL
Solution. buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of
hydroxy naphthol blue. Continue the titration to a blue
IDENTIFICATION endpoint. Each mL of 0.05 M edetate disodium is equivalent
A. It absorbs carbon dioxide from the air, and a film of to 10.91 mg of C6H10CaO6.
calcium carbonate forms on the surface of the liquid. Acceptance criteria: 98.0%101.0%
B. When heated, it becomes turbid, owing to the
separation of calcium hydroxide. IMPURITIES
C. IDENTIFICATION TESTSGENERAL, Calcium 191: Meets the Inorganic Impurities
requirements HEAVY METALS 231: NMT 20 ppm
Sample solution: Dissolve 1 g in 2.5 mL of 1 N acetic
ASSAY acid, and dilute with water to 25 mL.
PROCEDURE LIMIT OF MAGNESIUM AND ALKALI SALTS
Sample solution: To 100 mL of Topical Solution, add 50 Sample solution: Mix 1.0 g with 40 mL of water, carefully
mL of water, 15 mL of 1 N sodium hydroxide, and 300 mg add 1 mL of hydrochloric acid, and heat the solution to
of hydroxy naphthol blue. boiling.
Analysis: Titrate with 0.05 M edetate disodium VS to a blue Analysis: Rapidly add 40 mL of oxalic acid TS, and stir
endpoint. Each mL of 0.05 M edetate disodium is equivalent vigorously until precipitation is well established. Add
to 3.705 mg of Ca(OH)2. immediately to the warm mixture 2 drops of methyl red TS
Acceptance criteria: NLT 140 mg of Ca(OH)2 in each 100 and then 6 N ammonium hydroxide, dropwise, until the
mL of solution
mixture is just alkaline. Cool to room temperature, transfer ASSAY

to a 100-mL graduated cylinder, dilute with water to 100 PROCEDURE
mL, mix, and allow to stand for 4 h or overnight. Filter, Sample solution: Transfer an equivalent to 350 mg of
and to 50 mL of the clear filtrate in a platinum dish add C6H10CaO6 5H2O, from finely powdered Tablets (NLT 20),
0.5 mL of sulfuric acid. Evaporate the mixture on a steam to a suitable container, and add 150 mL of water and 2 mL
bath to a small volume. Carefully heat over a free flame to of 3 N hydrochloric acid. Stir, using a magnetic stirrer, for
dryness, and continue heating to complete decomposition 35 min.
and volatilization of ammonium salts. Finally, ignite the Analysis: While stirring, add 30 mL of 0.05 M edetate
residue to constant weight. disodium VS from a 50-mL buret. Add 15 mL of 1 N sodium
Acceptance criteria: The weight of the residue does not hydroxide and 300 mg of hydroxy naphthol blue, and
exceed 5.0 mg (1.0%). continue the titration to a blue endpoint. Each mL of 0.05
M edetate disodium is equivalent to 15.42 mg of C6H10CaO6
SPECIFIC TESTS 5H2O.
ACIDITY Acceptance criteria: 94.0%106.0%
Analysis: Titrate 20 mL of Sample solution with 0.10 N PERFORMANCE TESTS
sodium hydroxide, using phenolphthalein TS as the DISSOLUTION 711, Procedure for a Pooled Sample
indicator. Medium: Water; 500 mL
Acceptance criteria: NMT 0.50 mL is required for Apparatus 1: 100 rpm
neutralization (0.45% as lactic acid). Time: 45 min
LOSS ON DRYING 731 Analysis: Determine the amount of C6H10CaO6 5H2O
Sample: 12 g dissolved, as directed in the Assay, making any necessary
Analysis: Distribute the Sample evenly in a suitable modifications.
weighing dish to a depth of NMT 3 mm, and dry at 120 Tolerances: NLT 75% (Q) of the labeled amount of
for 4 h. C6H10CaO6 5H2O
Acceptance criteria: The pentahydrate loses between UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
22.0% and 27.0% of its weight; the trihydrate loses
between 15.0% and 20.0% of its weight; the monohydrate ADDITIONAL REQUIREMENTS
loses between 5.0% and 8.0% of its weight; and the dried PACKAGING AND STORAGE: Preserve in tight containers.
form loses NMT 3.0% of its weight. LABELING: The quantity of Calcium Lactate stated in the
VOLATILE FATTY ACID labeling is in terms of calcium lactate pentahydrate.
Sample: 500 mg
Analysis: Stir the Sample with 1 mL of sulfuric acid, and
warm. Calcium Lactobionate
Acceptance criteria: The mixture does not emit an odor of
volatile fatty acid. (Comment on this Monograph)id=m11827=Calcium
Lactobionate=Ca-Chl-Monos.pdf)
LABELING: The label indicates whether it is the dried form or
is hydrous; if the latter, the label indicates the degree of
hydration. Where the quantity of Calcium Lactate is
indicated in the labeling of any solution containing Calcium
Lactate, this shall be understood to be in terms of calcium
lactate pentahydrate (C6H10CaO6 5H2O).
USP REFERENCE STANDARDS 11 C24H42CaO24 2H2O 790.68
USP Calcium Lactate RS D-Gluconic acid, 4-O--D-galactopyranosyl-, calcium salt (2:1),
dihydrate;
Lactobionic acid, calcium salt (2:1), dihydrate;
Calcium lactobionate (1:2), dihydrate [110638-68-1].
Calcium Lactate Tablets
(Comment on this Monograph)id=m11820=Calcium Lactate DEFINITION
Tablets=Ca-Chl-Monos.pdf) Calcium Lactobionate contains NLT 96.0% and NMT 102.0% of
C24H42CaO24 2H2O.
DEFINITION
Calcium Lactate Tablets contain NLT 94.0% and NMT 106.0% IDENTIFICATION
of the labeled amount of calcium lactate (C6H10CaO6 5H2O). A. IDENTIFICATION TESTSGENERAL, Calcium 191: The
[NOTEAn equivalent amount of Calcium Lactate with less Sample solution meets the requirements.
water of hydration may be used in place of C6H10CaO6 5H2O Sample solution: 20 mg/mL
in preparing Calcium Lactate Tablets.] B. INFRARED ABSORPTION 197K
C. PROCEDURE
IDENTIFICATION Standard solution: 10 mg/mL of USP Calcium Lactobionate
A. IDENTIFICATION TESTSGENERAL, Calcium 191: Sample RS
solution meets the requirements Sample solution: 10 mg/mL
Sample solution: A filtered solution of Tablets, equivalent Chromatographic system
to 50 mg/mL of calcium lactate Adsorbent: 0.25-mm layer of chromatographic silica gel
B. IDENTIFICATION TESTSGENERAL, Lactate 191: Sample Application volume: 5 L
solution meets the requirements Developing solvent system: Alcohol, ethyl acetate,
Sample solution: A filtered solution of Tablets, equivalent ammonium hydroxide, and water (5:1:1:3)
to 50 mg/mL of calcium lactate
Spray reagent: Dissolve 2.5 g of ammonium molybdate in Calcium Levulinate

50 mL of 2 N sulfuric acid in a 100-mL volumetric flask. (Comment on this Monograph)id=m11850=Calcium
Add 1.0 g of ceric sulfate, swirl to dissolve, dilute with 2 N Levulinate=Ca-Chl-Monos.pdf)
sulfuric acid to volume, and mix.
Analysis
Apply the Samples to the plate. Dry the plate in a current
of cool air. Place the plate in a suitable chamber lined
with filter paper and previously equilibrated with the
Developing solvent system. Develop until the solvent front
has moved about three-fourths of the length of the plate.
Remove the plate from the chamber, and dry at 100 for C10H14CaO6 2H2O 306.32
20 min. Allow to cool, and spray with the Spray reagent. Pentanoic acid, 4-oxo-, calcium salt (2:1), dihydrate;
Heat the plate at 110 for about 10 min. Calcium levulinate (1:2) dihydrate [5743-49-7];
Acceptance criteria: The principal spot of the Sample Anhydrous 270.30
solution corresponds in color, size, and RF value to that of [591-64-0].
DEFINITION
ASSAY Calcium Levulinate contains NLT 97.5% and NMT 100.5% of
PROCEDURE C10H14CaO6, calculated on the dried basis.
Sample solution: Dissolve 0.8 g of Calcium Lactobionate in
a mixture of water and 3 N hydrochloric acid (150:2). IDENTIFICATION
Analysis: While stirring with a magnetic stirrer, add 15 mL A. IDENTIFICATION TESTSGENERAL, Calcium 191
of 0.05 M edetate disodium VS from a 50-mL buret. Add 15 Sample solution: 100 mg/mL
mL of 1 N sodium hydroxide and 300 mg of hydroxy Acceptance criteria: Meets the requirements of the tests
naphthol blue, and continue the titration to a blue B. PROCEDURE
endpoint. Each mL of 0.05 M edetate disodium is equivalent Sample solution: 0.5 g in 5 mL of water
to 39.53 mg of C24H42CaO24 2H2O. Analysis: To Sample solution add 5 mL of 1 N sodium
Acceptance criteria: 96.0%102.0% hydroxide, and filter. To the filtrate add 5 mL of iodine TS.
Acceptance criteria: A precipitate of iodoform is produced.
IMPURITIES
Inorganic Impurities ASSAY
HALIDES: A 1.2-g portion, tested as directed under Chloride PROCEDURE
and Sulfate 221, Chloride, shows no more turbidity than Sample solution: 600 mg of Calcium Levulinate in 150 mL
corresponds to 0.7 mL of 0.020 N hydrochloric acid of water containing 2 mL of 3 N hydrochloric acid
(0.04%). Analysis: While stirring with a magnetic stirrer, add 30 mL
SULFATE of 0.05 M edetate disodium VS from a 50-mL buret,
(See Chloride and Sulfate 221, Sulfate.) followed by 15 mL of 1 N sodium hydroxide and 300 mg of
A 2.0-g portion dissolved in boiling water shows no more hydroxy naphthol blue, and continue the titration to the
sulfate than corresponds to 1 mL of 0.020 N sulfuric acid blue endpoint. Each mL of 0.05 M edetate disodium is
(0.05%). equivalent to 13.51 mg of C10H14CaO6.
HEAVY METALS 231 Acceptance criteria: 97.5%100.5%
Sample solution: 1 g in 4 mL of 1.2 N hydrochloric acid
Add water to make 25 mL, warm gently until dissolved, IMPURITIES
and cool to room temperature. Inorganic Impurities
Acceptance criteria: NMT 20 ppm CHLORIDE AND SULFATE, Chloride 221: A 1.0-g portion
Organic Impurities shows no more chloride than corresponds to 1.0 mL of
PROCEDURE: REDUCING SUBSTANCES 0.020 N hydrochloric acid (0.07%).
Sample solution: Transfer 1.0 g to a 250-mL conical flask, CHLORIDE AND SULFATE, Sulfate 221: A 2.0-g portion shows
dissolve in 20 mL of water, and add 25 mL of alkaline no more sulfate than corresponds to 1.0 mL of 0.020 N
cupric citrate TS. Cover the flask, boil gently for 5 min, and sulfuric acid (0.05%).
cool rapidly to room temperature. Add 25 mL of 0.6 N HEAVY METALS 231: NMT 20 ppm
acetic acid, 10.0 mL of 0.1 N iodine VS, and 10 mL of 3 N Organic Impurities
hydrochloric acid. PROCEDURE: REDUCING SUGARS
Analysis: Titrate with 0.1 N sodium thiosulfate VS, adding Sample: 0.50 g
3 mL of starch TS as the endpoint is approached. Perform a Analysis: Dissolve the Sample in 10 mL of water, add 2 mL
blank determination, omitting the specimen, and note the of 3 N hydrochloric acid, boil for about 2 min, and cool.
difference in volumes required. Each mL of the difference in Add 5 mL of sodium carbonate TS, allow to stand for 5
volume of 0.1 N sodium thiosulfate consumed is equivalent min, dilute with water to 20 mL, and filter. Add 5 mL of
to 2.7 mg of reducing substances (as dextrose). the clear filtrate to 2 mL of alkaline cupric tartrate TS, and
Acceptance criteria: NMT 1.0% boil for 1 min.
Acceptance criteria: No red precipitate is formed
SPECIFIC TESTS immediately.
OPTICAL ROTATION, Specific Rotation 781S: +22.0 to
+26.5 SPECIFIC TESTS
Sample solution: 100 mg/mL MELTING RANGE OR TEMPERATURES, Class I 741: 119125
PH 791: 5.47.4, in a solution (1 in 20) PH 791: 7.08.5, in a solution (1 in 10)
LOSS ON DRYING 731: Dry it at a pressure not exceeding 5
ADDITIONAL REQUIREMENTS mm of mercury at 60 for 5 h: it loses 10.5%12.0% of its
PACKAGING AND STORAGE: Preserve in well-closed containers. weight.
USP Calcium Lactobionate RS
ADDITIONAL REQUIREMENTS Calcium Pantothenate

PACKAGING AND STORAGE: Preserve in well-closed containers. (Comment on this Monograph)id=m11910=Calcium
Pantothenate=Ca-Chl-Monos.pdf)
Calcium Levulinate Injection

(Comment on this Monograph)id=m11860=Calcium Levulinate
Injection=Ca-Chl-Monos.pdf)
DEFINITION
Calcium Levulinate Injection is a sterile solution of Calcium
Levulinate in Water for Injection. It contains NLT 95.0% and C18H32CaN2O10 476.53
NMT 105.0% of the labeled amount of calcium levulinate -Alanine, N-(2,4-dihydroxy-3,3-dimethyl-1-oxobutyl)-, calcium
(C10H14CaO6 2H2O). salt (2:1), (R)-;
IDENTIFICATION Calcium D-pantothenate (1:2) [137-08-6].
A. IDENTIFICATION TESTSGENERAL, Calcium 191 DEFINITION
Sample solution: 100 mg/mL Calcium Pantothenate is the calcium salt of the dextrorotatory
Acceptance criteria: Meets the requirements of the tests for isomer of pantothenic acid. It contains NLT 5.7% and NMT
Calcium. 6.0% of nitrogen (N), and NLT 8.2% and NMT 8.6% of
B. PROCEDURE calcium (Ca), both calculated on the dried basis.
Sample solution: 0.5 g in 5 mL of water
Analysis: Add 5 mL of 1 N sodium hydroxide, and filter. To IDENTIFICATION
the filtrate add 5 mL of iodine TS. A. INFRARED ABSORPTION 197K: Meets the requirements
Acceptance criteria: A precipitate of iodoform is produced. B. IDENTIFICATION TESTSGENERAL, Calcium 191
ASSAY Acceptance criteria: Meets the requirements
PROCEDURE
Sample solution: Transfer a volume of Injection, equivalent ASSAY
to 600 mg of calcium levulinate, to a 400-mL beaker, and CONTENT OF CALCIUM
add 2 mL of hydrochloric acid. Sample solution: 800 mg of Calcium Pantothenate in 150
Analysis: While stirring with a magnetic stirrer, add 30 mL mL of water containing 2 mL of 3 N hydrochloric acid. Add
of 0.05 M edetate disodium VS from a 50-mL buret, 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy
followed by 15 mL of 1 N sodium hydroxide and 300 mg of naphthol blue.
hydroxy naphthol blue, and continue the titration to the Analysis: Titrate with 0.05 M edetate disodium VS until the
blue endpoint. Each mL of 0.05 M edetate disodium is solution is a distinct blue in color. Each mL of 0.05 M
equivalent to 15.32 mg of C10H14CaO6.2H2O. edetate disodium is equivalent to 2.004 mg of Ca.
NITROGEN DETERMINATION, Method I 461: Proceed as
SPECIFIC TESTS directed, except to weigh 500 mg.
BACTERIAL ENDOTOXINS TEST 85: NMT 35.70 USP Acceptance criteria: 5.7%6.0%
Endotoxin Units/mg of calcium levulinate
PARTICULATE MATTER IN INJECTIONS 788: Meets the IMPURITIES
requirements for small-volume injections Inorganic Impurities
PH 791: 6.08.0 HEAVY METALS 231: NMT 20 ppm
OTHER REQUIREMENTS: It meets the requirements under Sample solution: 1.0 g in 25 mL of water
Injections 1. Organic Impurities
PROCEDURE: ORDINARY IMPURITIES 466
ADDITIONAL REQUIREMENTS Sample solution: Water
PACKAGING AND STORAGE: Preserve in single-dose containers, Standard solution: Water. Use USP Beta Alanine RS, in
preferably of Type I glass. place of USP Calcium Pantothenate RS, as the Standard.
LABELING: The label states the total osmolar concentration in Eluant: Alcohol and water (65:35)
mOsmol/L. Where the contents are less than 100 mL, or Visualization: 4
where the label states that the Injection is not for direct Acceptance criteria: NMT 1.0%
injection but is to be diluted before use, the label
alternatively may state the total osmolar concentration in SPECIFIC TESTS
mOsmol/mL. OPTICAL ROTATION, Specific Rotation 781S: +25.0 to
USP REFERENCE STANDARDS 11 +27.5
USP Endotoxin RS
Sample solution: 50 mg/mL PERFORMANCE TESTS

ALKALINITY DISSOLUTION, Procedure for a Pooled Sample 711
Sample solution: Dissolve 1.0 g in 15 mL of carbon Medium: Water; 900 mL
dioxide-free water in a small flask. Apparatus 2: 50 rpm
Analysis: As soon as the solution is complete, add 1.0 mL Time: 45 min
of 0.10 N hydrochloric acid, then add 0.05 mL of Sample solution: Sample per Dissolution 711. Dilute with
phenolphthalein TS. Medium to a concentration that is similar to that of the
Acceptance criteria: No pink color is produced within 5 s. Standard solution.
LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT Standard solution: 0.6 mg/mL of USP Calcium
5.0% of its weight. Pantothenate RS in Medium
Mobile phase: Water and phosphoric acid (1000:1)
PACKAGING AND STORAGE: Preserve in tight containers. (See Chromatography 621, System Suitability.)
USP Beta Alanine RS Detector: UV 210 nm
USP Calcium Pantothenate RS Column: 3.9-mm 15-cm; packing L1
Calcium Pantothenate Tablets System suitability
(Comment on this Monograph)id=m11940=Calcium Suitability requirements
Pantothenate Tablets=Ca-Chl-Monos.pdf) Relative standard deviation: NMT 3.0%
Analysis
DEFINITION Samples: Sample solution (filtered) and Standard solution
Calcium Pantothenate Tablets contain NLT 95.0% and NMT Calculate the quantity dissolved as a percentage of the
115.0% of the labeled amount of the dextrorotatory isomer of labeled amount of C18H32CaN2O10 in the portion of Tablets
calcium pantothenate (C18H32CaN2O10). taken:
IDENTIFICATION
A. IDENTIFICATION TESTSGENERAL, Calcium 191 Result = (rU/rS) (CS/CU ) 100
Sample solution: Digest a quantity of powdered Tablets, rU = peak response of calcium pantothenate from
equivalent to 150 mg of calcium pantothenate, with 15 mL the Sample solution
of 1 N sodium hydroxide, and filter. rS = peak response of calcium pantothenate from the
B. PROCEDURE Standard solution
Sample solution: 5 mL of the filtrate obtained in CS = concentration of the Standard solution (mg/mL)
Identification A CU = concentration of the Sample solution (mg/mL)
Analysis: Add 5 mL of 1 N hydrochloric acid and 2 drops of Acceptance criteria: NLT 75% (Q) of the labeled amount
ferric chloride TS to the Sample solution. of C18H32CaN2O10
Acceptance criteria: A strong yellow color is produced. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
CALCIUM PANTOTHENATE ASSAY 91 PACKAGING AND STORAGE: Preserve in tight containers.
Sample solution: Weigh and finely powder NLT 20 Tablets. LABELING: Label the Tablets to indicate the content of
Weigh a portion of powder, equivalent to 25 mg of calcium dextrorotatory calcium pantothenate.
pantothenate from powdered Tablets, and transfer, with the USP REFERENCE STANDARDS 11
aid of about 50 mL of water, to a 1000-mL volumetric flask. USP Calcium Pantothenate RS
Add 2 mL of 1 N acetic acid and 100 mL of sodium acetate
solution (1 in 60), then dilute with water to volume. Make
further accurate dilutions, with water, to a concentration
between 0.01 g/mL and 0.04 g/mL of calcium Racemic Calcium Pantothenate
pantothenate. (Comment on this Monograph)id=m11970=Racemic Calcium
Acceptance criteria: 95.0%115.0% Pantothenate=Ca-Chl-Monos.pdf)
OTHER COMPONENTS C18H32CaN2O10 476.53
CONTENT OF CALCIUM -Alanine, N-(2,4-dihydroxy-3,3-dimethyl-1-oxobutyl)-, calcium
Sample solution: Weigh and finely powder NLT 20 Tablets. salt (2:1), ()-;
Weigh a portion of powder, equivalent to 500 mg of Calcium DL-pantothenate (1:2) [6381-63-1].
calcium pantothenate, transfer to a suitable crucible, and
ignite, gently at first, until free from carbon. Cool the DEFINITION
crucible, add 10 mL of water, and dissolve the residue by Racemic Calcium Pantothenate is a mixture of the calcium salts
adding sufficient 3 N hydrochloric acid, dropwise, to achieve of the dextrorotatory and levorotatory isomers of pantothenic
complete solution. Transfer the solution to a suitable acid. It contains NLT 5.7% and NMT 6.0% of nitrogen (N),
container, dilute with water to 150 mL, and add 15 mL of 1 and NLT 8.2% and NMT 8.6% of calcium (Ca), both
N sodium hydroxide and 300 mg of hydroxy naphthol blue. calculated on the dried basis.
Analysis: Titrate with 0.05 M edetate disodium VS until the [NOTEThe physiological activity of Racemic Calcium
solution is deep blue in color. Each mL of 0.05 M edetate Pantothenate is approximately one-half that of Calcium
disodium is equivalent to 2.004 mg of Ca. Pantothenate.]
Acceptance criteria: 7.9%9.7% of the weight of
C18H32CaN2O10 in the Tablets, as determined by the Assay
ASSAY DEFINITION
CONTENT OF CALCIUM Dibasic Calcium Phosphate Dihydrate contains two molecules of
Sample: 800 mg of calcium pantothenate water of hydration. It contains NLT 98.0% and NMT 105.0%
Analysis: Dissolve in 150 mL of water containing 2 mL of 3 of dibasic calcium phosphate dihydrate (CaHPO4 2H2O).
N hydrochloric acid. Add 15 mL of 1 N sodium hydroxide
and 300 mg of hydroxy naphthol blue. Titrate with 0.05 M IDENTIFICATION
edetate disodium VS until the solution is a distinct blue in A. PROCEDURE
color. Each mL of 0.05 M edetate disodium is equivalent to Sample solution: Dissolve 100 mg by warming in 10 mL of
2.004 mg of Ca. 2 N hydrochloric acid.
Acceptance criteria: 8.2%8.6% Analysis: Add 2.5 mL of ammonia TS dropwise, with
NITROGEN DETERMINATION, Method I 461: Proceed as shaking, and then add 5 mL of ammonium oxalate TS.
directed, except to weigh accurately 500 mg. Acceptance criteria: A white precipitate is formed.
B. PROCEDURE
IMPURITIES Solution A: 106 mg/mL of ammonium molybdate in water
Inorganic Impurities (10%)
HEAVY METALS 231: NMT 20 ppm [NOTEPrepare before use.]
Sample solution: 1.0 g in 25 mL of water Sample solution: 100 mg of Dibasic Calcium Phosphate
Dihydrate in 5 mL of diluted nitric acid
SPECIFIC TESTS Analysis: Warm the Sample solution to 70, and add 2 mL
OPTICAL ROTATION, Specific Rotation 781S: 0.05 to of Solution A.
+0.05 Acceptance criteria: A yellow precipitate of ammonium
Sample solution: 50 mg/mL, in water phosphomolybdate is formed.
ALKALINITY: Dissolve 1.0 g in 15 mL of carbon dioxide-free
water in a small flask. As soon as solution is complete, add ASSAY
1.6 mL of 0.10 N hydrochloric acid, then add 0.05 mL of PROCEDURE
phenolphthalein TS, and mix: no pink color is produced Buffer: 53.5 g of ammonium chloride in water. Add 570
within 5 s. mL of ammonia water, stronger. Dilute with water to 1000
LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT mL.
5.0% of its weight. Sample solution: Transfer 400 mg of Dibasic Calcium
OTHER REQUIREMENTS: It responds to the Identification Phosphate Dihydrate to a 200-mL volumetric flask, dissolve
procedures under Calcium Pantothenate. in 12 mL of diluted hydrochloric acid with the aid of gentle
heat, if necessary, and dilute with water to volume.
ADDITIONAL REQUIREMENTS Analysis: Combine 20.0 mL of Sample solution with 25.0
PACKAGING AND STORAGE: Preserve in tight containers. mL of 0.02 M edetate disodium VS, 50 mL of water, and 5
LABELING: Label solutions containing it in terms of the mL of Buffer. Add 25 mg of eriochrome black Tsodium
equivalent amount of dextrorotatory calcium pantothenate. chloride. Titrate the excess edetate disodium with 0.02 M
USP REFERENCE STANDARDS 11 zinc sulfate VS. Perform a blank determination in the same
USP Calcium Pantothenate RS manner. Each mL of 0.02 M edetate disodium is equivalent
to 3.442 mg of CaHPO4 2H2O.
Dibasic Calcium Phosphate Dihydrate IMPURITIES
(Comment on this Monograph)id=m12000=Dibasic Calcium Inorganic Impurities
Phosphate Dihydrate=Ca-Chl-Monos.pdf) LOSS ON IGNITION 733: Ignite 1 g at 800 to 825 to
Pharmacopeial Discussion Group Sign-Off Document constant weight: it loses 24.5%26.5% of its weight.
CHLORIDE AND SULFATE, Chloride 221: To 0.20 g add 20 mL
Attribute JP EP USP of water and 13 mL of diluted nitric acid, and warm gently,
if necessary, until no more dissolves. Dilute to 100 mL, and
Definition + + + filter, if necessary. To 50 mL of this solution add 1 mL of
Identification A + + + silver nitrate TS: the turbidity does not exceed that
Identification B + + + produced by 0.70 mL of 0.010 N hydrochloric acid (0.25%).
CHLORIDE AND SULFATE, Sulfate 221: Dissolve 0.5 g in 5 mL
Acid-insoluble substances + + + of water and 5 mL of diluted hydrochloric acid, dilute with
Chloride + + + water to 100 mL, and filter, if necessary. To 20 mL of the
Sulfate + + + filtrate add 1 mL of diluted hydrochloric acid, and dilute
with water to 50 mL. Add 1 mL of barium chloride TS: the
Carbonate + + + turbidity does not exceed that produced by 1.0 mL of 0.010
Barium + + + N sulfuric acid (0.5%).
Loss on ignition + + + ARSENIC, Method I 211: NMT 3 ppm
Sample solution: 1.0 g in 25 mL of 3 N hydrochloric acid,
Assay + + + diluting with water to 55 mL: the resulting solution meets
the requirements of the test, the addition of 20 mL of 7 N
Legend: + will adopt and implement; - will not stipulate. sulfuric acid specified under Analysis being omitted.
Nonharmonized attributes: Packaging and storage, Heavy BARIUM: Heat to boiling 0.50 g with 10 mL of water, and
metals, Limit of fluoride, Iron add 1 mL of hydrochloric acid dropwise, stirring after each
Specific local attributes: Identification C (EP), Lead (USP), addition. Allow to cool; filter, if necessary; and to the filtrate
Description (JP) add 2 mL of potassium sulfate TS: no turbidity is produced
CaHPO4 2H2O within 10 min.
Phosphoric acid, calcium salt (1:1);
Calcium phosphate, dihydrate (1:1) [7789-77-7].
HEAVY METALS, Method I 231: NMT 30 ppm Anhydrous Dibasic Calcium Phosphate
Sample solution: Warm 1.3 g with 3 mL of 3 N (Comment on this Monograph)id=m12005=Anhydrous Dibasic
hydrochloric acid until no more dissolves, dilute with water Calcium Phosphate=Ca-Chl-Monos.pdf)
to 50 mL, and filter. Pharmacopeial Discussion Group Sign-Off Document
LIMIT OF FLUORIDE: NMT 50 ppm
Solution A: 294 mg/mL of sodium citrate dihydrate in Attribute JP EP USP
water Definition + + +
Standard stock solution: 1.1052 mg/mL of USP Sodium Identification A + + +
Fluoride RS in water
Standard solution: Transfer 20.0 mL of the Standard stock Identification B + + +
solution to a 100-mL volumetric flask containing 50 mL of Acid-insoluble substances + + +
Buffer, and dilute with water to volume. [NOTEEach mL of Chloride + + +
this solution contains 100 g of fluoride ion.]
Sample: 2 g Sulfate + + +
Sample solution: Transfer the Sample to a beaker Carbonate + + +
containing a plastic-coated stirring bar, add 20 mL of water Barium + + +
and 2.0 mL of hydrochloric acid, and stir until dissolved.
Add 50.0 mL of Solution A and sufficient water to make Loss on ignition + + +
100 mL. Assay + + +
Electrode system: Use a fluoride-specific, ion-indicating
electrode and a silversilver chloride reference electrode Legend: + will adopt and implement; - will not stipulate.
connected to a pH meter capable of measuring potentials Nonharmonized attributes: Packaging and storage, Heavy
with a minimum reproducibility of 0.2 mV (see pH 791). metals, Limit of fluoride, Iron
Standard response line: Transfer 50.0 mL of Solution A Specific local attributes: Identification C (EP), Lead (USP),
and 2.0 mL of hydrochloric acid to a beaker, and add Description (JP)
water to make 100 mL. Add a plastic-coated stirring bar, CaHPO4 136.06
insert the electrodes into the solution, stir for 15 min, and Phosphoric acid, calcium salt (1:1);
read the potential, in mV. Continue stirring, and at 5-min Calcium phosphate (1:1) [7757-93-9].
intervals add 100, 100, 300, and 500 L of Standard
solution, reading the potential 5 min after each addition. DEFINITION
Plot the logarithms of the cumulative fluoride ion Anhydrous Dibasic Calcium Phosphate contains NLT 98.0% and
concentrations (0.1, 0.2, 0.5, and 1.0 g/mL) versus NMT 103.0% of anhydrous dibasic calcium phosphate
potential, in mV. (CaHPO4).
Analysis: Rinse and dry the electrodes, insert them into
the Sample solution, stir for 5 min, and read the potential, IDENTIFICATION
in mV. From the measured potential and the Standard A. PROCEDURE
response line determine the concentration, C, in g/mL, of Sample solution: Dissolve 100 mg by warming in 10 mL of
fluoride ion in the Sample solution. 2 N hydrochloric acid.
Calculate the percentage of fluoride in the sample taken by Analysis: Add 2.5 mL of ammonia TS dropwise, with
multiplying C by 0.005. shaking, and then add 5 mL of ammonium oxalate TS.
Organic Impurities Acceptance criteria: A white precipitate is formed.
PROCEDURE 1: CARBONATE: Mix 1.0 g with 5 mL of carbon B. PROCEDURE
dioxide-free water, and immediately add 2 mL of Solution A: 106 mg/mL of ammonium molybdate in water
hydrochloric acid: no effervescence occurs. [NOTEPrepare before use.]
PROCEDURE 2: LIMIT OF ACID-INSOLUBLE SUBSTANCES Sample solution: 100 mg of sample in 5 mL of diluted
Sample solution: Dissolve 5.0 g with a mixture of 40 mL nitric acid
of water and 10 mL of hydrochloric acid by boiling gently Analysis: Warm the solution to 70, and add 2 mL of
for 5 min. Solution A.
Analysis: After cooling, collect the insoluble substance on Acceptance criteria: A yellow precipitate of ammonium
ashless filter paper, and wash with water until the last phosphomolybdate is formed.
washing does not give a reaction for chloride (no turbidity
results from the addition of silver nitrate TS). Ignite to ASSAY
incinerate completely the residue and ashless filter paper PROCEDURE
for assay at 600 50. Solution A: To 53.5 g of ammonium chloride in a 1000-mL
Acceptance criteria: The weight of the residue does not volumetric flask, add sufficient water to dissolve, followed by
exceed 10 mg: NMT 0.2% of acid-insoluble substances is 570 mL of ammonium hydroxide, and dilute with water to
found. volume.
Sample solution: 400 mg of Anhydrous Dibasic Calcium
ADDITIONAL REQUIREMENTS Phosphate into a 200-mL volumetric flask, dissolve in 12 mL
PACKAGING AND STORAGE: Preserve in well-closed containers. of 1.21M hydrochloric acid with the aid of gentle heat, if
No storage requirements specified. necessary, and dilute with water to volume.
USP Sodium Fluoride RS
Analysis: Combine 20.0 mL of Sample solution with 80.0 Analysis: Rinse and dry the electrodes, insert them into
mL of 0.02 M edetate disodium VS, Solution A, and water the Sample solution, stir for 5 min, and read the potential,
(5:1:10), and add 25 mg of eriochrome black Tsodium in mV. From the measured potential and the Standard
chloride. Titrate the excess edetate disodium with 0.02 M response line determine the concentration, C, in g/mL, of
zinc sulfate VS. Perform a blank determination in the same fluoride ion in the Sample solution.
manner. Each mL of 0.02 M edetate disodium is equivalent Calculate the percentage of fluoride in the sample taken by
to 2.721 mg of CaHPO4. multiplying C by 0.005.
Acceptance criteria: 98.0%103.0% Organic Impurities
PROCEDURE 1: CARBONATE: Mix 1.0 g with 5 mL of carbon
IMPURITIES dioxide-free water, and immediately add 2 mL of
Inorganic Impurities hydrochloric acid: no effervescence occurs.
LOSS ON IGNITION 733: It loses 6.6%8.5% of its weight. PROCEDURE 2: LIMIT OF ACID-INSOLUBLE SUBSTANCES
Sample: 1 g Sample solution: Dissolve 5.0 g in a mixture of 40 mL of
Ignition temperature range: 800825 to constant water and 10 mL of hydrochloric acid by boiling gently for
weight 5 min.
CHLORIDE AND SULFATE, Chloride 221: To 0.20 g add 20 mL Analysis: After cooling, collect the insoluble substance on
of water and 13 mL of diluted nitric acid, and warm gently, ashless filter paper, and wash with water until the last
if necessary, until no more dissolves. Dilute to 100 mL, and washing does not give a reaction for chloride (no turbidity
filter, if necessary. To 50 mL of this solution add 1 mL of results from the addition of silver nitrate TS). Ignite to
silver nitrate TS: the turbidity does not exceed that incinerate completely the residue and ashless filter paper
produced by 0.70 mL of 0.010 N hydrochloric acid (0.25%). for assay at 600 50.
CHLORIDE AND SULFATE, Sulfate 221: Dissolve 0.5 g in 5 mL Acceptance criteria: The weight of the residue does not
of water and 5 mL of diluted hydrochloric acid, dilute with exceed 10 mg: NMT 0.2% of acid-insoluble substances is
water to 100 mL, and filter, if necessary. To 20 mL of the found.
filtrate add 1 mL of diluted hydrochloric acid, and dilute
with water to 50 mL. Add 1 mL of barium chloride TS: the ADDITIONAL REQUIREMENTS
turbidity does not exceed that produced by 1.0 mL of 0.010 PACKAGING AND STORAGE: Preserve in well-closed containers.
N sulfuric acid (0.5%). No storage requirements specified.
ARSENIC, Method I 211: NMT 3 ppm USP REFERENCE STANDARDS 11
Sample solution: 1.0 g in 25 mL of 3 N hydrochloric acid, USP Sodium Fluoride RS
diluting with water to 55 mL: the resulting solution meets
the requirements of the test, the addition of 20 mL of 7 N
sulfuric acid specified under Analysis being omitted.
BARIUM: Heat to boiling 0.50 g with 10 mL of water, and Dibasic Calcium Phosphate Tablets
add 1 mL of hydrochloric acid dropwise, stirring after each (Comment on this Monograph)id=m12030=Dibasic Calcium
addition. Allow to cool; filter, if necessary; and to the filtrate Phosphate Tablets=Ca-Chl-Monos.pdf)
add 2 mL of potassium sulfate TS: no turbidity is produced
within 10 min. DEFINITION
HEAVY METALS, Method I 231: NMT 30 ppm Dibasic Calcium Phosphate Tablets contain NLT 92.5% and
Sample solution: Warm 1.3 g with 3 mL of 3 N NMT 107.5% of the labeled amount of CaHPO4 2H2O.
hydrochloric acid until no more dissolves, dilute with water [NOTEAn equivalent amount of Dibasic Calcium Phosphate
to 50 mL, and filter. with less water of hydration may be used in place of CaHPO4
LIMIT OF FLUORIDE: 50 ppm 2H2O in preparing Dibasic Calcium Phosphate Tablets.]
Solution A: 294 mg/mL of sodium citrate dihydrate in IDENTIFICATION
water A. IDENTIFICATION TESTSGENERAL, Calcium 191: Filtered
Standard stock solution: 1.1052 mg/mL of USP Sodium portion of the Sample solution from the Assay meets the
Fluoride RS in water requirements.
Standard solution: Combine 20.0 mL of Standard stock B. IDENTIFICATION TESTSGENERAL, Phosphate 191: Filtered
solution and 50 mL of Solution A in a 100-mL volumetric portion of the Sample solution from the Assay, neutralized
flask, and dilute with water to volume. [NOTEEach mL of with ammonium hydroxide, meets the requirements.
this solution contains 100 g of fluoride ion.] ASSAY
Electrode system: Use a fluoride-specific, ion-indicating PROCEDURE
electrode and a silversilver chloride reference electrode Sample solution: Transfer an equivalent to 1 g of dibasic
connected to a pH meter capable of measuring potentials calcium phosphate (dihydrate), from powdered Tablets (NLT
with a minimum reproducibility of 0.2 mV (see pH 791). 20), to a 100-mL volumetric flask containing 15 mL of
Standard response line: Transfer 50.0 mL of Solution A hydrochloric acid and 10 mL of water. Heat on a steam
and 2.0 mL of hydrochloric acid to a beaker, and add bath, with occasional mixing, to dissolve the dibasic calcium
water to make 100 mL. Add a plastic-coated stirring bar, phosphate, but not longer than 30 min. Cool, add water to
insert the electrodes into the solution, stir for 15 min, and volume, and mix. If the solution is not clear, filter,
read the potential, in mV. Continue stirring, and at 5-min discarding the first 10 mL of the filtrate.
intervals add 100, 100, 300, and 500 L of Standard Analysis: Transfer 25.0 mL of the Sample solution to a 250-
solution, reading the potential 5 min after each addition. mL beaker equipped with a magnetic stirrer. With constant
Plot the logarithms of the cumulative fluoride ion stirring, add, in the order named, 0.5 mL of
concentrations (0.1, 0.2, 0.5, and 1.0 g/mL) versus triethanolamine, 300 mg of hydroxy naphthol blue, and,
potential, in mV. from a 50-mL buret, 23 mL of 0.05 M edetate disodium VS.
Sample: 2.0 g Anhydrous Dibasic Calcium Phosphate Add 7.7 M sodium hydroxide solution until the initial red
Sample solution: Transfer the Sample to a beaker color changes to clear blue. Continue to add it dropwise
containing a plastic-coated stirring bar, add 20 mL of water until the color changes to violet, and add an additional 0.5
and 2.0 mL of hydrochloric acid, and stir until dissolved. mL. [NOTEThe pH is 12.312.5.] Continue the titration
Add 50.0 mL of Solution A and sufficient water to make dropwise with the 0.05 M edetate disodium VS to the
100 mL.
appearance of a clear blue endpoint that persists for NLT 60 Exercise care to avoid any loss of particles.] Add 35 mL of
s. Each mL of 0.05 M edetate disodium is equivalent to 0.1 N hydrochloric acid, and shake for 30 min. Centrifuge,
8.604 mg of CaHPO4 2H2O. decanting and discarding the supernatant. Repeat the
Acceptance criteria: 92.5%107.5% foregoing steps, using water instead of acid. Add 35 mL of a
sodium bicarbonate solution (15 in 1000), and shake,
PERFORMANCE TESTS venting as necessary to release any carbon dioxide liberated.
DISSOLUTION 711 Shake for 1 h, centrifuge, and decant the supernatant. Add
Medium: 0.1 N hydrochloric acid; 900 mL 35 mL of sodium bicarbonate solution (15 in 1000), and
Apparatus 2: 75 rpm shake for 1 h. Allow the tube and contents to stand
Time: 45 min overnight or until the contents have settled, and centrifuge.
Analysis: Determine the amount of CaHPO4 2H2O Withdraw the supernatant, and weigh the tube and
dissolved, employing atomic absorption spectrophotometry contents.
at a wavelength of 422.7 nm in filtered portions of the Calculate the weight of sodium bicarbonate solution
solution under test, suitably diluted with Medium if absorbed.
necessary, in comparison with a standard solution having a Acceptance criteria: NLT 35.0 g of the sodium bicarbonate
known concentration of calcium in Medium. solution is absorbed by 1.0 g of Calcium Polycarbophil,
Tolerances: NLT 75% (Q) of the labeled amount of CaHPO4 calculated on the dried basis.
2H2O
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS
LABELING: The quantity of dibasic calcium phosphate stated
in the labeling is in terms of CaHPO4 2H2O. Calcium Saccharate
(Comment on this Monograph)id=m12090=Calcium
Saccharate=Ca-Chl-Monos.pdf)
Calcium Polycarbophil
Polycarbophil=Ca-Chl-Monos.pdf)
Calcium polycarbophil [9003-97-8].
DEFINITION
Calcium Polycarbophil is the calcium salt of polyacrylic acid
cross-linked with divinyl glycol. C6H8CaO8 4H2O 320.26
D-Glucaric acid, calcium salt (1:1) tetrahydrate;
IDENTIFICATION Calcium D-glucarate (1:1), tetrahydrate [5793-89-5].
When tested as directed in the test for Absorbing Power, it
absorbs about 35 times its original weight. DEFINITION
Calcium Saccharate is the calcium salt of D-saccharic acid. It
OTHER COMPONENTS contains NLT 98.5% and NMT 102.0% of C6H8CaO8 4H2O.
CONTENT OF CALCIUM
Sample: 2 g IDENTIFICATION
Analysis: Place the Sample in a tared crucible, cover the A. IDENTIFICATION TESTSGENERAL, Calcium 191: Dissolve
crucible, leaving the lid slightly ajar, and place it in a muffle 0.2 g in 10 mL of water by the addition of 2 mL of
furnace. Heat to 600 over 2 h, increase the temperature to hydrochloric acid.
1000 over 1 h, and maintain at 1000 for 1 h. Allow to B. INFRARED ABSORPTION 197M
cool slowly. Dissolve the residue in dilute hydrochloric acid
(1 in 5), quantitatively transfer with the aid of dilute ASSAY
hydrochloric acid (1 in 5) to a 100-mL volumetric flask, and PROCEDURE
dilute with dilute hydrochloric acid (1 in 5) to volume. Pipet Sample solution: Dissolve 600 mg of Calcium Saccharate in
15 mL of this solution into a 250-mL beaker, and add, while 150 mL of water with the aid of a sufficient volume of
stirring with a magnetic stirrer, 100 mL of water, 20.0 mL of hydrochloric acid. While stirring, preferably with a magnetic
0.05 M edetate disodium VS, and 300 mg of hydroxy stirrer, add about 30 mL of 0.05 M edetate disodium VS
naphthol blue. Adjust with 1 N sodium hydroxide solution from a 50-mL buret. Add 15 mL of 1 N sodium hydroxide
to a pH of 9.09.5. Adjust with about 10 mL of 2 N sodium and 300 mg of hydroxy naphthol blue.
hydroxide to a pH of 12.4. Titrate with 0.05 M edetate Analysis: Continue the titration to a blue endpoint. Each
disodium VS to a persistent blue endpoint. Each mL of 0.05 mL of 0.05 M edetate disodium is equivalent to 16.01 mg of
M edetate disodium is equivalent to 2.004 mg of Ca. C6H8CaO8 4H2O.
Acceptance criteria: NLT 18.0% and NMT 22.0%, Acceptance criteria: 98.5%102.0%
calculated on the dried basis IMPURITIES
LOSS ON DRYING 731: Dry a sample in vacuum at 130 for CHLORIDE AND SULFATE, Chloride 221: A 0.50-g portion
4 h: it loses NMT 10.0% of its weight. dissolved in 10 mL of water by the addition of 2 mL of nitric
ABSORBING POWER acid shows no more chloride than corresponds to 0.50 mL
Sample: 250 mg of 0.020 N hydrochloric acid (0.07%).
Analysis: Transfer the Sample to a tared 50-mL centrifuge CHLORIDE AND SULFATE, Sulfate 221: A 0.50-g portion
tube fitted with a tight closure. Add 35 mL of 0.1 N dissolved in 10 mL of water by the addition of 2 mL of
hydrochloric acid to the tube, seal the tube, and shake by hydrochloric acid shows no more sulfate than corresponds
mechanical means for 30 min. Centrifuge at 2000 rpm for to 0.6 mL of 0.020 N sulfuric acid (0.12%).
20 min, and decant and discard the supernatant. [NOTE
USP 32 Official Monographs / Camphor 45
HEAVY METALS, Method II 231: NMT 20 ppm crystals with 75 mL of 25% alcohol: the crystals have a
Organic Impurities clean, white, glossy appearance. If not, recrystallize by
PROCEDURE: SUCROSE AND REDUCING SUGARS: Dissolve 0.5 g in dissolving the crystals in about 50 mL of alcohol. Add about
10 mL of water with the addition of 2 mL of hydrochloric 1 g of charcoal, stir, filter, and continue as directed above,
acid, and boil the solution for about 2 min. Cool, add 15 beginning with Add water dropwise. Dry the crystals in a
mL of sodium carbonate TS, allow to stand for 5 min, and vacuum at 50 for 2 h. [NOTEIf the melting point is low,
filter. Add 5 mL of the clear filtrate to about 2 mL of alkaline additional drying or recrystallization may be necessary.]
cupric tartrate TS, and boil for 1 min: no red precipitate is
formed immediately. ASSAY
PROCEDURE
SPECIFIC TESTS Sample solution: Boil 40.0 mL of 0.1 N hydrochloric acid
OPTICAL ROTATION, Specific Rotation 781S: +18.5 to VS with 600 mg of Calcium Undecylenate for 10 min, or
+22.5 until the undecylenic acid layer is clear, adding water, as
Sample solution: 60 mg/mL, in 4.8 N hydrochloric acid necessary, to maintain the original volume. Transfer the
that has been allowed to stand for 1 h mixture, with the aid of water, to a separator. Dilute with
water to about 75 mL, and extract with two 100-mL
ADDITIONAL REQUIREMENTS portions of solvent hexane. Wash the combined extracts
PACKAGING AND STORAGE: Preserve in well-closed containers. with water until the last washing is neutral to litmus, and
USP REFERENCE STANDARDS 11 add the washings to the original water layer. Cool, and add
USP Calcium Saccharate RS 3 drops of methyl orange TS.
Analysis: Titrate the excess hydrochloric acid with 0.1 N
sodium hydroxide VS. Perform a blank determination (see
Calcium Undecylenate Titrimetry 541, Residual Titrations).
Undecylenate=Ca-Chl-Monos.pdf) IMPURITIES
Organic Impurities
PROCEDURE: LIMIT OF FREE UNDECYLENIC ACID
Sample solution: 10g of Calcium Undecylenate in 250 mL
of solvent hexane
Mix for 2 h using a magnetic stirrer.
Analysis: Filter the Sample solution, evaporate with the aid
of a current of air to about 20 mL, and add 100 mL of
neutralized alcohol. Add 3 drops of phenolphthalein TS,
C22H38O4Ca 406.61 and titrate with 0.1 N sodium hydroxide VS.
10-Undecenoic acid, calcium (2+) salt; Acceptance criteria: NMT 0.1%
Calcium 10-undecenoate [1322-14-1].
SPECIFIC TESTS
DEFINITION LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses
Calcium Undecylenate contains NLT 98.0% and NMT 102.0% between 2.0% and 5.7% of its weight.
of C22H38O4Ca (calcium undecylenate), calculated on the dried PARTICLE SIZE DISTRIBUTION ESTIMATION BY ANALYTICAL SIEVING,
basis. Method I 786: Test in accordance with this procedure,
except use NMT 25 g and use a single No. 100 sieve that is
IDENTIFICATION to be shaken for NLT 30 min or until sifting is practically
A. A filtered solution (1 in 20) in 3 N hydrochloric acid complete: NLT 99.0% of it passes through a No. 100 sieve.
meets the requirements of the test for Identification Tests
General, Calcium 191. ADDITIONAL REQUIREMENTS
B. MELTING RANGE OR TEMPERATURE, Class Ia 741: The PACKAGING AND STORAGE: Preserve in well-closed containers.
crystals so obtained melt between 66 and 67.5.
Sample solution: Suspend calcium undecylenate 10 g in 40
mL of water in a 250-mL separator. Cautiously and slowly
add 10 mL of hydrochloric acid, while swirling. Insert the Camphor
stopper, and shake. (Comment on this Monograph)id=m12230=Camphor=Ca-Chl-
[NOTEThe separator will become quite warm, and Monos.pdf)
pressure must be carefully and frequently relieved through
the stopcock. If a curdy, white material remains after 5
min of shaking, add additional hydrochloric acid, 1 mL at
a time, and shake until a clear oily phase is formed.]
Analysis: Allow the phases to separate, drain, and discard
the bottom aqueous layer. Drain and discard the middle oily
layer, if present. Filter the top layer of undecylenic acid
through a pledget of cotton, noting the volume obtained.
To the filtrate add an equal volume of aniline. Reflux for 1 h, C10H16O 152.23
swirling the flask occasionally. Allow to cool, and pour 60 Bicyclo[2.2.1]heptane-2-one, 1,7,7-trimethyl-;
mL of alcohol through the condenser into the flask. Remove Camphor;
the flask from the condenser, add 1 g of charcoal, and stir. 2-Bornanone [76-22-2].
Filter the slurry. Add water dropwise until a few crystals
form or the solution becomes slightly cloudy. [NOTEIf too DEFINITION
much water is added, an oil will form. Add alcohol dropwise Camphor is a ketone of Cinnamomum camphora (Linne) Nees et
until the oil dissolves.] Allow the mixture to stand or Ebermaier (Fam. Lauraceae) (Natural Camphor) or produced
refrigerate until crystals are formed. Collect the crystals on a synthetically (Synthetic Camphor).
filter paper inserted in a porous glass filter funnel. Wash the
46 Camphor / Official Monographs USP 32
IMPURITIES water is removed, dry the crucible and precipitate at 80

Organic Impurities for 2 h, cool, and weigh. The weight of the precipitate so
PROCEDURE: LIMIT OF NONVOLATILE RESIDUE: Heat 2.0 g in a obtained, multiplied by 0.4581, represents the weight of
tared dish on a steam bath until sublimation is complete. C10H16O in the specimen taken.
Dry the residue at 120 for 3 h, cool, and weigh: the weight Acceptance criteria: 9.0 g11.0 g
of the residue does not exceed 1.0 mg (0.05%).
Inorganic Impurities OTHER COMPONENTS
PROCEDURE: HALOGENS ALCOHOL DETERMINATION, Method II 611: 80.0%87.0% of
Sample solution: Mix 100 mg of finely divided Camphor C2H5OH, the dilution to approximately 2% alcohol being
with 200 mg of sodium peroxide in a clean, dry, hard glass made with methanol instead of with water
test tube of about 25 mm internal diameter and 20 cm
length. Suspend the tube at an angle of about 45 by ADDITIONAL REQUIREMENTS
means of a clamp placed at the upper end, and gently PACKAGING AND STORAGE: Preserve in tight containers.
heat the tube, starting near the upper end, but not heating
the clamp, and gradually bringing the heat toward the
lower part of the tube until incineration is complete. Capecitabine
Analysis: Dissolve the residue in 25 mL of warm water,
acidify with nitric acid, and filter the solution into a (Comment on this Monograph)id=m12330=Capecitabine=Ca-
comparison tube. Wash the test tube and the filter with Chl-Monos.pdf)
two 10-mL portions of hot water, adding the washings to
the filtered solution. To the filtrate, add 0.50 mL of 0.10 N
silver nitrate, dilute with water to 50 mL, and mix.
Acceptance criteria: The turbidity does not exceed that
produced in a blank test with the same quantities of the
same reagents and 0.050 mL of 0.020 N hydrochloric acid
(0.035%).
SPECIFIC TESTS C15H22FN3O6 359.35
MELTING RANGE OR TEMPERATURE 741: 174179 Carbamic acid, [1-(5-deoxy--D-ribofuranosyl)-5-fluoro-1,2-
OPTICAL ROTATION, Specific Rotation 781S: +41 to +43, dihydro-2-oxo-4-pyrimidinyl]-, pentyl ester;
for natural Camphor Pentyl 1-(5-deoxy--D-ribofuranosyl)-5-fluoro-1,2-dihydro-2-
Sample solution: 100 mg/mL, in alcohol oxo-4-pyrimidinecarbamate [154361-50-9].
[NOTESynthetic Camphor is optically inactive.] DEFINITION
APPEARANCE OF SOLUTION: A 1 in 10 solution in solvent Capecitabine contains NLT 98.0% and NMT 102.0% of
hexane is clear. C15H22FN3O6, calculated on the anhydrous and solvent-free
ADDITIONAL REQUIREMENTS basis.
PACKAGING AND STORAGE: Preserve in tight containers, and IDENTIFICATION
avoid exposure to excessive heat. A. INFRARED ABSORPTION 197K
LABELING: Label it to indicate whether it is of natural sources Sample solution: 2 mg of sample in 300 mg of potassium
or is prepared synthetically. bromide
Camphor Spirit obtained in the Assay.
(Comment on this Monograph)id=m12240=Camphor Spirit=Ca- ASSAY
Chl-Monos.pdf) PROCEDURE
Diluent: Methanol, acetonitrile, and water (7:1:12)
DEFINITION Solution A: 0.1% mixture of acetic acid in water
Camphor Spirit is an alcohol solution containing, in each 100 Solution B: Methanol, acetonitrile, and Solution A (7:1:12)
mL, NLT 9.0 g and NMT 11.0 g of camphor (C10H16O). Solution C: Methanol, acetonitrile, and Solution A (16:1:3)
Camphor 100 g
Alcohol A sufficient quantity Time Solution B Solution C
To make 1000 mL (min) (%) (%)
0 100 0
Dissolve the camphor in about 800 mL of the alcohol, and add 5 100 0
alcohol to make 1000 mL. Filter, if necessary.
20 49 51
ASSAY 30 49 51
PROCEDURE
31 100 0
Transfer 2.0 mL of Camphor Spirit to a suitable pressure
bottle containing 50 mL of freshly prepared 40 100 0
dinitrophenylhydrazine TS. Close the pressure bottle,
immerse it in a water bath, and maintain at about 75 for System suitability solution: 0.6 g/mL of USP Capecitabine
16 h. Cool to room temperature, and transfer the contents RS, USP Capecitabine Related Compound A RS, USP
to a beaker with the aid of 100 mL of 3 N sulfuric acid. Capecitabine Related Compound B RS, and USP
Allow to stand at room temperature NLT 12 h, transfer the Capecitabine Related Compound C RS in Diluent
precipitate to a tared filter crucible, and wash with 100 mL Standard solution: 0.6 mg/mL of USP Capecitabine RS in
of 3 N sulfuric acid followed by 75 mL of cold water in Diluent
divided portions. Continue the suction until the excess
USP 32 Official Monographs / Capecitabine 47
Sample solution: 0.6 mg/mL of Capecitabine in Diluent Acceptance criteria

Chromatographic system Individual impurities: See Impurity Table.
(See Chromatography 621, System Suitability.) Total impurities: NMT 1.5%
Mode: LC
Detector: UV 250 nm with a refrigerated autosampler at Impurity Table
5
Column: 4.6-mm 25-cm; 5-m packing L1 Relative Relative Acceptance
Column temperature: 40 Retention Response Criteria,
Flow rate: 1 mL/min Name Time Factor NMT (%)
Injection size: 10 L Capecitabine related 0.18 1.05 0.3
System suitability compound A
Samples: System suitability solution and Standard solution Capecitabine related 0.19 0.81 0.3
[NOTEFor the purpose of peak identification, the compound B
approximate relative retention times are given in the
Impurity Table. The relative retention times are measured 2,3-Di-O-acetyl-5-deoxy-5- 0.36 0.89 0.1
with respect to capecitabine.] fluorocytidine
Suitability requirements 5-Deoxy-5-fluoro-N4-(2- 0.95 1.01 0.5
Resolution: NLT 1.0 between capecitabine related methyl-1-
compound A and capecitabine related compound B in the butyloxycarbonyl)cytidine +
System suitability solution 5-Deoxy-5-fluoro-N4-(3-
Tailing factor: NMT 1.5 in the Standard solution methyl-1-
Relative standard deviation: NMT 2.0% in the Standard butyloxycarbonyl)cytidine
solution Capecitabine 1.00 1.00
Analysis
Samples: Standard solution and Sample solution [1-[5-Deoxy-3-O-(5-deoxy-- 1.06 1.00 0.3
Calculate the percentage of C15H22FN3O6 in the portion of D-ribofuranosyl)--D-
Capecitabine: ribofuranosyl]-5-fluoro-2-
oxo-1,2-
Result = (rU/rS) (CS/CU) 100 dihydropyrimidin-4-yl]-
carbamic acid pentyl ester
rU = peak response from the Sample solution [1-[5-Deoxy-2-O-(5-deoxy-- 1.09 1.00 0.2
rS = peak response from the Standard solution D-ribofuranosyl)--D-
CS = concentration of USP Capecitabine RS in the ribofuranosyl]-5-fluoro-2-
Standard solution (mg/mL) oxo-1,2-
CU = concentration of Capecitabine in the Sample dihydropyrimidin-4-yl]-
solution (mg/mL) carbamic acid pentyl ester
Capecitabine related 1.11 0.91 0.3
IMPURITIES compound C
Inorganic Impurities [1-[5-Deoxy-3-O-(5-deoxy-- 1.20 1.00 0.3
RESIDUE ON IGNITION 281: NMT 0.1% D-ribofuranosyl)--D-
HEAVY METALS, Method II 231: NMT 20 ppm ribofuranosyl]-5-fluoro-2-
Organic Impurities oxo-1,2-
PROCEDURE dihydropyrimidin-4-yl]-
Diluent, Solution B, Solution C, System Suitability carbamic acid pentyl ester
solution, Standard solution, Sample solution, and
2,3-Di-O-acetyl-5-deoxy-5- 1.37 0.85 0.1
fluoro-N4-
Assay.
(pentyloxycarbonyl)cytidine
Analysis
Samples: Standard solution and Sample solution Individual unspecified 1.00 0.1
Calculate the percentage of each impurity in the portion impurity
of Capecitabine taken: Total unspecified impurities 0.5
Result = (rU/rS) (CS/CU) F 100
SPECIFIC TESTS
rU = peak response for each impurity from the OPTICAL ROTATION, Specific Rotation 781S: +96.0 to
Sample solution +100.0
rS = peak response for capecitabine from the Sample solution: 10 mg/mL, on the anhydrous and
Standard solution solvent-free basis, in methanol, at 20
CS = concentration of USP Capecitabine RS in the WATER DETERMINATION, Method Ic 921: NMT 0.3%
CU = concentration of capecitabine in the Sample ADDITIONAL REQUIREMENTS
solution (mg/mL) PACKAGING AND STORAGE: Preserve in tight containers. Store
F = relative response factor for an impurity, from at controlled room temperature.
the Impurity Table
48 Capecitabine / Official Monographs USP 32
USP REFERENCE STANDARDS 11 Impurity Table. The relative retention times are measured
USP Capecitabine RS with respect to capecitabine.]
USP Capecitabine Related Compound A RS Suitability requirements
USP Capecitabine Related Compound B RS Resolution: NLT 1.0 between capecitabine related
USP Capecitabine Related Compound C RS compound A and capecitabine related compound B in the
Tailing factor: NMT 1.5 in the Standard solution
Relative standard deviation: NMT 2.0% in the Standard
Capecitabine Tablets solution
(Comment on this Monograph)id=m12335=Capecitabine Analysis
Tablets=Ca-Chl-Monos.pdf) Samples: Standard solution and Sample solution
Calculate the percentage of C15H22FN3O6 in the portion of
DEFINITION Tablets taken:
Capecitabine Tablets contain NLT 93.0% and NMT 105.0% of
the labeled amount of capecitabine (C15H22FN3O6). Result = (rU/rS) (CS/CU) 100
IDENTIFICATION rU = peak response from the Sample solution
A. INFRARED ABSORPTION 197K rS = peak response from the Standard solution
Analytical wavelength: 15001760 cm1 CS = concentration of USP Capecitabine RS in the
Sample: Grind one Tablet to a fine powder with a mortar Standard solution (mg/mL)
and pestle. Mix 1 mg of this sample with 300 mg of CU = nominal concentration of capecitabine in the
potassium bromide. Sample solution (mg/mL)
B. The retention time of the major peak of the Sample Acceptance criteria: 93.0%105.0%
obtained in the Assay. PERFORMANCE TESTS
DISSOLUTION 711
ASSAY Medium: Water; 900 mL, degassed
Diluent: Methanol, acetonitrile, and water (7:1:12) Time: 30 min
Acetic acid solution: 0.1% mixture of acetic acid in water Sample solution: Sample per Dissolution 711. Dilute with
Solution A: Methanol, acetonitrile, and Acetic acid solution Medium to a concentration that is similar to that of the
(7:1:12) Standard solution. Pass a portion of the solution under test
Solution B: Methanol, acetonitrile, and Acetic acid solution through a 0.45-m fiberglass filter.
(16:1:3) Standard solutions:
System suitability solution: Includes 0.6 g/mL of USP For Tablets labeled to contain 150 mg: 17 mg of USP
Capecitabine RS, 0.6 g/mL of USP Capecitabine Related Capecitabine RS in 100 mL of Medium
Compound A RS, 0.6 g/mL of USP Capecitabine Related For Tablets labeled to contain 500 mg: 28 mg of USP
Compound B RS, and 0.6 g/mL of USP Capecitabine Capecitabine RS in 50 mL of Medium
Related Compound C RS in Diluent Analysis: Determine the amount of C15H22FN3O6 dissolved
Mobile phase: See the gradient table below. by employing UV absorption at the wavelength of maximum
absorbance at 304 nm (for Tablets labeled to contain 150
Time Solution A Solution B mg) and at 325 nm (for Tablets labeled to contain 500 mg)
(min) (%) (%) on portions of the Sample solution, suitably diluted with
0 100 0
Medium, if necessary, in comparison with the appropriate
Standard solution, using a 1-mm quartz cell. Calculate the
5 100 0 percentage of C15H22FN3O6 dissolved in each Tablet:
20 49 51
Result = (AU/AS) CS (V/L) 100
30 49 51
31 100 0 AU = absorbance of the Sample solution
40 100 0 AS = absorbance of the Standard solution
CS = concentration of capecitabine in the Standard
Standard solution: 0.6 mg/mL of USP Capecitabine RS in solution (mg/mL)
Diluent V = volume of medium, 900 mL
Sample solution: Equivalent to 0.6 mg/mL of Capecitabine, L = label claim (mg/Tablet)
from powdered Tablets (NLT 20), in Diluent [NOTEPass Tolerances: NLT 80% (Q) of the labeled amount of
through a PVDF 0.45-m membrane filter, and use the C15H22FN3O6
filtrate.] UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Chromatographic system IMPURITIES
(See Chromatography 621, System Suitability.) Organic Impurities
Mode: LC PROCEDURE
Detector: UV 250 nm Diluent, Acetic acid solution, Solution A, Solution B,
Column: 4.6-mm 25-cm; 5-m packing L1 System suitability solution, Mobile phase, Standard
Flow rate: 1 mL/min
Injection size: 10 L, using a refrigerated autosampler at
5
System suitability
[NOTEFor the purpose of peak identification, the
approximate relative retention times are given in the
USP 32 Official Monographs / Capreomycin 49
solution, Sample solution, and Chromatographic C25H44N14O7 652.71

system; Proceed as in the Assay. Capreomycin 1A (free base);
Analysis 3,6-Diamino-N-({(2S,5S,11S,15S,Z)-15-amino-2-
Samples: Standard solution and Sample solution (hydroxymethyl)-11-[(R)- iminohexahydropyrimidin-4-yl]
Calculate the percentage of each impurity in the portion -3,6,9,12,16-pentaoxo-8-(ureidomethylene)-1,4,7,10,13-
of Tablets taken: pentaazacyclohexadecan-5-yl}methyl)hexanamide
[37290-35-6].
Result = (rU/rS) (CS/CU) RRF 100 Capreomycin 1B (free base);
3,6-Diamino-N-({(2S,5S,11S,15S,Z)-15-amino-11-[(R)-
rU = peak response for each impurity from the iminohexahydropyrimidin-4-yl]-2-methyl-3,6,9,12,16-
Sample solution pentaoxo-8-(ureidomethylene)-1,4,7,10,13-
rS = peak response for capecitabine from the pentaazacyclohexadecan-5-yl}methyl)hexanamide
Standard solution [33490-33-4].
CS = concentration of USP Capecitabine RS in the
Standard solution (mg/mL) DEFINITION
CU = concentration of capecitabine in the portion Capreomycin Sulfate is the disulfate salt of capreomycin, a
taken for the Sample solution as determined in polypeptide mixture produced by the growth of Streptomyces
the Assay (mg/mL) capreolus, suitable for parenteral use. It has a potency
RRF = relative response factor for each impurity, from equivalent to NLT 700 g and NMT 1050 g of capreomycin/
the Impurity Table mg.
Acceptance criteria
Individual impurities: See Impurity Table. IDENTIFICATION
Total impurities: NMT 2.0% A. IDENTIFICATION TESTSGENERAL, Sulfate 191
Impurity Table
ASSAY
PROCEDURE: Proceed as directed in AntibioticsMicrobial
Relative Relative Acceptance Assays 81, for Capreomycin Sulfate.
Retention Response Criteria, Acceptance criteria: 7001050 g/mg
Capecitabine related 0.18 1.05 1.0
IMPURITIES
compound A
Capecitabine related 0.19 0.81 1.0 being moistened with 2 mL of nitric acid and 5 drops of
compound B sulfuric acid
Capecitabine 1.00 1.00 HEAVY METALS, Method II 231: NMT 30 ppm
Capecitabine related 1.11 0.91 0.5 SPECIFIC TESTS
compound C CAPREOMYCIN I CONTENT
Individual unspecified 1.00 0.1 Solution A: 0.5 mg/mL of ammonium bisulfate in water.
impurity Pass through a filter having a porosity of 0.5 m or less.
Total unspecified 0.5 Mobile phase: Methanol and Solution A (9:11)
impurities System suitability solution: 0.25 mg/mL of USP
Capreomycin Sulfate RS in water
Sample solution: 0.25 mg/mL of Capreomycin Sulfate in
ADDITIONAL REQUIREMENTS water
PACKAGING AND STORAGE: Preserve in tight containers. Store Chromatographic system
at controlled room temperature. (See Chromatography 621, System Suitability.)
USP Capecitabine RS Detector: UV 268 nm
USP Capecitabine Related Compound A RS Column: 4.6-mm 15-cm; packing L10 with a 3.5%
USP Capecitabine Related Compound B RS carbon loading
USP Capecitabine Related Compound C RS Flow rate: 1.5 mL/min
System suitability
Capreomycin Sulfate Suitability requirements
(Comment on this Monograph)id=m12358=Capreomycin Resolution: NMT 1.5 for the major peaks (capreomycin
Sulfate=Ca-Chl-Monos.pdf) IA and capreomycin IB), System suitability solution
Tailing factor: NMT 2.5 for the major peaks
(capreomycin IA and capreomycin IB), System suitability
solution
Analysis
Calculate the percentage of capreomycin I in the portion of
Capreomycin Sulfate taken:
C25H44N14O8 668.71 (rIA + rIB)/rT 100

Capreomycin, sulfate [1405-37-4].
50 Capreomycin / Official Monographs USP 32
rIA = peak area of capreomycin IA Tailing factor: NMT 2.5 for the major peaks (capreomycin
rIB = peak area of capreomycin IB IA and capreomycin IB) for the System suitability solution
rT = total of the peak areas for the peaks in the Analysis
chromatogram Sample: Sample solution
Acceptance criteria: NLT 90.0% Calculate the percentage of capreomycin I in the
PH 791: 4.57.5 Capreomycin Sulfate:
LOSS ON DRYING 731: Dry 100 mg in a vacuum at a (rIA + rIB)/rT 100
pressure not exceeding 5 mm of mercury at 100 for 4 h: it
loses NMT 10.0% of its weight. rIA = peak area for capreomycin IA
BACTERIAL ENDOTOXINS TEST 85: Where it is intended for rIB = peak area for capreomycin IB
use in preparing injectable dosage forms: NMT 0.35 USP rT = total of the peak areas for the peaks in the
Endotoxin Unit/mg of capreomycin chromatogram
OTHER REQUIREMENTS: Where the label states that Acceptance criteria: The capreomycin I content is NLT
Capreomycin Sulfate is sterile, it meets the requirements 90.0%
under Injections 1. PH 791: 4.57.5
ADDITIONAL REQUIREMENTS LOSS ON DRYING 731: Dry 100 mg in a vacuum at a
PACKAGING AND STORAGE: Preserve in tight containers. pressure not exceeding 5 mm of mercury at 100 for 4 h: it
LABELING: Where it is intended for use in preparing loses NMT 10.0% of its weight.
injectable dosage forms, the label states that it is sterile or BACTERIAL ENDOTOXINS TEST 85: NMT 0.35 USP Endotoxin
must be subjected to further processing during the Unit/mg of capreomycin
preparation of injectable dosage forms. CONSTITUTED SOLUTION: At the time of use, it meets the
USP REFERENCE STANDARDS 11 requirements for Injections 1, Constituted Solutions.
USP Capreomycin Sulfate RS OTHER REQUIREMENTS: It also meets the requirements under
USP Endotoxin RS Injections 1 and Identification TestsGeneral 191, Sulfate.
PACKAGING AND STORAGE: Preserve in Containers for Sterile
Capreomycin for Injection Solids as described under Injections 1.
(Comment on this Monograph)id=m12362=Capreomycin for USP REFERENCE STANDARDS 11
Injection=Ca-Chl-Monos.pdf) USP Capreomycin Sulfate RS
USP Endotoxin RS
DEFINITION
Capreomycin for Injection contains an amount of Capreomycin
Sulfate equivalent to NLT 90.0% and NMT 115.0% of the
labeled amount of capreomycin. Capsaicin
(Comment on this Monograph)id=m12365=Capsaicin=Ca-Chl-
ASSAY Monos.pdf)
PROCEDURE: Proceed as directed in AntibioticsMicrobial
Assays 81, for Capreomycin for injection.
IMPURITIES
being moistened with 2 mL of nitric acid and 5 drops of
sulfuric acid C18H27NO3 305.41
HEAVY METALS, Method II 231: NMT 30 ppm 6-Nonenamide, (E)-N-[(4-Hydroxy-3-methoxy-phenyl)methyl]-8-
methyl;
SPECIFIC TESTS (E)-8-Methyl-N-vanillyl-6-nonenamide [404-86-4].
CAPREOMYCIN I CONTENT
Solution A: 0.5 mg/mL of ammonium bisulfate in water. DEFINITION
Pass through a filter having a porosity of 0.5 m or less. Capsaicin contains NLT 90.0% and NMT 110.0% of the labeled
Mobile phase: Methanol and Solution A (9:11) percentage of total capsaicinoids. The content of capsaicin
System suitability solution: 0.25 mg/mL of USP (C18H27NO3) is NLT 55%, the sum of the contents of capsaicin
Capreomycin Sulfate RS in water and dihydrocapsaicin (C18H29NO3) is NLT 75%, and the
Sample solution: 0.25 mg/mL of Capreomycin for Injection content of other capsaicinoids is NMT 15%, all calculated on
in water the dried basis.
(See Chromatography 621, System Suitability.) [CAUTIONHandle Capsaicin with care. Prevent inhalation of
Mode: LC particles of it, and prevent its contact with any part of the
Detector: UV 268 nm body.]
Column: 4.6-mm 15-cm; packing L10 with a 3.5%
carbon loading IDENTIFICATION
Flow rate: 1.5 mL/min A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
Injection size: 20 L Adsorbent: 0.25-mm layer of chromatographic silica gel
System suitability mixture
Sample: System suitability solution Standard solution: 1 mg/mL of USP Capsaicin RS in
Resolution: NMT 1.5 between for the major peaks methanol
(capreomycin IA and capreomycin IB) for the System
USP 32 Official Monographs / Capsicum 51
Sample solution: 1 mg/mL of Capsaicin in methanol CU = concentration of capsaicin taken to prepare the
Application volume: 10 L Sample solution (mg/mL)
Developing solvent system: Ether and methanol (19:1) Acceptance criteria: NLT 55% of capsaicin is found, and
Spray reagent: 0.5% solution of 2,6-dibromoquinone- the sum of the percentage of capsaicin found and the
chlorimide in methanol percentage of dihydrocapsaicin found is NLT 75%.
Analysis Using the chromatograms obtained from Standard solution A
Samples: Standard solution and Sample solution and the Sample solution, calculate the percentage of other
Analysis: Develop in the solvent system until the solvent capsaicinoids in the portion of Capsaicin taken:
front has moved three-fourths of the length of the plate.
Remove the plate from the chamber, and allow it to air- (rT/rS) (CS/CU) 100
dry. Spray the plate with Spray reagent, and allow to stand
in a chamber containing ammonia fumes. Examine the rT = sum of the peak responses of the capsaicinoids,
chromatograms. other than capsaicin and dihydrocapsaicin, from
Acceptance criteria: The blue color and the RF value of the Sample solution
the principal spot from the Sample solution correspond to rS = peak response for capsaicin from Standard
those properties of the principal spot from the Standard solution A
solution. CS = concentration of USP Capsaicin RS in Standard
solution A (mg/mL)
ASSAY CU = concentration of capsaicin taken to prepare the
CONTENT OF CAPSAICIN DIHYDROCAPSAICIN, AND OTHER Sample solution (mg/mL)
CAPSAICINOIDS Acceptance criteria: NMT 15%
Mobile phase: Acetonitrile and 0.015 M phosphoric acid
(2:3) IMPURITIES
Standard solution A: 0.1 mg/mL of USP Capsaicin RS in Inorganic Impurities
methanol RESIDUE ON IGNITION 281: NMT 1.0%
Standard solution B: 0.025 mg/mL of USP
Dihydrocapsaicin RS in methanol SPECIFIC TESTS
Sample solution: 0.1 mg/mL of Capsaicin in methanol MELTING RANGE OR TEMPERATURE 741: 5766; the range
Chromatographic system between beginning and end of melting does not exceed 5.
(See Chromatography 621, System Suitability.) LOSS ON DRYING 731: Dry it in a vacuum over phosphorus
Mode: LC pentoxide at 40 for 5 h: it loses NMT 1.0% of its weight.
Detector: UV 281 nm
Column: 4.6-mm 25-cm; 5-m packing L11 ADDITIONAL REQUIREMENTS
Column temperature: 30 PACKAGING AND STORAGE: Preserve in tight containers,
[NOTEAdjust the flow rate to obtain a retention time of protected from light, and store in a cool place.
20 min for the main capsaicin peak.] LABELING: Label it to state the percentage content of total
Injection size: 20 L capsaicinoids.
Sample: Standard solution A USP Capsaicin RS
Suitability requirements USP Dihydrocapsaicin RS
Relative standard deviation: NMT 2
Analysis
Sample solution
Capsicum
Analysis: Record the chromatogram for a period of time (Comment on this Monograph)id=m12368=Capsicum=Ca-Chl-
that is twice that of the retention time of capsaicin, and Monos.pdf)
measure the areas of the responses for all of the peaks. DEFINITION
Calculate the percentage of C18H27NO3 in the portion of Capsicum is the dried ripe fruit of Capsicum frutescens Linne,
Capsaicin taken: known in commerce as African Chillies, or of Capsicum annuum
(rU/rS) (CS/CU) 100 Linne var. connoides Irish, known in commerce as Tabasco
Pepper, or Capsicum annuum var. longum Sendt, known in
rU = peak response for capsaicin from the Sample commerce as Louisiana Long Pepper, or of a hybrid between
solution the Honka variety of Japanese Capsicum and the Old Louisiana
rS = peak response for capsaicin from Standard Sport Capsicum known in commerce as Louisiana Sport
solution A Pepper (Fam. Solanaceae).
CS = concentration of USP Capsaicin RS in Standard SPECIFIC TESTS
solution A (mg/mL) BOTANIC CHARACTERISTICS
CU = concentration of capsaicin taken to prepare the Unground Capsicum: This occurs as oblong-conical fruits,
Sample solution (mg/mL) often curved (Louisiana Long Pepper), usually laterally
Calculate the percentage of C18H29NO3 in the portion of compressed, 1025 mm in length and 48 mm in diameter
Capsaicin taken: (African Chillies), or up to 15 cm in length and 2.5 cm in
(rU/rS) (CS/CU) 100 diameter (Louisiana Long Pepper), or up to 5.5 cm in length
and up to 13 mm in diameter (Louisiana Sport Pepper), or
rU = peak response for dihydrocapsaicin from the up to 4 cm in length and up to 9 mm in diameter (Tabasco
Sample solution Pepper). The fruit is two to three locular, the dissepiments
rS = peak response for dihydrocapsaicin from being united at the base to a conical, central placenta. The
Standard solution B pericarp is thin and membranous, its outer surface dark
CS = concentration of USP Dihydrocapsaicin RS in reddish brown to dusky yellowish orange, glabrous,
Standard solution B (mg/mL) shrivelled, its inner surface striate with two to three distinct
52 Capsicum / Official Monographs USP 32
longitudinal ridges representing the parietal placentae; the ADDITIONAL REQUIREMENTS

seeds are light brown to weak yellowish orange, PACKAGING AND STORAGE: Preserve in well-closed containers.
suborbicular or irregular, flattened, from 2 to 4 mm in A few drops of chloroform may be added from time to time
diameter, with a thickened edge and a prominent, pointed to prevent attack by insects.
micropyle. The calyx is gamosepalous, inferior, five-toothed, LABELING: Label each container to indicate which variety of
and sometimes attached to a long, straight peduncle. Capsicum is contained therein.
Capsicum has a characteristic odor, and is sternutatory.
Histology: The epicarp of Capsicum consists of mostly
quadrangular or rectangular cells up to 80 m in length and
up to 20 m deep, arranged in regular rows, with thickened Capsicum Oleoresin
and cutinized outer and radial walls, the surface of the (Comment on this Monograph)id=m12370=Capsicum
cuticle finely striated, the radial walls somewhat wavy Oleoresin=Ca-Chl-Monos.pdf)
(African Chillies), or of polygonal, quadrangular, triangular,
or irregular cells up to 76 m in length and up to 30.5 m DEFINITION
deep (Tabasco Pepper), or up to 125 m in length and up Capsicum Oleoresin is an alcoholic extract of the dried ripe
to 38 m deep (Louisiana Long Pepper), or up to 76 m in fruits of Capsicum annum var. minimum and small fruited
length and up to 38 m deep (Louisiana Sport Pepper), with varieties of C. frutescens (Solanaceae). It contains NLT 8.0% of
cuticularized outer and radial walls, the latter usually total capsaicins [capsaicin (C18H27NO3), dihydrocapsaicin
prominently beaded. The mesocarp consists of thin-walled (C18H29NO3), and nordihydrocapsaicin (C17H27NO3)].
parenchyma (African Chillies), or an outer hypodermis of [CAUTIONCapsicum Oleoresin is a powerful irritant, and even
tangentially elongated collenchymatous cells (Louisiana Long in minute quantities produces an intense burning sensation
Pepper and Tabasco Pepper), or of from one to three rows when it comes in contact with the eyes and tender parts of
of hypodermal cells with cuticularized walls (Louisiana Sport the skin. Care should be taken to protect the eyes and to
Pepper), a broad middle zone of thin-walled parenchyma prevent contact of the skin with Capsicum Oleoresin.]
containing yellow to red chromoplasts, oil droplets, and
elaioplasts, occasionally microcrystals, and traversed by IDENTIFICATION
vascular bundles, and an inner zone consisting of a layer of A. PROCEDURE
giant cells. The endocarp consists of a layer of elongated Sample solution: 0.5 g of Capsicum Oleoresin in 5 mL of
cells, some of them very thin-walled and containing water and 10 mL of a mixture of water, 0.2 M potassium
chromoplasts and others in large oval areas with thickened, chloride, and 0.2 N hydrochloric acid (135:50:13), and mix.
beaded, lignified walls. Epidermal cells of the seed are Add 5.0 mL of 0.5 M sodium nitrite and 5.0 mL of 0.02 M
irregular in outline and up to 342 m in length, and have sodium tungstate, and mix.
very sinuous, contorted, lignified walls, the cells from the Analysis: Heat the Sample solution at 5560 for 15 min,
edge of the seed being much thicker walled than those from allow to cool, and filter. To the filtrate add 10 mL of 1 N
the flat surface of the seed. The embryo is curved and sodium hydroxide.
embedded in the endosperm, the latter consisting of small- Acceptance criteria: A bright yellow color is produced
celled parenchyma containing fixed oil droplets and (presence of capsaicin).
aleurone grains. ASSAY
Powdered Capsicum: This is a dark orange or dark reddish PROCEDURE
orange to strong yellowish brown powder. It shows Mobile phase: Methanol and 2% acetic acid (14:11). Pass
numerous fragments of thin-walled parenchyma containing through a 0.5-m or finer porosity filter.
oil globules and orange, red, or yellow chromoplasts; and Standard solution: 0.5 mg/mL of USP Capsaicin RS in
fragments of epicarp with either striated, rectangular cells methanol. Pass a portion of this solution through a 0.2-m
arranged in parallel series (African Chillies), or with porosity filter, and use the filtrate as the Standard solution.
polygonal, triangular, or irregular cells, with or without Sample solution: 10 mg/mL of Capsicum Oleoresin in
beaded walls. The endocarp contains stone cells with slightly methanol. Pass a portion of this solution through a 0.2-m
wavy, lignified walls and broad lumina. Numerous fragments porosity filter, and use the filtrate as the Sample solution.
of spermoderm composed of stone cells are present, Chromatographic system
showing in surface view, deeply sinuate, greatly thickened (See Chromatography 621, System Suitability.)
and lignified vertical walls containing numerous pore canals. Mode: LC
Fragments of small-celled parenchyma of the endosperm Detector: UV 280 nm
containing fixed oil and aleurone grains, the latter up to 5.5 Column: 3.9-mm 30-cm; packing L1
m in diameter, are also present, as well as occasional Flow rate: 2 mL/min
fibrovascular elements and calyx tissues. Injection size: 10 L
ARTICLES OF BOTANICAL ORIGIN, Acid-Insoluble Ash 561: System suitability
NMT 1.25% Samples: Standard solution and Sample solution
ARTICLES OF BOTANICAL ORIGIN, Foreign Organic Matter 561: [NOTEThe relative retention times for
NMT 1%, other than stems and calyces, the proportion of nordihydrocapsaicin, capsaicin, and dihydrocapsaicin are
which does not exceed 3% about 0.9, 1.0, and 1.6, respectively.]
NONVOLATILE ETHER-SOLUBLE EXTRACTIVE: Dry a sample of Suitability requirements
Capsicum, taken as directed under Articles of Botanical Origin Resolution: NLT 1.2 between nordihydrocapsaicin and
561, Sampling, over phosphorus pentoxide for NLT 12 h. capsaicin in the Sample solution
Extract 2.0 g of dried Capsicum with anhydrous ether for 20 Tailing factor: NMT 2.0 in the Standard solution
h in a continuous extraction apparatus. Transfer the ether Relative standard deviation: NMT 2.0% in the Standard
solution to a tared porcelain dish, and allow it to evaporate solution
spontaneously. Dry the residue, still in the tared dish, over
phosphorus pentoxide for 18 h, and weigh to obtain the
weight of the total ether extractive. Then heat the dish
gradually up to 105, until a constant weight is obtained:
NLT 12% of nonvolatile ether-soluble extractive is found.
USP 32 Official Monographs / Captopril 53
Analysis System suitability solution: 10 g/mL each of USP

Samples: Standard solution and Sample solution Captopril RS, USP Captopril Disulfide RS, and 3-
Calculate the percentage of total capsaicins in the portion acetylthio-2-methylpropanoic acid in methanol
of Capsicum Oleoresin: Standard solution: 10 g/mL of USP Captopril Disulfide
RS in methanol [NOTEUse low-actinic glassware.]
Result = (rU/rS) (CS/CU) P 100 Sample solution: 2 mg/mL of Captopril in methanol. Use
the solution promptly. [NOTEUse low-actinic glassware.]
rU = sum of the peak areas for nordihydrocapsaicin, Chromatographic system
capsaicin, and dihydrocapsaicin from the (See Chromatography 621, System Suitability.)
rS = peak area from the Standard solution Detector: UV 220 nm
CS = concentration of USP Capsaicin RS in the Column: 3.9-mm 30-cm; packing L1
Standard solution (mg/mL) Flow rate: 1 mL/min
CU = concentration of Capsicum Oleoresin in the Injection size: 20 L
Sample solution (mg) System suitability
P = designated purity, in percentage of USP Sample: System suitability solution
Capsaicin RS [NOTEThe relative retention times for captopril, 3-
Acceptance criteria: NLT 8.0% acetylthio-2-methylpropanoic acid, and captopril
disulfide are 0.32, 0.42, and 1.0, respectively.]
PACKAGING AND STORAGE: Preserve in tight containers. Resolution: NLT 3.0 between captopril and 3-
LABELING: Label it to indicate that if separation occurs, it acetylthio-2-methylpropanoic acid
should be warmed and mixed before use. Analysis
USP REFERENCE STANDARDS 11 Samples: Standard solution and Sample solution
USP Capsaicin RS Calculate the percentage of captopril disulfide in the
portion of Captopril taken:
Captopril Result = (rU/rS) (CS/CU) 100

(Comment on this Monograph)id=m12380=Captopril=Ca-Chl- rU = peak area for captopril disulfide from the Sample
Monos.pdf) solution
rS = peak area for captopril disulfide from the
Standard solution
CS = concentration of USP Captopril Disulfide RS in
the Standard solution (g/mL)
CU = concentration of captopril in the Sample solution
(g/mL)
Acceptance criteria: NMT 1.0% of captopril disulfide.
Compare the peak responses from the Sample solution,
C9H15NO3S 217.29 excluding those of the solvent, captopril, and captopril
L-Proline, 1-[(2S)-3-mercapto-2-methyl-1-oxopropyl]-;
disulfide, with the main peak response from the Standard
1-[(2S)-3-Mercapto-2-methylpropionyl]-L-proline [62571-86-2]. solution: the peak response of each impurity does not
DEFINITION exceed 40% of the main peak response from the Standard
Captopril contains NLT 97.5% and NMT 102.0% of C9H15NO3S, solution (0.2%), and the sum of the impurity peak
calculated on the dried basis. responses does not exceed the main peak response from
the Standard solution (0.5%).
IDENTIFICATION
SPECIFIC TESTS
A. INFRARED ABSORPTION 197K OPTICAL ROTATION, Specific Rotation 781S: 125 to 134
ASSAY Sample solution: 10 mg/mL, in dehydrated alcohol
PROCEDURE LOSS ON DRYING 731: Dry it in a vacuum at 60 for 3 h: it
0.1 N Potassium iodate titrant: 3.567 mg/mL of potassium loses NMT 1.0% of its weight.
iodate, previously dried at 110 to constant weight, in water ADDITIONAL REQUIREMENTS
Sample solution: 300 mg of Captopril in 100 mL of water PACKAGING AND STORAGE: Preserve in tight containers.
in a suitable glass-stoppered flask. Add 10 mL of 3.6 N USP REFERENCE STANDARDS 11
sulfuric acid, 1 g of potassium iodide, and 2 mL of starch TS. USP Captopril RS
Analysis: Titrate with 0.1 N Potassium iodate titrant to a USP Captopril Disulfide RS
faint blue endpoint that persists for NLT 30 s. Perform a
blank determination (see Titrimetry 541) and make any
necessary correction. Each mL of 0.1 N Potassium iodate
titrant is equivalent to 21.73 mg of C9H15NO3S. Captopril Oral Solution
Acceptance criteria: 97.5%102.0% (Comment on this Monograph)id=m1599=Captopril Oral
IMPURITIES Solution=Ca-Chl-Monos.pdf)
RESIDUE ON IGNITION 281: NMT 0.2% Captopril Oral Solution contains NLT 90.0% and NMT 110.0%
HEAVY METALS, Method II 231: NMT 30 ppm of the labeled amount of captopril (C9H15NO3S). Prepare
Organic Impurities Captopril Oral Solution 0.75 mg/mL as follows. (See
PROCEDURE Pharmaceutical CompoundingNonsterile Preparations 795.
Mobile phase: 9 in 100 solution of tetrahydrofuran in See also Captopril Oral Suspension.)
methanol and 7.4 mM phosphoric acid (33:67)
54 Captopril / Official Monographs USP 32
Captopril powder 75 mg Prepare Captopril Oral Suspension 0.75 mg/mL as follows.

Vehicle for Oral Solution A sufficient quantity (See Pharmaceutical CompoundingNonsterile Preparations
(regular or sugar-free), NF 795. See also Captopril Oral Solution.)
To make 100 mL
Captopril 75 mg
Add Captopril powder and about 10 mL of Vehicle to a mortar, Vehicle: a mixture of Vehicle for Oral Solution
A sufficient
and mix. Add the Vehicle in small portions almost to volume, (regular or sugar-free), NF, and Vehicle for
quantity
and mix thoroughly after each addition. Transfer the contents Oral Suspension, NF (1:1)
of the mortar, stepwise and quantitatively, to a calibrated To make 100 mL
bottle. Add enough Vehicle to bring to final volume, and mix
well. If using Tablets, place the required number of Captopril Tablets
ASSAY in a suitable mortar, and comminute to a fine powder; or add
PROCEDURE Captopril powder to the mortar. Add about 10 mL of the
Mobile phase: Methanol and water (11:9) containing 0.5 Vehicle, and mix to a uniform paste. Add the Vehicle in small
mL of phosphoric acid portions, and mix thoroughly after each addition. Transfer the
Standard solution: 7.5 g/mL of USP Captopril RS in water contents of the mortar, stepwise and quantitatively, to a
Sample solution: Agitate the container of Oral Solution for calibrated bottle. Add sufficient Vehicle to bring to final
30 min on a rotating mixer, remove a 5 mL sample, and volume, and mix well.
store in a clear glass vial at 70 until analyzed. At the time ASSAY
of analysis, remove the sample from the freezer, allow it to PROCEDURE
reach room temperature, and mix with a vortex mixer for Mobile phase: Methanol and water (11:9) containing 0.5
30 s. Pipet 1.0 mL of the sample into a 100-mL volumetric mL of phosphoric acid
flask, and dilute with Mobile phase to volume. Standard solution: 7.5 g/mL of USP Captopril RS in water
Chromatographic system Sample solution: Agitate the container of Oral Suspension
(See Chromatography 621, System Suitability.) for 30 min on a rotating mixer, remove a 5mL sample, and
Mode: LC store in a clear glass vial at 70 until analyzed. At the time
Detector: UV 220 nm of analysis, remove the sample from the freezer, allow it to
Column: 4.6-mm 25-cm; 5-m packing L1 reach room temperature, and mix with a vortex mixer for
Flow rate: 1 mL/min 30 s. Pipet 1.0 mL of the sample into a 100-mL volumetric
Injection size: 20 L flask, and dilute with Mobile phase to volume.
[NOTEThe retention time for Captopril is 5 min.] Mode: LC
Suitability requirements Detector: UV 220 nm
Relative standard deviation: NMT 0.9% Column: 4.6-mm 25-cm; 5-m packing L1
Calculate the percentage of C9H15NO3S in the volume of System suitability
Oral Solution taken (mg/mL): Sample: Standard solution
[NOTEThe retention time for Captopril is about 5.0 min.]
Result = (rU/rS)(CS/CU) 100 Suitability requirements
rU = peak response from the Sample solution Relative standard deviation: NMT 0.9%
rS = peak response from the Standard solution Analysis
CS = concentration of USP Captopril RS in the Samples: Standard solution and Sample solution
Standard solution (g/mL) Calculate the percentage of C9H15NO3S in the volume of
CU = nominal concentration of captopril in the Sample Oral Suspension taken:
solution (g/mL) Result = (rU/rS) (CS/CU) 100
PH 791: 3.84.3 rS = peak response from the Standard solution
CS = concentration of USP Captopril RS in the
ADDITIONAL REQUIREMENTS Standard solution (g/mL)
PACKAGING AND STORAGE: Preserve in tight, light-resistant CU = nominal concentration of captopril in the Sample
containers. Store in a cold place. solution (g/mL)
LABELING: Label to state the beyond-use date. Assay acceptance criteria: 90.0%110.0%
BEYOND-USE DATE: 7 days after the day on which it was
compounded SPECIFIC TESTS
USP REFERENCE STANDARDS 11 PH 791: 3.84.3
USP Captopril RS ADDITIONAL REQUIREMENTS
containers. Store in a cold place.
Captopril Oral Suspension LABELING: Label it to state that it is to be well shaken, and
to state the beyond-use date.
(Comment on this Monograph)id=m1382=Captopril Oral BEYOND-USE DATE: 7 days after the day on which it was
Suspension=Ca-Chl-Monos.pdf) compounded
Captopril Oral Suspension contains NLT 90.0% and NMT USP Captopril RS
110.0% of the labeled amount of Captopril (C9H15NO3S).
USP 32 Official Monographs / Captopril 55
Captopril Tablets Acceptance criteria: 90.0%110.0%

(Comment on this Monograph)id=m12387=Captopril PERFORMANCE TESTS
Tablets=Ca-Chl-Monos.pdf) DISSOLUTION 711
DEFINITION [NOTECompletely deaerate the Medium to minimize
Captopril Tablets contain NLT 90.0% and NMT 110.0% of the exposure of captopril to air, and analyze the samples
labeled amount of captopril (C9H15NO3S). immediately.]
IDENTIFICATION Apparatus 1: 50 rpm
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Time: 20 min
Standard solution: 4 mg/mL, in methanol Detector: UV 205 nm
Sample solution: Equivalent to 100 mg of captopril from a Sample solutions: Sample per Dissolution 711. Dilute with
portion of powdered Tablets in 25 mL of methanol in a Medium to concentration similar to that of the Standard
conical flask solution.
Stir for 30 min using a magnetic stirrer, and centrifuge. Use Standard solution: USP Captopril RS in Medium
the clear supernatant. Tolerances: NLT 80% (Q) of the labeled amount of
Application volume: 50 L, as streaks C9H15NO3S
Developing solvent system: Toluene, glacial acetic acid, UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
and methanol (75:25:1)
Analysis: Proceed as directed in the chapter. Locate the IMPURITIES
spots on the plate by lightly spraying with a freshly Organic Impurities
prepared mixture of 1 volume of ammonium hydroxide and PROCEDURE: LIMIT OF CAPTOPRIL DISULFIDE
6 volumes of a solution of 0.04% 5,5-dithiobis(2- [NOTEProtect solutions from exposure to air. Use within 8
nitrobenzoic acid) in methanol. h of preparation.]
Mobile phase: Proceed as directed in the Assay.
ASSAY System suitability solution: 1 mg/mL of USP Captopril RS
PROCEDURE and 0.05 mg/mL of USP Captopril Disulfide RS in Mobile
[NOTEProtect solutions from exposure to air. Use within 8 phase
h of solution.] Standard solution: 0.05 mg/mL of USP Captopril Disulfide
Mobile phase: Methanol, phosphoric acid, and water RS in Mobile phase
(550:0.5:450) Sample solution: Equivalent to 25 mg of captopril from
Standard solution: 1 mg/mL of USP Captopril RS and 0.05 NLT 20 powdered Tablets in 25.0 mL of Mobile phase in a
mg/mL of USP Captopril Disulfide RS in Mobile phase centrifuge tube
Sample solution: 1 mg/mL of captopril from NLT 20 Sonicate for 15 min, and centrifuge. Use the clear
Tablets in Mobile phase to fill the flask to about half of its supernatant.
capacity. Chromatographic system
Sonicate for 15 min. Dilute with Mobile phase to volume, Mode: LC
shake by mechanical means for 15 min, and filter. Detector: UV 220 nm
Chromatographic system Column: 4.6-mm 25-cm; packing L1 with about 15%
(See Chromatography 621, System Suitability.) hydrocarbon load
Mode: LC Flow rate: 1 mL/min
Column: 4.6-mm 25-cm; packing L1 with about 15% System suitability
hydrocarbon load Samples: System suitability solution and Standard solution
Flow rate: 1 mL/min [NOTEThe relative retention times for captopril and
Injection size: 20 L captopril disulfide are 0.5 and 1.0, respectively.]
Sample: Standard solution Resolution: NLT 2.0 between captopril and captopril
[NOTEThe relative retention times for captopril and disulfide in the System suitability solution
captopril disulfide are 0.5 and 1.0, respectively.] Relative standard deviation: NMT 2.0%
Suitability requirements Analysis: Separately inject equal volumes of the Standard
Resolution: NLT 2.0 between captopril and captopril solution and the Sample solution into the chromatograph,
disulfide record the chromatograms, and measure the responses for
Relative standard deviation: NMT 2.0% the major peaks.
Analysis: Separately inject equal volumes of the Standard Calculate the percentage of captopril disulfide in the
solution and the Sample solution into the chromatograph, portion of Tablets taken:
record the chromatograms, and measure the responses for
the major peaks. Result = (rU/rS) (CS/CU) 100
Calculate the percentage of C9H15NO3S in the portion of
Tablets taken: rU = peak response for captopril disulfide from the
Sample solution
Result = (rU/rS) (CS/CU) 100 rS = peak response for captopril disulfide from the
Standard solution
rU = peak response from the Sample solution CS = concentration of USP Captopril Disulfide RS in
rS = peak response from the Standard solution the Standard solution (mg/mL)
CS = concentration of USP Captopril RS in the CU = nominal concentration of captopril in the Sample
Standard solution (mg/mL) solution (mg/mL)
CU = nominal concentration of captopril in the Sample
solution (mg/mL)
56 Captopril / Official Monographs USP 32
Acceptance criteria: NMT 3.0% CU = nominal concentration of captopril and

hydrochlorothiazide in the Sample solution
PACKAGING AND STORAGE: Preserve in tight containers. Acceptance criteria: 90.0%110.0%
USP Captopril RS PERFORMANCE TESTS
USP Captopril Disulfide RS DISSOLUTION 711
Apparatus 1: 50 rpm
Times: 20 min for captopril; 30 min for hydrochlorothiazide
Captopril and Hydrochlorothiazide Analysis: Determine the amounts of C9H15NO3S and
Tablets C7H8ClN3O4S2 dissolved, using the procedure set forth in the
Assay. Use filtered portions of the solution under test,
(Comment on this Monograph)id=m12398=Captopril and making any necessary volumetric adjustments with Medium,
Hydrochlorothiazide Tablets=Ca-Chl-Monos.pdf) and compare with a Standard solution having known
DEFINITION concentrations of USP Captopril RS and USP
Captopril and Hydrochlorothiazide Tablets contain NLT 90.0% Hydrochlorothiazide RS in the same Medium.
and NMT 110.0% of the labeled amounts of captopril Acceptance criteria: NLT 80% (Q) of the labeled amount
(C9H15NO3S) and hydrochlorothiazide (C7H8ClN3O4S2). of C9H15NO3S and NLT 60% (Q) of the labeled amount of
C7H8ClN3O4S2
IDENTIFICATION UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
The retention time of the Sample solution corresponds to that
of the Standard solution, as obtained in the Assay. IMPURITIES
Organic Impurities
ASSAY PROCEDURE 1: LIMIT OF CAPTOPRIL DISULFIDE
PROCEDURE Mobile phase: Methanol, phosphoric acid, and water
Mobile phase: Methanol, phosphoric acid, and water (450:0.5:550)
(250:0.5:750) System suitability solution: 0.0075 mg/mL of USP
System suitability solution: 0.3 mg/mL each of USP Captopril RS and USP Hydrochlorothiazide RS, and 0.015
Captopril RS, USP Hydrochlorothiazide RS, and USP mg/mL of USP Captopril Disulfide RS, in Mobile phase
Benzothiadiazine Related Compound A RS in Mobile phase Standard solution: 15 g/mL of USP Captopril Disulfide
Standard solution: 0.3 mg/mL of USP Hydrochlorothiazide RS in Mobile phase
RS and about 0.3J mg/mL of USP Captopril RS, J being the Sample solution: Equivalent to 25 mg of captopril from
ratio of the labeled amount of captopril (mg) to the labeled NLT 20 powdered Tablets
amount of hydrochlorothiazide/Tablet in Mobile phase (mg) Add about 20 mL of Mobile phase, and sonicate for 15 min,
Sample solution: Equivalent to 15 mg of with occasional shaking. Dilute with Mobile phase to
hydrochlorothiazide from NLT 20 powdered Tablets, in volume, and centrifuge, made to 50 mL. Use the clear
Mobile phase supernatant.
Sonicate for 15 min with occasional shaking, and centrifuge, Chromatographic system
made to 50 mL. (See Chromatography 621, System Suitability.)
Chromatographic system Mode: LC
(See Chromatography 621, System Suitability.) Detector: UV 210 nm
Mode: LC Column: 4.6-mm 30-cm; packing L11
Detector: UV 210 nm Flow rate: 2 mL/min
Column: 4.6-mm 30-cm; packing L11 Injection size: 20 L
Flow rate: 1.5 mL/min System suitability
Injection size: 20 L Samples: System suitability solution and Standard solution
System suitability [NOTEThe relative retention times for captopril and
Samples: System suitability solution and Standard solution captopril disulfide are 0.3 and 1.0, respectively.]
[NOTEThe relative retention times for benzothiadiazine Suitability requirements
related compound A, hydrochlorothiazide, and captopril Resolution: NLT 4.0 between captopril and captopril
are 0.4, 0.5, and 1.0, respectively.] disulfide. Both peaks are resolved from
Suitability requirements hydrochlorothiazide, System suitability solution.
Resolution: NLT 1.7 between the void volume and Relative standard deviation: NMT 3.0%, Standard
benzothiadiazine related compound; NLT 1.8 between solution
benzothiadiazine related compound A and Analysis
hydrochlorothiazide; and NLT 2.0 between captopril and Samples: Standard solution and Sample solution
hydrochlorothiazide Calculate the percentage of captopril disulfide in the
Relative standard deviation: NMT 3.0% portion of Tablets taken:
Analysis
Samples: Standard solution and Sample solution Result = (rU/rS) (CS/CU) 100
Calculate the percentage of C9H15NO3S and C7H8ClN3O4S2 in
the portion of Tablets taken: rU = captopril disulfide peak response from the
Sample solution
Result = (rU/rS) (CS/CU) 100 rS = captopril disulfide peak response from the
Standard solution
rU = peak response of the corresponding analyte from CS = concentration of USP Captopril Disulfide RS in
the Sample solution the Standard solution (g/mL)
rS = peak response of the corresponding analyte from CU = nominal concentration of captopril disulfide in
the Standard solution the Sample solution, based on the labeled
CS = concentration of the appropriate USP Reference amount/Tablet (g/mL)
USP 32 Official Monographs / Carbachol 57
Acceptance criteria: NMT 3.0% perceptible when the mixture cools. Decant the supernatant,
PROCEDURE 2: LIMIT OF BENZOTHIADIAZINE RELATED COMPOUND and add to the precipitate 3 mL of 3 N hydrochloric acid.
A Acceptance criteria: Effervescence is produced.
Mobile phase, System suitability solution, and C. IDENTIFICATION TESTSGENERAL, Calcium 191: 50 mg/mL
Chromatographic system: Proceed as directed in the solution
Assay. D. PROCEDURE
Standard solution: 10 g/mL of USP Benzothiadiazine Sample solution: 100 mg/mL, in water
Related Compound A RS in Mobile phase Analysis: To 1 mL of the Sample solution, add 3 mL of gold
Sample solution: Use the Sample solution as directed in chloride solution (1 in 10): a precipitate of yellow crystals of
the Assay. the aurichloride is formed. The precipitate, upon
Analysis recrystallization from 5 mL of hot water, separates in
Samples: Standard solution and Sample solution glistening scale-like crystals. Dry at 105 for 1 h.
Calculate the percentage of benzothiadiazine related Acceptance criteria: The crystals melt between 183 and
compound A in the portion of Tablets taken: 185.
PROCEDURE
rU = peak response for benzothiadiazine related Sample: 400 mg of Carbachol
compound A from the Sample solution Analysis: Dissolve the Sample in a mixture of 10 mL of
rs = peak response for benzothiadiazine related glacial acetic acid and 10 mL of mercuric acetate TS. Add 2
compound A from the Standard solution drops of crystal violet TS, and titrate with 0.1 N perchloric
CS = concentration of USP Benzothiadiazine Related acid VS. Perform a blank determination, and make any
Compound A RS in the Standard solution necessary correction (see Titrimetry 541). Each mL of 0.1 N
(g/mL) perchloric acid is equivalent to 18.27 mg of C6H15ClN2O2.
CU = nominal concentration of benzothiadiazine in the Acceptance criteria: 99.0%101.0%
Sample solution, based on the labeled
amount/Tablet (g/mL) IMPURITIES
Acceptance criteria: NMT 1.0% Inorganic Impurities
PACKAGING AND STORAGE: Preserve in tight containers. PROCEDURE: ORDINARY IMPURITIES 466
USP REFERENCE STANDARDS 11 Standard solvent and Sample solvent: Methanol and
USP Benzothiadiazine Related Compound A RS water (4:1)
USP Captopril RS Eluant: Alcohol
USP Captopril Disulfide RS Visualization: 16
USP Hydrochlorothiazide RS
SPECIFIC TESTS
MELTING RANGE OR TEMPERATURE 741: 200204, with
some decomposition
Carbachol LOSS ON DRYING 731: Dry 1 g at 105 for 2 h: it loses NMT
(Comment on this Monograph)id=m12480=Carbachol=Ca-Chl- 2.0% of its weight.
Monos.pdf)
USP Carbachol RS
Carbachol Intraocular Solution

C6H15ClN2O2 182.65 (Comment on this Monograph)id=m12490=Carbachol
Ethanaminium, 2-[(aminocarbonyl)oxy]-N, N,N-trimethyl-, Intraocular Solution=Ca-Chl-Monos.pdf)
chloride;
Choline chloride, carbamate [51-83-2]. DEFINITION
Carbachol Intraocular Solution is a sterile solution of Carbachol
DEFINITION in an aqueous medium. It contains NLT 90.0% and NMT
Carbachol contains NLT 99.0% and NMT 101.0% of 115.0% of the labeled amount of carbachol (C6H15ClN2O2). It
C6H15ClN2O2, calculated on the dried basis. contains no preservatives or antimicrobial agents.
A. PROCEDURE A. PROCEDURE
Sample solution: 1 mg/mL Solution A: 2 mg/mL of hexanitrodiphenylamine in 0.1 N
Analysis: To 5 mL of Sample solution, add 5 mL of sodium hydroxide
ammonium reineckate solution (1 in 30), and shake Sample solution: 100 g/mL of carbachol from Intraocular
vigorously for 1 min. Solution in water
Acceptance criteria: A red precipitate is formed; it is Analysis: Transfer 5 mL of the Sample solution to a 125-mL
soluble in acetone. separator. To another separator, add 5 mL of water to
B. PROCEDURE provide a blank. To each separator, add 1.0 mL of 1 N
Sample solution: 50 mg/mL in alcoholic potassium sodium hydroxide and 2.0 mL of Solution A. Mix, add 15 mL
hydroxide TS of methylene chloride to each separator, shake for 1 min,
Analysis: Boil 10 mL of Sample solution gently for 12 min: and allow the layers to separate.
a white precipitate is formed, and an amine odor is
58 Carbachol / Official Monographs USP 32
Acceptance criteria: A deep amber color is produced in the Carbachol Ophthalmic Solution
methylene chloride layer from the Sample solution. (Comment on this Monograph)id=m12500=Carbachol
ASSAY Ophthalmic Solution=Ca-Chl-Monos.pdf)
Solution A: Dilute 1 volume of sodium hypochlorite Carbachol Ophthalmic Solution is a sterile solution of Carbachol
solution with water to 15 volumes, allow to stand for 30 in an isotonic, aqueous medium. It contains NLT 95.0% and
min, and mix equal volumes of the resulting solution and 1 NMT 105.0% of the labeled amount of carbachol
N sodium hydroxide. [NOTEPrepare fresh daily.] (C6H15ClN2O2). It may contain suitable preservatives and
Standard solution: 100 g/mL of USP Carbachol RS in antimicrobial agents.
water
Sample solution: 100 g/mL of carbachol from a volume IDENTIFICATION
of Intraocular Solution in water A. PROCEDURE
Blank: Water Sample solution: 1 mg/mL of carbachol from Ophthalmic
Spectrometric conditions Solution in water
Mode: UV-Vis Analysis: Add 5 mL of ammonium reineckate solution (1 in
Cell: 1 cm 30), and shake vigorously for 1 min.
Analytical wavelength: 590 nm Acceptance criteria: A red precipitate is formed; it is
Analysis soluble in acetone.
Samples: Standard solution, Sample solution, and Blank B. PROCEDURE
Transfer 2.0-mL portions of the Standard solution, Sample Sample solution: Evaporate the equivalent of 500 mg of
solution, and Blank to separate 50-mL conical flasks. To carbachol from a volume of Ophthalmic Solution, on a
each flask, add 1.0 mL of 0.1 N hydrochloric acid, and steam bath, to dryness.
mix. Treat each as follows. Add 4.0 mL of Solution A, Analysis: To the residue, add 10 mL of alcoholic potassium
rinsing the inner walls of the flask with small portions of hydroxide TS, and boil gently for 12 min: a white
water. Mix, and allow to stand for 15 min. Add 2.0 mL of precipitate is formed, and an amine odor is perceptible
5 mg/mL phenol solution, rinsing the walls of the flask when the mixture cools. Decant the supernatant, and add to
with the solution and with additional small portions of the precipitate 3 mL of 3 N hydrochloric acid.
water. Mix, and allow to stand for 5 min. Add 2.0 mL of Acceptance criteria: Effervescence is produced.
3.5 N hydrochloric acid, washing the sides of the flask C. PROCEDURE
upon addition. Rinse the flask sparingly with 0.1 N Sample solution: Evaporate the equivalent of 100 mg of
hydrochloric acid to ensure complete acidification of all carbachol from Ophthalmic Solution, on a steam bath, to
contents, and mix. Add 1.0 mL of 3 mg/mL potassium dryness.
iodide solution, mix, and allow to stand for 5 min. Add Analysis: To the residue, add 3 mL of gold chloride solution
3.0 mL of starch TS, mix, transfer the solutions to 50-mL (1 in 10): a precipitate of yellow crystals of the aurichloride
volumetric flasks with the aid of several small portions of is formed. The precipitate, upon recrystallization from 5 mL
water, and dilute each solution with water to volume. of hot water, separates in glistening scale-like crystals. Dry at
Concomitantly determine the absorbances of the solutions 105 for 1 h.
from the Sample solution and the Standard solution against Acceptance criteria: The crystals melt between 183and
the Blank. 185.
Calculate the percentage of C6H15ClN2O2 in each mL of
Intraocular Solution taken: ASSAY
PROCEDURE
Result = (AU/AS) (CS/CU) 100 Solution A: Dilute 1 volume of sodium hypochlorite TS
with water to 15 volumes, allow to stand for 30 min, and
AU = absorbance of the Sample solution mix equal volumes of the resulting solution and 1 N sodium
AS = absorbance of the Standard solution hydroxide. [NOTEPrepare fresh daily. ]
CS = concentration of USP Carbachol RS in the Standard solution: 100 g/mL of USP Carbachol RS in
Standard solution (g/mL) water
CU = concentration of the Sample solution (g/mL) Sample solution: 100 g/mL of carbachol from a volume
Acceptance criteria: 90.0%115.0% of Ophthalmic Solution in water
Blank: Water
SPECIFIC TESTS Spectrometric conditions
STERILITY TESTS 71: Meets the requirements Mode: UV-Vis
PH 791: 5.07.5 Cell: 1 cm
ADDITIONAL REQUIREMENTS Analytical wavelength: 590 nm
PACKAGING AND STORAGE: Preserve in tight containers, at Analysis
controlled room temperature, and protect from freezing. Samples: Standard solution, Sample solution, and Blank
LABELING: Label it to indicate that it is for single-dose Transfer 2.0-mL portions of the Standard solution, Sample
intraocular use only, and that the unused portion is to be solution, and Blank to separate 50-mL conical flasks. To
discarded. each flask, add 1.0 mL of 0.1 N hydrochloric acid, and
USP REFERENCE STANDARDS 11 mix. Treat each as follows. Add 4.0 mL of Solution A,
USP Carbachol RS rinsing the inner walls of the flask with small portions of
USP 32 Official Monographs / Carbamazepine 59
water, mix, and allow to stand for 15 min. Add 2.0 mL of Related Compound A RS from System suitability stock solution
5 mg/mL phenol solution, rinsing the walls of the flask in methanol and water (1:1)
with the solution and with additional small portions of Standard stock solution: 2 mg/mL of USP Carbamazepine
water. Mix, and allow to stand for 5 min. Add 2.0 mL of RS in methanol
3.5 N hydrochloric acid, washing the sides of the flask Standard solution: 0.2 mg/mL of USP Carbamazepine RS
upon addition. Rinse the flask sparingly with 0.1 N from Standard stock solution in methanol and water (1:1)
hydrochloric acid to ensure complete acidification of all Sample stock solution: 2 mg/mL of Carbamazepine in
contents, and mix. Add 1.0 mL of 3 mg/mL potassium methanol
iodide solution, mix, and allow to stand for 5 min. Add Sample solution: 0.2 mg/mL of Carbamazepine from
3.0 mL of starch TS, mix, transfer the solutions to 50-mL Sample stock solution in methanol and water (1:1)
volumetric flasks with the aid of several small portions of Chromatographic system
water, and dilute each solution with water to volume. (See Chromatography 621, System Suitability.)
Concomitantly determine the absorbances of the solutions Mode: LC
from the Sample solution and the Standard solution against Detector: UV 230 nm
the Blank. Column: 4.6-mm 25-cm; packing L10
Calculate the percentage of C6H15ClN2O2 in each mL of Flow rate: 1.5 mL/min
Ophthalmic Solution taken: Injection size: 20 L
System suitability
Result = (AU/AS) (CS/CU) 100 Samples: System suitability solution and Standard solution
AU = absorbance of the Sample solution Resolution: NLT 1.70 between carbamazepine related
AS = absorbance of the Standard solution compound A and carbamazepine from the System
CS = concentration of USP Carbachol RS in the suitability solution
Standard solution (g/mL) Relative standard deviation: NMT 2.0% from the
CU = concentration of the Sample solution (g/mL) Standard solution
SPECIFIC TESTS Calculate the percentage of C15H12N2O in the portion of
STERILITY TESTS 71: Meets the requirements Carbamazepine taken:
PH 791: 5.07.0
USP Carbachol RS CS = concentration of USP Carbamazepine RS in the
CU = concentration of Carbamazepine in the Sample
Carbamazepine solution (mg/mL)
(Comment on this
Monograph)id=m12530=Carbamazepine=Ca-Chl-Monos.pdf) IMPURITIES
RESIDUE ON IGNITION 281: NMT 0.1%, a 2.0-g sample
being used
CHLORIDE AND SULFATE, Chloride 221: Boil 1.0 g with 20.0
mL of water for 10 min, cool, again adjust the volume, and
filter: a 10.0-mL portion of the filtrate shows no more
chloride than corresponds to 0.10 mL of 0.020 N
hydrochloric acid (0.014%).
C15H12N2O 236.27 HEAVY METALS, Method II 231: NMT 10 ppm
Dibenz[b,f]azepine, 5-carboxamide; Organic Impurities
5H-Dibenz[b,f]azepine-5-carboxamide [298-46-4]. PROCEDURE
DEFINITION Mobile phase and System suitability solution: Proceed as
Carbamazepine contains NLT 98.0% and NMT 102.0% of directed in the Assay.
C15H12N2O, calculated on the dried basis. Standard stock solution: 0.02 mg/mL of USP
Carbamazepine RS, 0.02 mg/mL of USP Carbamazepine
IDENTIFICATION Related Compound A RS, and 0.02 mg/mL of USP
A. INFRARED ABSORPTION 197M Carbamazepine Related Compound B RS in methanol
Standard solution: Dilute 5.0 mL of Standard stock
ASSAY solution with methanol and water (1:1) to 100.0 mL.
PROCEDURE Sample stock solution: 2 mg/mL of carbamazepine in
Mobile phase: Methanol, tetrahydrofuran, and water methanol
(12:3:85). Add 0.22 mL of formic acid to each L, mix, and Sample solution: Transfer 25.0 mL of Sample stock solution
then add 0.5 mL of triethylamine/L. to a 50-mL volumetric flask. Add 20 mL of water, shake,
System suitability stock solution: 0.1 mg/mL of USP allow to cool, and dilute with water to volume.
Carbamazepine RS and 0.5 mg/mL of USP Carbamazepine Chromatographic system
Related Compound A RS in methanol (See Chromatography 621, System Suitability.)
Carbamazepine RS and 0.05 mg/mL of USP Carbamazepine
60 Carbamazepine / Official Monographs USP 32
Mode: LC USP Carbamazepine Related Compound A RS

Detector: UV 230 nm USP Carbamazepine Related Compound B RS
System suitability Carbamazepine Oral Suspension
Sample: System suitability solution (Comment on this Monograph)id=m12550=Carbamazepine
Suitability requirements Oral Suspension=Ca-Chl-Monos.pdf)
Resolution: NLT 1.70 between carbamazepine related
compound A and carbamazepine DEFINITION
Relative standard deviation: NMT 2.0% Carbamazepine Oral Suspension contains NLT 90.0% and NMT
Analysis 110.0% of the labeled amount of carbamazepine (C15H12N2O).
Calculate the percentage of carbamazepine related IDENTIFICATION
compound A and carbamazepine related compound B in A. INFRARED ABSORPTION 197S
the portion of Carbamazepine: Sample solution: Place 5 mL of Oral Suspension in a
separator containing 20 mL of 0.1 N sodium hydroxide, and
Result = (ri/rS) (CS/CU) 100 extract with 25 mL of chloroform. Pass the extract through
anhydrous sodium sulfate supported on filter paper into a
ri = peak response for either carbamazepine related beaker. Wash the anhydrous sodium sulfate with 10 mL of
compound A or carbamazepine related chloroform, and add the washing to the extract. Evaporate
compound B from the Sample solution the chloroform extract to dryness with the aid of a stream of
rS = peak response for the corresponding peak from nitrogen. Dissolve the residue in 10 mL of methylene
the Standard solution chloride.
CS = concentration of carbamazepine related
compound A or carbamazepine related ASSAY
compound B in the Standard solution (mg/mL) PROCEDURE
CU = concentration of the Sample solution (mg/mL) Mobile phase: Methanol, tetrahydrofuran, and water
Calculate the percentage of all other impurities found in (12:3:85). Add 0.22 mL of formic acid to each L, mix, and
the portion of Carbamazepine: add 0.5 mL of triethylamine/L.
Result = (ri/rS) (CS/CU) 100 Carbamazepine RS and 0.5 mg/mL of USP Carbamazepine
Related Compound A RS in methanol
ri = peak response for any other impurity System suitability solution: 0.01 mg/mL of USP
rS = peak response for carbamazepine from the Carbamazepine RS and 0.05 mg/mL of USP Carbamazepine
Standard solution Related Compound A RS from System suitability stock solution
CS = concentration of Carbamazepine in the Standard in methanol and water (1:1). Combine equal amounts of
solution (mg/mL) this solution and the Sample solution, accurately measured.
CU = concentration of the Sample solution (mg/mL) Standard stock solution: 2 mg/mL of USP Carbamazepine
Acceptance criteria RS in methanol
Individual impurities: NMT 0.2% of any individual Standard solution: 0.2 mg/mL of USP Carbamazepine RS
impurity from Standard stock solution, diluted with methanol and
Total impurities: NMT 0.5% (including carbamazepine water (1:1)
related compound A and carbamazepine related compound Sample stock solution: Equivalent to 200 mg of
B) carbamazepine from a volume of freshly mixed Oral
Suspension, in a 100-mL volumetric flask. Add 70 mL of
SPECIFIC TESTS methanol, shake by mechanical means for 30 min, sonicate
X-RAY DIFFRACTION 941: The X-ray diffraction pattern for 2 min, dilute with methanol to volume, and mix. Allow
conforms to that of USP Carbamazepine RS, similarly the solution to stand for 10 min.
determined. Sample solution: Dilute 10.0 mL of the clear Sample stock
ACIDITY solution with methanol to 100.0 mL.
Sample solution: 50 mg/mL of Carbemazepine in water Chromatographic system
[NOTESolution is prepared by mixing for 15 min and (See Chromatography 621, System Suitability.)
filtering.] Mode: LC
Analysis: To a 10.0-mL aliquot of Sample solution, add 1 Detector: UV 230 nm
drop of phenolphthalein TS, and titrate with 0.01 N sodium Column: 4.6-mm 25-cm; packing L10
hydroxide VS. Perform a blank determination, and make any Flow rate: 1.5 mL/min
necessary correction. NMT 1.0 mL of 0.010 N sodium Injection size: 20 L
hydroxide is required for each 1.0 g of Carbamazepine. System suitability
ALKALINITY: To a 10.0-mL aliquot of Sample solution from Samples: System suitability solution and Standard solution
Acidity, add 1 drop of methyl red TS, and titrate with 0.01 N Suitability requirements
hydrochloric acid VS. Perform a blank determination, and Resolution: NLT 1.70 between carbamazepine related
make any necessary correction. NMT 1.0 mL of 0.010 N compound A and carbamazepine from the System
hydrochloric acid is required for each 1.0 g of suitability solution
Carbamazepine. Relative standard deviation: NMT 2.0% Standard
LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT solution
0.5% of its weight.
USP Carbamazepine RS
Analysis volumetric flask, add 40 mL of methanol, and sonicate for

Samples: Standard solution and Sample solution 15 min. Allow to cool to room temperature, dilute with
Calculate the percentage of C15H12N2O in the portion of methanol to volume, mix, and filter, discarding the first 10
Oral Suspension taken: mL of the filtrate.
Sample solution: 0.2 mg/mL of carbamazepine from
Result = (rU/rS) (CS/CU) 100 Sample stock solution in methanol and water (1:1)
rU = peak response from the Sample solution (See Chromatography 621, System Suitability.)
rS = peak response from the Standard solution Mode: LC
CS = concentration of USP Carbamazepine RS in the Detector: UV 230 nm
Standard solution (mg/mL) Column: 4.6-mm 25-cm; packing L10
CU = nominal concentration of carbamazepine in the Flow rate: 1.5 mL/min
Sample solution (mg/mL) Injection size: 20 L
Acceptance criteria: 90.0%110.0% System suitability
MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED Resolution: NLT 1.70 between carbamazepine related
MICROORGANISMS 62: The total bacterial count does not compound A and carbamazepine from the System
exceed 100 cfu/g, and the tests for Salmonella species and suitability solution
Escherichia coli are negative. Relative standard deviation: NMT 2.0% from the
PERFORMANCE TESTS Standard solution
UNIFORMITY OF DOSAGE UNITS 905: For Oral Suspension Analysis
packaged in single-unit containers: Meets the requirements Samples: Standard solution and Sample solution
DELIVERABLE VOLUME 698: For Oral Suspension packaged in Calculate the percentage of C15H12N2O in the portion of
multiple-unit containers: Meets the requirements. It is a Tablets taken:
performance test.
PACKAGING AND STORAGE: Preserve in tight, light-resistant rU = peak response from the Sample solution
containers, protected from freezing and from excessive heat. rS = peak response from the Standard solution
USP REFERENCE STANDARDS 11 CS = concentration of USP Carbamazepine RS in the
USP Carbamazepine RS Standard solution (mg/mL)
USP Carbamazepine Related Compound A RS CU = nominal concentration of carbamazepine in the
Carbamazepine Tablets PERFORMANCE TESTS

DISSOLUTION 711
(Comment on this Monograph)id=m12560=Carbamazepine For products labeled as 100-mg chewable tablets
Tablets=Ca-Chl-Monos.pdf) Test 1: If the product complies with this test, the labeling
DEFINITION indicates that it meets USP Dissolution Test 1.
Carbamazepine Tablets contain NLT 92.0% and NMT 108.0% Medium: Water containing 1% sodium lauryl sulfate; 900
of the labeled amount of carbamazepine (C15H12N2O). mL
Apparatus 2: 75 rpm
IDENTIFICATION Time: 60 min
A. INFRARED ABSORPTION 197M Detector: UV 288 nm
Sample solution: Boil an equivalent of 250 mg of Sample solution: Sample per Dissolution 711. Dilute
carbamazepine, from powdered Tablets, in 15 mL of with Medium to a concentration that is similar to the
acetone for 5 min. Filter while hot, using two 5-mL portions Standard solution.
of hot acetone to effect transfer. Evaporate the filtrate with Standard solution: USP Carbamazepine RS in Medium
the aid of nitrogen to 5 mL, and cool in an ice bath until [NOTEA volume of methanol not exceeding 1% of the
crystals are formed. Filter the crystals, wash with 3 mL of final total volume of the Standard solution may be used to
cold acetone, and dry in a vacuum at 70 for 30 min. dissolve the carbamazepine.]
ASSAY C15H12N2O. Use Acceptance Table 1, with the following
PROCEDURE exceptions: at S2, no unit is less than Q 5%; at S3, no
Mobile phase: Methanol, tetrahydrofuran, and water unit is less than Q 10%; and NMT 2 of the 24 units are
(12:3:85). Add 0.22 mL of formic acid to each L, mix, and less than Q 5%.
add 0.5 mL of triethylamine/ L. For products labeled as 200-mg tablets
System suitability stock solution: 0.1 mg/mL of USP Test 2: If the product complies with this test, the labeling
Carbamazepine RS and 0.5 mg/mL of USP Carbamazepine indicates that it meets USP Dissolution Test 2.
Related Compound A RS in methanol Medium: Water containing 1% sodium lauryl sulfate; 900
System suitability solution: 0.01 mg/mL of USP mL
Carbamazepine RS and 0.05 mg/mL of USP Carbamazepine Apparatus 2: 75 rpm
Related Compound A RS from System suitability stock solution Detector: UV 288 nm
in methanol and water (1:1) Sample solution: Sample per Dissolution 711. Dilute
Standard stock solution: 2 mg/mL of USP Carbamazepine with Medium to a concentration that is similar to that of
RS in methanol the Standard solution.
Standard solution: 0.2 mg/mL of USP Carbamazepine RS
from Standard stock solution in methanol and water (1:1)
Sample stock solution: Transfer an equivalent to 100 mg of
carbamazepine from NLT 20 powdered Tablets to a 50-mL
Standard solution: USP Carbamazepine RS in Medium Carbamazepine Extended-Release

[NOTEA volume of methanol not exceeding 1% of the Tablets
final total volume of the Standard solution may be used to
dissolve the carbamazepine.] (Comment on this Monograph)id=m12565=Carbamazepine
Times: See the table below. Extended-Release Tablets=Ca-Chl-Monos.pdf)
Tolerances: See the table below. DEFINITION
Carbamazepine Extended-Release Tablets contain NLT 90.0%
Times Tolerances and NMT 110.0% of the labeled amount of carbamazepine
(min) (% dissolved [Q]) (C15H12N2O).
15 45%75%
IDENTIFICATION
60 NLT 75% A. ULTRAVIOLET ABSORPTION 197U
Sample solution: Use the Sample solution from the test for
Use Acceptance Table 2, with the following exceptions: at Uniformity of Dosage Units.
15 minat L2, no unit is more than 5% outside the B. The retention time of the major PEAK of the Sample
stated range; at L3, no unit is more than 10% outside the solution corresponds to that of the Standard solution, as
stated range; and NMT 2 of the 24 units are more than obtained in the Assay.
5% outside the stated range. At 60 minat L2, no unit is
less than Q 5%; at L3, no unit is less than Q 10%; ASSAY
and NMT 2 of the 24 units are less than Q 5%. PROCEDURE
Test 3: If the product complies with this test, the labeling Mobile phase: Methanol, methylene chloride, and water
indicates that it meets USP Dissolution Test 3. (90:9:120)
Medium: Water containing 1% sodium lauryl sulfate; 900 Internal standard solution: 600 g/mL of phenytoin in
mL methanol
Apparatus 2: 75 rpm Standard stock solution: 200 g/mL of USP
Detector: UV 288 nm Carbamazepine RS in methanol
Sample solutions: Sample per Dissolution 711. Dilute Standard solution: 100 g/mL of USP Carbamazepine RS
with Medium to a concentration that is similar to that of from Standard stock solution in Internal standard solution
the Standard solution. System suitability solution: Internal standard solution and
Standard solution: USP Carbamazepine RS in Medium Standard solution (1:1)
[NOTEA volume of methanol not exceeding 1% of the Sample stock solution: Finely powder 10 Tablets, and
final total volume of the Standard solution may be used to quantitatively transfer the powder, with the aid of alcohol,
dissolve the carbamazepine.] to a volumetric flask of a volume such that when the
Times: See the table below. solution is diluted to volume, a final concentration estimated
Tolerances: See the table below. to be 4 mg/mL of carbamazepine is obtained. Shake by
mechanical means for 60 min. Sonicate for 15 min, and
Test 3 Table
dilute with methanol to volume. Allow to stand for 1015
min, then filter a portion of the supernatant, and use the
Times Tolerances clear filtrate.
(min) (% dissolved [Q]) Sample solution A: 0.2 mg/mL from Standard stock solution
15 60%85% in methanol
Sample solution: Sample solution A and Internal standard
60 NLT 75%
solution (1:1)
Use Acceptance Table 2, with the following exceptions: at (See Chromatography 621, System Suitability.)
15 minat L2, no unit is more than 5% outside the Mode: LC
stated range; at L3, no unit is more than 10% outside the Detector: UV 230 nm
stated range; and NMT 2 of the 24 units are more than Analytical Column: 3.9-mm 30-cm; packing L1
5% outside the stated range. At 60 minat L2, no unit is [NOTEWash the column with 50100 mL of methanol
less than Q 5%; at L3, no unit is less than Q 10%; before and after use.]
and NMT 2 of the 24 units are less than Q 5%. Guard column: 4.6-mm 30-mm; 7-m packing L7
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Flow rate: 2 mL/min
WATER DETERMINATION, Method III 921: Dry 1.5 g of the System suitability
powdered Tablets (NLT 20) at 120 for 2 h: it loses NMT Sample: System suitability solution
5.0% of its weight. [NOTEThe relative retention times for phenytoin and
carbamazepine are about 0.8 and 1.0, respectively.]
PACKAGING AND STORAGE: Preserve in tight containers, Resolution: NLT 2.8 between phenytoin and
preferably of glass. Dispense Carbamazepine Tablets in a carbamazepine
container labeled Store in a dry place. Protect from Relative standard deviation: NMT 2.0%
moisture. Analysis
LABELING: The labeling indicates the Dissolution test with Samples: Standard solution and Sample solution
which the product complies. Calculate the percentage of C15H12N2O in each Tablet
USP REFERENCE STANDARDS 11 taken:
USP Carbamazepine Related Compound A RS Result = (RU/RS) (CS/CU) 100
RU = peak response ratio of carbamazepine to the Suitability requirements

internal standard from the Sample solution Resolution: NLT 2.8 between phenytoin and
RS = peak response ratio of carbamazepine to the carbamazepine
internal standard from the Standard solution Relative standard deviation: NMT 2.0%
CS = concentration of USP Carbamazepine RS in the Analysis
Standard solution (g/mL) Samples: Standard solution and Sample solution
CU = nominal concentration of carbamazepine in the Calculate the percentage of each impurity in the portion
Sample solution (g/mL) of Tablets taken:
IMPURITIES
Organic Impurities rU = peak response of each impurity from the Sample
PROCEDURE 1 solution
Standard solution: 5 g/mL of methanol and methylene rS = peak response for carbamazepine from the
chloride in dimethylformamide Standard solution
Sample solution: Powder 10 Tablets, and transfer to a 50- CS = concentration of USP Carbamazepine RS in the
mL volumetric flask. Add 30 mL of dimethylformamide, Standard solution (mg/mL)
and shake by mechanical means for 60 min. Sonicate for 2 CU = nominal concentration of carbamazepine in the
min, and dilute with dimethylformamide to volume. Sample solution (mg/mL)
Centrifuge a portion at about 2500 rpm for 20 min, and Acceptance criteria: See Impurity Table
use the clear supernatant. PROCEDURE 3
Chromatographic system Mobile phase: Methanol, acetonitrile, and water (7:3:10)
(See Chromatography 621, System Suitability.) System suitability solution: 12.5 g/mL of iminostilbene
Mode: GC and 5.0 g/mL of USP Carbamazepine RS in methanol
Detector: Flame ionization Standard solution: Use the Standard solution from Organic
Column: 2-mm 3-m glass; 0.2% phase G39 on support Impurities, Procedure 1.
S7 Sample solution: Use the Sample stock solution from the
Temperature Assay.
Injector: 170 Chromatographic system
Detector: 300 (See Chromatography 621, System Suitability.)
Column: 75 for 10 min, increase 20/min to 155 and Mode: LC
maintain at 155 for 30 min. Detector: UV 230 nm
Carrier gas: Helium Column: 3.9-mm 30-cm; packing L1
Flow rate: 10 mL/min [NOTEWash the column with 50100 mL of methanol
Injection volume: 2 L before and after use.]
Analysis Guard column: 4.6-mm 30-mm; 7-m packing L7
Samples: Standard solution and Sample solution Flow rate: 2 mL/min
Calculate the amount of methylane chloride and methanol, Injection size: 10 L
in g, in each Tablet taken: System suitability
5 CS rU/rS [NOTEThe relative retention times for carbamazepine
and iminostilbene are about 0.3 and 1.0, respectively.]
CS = concentration (g/mL) of methylene chloride or Suitability requirements
methanol in the Standard solution Resolution: NLT 10.0 between carbamazepine and
rU = peak response of the respective analyte in the iminostilbene
Sample solution Relative standard deviation: NMT 2.0%
rS = peak response of the respective analyte in the Analysis
Standard solution Samples: Standard solution and Sample solution
Acceptance criteria: NMT 23 g/Tablet of methylene Calculate the percentage of each impurity in the portion
chloride; NMT 100 g/Tablet of methanol of Tablets:
PROCEDURE 2
Mobile phase: Proceed as in the Assay. Result = (rU/rS) (CS/CU) 100
System suitability solution: 60 g/mL of phenytoin and
20 g/mL of USP Carbamazepine RS in methanol rU = peak response of each impurity from the Sample
Standard solution: 4 g/mL of USP Carbamazepine RS in solution
methanol rS = peak response for carbamazepine from the
Sample solution: Use the Sample stock solution from the Standard solution
Assay. CS = concentration of USP Carbamazepine RS in the
Chromatographic system Standard solution (mg/mL)
(See Chromatography 621, System Suitability.) CU = nominal concentration of carbamazepine in the
Mode: LC Sample solution (mg/mL)
Detector: UV 230 nm Acceptance criteria: See Impurity Table.
Analytical column: 3.9-mm 30-cm; packing L1
[NOTEWash the column with 50100 mL of methanol Impurity Table
before and after use.]
Guard column: 4.6-mm 30-mm; 7-m packing L7 Acceptance Criteria
Flow rate: 2 mL/min Impurity (NMT)
Injection size: 10 L Any individual impurity 0.2%
System suitability Sum of all impurities (Procedure 1 + 0.5%
Sample: System suitability solution Procedure 2)
[NOTEThe relative retention times for phenytoin and
carbamazepine are about 0.8 and 1.0, respectively.]
PERFORMANCE TESTS Carbamide Peroxide

DISSOLUTION 711 (Comment on this Monograph)id=m12585=Carbamide
Medium: Water; 900 mL or 1800 mL for 400-mg Tablets Peroxide=Ca-Chl-Monos.pdf)
Time: 3, 6, 12, and 24 h
Detector: UV 284 nm
Sample solution: Sample per Dissolution 711. Dilute with
Medium to a concentration that is similar to that of the
Standard solution.
Standard solution: USP Carbamazepine RS in Medium
Tolerances: The percentages (Q) of the labeled amount of
C15H12N2O dissolved at the times specified conform to CH6N2O3 94.07
Acceptance Table 2. Urea compound with hydrogen peroxide (1:1) [124-43-6].
Time
DEFINITION
(h) Amount Dissolved
Carbamide Peroxide contains NLT 96.0% and NMT 102.0% of
CH6N2O3.
3 10%35%
6 35%65% IDENTIFICATION
A. Combine 1 mL of a 100 mg/mL Carbamide Peroxide
12 65%90% solution with 1 mL of nitric acid: a white, crystalline
24 NLT 75% precipitate is formed.
B. IDENTIFICATION TESTSGENERAL, Peroxide 191: 100
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements mg/mL Carbamide Peroxide solution
for Content Uniformity
Standard solution: 10 g/mL of USP Carbamazepine RS in ASSAY
methanol PROCEDURE
Sample solution: Finely powder 1 Tablet, and quantitatively Sample solution: Transfer 100 mg of Carbamide Peroxide
transfer the powder, with the aid of methanol, to a 100-mL to a 500-mL iodine flask with the aid of 25 mL of water,
volumetric flask. Add about 70 mL of methanol, and shake add 5 mL of glacial acetic acid, and mix. Add 2 g of
by mechanical means for 60 min. Sonicate for 15 min, and potassium iodide and 1 drop of ammonium molybdate TS,
dilute with methanol to volume. Allow to stand for 1015 insert the stopper, and allow to stand in the dark for 10
min. Dilute a portion of the clear solution with methanol to min.
obtain a solution containing about 10 g/mL of Analysis: Titrate the liberated iodine in the Sample solution
carbamazepine. with 0.1 N sodium thiosulfate VS, adding 3 mL of starch TS
Blank: Methanol as the endpoint is approached. Each mL of 0.1 N sodium
Spectrometric conditions thiosulfate is equivalent to 4.704 mg of CH6N2O3.
Mode: UV Acceptance criteria: 96.0%102.0%
Samples: Standard solution, Sample solution, and Blank PACKAGING AND STORAGE: Preserve in tight, light-resistant
Calculate percentage of C15H12N2O in the Tablet: containers, and avoid exposure to excessive heat.

Carbamide Peroxide Topical Solution
AS = absorbance of the Standard solution (Comment on this Monograph)id=m12590=Carbamide
CS = concentration of USP Carbamazepine RS in the Peroxide Topical Solution=Ca-Chl-Monos.pdf)
CU = nominal concentration of the Sample solution DEFINITION
(g/mL) Carbamide Peroxide Topical Solution is a solution in anhydrous
glycerin of Carbamide Peroxide or of carbamide peroxide
prepared from hydrogen peroxide and Urea. It contains NLT
78.0% and NMT 110.0%, by weight, of the labeled amount of
SPECIFIC TESTS
CH6N2O3.
WATER DETERMINATION, Method Ia 921: NMT 5.0%
IDENTIFICATION
A. Mix 1 mL of Topical Solution with 1 mL of nitric acid: a
white, crystalline precipitate is formed.
USP 32 Official Monographs / Carbenicillin 65
B. IDENTIFICATION TESTSGENERAL, Peroxide 191 WATER DETERMINATION, Method I 921: NMT 6.0%
BACTERIAL ENDOTOXINS TEST 85: Where it is labeled for use
ASSAY in preparing injectable dosage forms, NMT 0.05 USP
PROCEDURE Endotoxin Unit/mg of carbenicillin
Sample solution: Transfer an equivalent to 100 mg of STERILITY TESTS 71: Where the label states that
carbamide peroxide from Topical Solution to a 500-mL Carbenicillin Disodium is sterile, it meets the requirements
iodine flask with the aid of 25 mL of water, add 5 mL of when tested as directed under Test for Sterility of the Product
glacial acetic acid, and mix. Add 2 g of potassium iodide to Be Examined, Membrane Filtration, 6 g being aseptically
and 1 drop of ammonium molybdate TS, and allow to stand dissolved in 200 mL of Fluid A.
in the dark for 10 min.
Analysis: Titrate the liberated iodine in the Sample solution ADDITIONAL REQUIREMENTS
with 0.1 N sodium thiosulfate VS, adding 3 mL of starch TS PACKAGING AND STORAGE: Preserve in tight containers.
as the endpoint is approached. Each mL of 0.1 N sodium LABELING: Where it is intended for use in preparing
thiosulfate is equivalent to 4.704 mg of CH6N2O3. injectable dosage forms, the label states that it is sterile or
Acceptance criteria: 78.0%110.0% must be subjected to further processing during the
SPECIFIC TESTS USP REFERENCE STANDARDS 11
SPECIFIC GRAVITY 841: 1.2451.272 USP Carbenicillin Monosodium Monohydrate RS
PH 791: 4.07.5 USP Endotoxin RS
containers, and avoid exposure to excessive heat. Carbenicillin for Injection
(Comment on this Monograph)id=m12710=Carbenicillin for
Carbenicillin Disodium DEFINITION
(Comment on this Monograph)id=m12700=Carbenicillin Carbenicillin for Injection contains an amount of Carbenicillin
Disodium=Ca-Chl-Monos.pdf) Disodium equivalent to NLT 90.0% and NMT 120.0% of the
labeled amount of carbenicillin (C17H18N2O6S).
IDENTIFICATION
IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the
requirements
ASSAY
PROCEDURE
C17H16N2Na2O6S (anhydrous) 422.36 Buffer: Use Buffer No. 1 as described in Antibiotics
4-Thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, 6- Microbial Assays 81.
[(carboxyphenylacetyl)amino]-3,3-dimethyl-7-oxo-, disodium Sample solution A (single-dose): Where it is packaged for
salt, [2S-(2,5,6)]-; dispensing and where the package is represented as being a
N-(2-Carboxy-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]- single-dose container, constitute Carbenicillin for Injection as
hept-6-yl)- 2-phenylmalonamic acid disodium salt directed in the labeling. Withdraw all of the withdrawable
[4800-94-6]. contents, and dilute quantitatively with Buffer to obtain a
solution having a convenient concentration of carbenicillin.
DEFINITION Sample solution B: Where the label states the quantity of
Carbenicillin Disodium has a potency equivalent to NLT 770 g carbenicillin in a given volume of constituted solution,
of carbenicillin (C17H18N2O6S)/mg, calculated on the anhydrous constitute Carbenicillin for Injection as directed in the
basis. labeling. Dilute a measured volume of the constituted
solution quantitatively with Buffer to obtain a solution having
IDENTIFICATION a convenient concentration of carbenicillin.
IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the Analysis: Proceed as directed under AntibioticsMicrobial
requirements Assays 81, using a measured volume of Sample solution
diluted quantitatively with Buffer to yield a Sample Dilution
ASSAY having a concentration assumed to be equal to the median
PROCEDURE dose level of the Standard.
Buffer: Use Buffer No. 1 as described in Antibiotics Acceptance criteria: 90.0%120.0%
Microbial Assays 81.
Sample solution: Dissolve a suitable quantity of PERFORMANCE TESTS
Carbenicillin Disodium in Buffer, and dilute quantitatively UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
with Buffer to obtain a solution having a convenient
concentration of carbenicillin. SPECIFIC TESTS
Analysis: Proceed as directed under AntibioticsMicrobial BACTERIAL ENDOTOXINS TEST 85: NMT 0.05 USP Endotoxin
Assays 81, using a volume of Sample solution diluted Unit/mg of carbenicillin
quantitatively with Buffer to yield a Sample Dilution having a STERILITY TESTS 71: Where the label states that
concentration assumed to be equal to the median dose level Carbenicillin Disodium is sterile, it meets the requirements
of the Standard. when tested as directed under Test for Sterility of the Product
Acceptance criteria: NLT 770 g/mL to Be Examined, Membrane Filtration, 6 g being aseptically
dissolved in 200 mL of Fluid A.
SPECIFIC TESTS PH 791: 6.58.0, in a solution constituted as directed in
PH 791: 6.58.0, in a solution containing 10 mg/mL of the labeling
carbenicillin
66 Carbenicillin / Official Monographs USP 32
PARTICULATE MATTER IN INJECTIONS 788: Meets the Mode: LC

requirements for small-volume injections. Detector: UV 210 nm
WATER DETERMINATION, Method I 921: NMT 6.0% Column: 4.6-mm 25-cm; packing L7
INJECTIONS 1: Meets the requirements for Constituted Flow rate: 2 mL/min
Solutions and Labeling. Injection size: 25 L
System suitability
PACKAGING AND STORAGE: Preserve as described in Injections Suitability requirements
1, Containers for Sterile Solids. Relative standard deviation: NMT 2.0%
USP Carbenicillin Monosodium Monohydrate RS Calculate the quantity, in g, of C17H18N2O6S in each mg of
USP Endotoxin RS Carbenicillin Indanyl Sodium taken:
Result = (rU/rS) (CS/CU) P
Carbenicillin Indanyl Sodium rU = peak response from the Sample solution
(Comment on this Monograph)id=m12750=Carbenicillin rS = peak response from the Standard solution
Indanyl Sodium=Ca-Chl-Monos.pdf) CS = concentration of USP Carbenicillin Indanyl
Sodium RS, calculated on the anhydrous basis,
in the Standard solution (g/mL)
CU = concentration of Carbenicillin Indanyl Sodium in
P = assigned potency of USP Carbenicillin Indanyl
Sodium RS (g/mg)
Acceptance criteria: 630769 g/mg
C26H25N2NaO6S 516.54 SPECIFIC TESTS
4-Thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, 6-[[3-[(2,3- PH 791: 5.08.0, 100 mg/mL
dihydro-1H-inden-5-yl)oxy]-1,3-dioxo-2- WATER DETERMINATION, Method I 921: NMT 2.0%
phenylpropyl]amino]-3,3-dimethyl-7-oxo-, monosodium salt,
[2S-(2,5,6)]-; ADDITIONAL REQUIREMENTS
1-(5-Indanyl)(2S,5R,6R)-N-(2-carboxy-3,3-dimethyl-7-oxo-4- PACKAGING AND STORAGE: Preserve in tight containers. For
thia-1-azabicyclo[3.2.0]hept-6-yl)-2-phenylmalonamate periods up to 18 months, store at controlled room
monosodium salt [26605-69-6]. temperature.
DEFINITION USP Carbenicillin Indanyl Sodium RS
Carbenicillin Indanyl Sodium has a potency equivalent to NLT
630 g and NMT 769 g of carbenicillin (C17H18N2O6S)/mg,
Carbenicillin Indanyl Sodium Tablets
IDENTIFICATION (Comment on this Monograph)id=m12780=Carbenicillin
A. INFRARED ABSORPTION 197K Indanyl Sodium Tablets=Ca-Chl-Monos.pdf)
B. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the
requirements DEFINITION
Carbenicillin Indanyl Sodium Tablets contain the equivalent of
ASSAY NLT 90.0% and NMT 120.0%of the labeled amount of
PROCEDURE carbenicillin (C17H18N2O6S).
Solution A: 0.302 mg/mL of tetrabutylammonium
phosphate and 13.4 mg/mL of dibasic sodium phosphate in IDENTIFICATION
water, adjusted with phosphoric acid to a pH of 3.8 before THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
final dilution Adsorbent: 0.25-mm layer of chromatographic silica gel
Mobile phase: Acetonitrile and Solution A (21:29). Allow to mixture
stand for 1 h, and if necessary, readjust with phosphoric Diluent: Acetone, ethyl acetate, pyridine, glacial acetic acid,
acid to a pH of 3.8 and water (200:100:25:1.5:75)
Diluent: Acetonitrile and 5 mM monobasic potassium Standard solution: 1 mg/mL of USP Carbenicillin Indanyl
phosphate (17:3) Sodium RS in Diluent
Standard solution: 250 g/mL of USP Carbenicillin Indanyl Sample stock solution: Equivalent to 10 mg/mL of
Sodium RS in Diluent carbenicillin from powdered Tablets in Diluent. Shake the
[NOTEThe Standard solution contains the equivalent of mixture for 5 min.
222 g/mL of carbenicillin.] Sample solution: 1 mg/mL of carbenicillin, from Sample
Sample solution: 250 g/mL of Carbenicillin Indanyl stock solution, in Diluent
Sodium in Diluent. Sonication may be necessary to dissolve. Application volume: 10 L
Chromatographic system Developing solvent system: Acetone, ethyl acetate,
(See Chromatography 621, System Suitability.) pyridine, glacial acetic acid, and water (400:300:25:2:75)
USP 32 Official Monographs / Carbenicillin 67
Spray reagent: Methanol, 10 mg/mL of ferric chloride in rU = peak response from the Sample solution
0.1 N hydrochloric acid, and 10 mg/mL of potassium rS = peak response from the Standard solution
ferricyanide solution (3:4:4) CS = concentration of USP Carbenicillin Indanyl
Analysis Sodium RS, calculated on the anhydrous basis,
Samples: Standard solution and Sample solution in the Standard solution (g/mL)
Develop until the solvent front has moved three-fourths of CU = nominal concentration of carbenicillin in the
the length of the plate. Remove the plate from the Sample solution (g/mL)
chamber, mark the solvent front, and heat the plate at P = assigned potency of USP Carbenicillin Indanyl
80 for 30 min. Allow the plate to cool, and expose it to Sodium RS (g/mg)
iodine vapors in a closed chamber for 30 s. Spray the Acceptance criteria: 90%120%
plate with Spray reagent.
Acceptance criteria: The principal spots from the Standard PERFORMANCE TESTS
solution and the Sample solution are blue on a yellow-green DISSOLUTION 711
background, and the RF value of the principal spot from the Medium: Water; 900 mL
Sample solution corresponds to that from the Standard Apparatus 1: 100 rpm
solution (RF value of 0.5). Time: 45 min
Detector: UV
ASSAY Analytical wavelengths: Maximum at 267 nm and
PROCEDURE minimum at 254 nm
Solution A: 0.302 mg/mL of tetrabutylammonium Standard solution: USP Carbenicillin Indanyl Sodium RS in
phosphate and 13.4 mg/mL of dibasic sodium phosphate in Medium
water, adjusted with phosphoric acid to a pH of 3.8 before Sample solution: Sample per Dissolution 711. Dilute with
final dilution Medium to a concentration that is similar to that of the
Mobile phase: Acetonitrile and Solution A (21:29). Allow to Standard solution.
stand for 1 h, and if necessary, readjust with phosphoric Analysis
acid to a pH of 3.8. Samples: Standard solution and Sample solution
Diluent: Acetonitrile and 5 mM monobasic potassium Calculate the percentage of C17H18N2O6S equivalent
phosphate (17:3) dissolved in the portion of Tablets taken:
Standard solution: 250 g/mL of USP Carbenicillin Indanyl
Sodium RS in Diluent Result = (AU/AS) x (CS/CU) x (Mr1/Mr2) x 100
[NOTEContains the equivalent of 222 g/mL of
carbenicillin.] AU = absorbance of A267 A254 for the Sample solution
Sample stock solution: Equivalent to 222 g/mL of AS = absorbance of A267 A254 for the Standard
carbenicillin from NLT 20 powdered Tablets in Diluent solution
[NOTESonication may be necessary to dissolve. Contains CS = concentration of USP Carbenicillin Indanyl
250 g/mL of carbenicillin indanyl sodium.] Sodium RS in the Standard solution (g/mL)
Chromatographic system CU = nominal concentration of carbenicillin in the
(See Chromatography 621, System Suitability.) Sample solution (g/mL)
Mode: LC Mr1 = molecular weight of carbenicillin, 378.40
Detector: UV 210 nm Mr2 = molecular weight of carbenicillin indanyl sodium,
Column: 4.6-mm 25-cm; packing L7 516.54
Flow rate: 2 mL/min Tolerances: NLT 75% (Q) of the labeled amount of
Injection size: 25 L C17H18N2O6S is dissolved.
System suitability UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Suitability requirements SPECIFIC TESTS
Relative standard deviation: NMT 2.0% WATER DETERMINATION, Method I 921: NMT 2.0%
Samples: Standard solution and Sample solution PACKAGING AND STORAGE: Preserve in tight containers.
Calculate the percentage of C17H18N2O6S in the portion of USP REFERENCE STANDARDS 11
Tablets taken: USP Carbenicillin Indanyl Sodium RS
68 Carbidopa / Official Monographs USP 32
Carbidopa Acceptance criteria: 98.0%102.0%

(Comment on this Monograph)id=m12810=Carbidopa=Ca-Chl- IMPURITIES
Monos.pdf) Inorganic Impurities
Organic Impurities
PROCEDURE: LIMIT OF METHYLDOPA AND CARBIDOPA RELATED
COMPOUND A
Mobile phase, System suitability solution, Standard
solution, Sample solution, Chromatographic system, and
C10H14N2O4 H2O 244.24 Suitability requirements: Proceed as directed in the
Benzenepropanoic acid, -hydrazino-3,4-dihydroxy--methyl-, Assay.
monohydrate, (S)-; Impurity standard solution: 2.5 g/mL of USP
(-)-L--Hydrazino-3,4-dihydroxy--methylhydrocinnamic acid Methyldopa RS and 2.5 g/mL of USP Carbidopa Related
monohydrate [38821-49-7]. Compound A RS in Mobile phase
Anhydrous 226.23 System suitability
[28860-95-9]. Samples: System suitability solution and Standard solution
[NOTEThe relative retention times for methyldopa,
DEFINITION carbidopa, and carbidopa related compound A are
Carbidopa contains NLT 98.0% and NMT 102.0% of about 0.8, 1.0, and 1.8, respectively.]
C10H14N2O4 H2O. Analysis
Samples: Sample solution and Impurity standard solution
IDENTIFICATION Calculate the percentage of methyldopa in the portion of
A. INFRARED ABSORPTION 197M Carbidopa taken:
B. The retention time of the major peak from the Sample
solution corresponds to that of the Standard solution, as Result = (rU/rS) (CS/CU) 100
rU = peak response of methyldopa or carbidopa
ASSAY related compound A from the Sample solution
PROCEDURE rS = peak response of methyldopa or carbidopa
Mobile phase: Alcohol and 0.05 M monobasic sodium related compound A from the Impurity
phosphate, adjusted with phosphoric acid to a pH of 2.7 standard solution
(1:19) CS = concentration of USP Methyldopa RS or USP
System suitability solution: 0.1 mg/mL of USP Carbidopa Carbidopa Related Compound A RS in the
RS and 0.1 mg/mL of USP Methyldopa RS in Mobile phase Impurity standard solution (g/mL)
Standard solution: 0.5 mg/mL of USP Carbidopa RS in CU = concentration of the Sample solution (g/mL)
Mobile phase Acceptance criteria: NMT 0.5% of methyldopa, and NMT
[NOTEUse gentle heat and ultrasonification, if necessary, 0.5% of carbidopa related compound A
to dissolve.]
Sample solution: 0.5 mg/mL of Carbidopa in Mobile phase SPECIFIC TESTS
Chromatographic system OPTICAL ROTATION, Specific Rotation 781S: 21.0 to
(See Chromatography 621, System Suitability.) 23.5, calculated as the monohydrate
Mode: LC Sample solution: 10 mg/mL, in 666.6 mg/mL of aluminum
Detector: UV 280 nm chloride solution that has been filtered and adjusted with
Column: 3.9-mm 30-cm; packing L1 0.25 N sodium hydroxide to a pH of 1.5
Flow rate: 1 mL/min LOSS ON DRYING 731
Injection size: 20 L Analysis: Heat 1 g in a suitable vacuum drying apparatus at
System suitability 100 and a pressure of NMT 5 mm of mercury to constant
Samples: System suitability solution and Standard solution weight. Cool and weigh.
[NOTEThe relative retention times for methyldopa and Acceptance criteria: It loses 6.9%7.9% of its weight.
carbidopa are about 0.8 and 1.0, respectively.]
Resolution: NLT 0.9 between methyldopa and carbidopa PACKAGING AND STORAGE: Preserve in well-closed, light-
in the System suitability solution resistant containers.
Relative standard deviation: NMT 1.5% in the Standard USP REFERENCE STANDARDS 11
solution USP Carbidopa RS
Analysis USP Carbidopa Related Compound A RS
Samples: Standard solution and Sample solution USP Methyldopa RS
Calculate the percentage of C10H14N2O4 H2O in the
portion of Carbidopa taken:
Carbidopa and Levodopa Tablets
(Comment on this Monograph)id=m12870=Carbidopa and
rU = peak response from the Sample solution Levodopa Tablets=Ca-Chl-Monos.pdf)
CS = concentration of USP Carbidopa RS, as the DEFINITION
monohydrate, in the Standard solution Carbidopa and Levodopa Tablets contain NLT 90.0% and NMT
(mg/mL) 110.0% of the labeled amounts of carbidopa (C10H14N2O4) and
CU = concentration of Carbidopa in the Sample levodopa (C9H11NO4).
solution (mg/mL)
USP 32 Official Monographs / Carbinoxamine 69
IDENTIFICATION CS = concentration of USP Carbidopa RS or USP

THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Levodopa RS in the Standard solution (mg/mL)
Adsorbent: 0.25-mm layer of chromatographic silica gel CU = nominal concentration of carbidopa or levodopa
Standard solution A: 0.1 mg/mL of USP Carbidopa RS in in the Sample solution (mg/mL)
0.05 N hydrochloric acid and methanol (1:1) Acceptance criteria: 90.0%110.0%
Standard solution B: 0.1 mg/mL of USP Levodopa RS in
0.05 N hydrochloric acid and methanol (1:1) PERFORMANCE TESTS
Sample solution: Equivalent to 10 mg of carbidopa, from DISSOLUTION 711
powdered Tablets, in a 100-mL volumetric flask containing Medium: 0.1 N hydrochloric acid; 750 mL
50 mL of 0.05 N hydrochloric acid. Agitate for 20 min, add Apparatus 1: 50 rpm
methanol to volume, and filter or centrifuge. Time: 30 min
Application volume: 20 L Analysis: Determine the amounts of carbidopa and
Developing solvent system: Acetone, chloroform, n- levodopa in filtered portions of the solution under test, in
butanol, glacial acetic acid, and water (12:8:8:8:7) comparison with a Standard solution having known
Spray reagent: 3 mg/mL of ninhydrin in n-butanol and concentrations of USP Carbidopa RS and USP Levodopa RS
glacial acetic acid (97:3) in Medium, as directed in the Assay.
Analysis Tolerances: NLT 80% (Q) of the labeled amounts of
Samples: Standard solution A, Standard solution B, and C10H14N2O4 and C9H11NO4 are dissolved.
Sample solution UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Develop, using the Developing solvent system, until the
solvent front has moved 15 cm. Air-dry, spray uniformly ADDITIONAL REQUIREMENTS
with 0.5 mL of Spray reagent, and heat at 105 for 10 PACKAGING AND STORAGE: Preserve in well-closed, light-
min. resistant containers.
Acceptance criteria: The Sample solution exhibits two spots USP REFERENCE STANDARDS 11
(reddish brown for levodopa and yellow-orange for USP Carbidopa RS
carbidopa) having RF values that correspond to those USP Levodopa RS
exhibited by the Standard solutions.
ASSAY Carbinoxamine Maleate
PROCEDURE
Solution A: 0.24 mg/mL of sodium 1-decanesulfonate in (Comment on this Monograph)id=m12900=Carbinoxamine
water Maleate=Ca-Chl-Monos.pdf)
Mobile phase: 11.04 g of monobasic sodium phosphate
and 950 mL of water in a beaker. Add 1.3 mL of Solution A,
and adjust with phosphoric acid to a pH of 2.8. Transfer to a
1-L volumetric flask, and dilute with water to volume.
Standard solution: 50 mg of USP Levodopa RS in a 100-
mL volumetric flask. Add a quantity of USP Carbidopa RS,
which is in a ratio with USP Levodopa RS that corresponds
to the ratio of carbidopa to levodopa in the Tablets. Add 10
mL of 0.1 N phosphoric acid. Warm gently to dissolve the C16H19ClN2O C4H4O4 406.86
standards, and dilute with water to volume. Ethanamine, 2-[(4-chlorophenyl)-2-pyridinylmethoxy]-N, N-
Sample solution: Equivalent to 50 mg of levodopa from dimethyl-, (Z)-2-butenedioate (1:1);
powdered NLT 20 Tablets, in a 100-mL volumetric flask. Add 2-[p-Chloro--[2-(dimethylamino)ethoxy]benzyl]pyridine
10 mL of 0.1 N phosphoric acid, and dilute with water to maleate (1:1) [3505-38-2].
volume. DEFINITION
Chromatographic system Carbinoxamine Maleate contains NLT 98.0% and NMT 102.0%
(See Chromatography 621, System Suitability.) of C16H19ClN2O C4H4O4, on the previously dried basis.
Mode: LC
Flow rate: 2 mL/min B. ULTRAVIOLET ABSORPTION 197U
[NOTEAdjust until the retention times for levodopa and Analytical wavelength: 260 nm
carbidopa are 4 min and 11 min, respectively.] Sample solution: 50 g/mL in methanol
Injection size: 20 L Acceptance criteria: Absorptivities, calculated on the dried
System suitability basis, do not differ by more than 3.0%.
Resolution: NLT 6 between levodopa and carbidopa PROCEDURE
Relative standard deviation: NMT 2.0% Sample solution: 8 mg/mL of Carbinoxamine Maleate,
Analysis previously dried in glacial acetic acid
Samples: Standard solution and Sample solution Analysis: To 50 mL of Sample solution, add 1 drop of crystal
Calculate the percentage of C10H14N2O4 and C9H11NO4 in violet TS, and titrate with 0.1 N perchloric acid VS to a blue-
the portion of Tablets taken: green endpoint. Perform a blank determination, and make
any necessary correction (see Titrimetry 541). Each mL of
Result = (rU/rS) (CS/CU) 100 0.1 N perchloric acid is equivalent to 20.34 mg of
C16H19ClN2O C4H4O4.
rU = peak response of carbidopa or levodopa from
the Sample solution
rS = peak response of carbidopa or levodopa from
the Standard solution
70 Carbinoxamine / Official Monographs USP 32

IMPURITIES PERFORMANCE TESTS
Inorganic Impurities DISSOLUTION 711
RESIDUE ON IGNITION 281: NMT 0.1% Medium: Water; 900 mL
Organic Impurities Apparatus 2: 50 rpm
PROCEDURE: ORDINARY IMPURITIES 466 Time: 45 min
Standard solution: Chloroform Detector: UV 260 nm
Sample solution: Chloroform Standard solution: USP Carbinoxamine Maleate RS in
Eluant: Cyclohexane, chloroform, and diethylamine Medium
(15:3:2) Sample solution: Sample per Dissolution 711. Dilute with
Visualization: 1 Medium to a concentration that is similar to that of the
Standard solution.
SPECIFIC TESTS Tolerances: NLT 75% (Q) of the labeled amount of
MELTING RANGE OR TEMPERATURE 741: 116121, C16H19ClN2O C4H4O4
determined after drying UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
PH 791: 4.65.1, in a 10-mg/mL solution Analysis for content uniformity
LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT Standard solution: 40 g/mL of USP Carbinoxamine
0.5% of its weight. Maleate RS in methanol and water (9:1)
Sample stock solution: Place 1 Tablet in a 100-mL
ADDITIONAL REQUIREMENTS volumetric flask, add 10.0 mL of water, and shake by
PACKAGING AND STORAGE: Preserve in tight, light-resistant mechanical means for 15 min. Dilute with methanol to
containers. volume, and filter, discarding the first 20 mL of the filtrate.
USP REFERENCE STANDARDS 11 Sample solution: Equivalent to 40 g/mL of
USP Carbinoxamine Maleate RS carbinoxamine maleate from the Sample stock solution in
methanol and water (9:1)
Blank: Methanol and water (9:1)
Carbinoxamine Maleate Tablets Spectrometric conditions
Mode: UV-Vis
(Comment on this Monograph)id=m12920=Carbinoxamine Cell: 1 cm
Maleate Tablets=Ca-Chl-Monos.pdf) Analytical wavelength: Peak maximum at about 260 nm
DEFINITION Analysis
Carbinoxamine Maleate Tablets contain NLT 93.0% and NMT Samples: Standard solution, Sample solution, and Blank
107.0% of the labeled amount of carbinoxamine maleate Calculate the percentage of C16H19ClN2O C4H4O4 in the
(C16H19ClN2O C4H4O4). portion of Tablets taken:
IDENTIFICATION Result = (AU/AS) (CS/CU) 100

ULTRAVIOLET ABSORPTION 197U
Standard solution: 0.02 mg/mL of USP Carbinoxamine AU = absorbance of the Sample solution
Maleate RS in 0.25 M sulfuric acid AS = absorbance of the Standard solution
Sample solution: Prepare a solution equivalent to 0.02 CS = concentration of USP Carbinoxamine Maleate RS
mg/mL of carbinoxamine maleate in 0.25 M sulfuric acid, in the Standard solution (g/mL)
from the Tablets, as directed under Salts of Organic CU = nominal concentration of carbinoxamine
Nitrogenous Bases 501. maleate in the Sample solution (g/mL)
Analytical wavelength: Peak maximum at 263 2 nm ADDITIONAL REQUIREMENTS
Acceptance criteria: The absorptivity of the Sample solution PACKAGING AND STORAGE: Preserve in tight, light-resistant
is within 7.0% of that of the Standard solution. containers.
PROCEDURE USP Carbinoxamine Maleate RS
Sample solution: Equivalent to 100 mg of carbinoxamine
maleate from NLT 30 powdered Tablets. In a separator, add
35 mL of water and 3 g of sodium bicarbonate. Extract with
five 20-mL portions of chloroform, filtering the extracts
Carbol-Fuchsin Topical Solution
through a pledget of cotton. Evaporate the combined (Comment on this Monograph)id=m12980=Carbol-Fuchsin
chloroform extracts on a steam bath just to dryness, dissolve Topical Solution=Ca-Chl-Monos.pdf)
the residue in 50 mL of glacial acetic acid, and add 1 drop Prepare Carbol-Fuchsin Topical Solution as follows:
of crystal violet TS.
Analysis: Titrate with 0.05 N perchloric acid VS to a blue- Basic Fuchsin 3g
green endpoint. Perform a blank determination, and make Phenol 45 g
any necessary correction (see Titrimetry 541). Each mL of
0.05 N perchloric acid is equivalent to 10.17 mg of
C16H19ClN2O C4H4O4.
USP 32 Official Monographs / Carbon 71
Resorcinol 100 g hydroxide solution to fill the pipet and capillary connection
Acetone 50 mL up to the stopcock. Fill the buret with the leveling water,
and draw it through the other stopcock opening in such a
Alcohol 100 mL manner that all gas bubbles are eliminated from the system.
Purified Water A sufficient quantity Draw the Sample into the buret. By raising the leveling
To make 100 mL bottle, force the measured specimen into the pipet. The
absorption may be facilitated by rocking the pipet or by
Dissolve Basic Fuchsin in a mixture of Acetone and Alcohol, and flowing the specimen between pipet and buret. Draw any
add to this solution Phenol and Resorcinol, previously dissolved residual gas into the buret, and measure its volume.
in 725 mL of Purified Water. Then add sufficient Purified Water Acceptance criteria: NMT 1.0 mL of gas remains (NLT
to make the product measure 1000 mL, and mix. 99.0%, by volume, of CO2).
OTHER COMPONENTS IMPURITIES

ALCOHOL DETERMINATION 611: 7.0%10.0% Organic Impurities
PROCEDURE 1: LIMIT OF AMMONIA
SPECIFIC TESTS Sample: 1050 50 mL of the gas obtained as directed in
SPECIFIC GRAVITY 841: 0.9901.050 the test for Nitrogen Dioxide (below)
Analysis: Proceed with Carbon Dioxide as directed in the
ADDITIONAL REQUIREMENTS test for Nitrogen Dioxide (below), except to pass 1050 50
PACKAGING AND STORAGE: Preserve in tight, light-resistant mL of this gas through an ammonia detector tube at the
containers. rate specified for the tube.
Acceptance criteria: The indicator change corresponds to
NMT 0.0025%.
PROCEDURE 2: LIMIT OF HYDROGEN SULFIDE
Carbon Dioxide Sample: 1050 50 mL, released from the vapor phase
(Comment on this Monograph)id=m13040=Carbon Analysis: Pass 1050 50 mL, released from the vapor
Dioxide=Ca-Chl-Monos.pdf) phase, through a hydrogen sulfide detector tube at the rate
CO2 44.01 specified for the tube.
Carbon dioxide; Acceptance criteria: The indicator change corresponds to
Carbon dioxide [124-38-9]. NMT 1 ppm.
PROCEDURE 3: LIMIT OF NITRIC OXIDE
DEFINITION Sample: 550 50 mL, released from the vapor phase
Carbon Dioxide contains NLT 99.0%, by volume, of CO2. Analysis: Pass 550 50 mL, released from the vapor
[NOTEThe following tests are designed to reflect the quality of phase, through a nitric oxidenitrogen dioxide detector
Carbon Dioxide in both its vapor and liquid phases, which are tube at the rate specified for the tube.
present in previously unopened cylinders. Reduce the Acceptance criteria: The indicator change corresponds to
container pressure by means of a regulator. Withdraw the NMT 2.5 ppm.
specimens for the tests with the least possible release of PROCEDURE 4: CARBON MONOXIDE
Carbon Dioxide consistent with proper purging of the Sample: 1050 50 mL, released from the vapor phase of
sampling apparatus. Measure the gases with a gas volume the contents of the container
meter downstream from the detector tubes in order to Analysis: Pass 1050 50 mL, released from the vapor
minimize contamination or change of the specimens. Perform phase of the contents of the container, through a carbon
tests in the sequence in which they are listed.] monoxide detector tube at the rate specified for the tube.
The various detector tubes called for in the respective tests are Acceptance criteria: The indicator change corresponds to
listed in Reagents, Indicators, and SolutionsReagent NMT 0.001%.
Specifications. PROCEDURE 5: NITROGEN DIOXIDE
Sample: 550 50 mL, obtained as directed in the Analysis
IDENTIFICATION Analysis: Arrange the container so that when its valve is
PROCEDURE opened, a portion of the liquid phase of the contents is
Sample: 100 5 mL, released from the vapor phase of the released through a piece of tubing of sufficient length to
contents of the container allow all of the liquid to vaporize during passage through
Analysis: Pass the Sample through a Carbon Dioxide it, and to prevent frost from reaching the inlet of the
detector tube at the rate specified for the tube. detector tube. Release into the tubing a flow of liquid
Acceptance criteria: The indicator change extends sufficient to provide 550 mL of the vaporized specimen
throughout the entire indicating range of the tube. plus any excess necessary to ensure adequate flushing of air
from the system. Pass 550 50 mL of this gas through a
ASSAY nitric oxide-nitrogen dioxide detector tube at the rate
PROCEDURE specified for the tube.
[NOTESampling for this Assay may be done from the vapor Acceptance criteria: The indicator change corresponds to
phase for convenience, but this method results in more NMT 2.5 ppm.
residual volume. If the specification of 1 mL is exceeded PROCEDURE 6: SULFUR DIOXIDE
from the vapor phase, a liquid specimen may be taken.] Sample: 1050 50 mL, obtained as directed in the test for
Sample: 100.0 mL of specimen taken from the liquid Nitrogen Dioxide (above)
phase, as directed in the test for Nitrogen Dioxide (below) Analysis: Proceed with Carbon Dioxide as directed in the
Analysis: Assemble a 100-mL gas buret, provided with a test for Nitrogen Dioxide (above), except to pass 1050 50
leveling bulb and two-way stopcock, and a gas absorption mL through a sulfur dioxide detector tube at the rate
pipet of suitable capacity by connecting the pipet to one of specified for the tube.
the buret outlets. Fill the buret with slightly acidified water Acceptance criteria: The indicator change corresponds to
(turned pink with methyl orange), and fill the pipet with NMT 5 ppm.
potassium hydroxide solution (1 in 2). By manipulation of
the leveling bulb and leveling water, draw the potassium
72 Carbon / Official Monographs USP 32
SPECIFIC TESTS column containing support S3 and of sufficient length to

WATER DETERMINATION, Method I 921 separate CO2 from N2 and CO (which co-elute) and CO2
Analysis: Flush the regulator that has been flushed with 5 L from NO2 at room temperature. A simple radioactivity
or more of the gas specimen. Pass 50 5 L, released from detector and a thermal conductivity detector (or equivalent)
the vapor phase, through a water vapor detector tube are required for the mass determination of carbon
connected to the regulator with a minimum length of metal monoxide. The radiochemical purity is NLT 98%. Mass
or polyethylene tubing. Measure the gas passing through analysis of the gasair mixture must demonstrate levels of
the detector tube with a gas flowmeter set at a flow rate of carbon monoxide less than 1.23 mmoles in the entire dose,
2 L/min. which is the upper limit for a single-bolus inhalation.
Acceptance criteria: The indicator change corresponds to
NMT 150 mg/cubic meter. ADDITIONAL REQUIREMENTS
PACKAGING AND STORAGE: Dispense the gas either
ADDITIONAL REQUIREMENTS continuously or batchwise, and preserve in a single-dose
PACKAGING AND STORAGE: Preserve in cylinders. container that is adequately shielded. It may also be trapped
either on activated charcoal at 196 or on a molecular sieve
at 72.
LABELING: The label must include the following: the time
Carbon Monoxide C 11 and date of calibration; the amount of 11C as carbon
(Comment on this Monograph)id=m13050=Carbon Monoxide monoxide expressed as total MBq (mCi) at time of
C 11=Ca-Chl-Monos.pdf) calibration; the expiration time and date; and the statement
Radioactive Material. The labeling indicates that in making
DEFINITION dosage calculations, correction is to be made for radioactive
Carbon Monoxide C 11 is a colorless, odorless, nonirritating decay, and states that the radioactive half-life of 11C is 20.39
gas, suitable for administration by inhalation, in which a min. Each container to hold 11CO shall be independently
portion of the molecules are labeled with radioactive 11C. It labeled to indicate lot number and/or batch number. The
contains NLT 90.0% and NMT 110.0% of the labeled amount labeling states that a microbiological filter (0.22 m) is to be
of 11C expressed in MBq (or in mCi) at the time indicated in in place to remove any possible particulate matter that could
the labeling. be carried through to the final product.
IDENTIFICATION
RADIONUCLIDE IDENTIFICATION
(See Radioactivity 821.) Flumazenil C 11 Injection
A. Its gamma-ray spectrum is identical to that of a (Comment on this Monograph)id=m13053=Flumazenil C 11
specimen of 11C in that it exhibits a positron annihilation Injection=Ca-Chl-Monos.pdf)
peak at 0.511 MeV and possibly a sum peak of 1.022 MeV,
dependent upon geometry and detector efficiency. DEFINITION
B. Radio-gas chromatography, using a molecular sieve Flumazenil C 11 Injection is a sterile solution, suitable for
chromatographic column, to determine the absence of intravenous administration, of Flumazenil in which a portion of
[13N]N2, using a suitable radioactivity detector and mass the molecules are labeled at the N-position with radioactive
detector 11C. It contains NLT 90.0% and NMT 110% of the labeled
amount of 11C, expressed in MBq (or mCi) at the time

ASSAY indicated in the labeling. Its specific activity is NLT 14.8 GBq
PROCEDURE (400 mCi)/mol. It may contain suitable buffers.
Analysis: Determine the radioactivity, in MBq (or mCi), by
use of a calibrated system as directed under Radioactivity IDENTIFICATION
821. RADIONUCLIDIC IDENTIFICATION: Its gamma-ray spectrum is
Acceptance criteria: 90.0%110.0% identical to that of a specimen of 11C in that it exhibits a
positron annihilation peak at 0.511 MeV and possibly a sum
SPECIFIC TESTS peak of 1.022 MeV, dependent upon geometry and detector
SPECIFIC ACTIVITY: Dependent upon the amount of efficiency (see Radioactivity 821).
radioactivity to be inhaled, but NMT 1.23 mmoles of carbon
monoxide/volume ASSAY
RADIONUCLIDIC PURITY: A multichannel analyzer is used to RADIOACTIVITY
count all radioactivity from 40 to 2,500 KeV to determine (See Radioactivity 821, Selection of a Counting Assembly.)
the absence of radiation, other than at 0.511 MeV and 1.022 Analysis: Using a suitable counting assembly, determine the
MeV, over a period of 4 h. Possible impurities could be 13N2 radioactivity, in MBq (or mCi)/mL, of Injection by use of a
[t1/2 = 9.97 min; this gamma-ray spectrum is calibrated system.
indistinguishable from 11C, (B+) 491 KeV],10C [t1/2 = 19 s, Acceptance criteria: 90.0%110%
718.3 KeV (100%)], 14O [t1/2 = 70 s, 2312.7 KeV (99.4%)].
[11C]CO should contain NMT 10% impurities at the time of IMPURITIES
inhalation. Organic impurities
RADIOCHEMICAL PURITY AND MASS DETERMINATION: [NOTE PROCEDURE: CHEMICAL PURITY
This pharmaceutical may be synthesized by different Solution A: 1.2 mg/mL of monobasic sodium phosphate
methods and may therefore contain different impurities. in deionized distilled water
Additional validated tests relevant to the synthetic procedure Mobile phase: Acetonitrile and Solution A (3:7)
may be necessary in order to ensure radiochemical purity of System suitability stock solution: 1 mg/mL of USP
the final product.] Confirm by radio-gas chromatography. Flumazenil RS in acetonitrile
The gas stream, either directly from the target, or after initial System suitability solution: 10 g/mL of USP Flumazenil
chemical processing, is directed to an injection loop valve of RS from System suitability stock in Mobile phase
a gas chromatograph and two precalibrated columns, a Sample solution: 0.1 mL/mL of Injection in Mobile phase
molecular sieve that allows separation of carbon monoxide Chromatographic system
from the different air components (O2, N2, and CH4), and a (See Chromatography 621, System Suitability.)
Mode: LC and except that it is not subject to the recommendation on

Detector: UV 254 nm Volume in Container.
Temperature: 20 ADDITIONAL REQUIREMENTS
Flow rate: 2 mL/min PACKAGING AND STORAGE: Preserve in single-dose or
Injection size: 20 L multiple-dose containers that are adequately shielded.
System suitability LABELING: Label it to include the following, in addition to
Sample: System suitability solution the information specified for Labeling under Injections 1:
Suitability requirements the time and date of calibration; the amount of 11C as [N-
Column efficiency: NLT 100 theoretical plates methyl-11C]Flumazenil, expressed as MBq (or mCi); the
Tailing factor: NMT 1.1 specific activity, expressed as MBq (or mCi)/mol; the
Relative standard deviation: NMT 3.2% concentration, expressed as MBq (or mCi)/mL, at the date
Analysis and time of calibration; the expiration date and time; the lot
Sample: Sample solution or batch number; the name and quantity of any added
Separately calculate the percentage of each impurity in preservative or stabilizer; and the statements, Radioactive
the portion of Injection taken: Material and Do not use if cloudy or if it contains
particulate matter. The labeling indicates that in making
Result = (rU/rS) 100 dosage calculations, correction is to be made for radioactive
decay, and states that the radioactive half-life of 11C is 20
rU = peak response for each impurity min.
rS = sum of the responses of all the peaks USP REFERENCE STANDARDS 11
Acceptance criteria: NMT 0.2% of any individual USP Endotoxin RS
impurity, and NMT 0.9% of total impurities USP Flumazenil RS
SPECIFIC TESTS
BACTERIAL ENDOTOXINS TEST 85: NMT 175/V USP
Endotoxin Unit/mL, in which V is the maximum Mespiperone C 11 Injection
recommended total dose, in mL, at the expiration date or (Comment on this Monograph)id=m13057=Mespiperone C 11
time Injection=Ca-Chl-Monos.pdf)
PH 791: 4.58.5
RADIONUCLIDIC PURITY: Using a multichannel analyzer, count
all radioactivity from 40 to 2,500 keV to determine the
absence of radiation, other than at 0.511 MeV and 1.022
MeV, over a period of 4 h. Determine the half-life (20 min)
by a suitable detector system.
RADIOCHEMICAL PURITY
Mobile phase and System suitability solution: Prepare as
directed in the test for Chemical Purity.
Chromatographic system and System suitability: Proceed DEFINITION
as directed in the test for Chemical Purity, except that the Mespiperone C 11 Injection is a sterile, isotonic solution,
liquid chromatograph is also equipped with a suitable suitable for intravenous administration, of 3-N-[11C]
collimated radiation detector (see Radioactivity 821). methylspiperone in which a portion of the molecules are
Analysis: Inject 20 L of the Injection into the labeled with radioactive 11C. It contains NLT 90.0% and NMT
chromatograph, record the chromatogram, and measure 110.0% of the labeled amount of 11C expressed in GBq (or
responses for the major peaks. mCi) at the time indicated in the labeling. Its specific activity is
Acceptance criteria: The radioactivity under the main peak NLT 18.5 GBq (500 mCi)/mol.
is NLT 98% of the total radioactivity measured.
SPECIFIC ACTIVITY IDENTIFICATION
Mobile phase and System suitability solution: Proceed as RADIONUCLIDIC IDENTIFICATION: Its gamma- ray spectrum is
directed in the test for Chemical Purity. identical to that of a specimen of 11C in that it exhibits a
Chromatographic system and System suitability: Proceed positron annihilation peak at 0.511 MeV and possibly a sum
as directed in the test for Chemical Purity, except that the peak of 1.022 MeV, dependent on geometry and detector
liquid chromatograph is also equipped with a suitable efficiency. (See Radioactivity 821.)
collimated radiation detector (see Radioactivity 821). ASSAY
Analysis: Calculate the specific activity, in MBq (or ASSAY FOR RADIOACTIVITY 821
mCi)/mol, of Injection taken: Analysis: Using a suitable counting assembly (see
Radioactivity 821, Selection of a Counting Assembly),
Result = (Mr CR PR)/(C 100) determine the radioactivity, in GBq (or mCi)/mL, of Injection
Mr = molecular weight of flumazenil, 303.29 by use of a calibrated system.
CR = radioactivity content as determined in the Assay Acceptance criteria: 90.0%110%
[MBq (or mCi)/mL] SPECIFIC TESTS
PR = radiochemical purity as determined in the test for BACTERIAL ENDOTOXINS TEST 85: NMT 175/V USP
Radiochemical Purity (%) Endotoxin Unit/mL of the Injection, in which V is the
C = concentration of flumazenil in the Injection as maximum recommended total dose, in mL, at the expiration
determined in the test for Chemical Purity time
(g/mL) PH 791: 4.57
Acceptance criteria: NLT 400 mCi/mol RADIONUCLIDIC PURITY: Using a multichannel analyzer, count
OTHER REQUIREMENTS: It meets the requirements under all radioactivity from 40 to 2500 KeV to determine the
Injections 1, except that the Injection may be distributed or absence of radiation, other than at 0.511 MeV and 1.022
dispensed prior to completion of Sterility Tests 71, the latter
test being started on the day following final manufacture,
MeV, over a period of 4 h. Determine the half-life (20 min) expressed as total GBq (or mCi) at time of calibration; the
by a suitable detector system. expiration time and date; the lot or batch number; and the
CHEMICAL PURITY statements, [CAUTIONRadioactive Material] and Do not
Mobile phase: Acetonitrile and 0.05 M monobasic use if cloudy or if it contains particulate matter. The
potassium phosphate (7:3) labeling indicates that in making dosage calculations
Standard solution: 0.1 mg/mL of 3-methylspiperone correction is to be made for radioactive decay, and states
hydrochloride in Mobile phase that the radioactive half-life of 11C is 20 min.
Sample solution: 0.1 mg/mL from a volume of Injection in USP REFERENCE STANDARDS 11
Mobile phase USP Endotoxin RS
Mode: LC
Detector: UV 254 nm, and a suitable radioactivity detector Methionine C 11 Injection
(see Radioactivity 821) (Comment on this Monograph)id=m13060=Methionine C 11
Column: 3.9-mm 30-cm; packing L1 Injection=Ca-Chl-Monos.pdf)
Injection size: 20 L DEFINITION
System suitability Methionine C 11 Injection is a sterile isotonic solution, suitable
Sample: Standard solution for intravenous administration of L[11C]methionine, in which a
Suitability requirements portion of the molecules are labeled with radioactive 11C. It
Column efficiency: NLT 100 theoretical plates contains NLT 90.0% and NMT 110.0% of the labeled amount
Tailing factor: NMT 1.1 of 11C expressed in MBq (or in mCi) at the time indicated in
Relative standard deviation: NMT 3.2% the labeling. It may contain preservatives and stabilizers.
Analysis
Separately calculate the percentage of each impurity in the RADIONUCLIDIC IDENTIFICATION: Its gamma-ray spectrum is
portion of the Injection taken: identical to that of a specimen of 11C in that it exhibits a
positron annihilation peak at 0.511 MeV and possibly a sum
Result = (ri/rs) 100 peak of 1.022 MeV, dependent upon geometry and detector
efficiency. (See Radioactivity 821.)
ri = peak response for each impurity
rs = sum of the responses of all the peaks ASSAY
Acceptance criteria ASSAY FOR RADIOACTIVITY 821
Individual impurities: NMT 0.2% Analysis: Using a suitable counting assembly, (see Selection
Total impurities: NMT 0.9% of a Counting Assembly) determine the radioactivity (see
RADIOCHEMICAL PURITY 821: Proceed as directed in the test Radioactivity 821), in GBq (Ci)/mL, of the Injection by use
for Chemical Purity, except that the liquid chromatograph is of a calibrated system.
also equipped with a suitable collimated radioactivity Acceptance criteria: 90.0%110%
detector. The radioactivity under the main peak is NLT 98% SPECIFIC TESTS
of the total radioactivity measured. BACTERIAL ENDOTOXINS TEST 85: NMT 175/V USP
SPECIFIC ACTIVITY Endotoxin Unit/mL of the Injection, in which V is the
Mobile phase and Standard solution: Proceed as directed maximum recommended total dose, in mL, at the expiration
in the test for Chemical Purity. time
Chromatographic system: Proceed as directed in the test PH 791: 6.08.0
for Chemical Purity. RADIONUCLIDIC PURITY: Using a suitable gamma-ray
Analysis: Calculate the specific activity, in MBq (or spectrometer, determine the absence of radiation other than
mCi)/mol, of Mespiperone C 11 Injection taken: at 0.511 MeV, over a period of 20 min. Determine the half-
life (20.41 min) by a suitable detector system.
Result = 3.11 (Cr Pr)/C CHEMICAL PURITY
Cr = radioactivity content, as determined in the Assay Mobile phase: 0.008 M copper acetate and 0.017 M L-
for Radioactivity [MBq (or mCi)/mL] proline, adjust with 0.030 M sodium acetate to a pH of 5
Pr = radiochemical purity, as determined in the test Standard solution: 0.1 mg/mL of DL-methionine in Mobile
for Radiochemical Purity (%) phase
C = concentration of 3-methylspiperone in the Sample solution: 0.1 mg/mL from a volume of Injection in
Injection as determined in the test for Chemical Mobile phase
Purity (g/mL) Chromatographic system
Acceptance criteria: NLT 18.5 GBq/mol (400 mCi/mol) (See Chromatography 621, System Suitability.)
OTHER REQUIREMENTS: It meets the requirements under Mode: LC
Injections 1, except that the Injection may be distributed or Detector: UV 254 nm
dispensed prior to completion of the test for Sterility, the Column: 4.6-mm 15-cm; packing L1
latter test being started on the day following final Flow rate: 0.5 mL/min
manufacture, and except that it is not subject to the Injection size: 10 L
recommendation on Volume in Container. System suitability
ADDITIONAL REQUIREMENTS [NOTEThe relative retention times for D-methionine and L-
PACKAGING AND STORAGE: Preserve in single-dose or in methionine are 1.0 and 2.4, respectively.]
multiple-dose containers that are adequately shielded.
LABELING: Label it to include the following: the time and
date of calibration; the amount of 11C as methylspiperone

Resolution: NLT 1.5 between the D- and L-enantiomers RADIONUCLIDIC IDENTIFICATION: Its gamma-ray spectrum is
Column efficiency: NLT 1400 theoretical plates identical to that of a specimen of 11C in that it exhibits a
Tailing factor: NMT 2.0 positron annihilation peak at 0.511 MeV and possibly a sum
Relative standard deviation: NMT 2.0% peak of 1.022 MeV, dependent upon geometry and detector
Analysis efficiency (see Radioactivity 821).
Calculate the percentage of D-methionine in the portion of ASSAY
Injection taken: ASSAY FOR RADIOACTIVITY 821
(See Radioactivity 821, Selection of a Counting Assembly.)
Result = (rU/rS) 100 Analysis: Using a suitable counting assembly, determine the
radioactivity, in MBq (or mCi)/mL, of Injection by use of a
rU = peak response for D-methionine calibrated system.
rS = sum of the responses of all the peaks Acceptance criteria: 90.0%110%
Acceptance criteria: NMT 4% of D-enantiomer is found
RADIOCHEMICAL PURITY 821: Proceed as directed under SPECIFIC TESTS
Chemical Purity. The radioactivity of the main peak is NLT RADIONUCLIDIC PURITY: Using a multichannel analyzer, count
98% of the total radioactivity measured. all radioactivity from 402500 keV to determine the absence
SPECIFIC ACTIVITY: NLT 37.0 GBq (1.0 Ci)/mmol of radiation, other than at 0.511 MeV and 1.022 MeV, over a
OTHER REQUIREMENTS: It meets the requirements under period of 4 h. Determine the half-life (20 min) by a suitable
Injections 1, except that the Injection may be distributed or detector system.
dispensed prior to completion of Sterility Tests 71 and CHEMICAL PURITY
Bacterial Endotoxins Test 85, these tests being started on the Mobile phase: Add 840 L of phosphoric acid to 500 mL
day of final manufacture, and except that it is not subject to of deionized distilled water in a 1000-mL volumetric flask.
the recommendation of Volume in Container. Add 270 mL of acetonitrile, and dilute with deionized
distilled water to volume.
ADDITIONAL REQUIREMENTS Standard stock solution: 1 mg/mL of raclopride (as the
PACKAGING AND STORAGE: Preserve in single-dose or tartrate salt) in water. Dilute a volume of the resulting
multiple-dose containers that are adequately shielded. solution with Mobile phase to obtain a concentration of 10
LABELING: Label it to include the following: the time and g/mL.
date of calibration; the amount of 11C as methionine Sample solution: Transfer a volume of Injection equivalent
expressed as total MBq (or mCi)/mL at time of calibration; to 37 MBq (1 mCi) of radioactivity from a volume of
the expiration time and date; the name and quantity of any Injection, with 10 parts of Mobile phase.
added preservative or stabilizer; and the statement Chromatographic system
[CAUTIONRadioactive Material]. The labeling indicates that (See Chromatography 621, System Suitability.)
in making dosage calculations, correction is to be made for Mode: LC
radioactive decay, and states that the radioactive half-life of Detector: UV 254 nm
11C is 20.4 min. Each container to hold 11C methionine shall Column: 3.9-mm 30-cm; packing L9
be independently labeled to indicate lot number and batch Flow rate: 2 mL/min
number. The labeling states that a microbiological filter Injection size: 20 L
(0.22 m) is to be in place to remove any possible System suitability
particulate matter that could be carried through to the final Sample: Standard solution
product. [NOTEThe relative retention times for o-
USP REFERENCE STANDARDS 11 desmethylraclopride and for raclopride are 0.75 and 1.0,
USP Endotoxin RS respectively.]
Resolution: NLT 1.5 between acetate and carbonate
Raclopride C 11 Injection Relative standard deviation: NMT 10.0%
(Comment on this Monograph)id=m13070=Raclopride C 11 Analysis
Injection=Ca-Chl-Monos.pdf) Samples: Standard solution and Sample solution
Measure the areas of the responses for the raclopride
peaks. Calculate the concentration, in g/mL, of
C15H20Cl2N2O3 in the portion of Injection taken:

C1411CH20Cl2N2O3 346.24 CS = concentration in the Standard solution (g/mL)
[97849-54-2]. CU = concentration in the Sample solution (g/mL)
Acceptance criteria: In the chromatogram of the Sample
DEFINITION solution, the area of the peak with a retention time of 6 min
Raclopride C 11 Injection is a sterile solution, suitable for (raclopride) is NLT 98% of the total area of all peaks.
intravenous administration, of raclopride, in which a portion of RADIOCHEMICAL PURITY
the molecules are labeled at the O-methyl position with Mobile phase: Prepare as directed in the test for Chemical
radioactive 11C. It contains NLT 90.0% and NMT 110% of the Purity.
labeled amount of 11C expressed in MBq (or mCi) at the time Standard solution: Prepare as directed in the test for
indicated in the labeling. Its specific activity is NLT 18.5 Gbq Chemical Purity.
(500 mCi)/mol. It may contain suitable buffers.
Chromatographic system: Proceed as directed in the test Sodium Acetate C 11 Injection

for Chemical Purity, except that the liquid chromatograph is (Comment on this Monograph)id=m13080=Sodium Acetate C
also equipped with a suitable collimated radiation detector 11 Injection=Ca-Chl-Monos.pdf)
(see Radioactivity 821).
Acceptance criteria: NLT 95% of the radioactivity under
the main peak [NOTEIts retention time is within 10% of
that obtained for the Standard solution, similarly
chromatographed.]
Endotoxin Unit/mL, in which V is the maximum
recommended total dose, in mL, at the expiration date or
time
PH 791: 4.57 DEFINITION
SPECIFIC ACTIVITY Sodium Acetate C 11 Injection is a sterile solution, suitable for
Mobile phase and Standard solution: Proceed as directed intravenous administration, of Sodium Acetate in which a
in the test for Chemical Purity. portion of the carboxyl molecules are labeled with radioactive
11C. It contains NLT 90.0% and NMT 110.0% of the labeled
Chromatographic system: Proceed as directed in the test
for Radiochemical Purity. amount of 11C expressed in MBq (or in Ci or mCi) at the
Analysis: Calculate the specific activity, in GBq (or time indicated in the labeling. It may contain suitable buffers.
mCi)/mol, of the Injection taken:
IDENTIFICATION
Result = (Mr Cr Pr)/(100 C) RADIOACTIVITY, Radionuclides Identification 821: Its gamma-
ray spectrum is identical to that of a specimen of 11C in that
Mr = molecular weight of raclopride, 347 it exhibits a positron annihilation peak at 0.511 MeV and
Cr = radioactivity content, as determined in the Assay possibly a sum peak of 1.022 MeV, depending upon
for Radioactivity [GBq (or mCi)/mL] geometry and detector efficiency.
Pr = radiochemical purity, as determined in the test
for Radiochemical Purity (%) ASSAY
C = concentration of raclopride in the Injection as PROCEDURE
determined in the test for Chemical Purity (See Radioactivity 821, Selection of a Counting Assembly.)
(g/mL) Analysis: Using a suitable counting assembly, determine the
Acceptance criteria: NLT 18.5 GBq/mol (500 mCi/mol) radioactivity in MBq (mCi)/mL of the Injection by use of a
OTHER REQUIREMENTS: It meets the requirements under calibrated system.
Injections 1, except that the Injection may be distributed or Acceptance criteria: 90.0%110% of the labeled amount
dispensed prior to completion of the test for Sterility, the of 11C
latter test being started on the day following final SPECIFIC TESTS
manufacture, and except that it is not subject to the RADIONUCLIDIC PURITY 821:
recommendation on Volume in Container. Use a multichannel analyzer to count all radioactivity from
ADDITIONAL REQUIREMENTS 402,500 keV to determine the absence of radiation, other
PACKAGING AND STORAGE: Preserve in single-dose or in than at 0.511 MeV and 1.022 MeV, over a period of 4 h.
multiple-dose containers that are adequately shielded. Determine the half-life by a suitable detector system.
LABELING: Label it to include the following, in addition to CHEMICAL PURITY
the information specified for Labeling under Injections 1: the Mobile phase Add 14 mL of 0.5 N sulfuric acid to 500 mL
time and date of calibration; the amount of 11C as [o- of water in a 1000-mL volumetric flask. Add 100 mL of
methyl-11C]raclopride, expressed as total megabecquerels (or acetonitrile, and dilute with water to volume.
millicuries); the specific activity, expressed as Standard solution: 1 mg/mL of sodium acetate in water.
megabecquerels (or millicuries)/mol; and the concentration, Dilute a portion of the resulting solution with Mobile phase
expressed as megabecquerels (or millicuries)/mL, at the date to obtain a concentration of 20 g/mL.
and time of calibration; the expiration date and time; the lot Sample solution: Dilute a volume of Injection equivalent to
or batch number; the name and quantity of any added 1 mCi of radioactivity with 10 parts of Mobile phase.
preservative or stabilizer; and the statements, [CAUTION Chromatographic system
Radioactive Material] and Do not use if cloudy or if it (See Chromatography 621, System Suitability.)
contains particulate matter. The labeling indicates that in
making dosage calculations correction is to be made for
radioactive decay, and states that the radioactive half-life of
11C is 20 min.

USP Endotoxin RS

Flow rate: 1 mL/min
System suitability Urea C 13
Sample: Standard solution (Comment on this Monograph)id=m13083=Urea C 13=Ca-Chl-
[NOTEThe relative retention times for acetate and Monos.pdf)
carbonate are about 0.8 and 1.0 min, respectively.]
Resolution: NLT 85 theoretical plates Urea C 13 contains NLT 99.0% and NMT 100.5% of 13CH4N2O.
Analysis ASSAY
Samples Standard solution and Sample solution PROCEDURE
Measure the responses for the acetate peaks. Mobile phase: Acetonitrile, methanol, and water (89:10:1)
Calculate the concentration, in g/mL, of sodium acetate in Biuret stock solution: 0.03 mg/mL of biuret in Mobile
the Injection: phase
System suitability solution: 25 mg of Urea in 1 mL Biuret
Result = (rU/rS) CS stock solution, and dilute with Mobile phase to 10 mL
Standard solution: 2 mg/mL of USP Urea C 12 RS in
rU = peak response of acetate from the Sample Mobile phase
solution Sample solution: 2 mg/mL of Urea C 13 in Mobile phase
rS = peak response of acetate from the System Chromatographic system
suitability solution (g/mL) (See Chromatography 621, System Suitability.)
CS = concentration of sodium acetate in the Standard Mode: LC
solution (g/mL) Detector: UV 200 nm
RADIOCHEMICAL PURITY Column: 4.6-mm 25-cm; 5-m packing L8
Mobile phase and Standard solution: Proceed as directed Flow rate: 0.8 mL/min
under Chemical Purity. Injection size: 20 L
Sample solution: Sodium Acetate C 11 Injection, undiluted System suitability
Chromatographic system and System suitability: Proceed Samples: System suitability solution and Standard solution
as directed under Chemical Purity except that the liquid Suitability requirements
chromatograph is also equipped with a suitable collimated Resolution: NLT 1.5 between urea and biuret
radiation detector (see Radioactivity 821). Relative standard deviation: NMT 1.0%
Analysis Analysis
Sample: Sample Solution (30 L) Samples: Standard solution and Sample solution
Measure the areas for the major peaks. Measure the areas for the major peaks.
Acceptance criteria: The radioactivity under the acetate C Calculate the percentage of 13CH4N2O in the portion of
11 peak is NLT 95% of the total area of all peaks observed. Urea C 13 taken:
[NOTEIts retention time is within 10% of that obtained
for the Standard solution, similarly chromatographed.] Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
Endotoxin Unit/mL, in which V is the maximum rU = peak response from the Sample solution
recommended total dose, in mL, at the expiration date or rS = peak response from the Standard solution
time CS = concentration of USP Urea C 12 RS in the
PH 791: 4.58.5
SPECIFIC ACTIVITY: NLT 3.7 GBq (100 mCi)/mol CU = concentration of Urea C 13 in the Sample
OTHER REQUIREMENTS: It meets the requirements under solution (mg/mL)
Injections 1, except that the Injection may be distributed or Mr1 = molecular weight of Urea C 13
dispensed prior to completion of the test for Sterility, the Mr2 = molecular weight of USP Urea C 12 RS
latter test being started on the day following final Acceptance criteria: 99.0%100.5%
manufacture, and except that it is not subject to the IMPURITIES
recommendation of Volume in Container. Inorganic Impurities
ADDITIONAL REQUIREMENTS RESIDUE ON IGNITION: NMT 0.1%
PACKAGING AND STORAGE: Preserve in single-dose or in HEAVY METALS
multiple-dose containers that are adequately shielded. Analysis: Dissolve 1.0 g in 20 mL of water, and add 5 mL
LABELING: Label it to include the following, in addition to of 0.1 N hydrochloric acid.
the information specified in Injections 1, Labeling: the time Acceptance criteria: NMT 20 ppm
and date of calibration; the amount of 11C as labeled sodium SPECIFIC TESTS
acetate expressed as total MBq (or mCi) and the LIMIT OF BIURET
concentration as megabecquerels/mL (or as millicuries/mL), Standard solution: 0.033 mg/mL of biuret
on the date and time of calibration; the expiration date and Sample solution: 33.3 mg/mL of Urea C 13
time; the lot or batch number; the name and quantity of Analysis: To the Sample solution and to 3 mL of the
any added preservative or stabilizer; an indication on the Standard solution, add 3 mL of sodium hydroxide solution
labeling that states, Do not use if cloudy or if it contains (10 in 100) and 3 drops of copper sulfate solution (0.5 in
particulate matter; and the statement [CAUTIONRadioactive 100), and allow to stand for 5 min.
Material.]. The labeling indicates that in making dosage Acceptance criteria: NMT 0.1% of biuret. Any reddish
calculations, correction is to be made for radioactive decay, violet color in the Sample solution is not more intense than
and also indicates that the radioactive half-life of 11C is 20 that from the Standard solution.
min.
ISOTOPIC PURITY Urea C 13 for Oral Solution

Sample solution: 12 mg/mL of Urea C 13 in methanol (Comment on this Monograph)id=m13085=Urea C 13 for Oral
Chromatographic system Solution=Ca-Chl-Monos.pdf)
(See Chromatography 621 and Mass Spectrometry 736.)
Mode: GC connected to a mass spectrometer DEFINITION
[NOTEThe mass spectrometer is operated in a single-ion Urea C 13 for Oral Solution is a dry powder prepared from
response mode. The electron energy is 70 eV.] Urea C 13. It contains NLT 90.0% and NMT 110.0% of the
Column: 0.25-mm 15-m capillary column coated with a labeled amount of urea C 13 (13CH4N2O). It contains no
0.1-m film of phase G47 preservatives.
Temperature
Injection port: 250 ASSAY
Detector: 200 PROCEDURE
Transfer line to the mass spectrometer: 265 Mobile phase: Acetonitrile, methanol, and water (89:10:1)
Carrier gas: Helium Biuret stock solution: 0.03 mg/mL of biuret in Mobile
Injection size: 1 L phase
Analysis System suitability solution: 25 mg of Urea in 1 mL Biuret
Sample: Sample solution stock solution, and dilute with Mobile phase to 10 mL
Record the total ion chromatogram, and combine all of the Standard solution: 2 mg/mL of USP Urea C 12 RS in
mass spectra scans across the entire major peak. Record Mobile phase
the peak intensities at mass-to-charge ratios of 60, 61, 62, Sample solution: 2 mg/mL of Urea C 13 in Mobile phase
and 63. Chromatographic system
Calculate the percentage of carbon that is C 13 in the (See Chromatography 621, System Suitability.)
portion of Urea C 13: Mode: LC
Detector: UV 200 nm
Result = [(I61 + I63)/(I60 + I61 + I63)] 100 Column: 4.6-mm 25-cm; 5-m packing L8
I60 = relative peak intensity at mass-to-charge ratio of Injection size: 20 L
60 System suitability
I61 = relative peak intensity at mass-to-charge ratio of Samples: System suitability solution and Standard solution
61 Suitability requirements
I63 = relative peak intensity at mass-to-charge ratio of Resolution: NLT 1.5 between urea and biuret
63 Relative standard deviation: NMT 1.0%
Acceptance criteria: NLT 99% is found Analysis
Calculate the percentage of oxygen that is O 18 in the Samples: Standard solution and Sample solution
portion of Urea C 13: Measure the areas for the major peaks.
Calculate the percentage of 13CH4N2O in the portion of
Result = [(I62 + I63)/(I60 + I61 + I62 + I63)] 100 Urea C 13 for Oral Solution taken:
I62 = relative peak intensity at a mass-to-charge ratio Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
of 62, and the other terms are as defined above
Acceptance criteria: NMT 15% is found rU = peak response from the Sample solution
MELTING RANGE OR TEMPERATURE: 132135 rS = peak response from the Standard solution
ALCOHOL-INSOLUBLE MATTER CS = concentration of USP Urea C 12 RS in the
Analysis: Dissolve 5.0 g in 50 mL of warm alcohol, and if Standard solution (mg/mL)
any insoluble residue remains, filter the solution on a tared CU = nominal concentration of Urea C 13 in the
filter, wash the residue, and the filter with 20 mL of warm Sample solution (mg/mL)
alcohol, and dry at 105 for 1 h. Mr1 = molecular weight of Urea C 13
Acceptance criteria: The weight of the residue does not Mr2 = molecular weight of USP Urea C 12 RS
exceed 2 mg (0.04%). Acceptance criteria: 90.0%110.0%
CHLORIDE AND SULFATE 221, Chloride: A 2.0-g portion
shows no more chloride than corresponds to 0.20 mL of PERFORMANCE TESTS
0.020 N hydrochloric acid (0.007%). UNIFORMITY OF DOSAGE UNITS: Meets the requirements for
CHLORIDE AND SULFATE 221, Sulfate: A 2.0-g portion shows packaged solids
no more sulfate than corresponds to 0.20 mL of 0.020 N
sulfuric acid (0.010%). SPECIFIC TESTS
OTHER REQUIREMENTS: It meets the requirements for MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED
Identification tests A and B under Urea. MICROORGANISMS 62 and TESTS FOR SPECIFIED
MICROORGANISMS 62: The total aerobic microbial count
ADDITIONAL REQUIREMENTS does not exceed 1000 cfu/g, the total combined molds and
PACKAGING AND STORAGE: Preserve in well-closed containers yeasts count does not exceed 100 cfu/g, and it meets the
at room temperature. requirements for the absence of Salmonella species and
USP REFERENCE STANDARDS 11 Eschericha coli.
USP Urea C 13 RS
USP 32 Official Monographs / Carboplatin 79
COMPLETENESS OF SOLUTION 641: Meets the requirements, a Identification solution: 40 mg/mL of urea
solution in carbon dioxide-free water containing 100 mg/mL Application volume: 20 L of the Sample solution and 4 L
being used of the Identification solution
OTHER REQUIREMENTS: It meets the requirements for Developing solvent system: n-Butanol saturated with water
Identification tests A and B under Urea. Chromatographic system
ADDITIONAL REQUIREMENTS Mode: TLC
PACKAGING AND STORAGE: Preserve in sterile, well-closed Analysis
containers. Samples: Sample solution and Identification solution
LABELING: Label it to indicate that the solution is to be Locate the spots on the plate by spraying with Ehrlichs
discarded if particulate matter is visible after reconstitution. reagent. Determine the radioactivity distribution with a
[NOTEIt is to be reconstituted with Sterile Purified Water.] suitable radiation detector (see Radioactivity 821), and
obtain the RF value.
Acceptance criteria: NLT 90% of the total radioactivity is in
Urea C 14 Capsules the 14C band, and the RF value of the principal spot from the
Sample solution corresponds to that from the Identification
(Comment on this Monograph)id=m13090=Urea C 14 solution.
Capsules=Ca-Chl-Monos.pdf)
LABELING: Label it to include the following: the amount of
14C, expressed in MBq (or Ci)/Capsule at the time of
calibration; the expiration date; the total

radioactivity/container; and the statement, Caution
Radioactive Material.
DEFINITION
Urea C 14 Capsules contain 14CH4N2O in which a portion of the
molecules are labeled with radioactive 14C to provide 0.04 Carboplatin
MBq (or 1 Ci) of radioactivity per capsule. It contains NLT (Comment on this Monograph)id=m13120=Carboplatin=Ca-
90.0% and NMT 110.0% of the labeled amount of 14C Chl-Monos.pdf)
expressed as MBq (or Ci).
IDENTIFICATION
RADIOACTIVITY, Identification and Assay of Radionuclides 821
A solution of 1 or more Capsules in 1 N hydrochloric acid
when tested using a liquid scintillation counter produces
beta emission having a 49 keV mean and a 156 keV max.
ASSAY
PROCEDURE C6H12N2O4Pt 371.25
Assay of Radioactivity 821 Platinum, diammine[1,1-cyclobutanedicarboxylato(2-)-
Analysis: Prepare a solution of 1 or more Capsules in 1 N O,O]-, (SP-4-2);
hydrochloric acid. Using a liquid scintillation counter, cis-Diammine(1,1-cyclobutanedicarboxylato)platinum
determine the radioactivity, in MBq (or mCi)/mL by use of [41575-94-4].
a calibrated system. DEFINITION
Acceptance criteria: 90.0%110.0% of the labeled amount Carboplatin contains NLT 98.0% and NMT 102.0% of
of 14C. C6H12N2O4Pt, calculated on the anhydrous basis.
PERFORMANCE TESTS
DISSOLUTION 711 [CAUTIONGreat care should be taken in handling Carboplatin
Medium: Simulated gastric fluid TS; 500 mL because it is a suspected carcinogen.]
Apparatus 1: 50 rpm IDENTIFICATION
Time: 10 min INFRARED ABSORPTION 197K: Meets the requirements
Analysis: Determine the background levels of 14C with a 1-
mL portion of the solution under test using a liquid ASSAY
scintillation counter. PROCEDURE
Tolerances: NLT 80% (Q) of the labeled amount of 14C is Mobile phase: Acetonitrile and water (87:13)
dissolved. Standard solution: 1 mg/mL of USP Carboplatin RS
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements [NOTEUse this solution within 2 h.]
Sample solution: 1 mg/mL of Carboplatin
SPECIFIC TESTS [NOTEUse this solution within 2 h.]
RADIONUCLIDIC PURITY 821 Chromatographic system
Analysis: Determine the radionuclidic purity of a solution of (See Chromatography 621, System Suitability.)
1 or more Capsules in water using a liquid scintillation
counter.
Acceptance criteria: NLT 99.9% of the radioactivity is
present as C 14.
RADIOCHEMICAL PURITY
Adsorbent: 0.25-mm layer of chromatographic cellulose
Sample solution: Open 2 Capsules and place them in a
suitable container, and add 8 mL of methanol.
80 Carboplatin / Official Monographs USP 32
Mode: LC Mobile phase: Acetonitrile, Solution A, and water (5:1:44)

Detector: UV 230 nm Standard solution A: 0.005 mg/mL of 1,1-
Column: 4.0-mm 30-cm; packing L8 cyclobutanedicarboxylic acid in Mobile phase
Flow rate: 2 mL/min Standard solution B: 1 mg/mL of USP Carboplatin RS
Injection size: 10 L System suitability solution: Mix 1.0 mL of Standard
System suitability solution A with 1.0 mL of Standard solution B.
Sample: Standard solution Sample solution: 1 mg/mL of Carboplatin in Mobile phase
Suitability requirements Chromatographic System
Capacity factor, k: NLT 3.0 (See Chromatography 621, System Suitability.)
Column efficiency: NLT 2500 theoretical plates Mode: LC
Tailing factor: NMT 2.5 Detector: UV 220 nm
Relative standard deviation: NMT 1.2% Column: 4.0-mm 30-cm; packing L1
Calculate the percentage of C6H12N2O4Pt in the portion of System suitability
Carboplatin taken: Sample: System suitability solution
Result = (rU/rS) (CS/CU) 100 [NOTEThe relative retention times are 0.65 for
carboplatin, and 1.0 for 1,1-cyclobutanedicarboxylic
rU = peak response from the Sample solution acid.]
rS = peak response from the Standard solution Resolution: NLT 2.5 between the carboplatin and 1,1-
CS = concentration of USP Carboplatin RS in the cyclobutanedicarboxylic acid peaks
Standard solution (mg/mL) Column efficiency: NLT 1500 theoretical plates,
CU = concentration of the Sample solution (mg/mL) determined from the cyclobutanedicarboxylic acid peak
Acceptance criteria: 98.0%102.0% Relative standard deviation: NMT 10%
Analysis
OTHER COMPONENTS Samples: Standard solution and Sample solution
PLATINUM CONTENT Calculate the percentage of 1,1-cyclobutanedicarboxylic
[NOTEThoroughly cleanse all glassware with nitric acid and acid in the portion of Carboplatin taken:
rinse with water to prevent mirroring of platinum
precipitate.] Result = (rU/rs) (CS/CU) 100
Sample solution: Transfer 0.25 g of Carboplatin to a 600-
mL beaker. Add 400 mL of water, and slowly dissolve by rU = peak response from the Sample solution
heating almost to the boiling point on a hot plate covered rS = peak response from the Standard solution
with an insulating pad, stirring frequently with a glass rod. CS = concentration of 1,1-cyclobutanedicarboxylic
Analysis: When the Sample solution is complete, remove the acid in the Standard solution (mg/mL)
insulating pad, and boil for about 10 min. Remove the CU = concentration of carboplatin in the Sample
beaker from the hot plate, allow to cool for 1 min without solution (mg/mL)
stirring, and filter through quantitative, fine-porosity, Acceptance criteria: NMT 0.5%
smooth, dense, ashless filter paper, collecting the filtrate in a PROCEDURE 2
600-mL beaker, and completing the transfer to the filter Mobile phase, Chromatographic system, and Analysis:
with hot water. Wash the filter with hot water. Place the Proceed as directed in the Assay.
beaker containing the combined filtrate and washings on a Standard solution: 2.5 g/mL of USP Carboplatin RS
hot plate, and evaporate to a volume of about 300 mL. Sample solution: Use the Sample solution, prepared under
Place a glass stirring rod in the beaker, and heat the solution Assay.
to boiling. Slowly add to the center of the beaker, by Analysis
dropwise additions, 10.0 mL of hydrazine hydrate, 85%. Samples: Standard solution and Sample solution
[CautionHydrazine is toxic.] Analysis: Separately inject 10 L of the Standard solution
Add 2 drops of 10 N sodium hydroxide, boil for 10 min to and the Sample solution into the chromatograph, record
coagulate the precipitate for ease of filtration, cool, and the chromatograms, and measure the peak responses.
filter through quantitative, medium-porosity, smooth, Acceptance criteria
ashless filter paper. Rinse the beaker with hot water, and [NOTEThe sum of the peak responses, excluding the
pour the rinsings onto the filter. Wipe the beaker and the carboplatin and 1,1-cyclobutanedicarboxylic acid
stirring rod with small pieces of the same kind of paper responses, from the Sample solution, is NMT 2 times the
used for this filtration, and place these and the filter carboplatin response from the Standard solution, and no
containing the precipitate in a No. 1 porcelain crucible, single peak response is greater than that of the
previously ignited to constant weight. Dry on a hot plate carboplatin peak from the Standard solution.]
covered with an insulating pad, slowly increase the heat to Individual impurities: NMT 0.25%
char, and ignite for 1 h at 800. Cool in a desiccator, and Total impurities: NMT 0.5%
weigh again.
Acceptance criteria: The weight of the platinum so SPECIFIC TESTS
obtained is between 52.0%53.0% of the carboplatin taken. CRYSTALLINITY 695: Meets the requirements
PH 791: 5.07.0, in a 10 mg/mL solution
IMPURITIES WATER DETERMINATION, Method I 921: NMT 0.5%, using
Organic Impurities anhydrous formamide as the solvent
PROCEDURE 1: LIMIT OF 1,1-CYCLOBUTANEDICARBOXYLIC ACID TRANSMITTANCE
Solution A: Dissolve 8.5 g of tetrabutylammonium Sample solution: 10 mg/mL of Carboplatin
hydrogen sulfate in 80 mL of water. Add 3.4 mL of Analysis: Determine the percent transmittance in 1-cm cells
phosphoric acid, and adjust with 10 N sodium hydroxide to at a wavelength of 440 nm, using water as the blank.
a pH of 7.55.
USP 32 Official Monographs / Carboplatin 81
Acceptance criteria: NLT 97% transmittance Chromatographic system

WATER-INSOLUBLE MATTER (See Chromatography 621, System Suitability.)
Sample solution: Dissolve 1 g of carboplatin in 100 mL of Mode: LC
water. Detector: UV 230 nm
Dissolve by stirring with a stirring bar for 30 min. With the Column: 4.0-mm 30-cm; packing L8
aid of suction, filter through a tared filtering crucible. Rinse Flow rate: 2 mL/min
the beaker with water, and transfer the rinsings to the Injection size: 10 L
crucible. Dry the crucible at 130 10 to constant weight. System suitability
Acceptance criteria: NMT 0.5% Sample: Standard solution
ADDITIONAL REQUIREMENTS Capacity factor, k: NLT 3.0
PACKAGING AND STORAGE: Preserve in tight containers, Column efficiency: NLT 2500 theoretical plates
protected from light. Tailing factor: NMT 2.5
USP Carboplatin RS Analysis
Calculate the percentage of C6H12N2O4Pt in the portion of
Carboplatin for Injection Carboplatin for Injection taken:
(Comment on this Monograph)id=m13125=Carboplatin for Result = (rU/rS) (CS/CU) 100
DEFINITION rS = peak response from the Standard solution
Carboplatin for Injection is a sterile, lyophilized mixture of CS = concentration of USP Carboplatin RS in the
Carboplatin and Mannitol. It contains NLT 90.0% and NMT Standard solution (mg/mL)
110.0% of the labeled amount of C6H12N2O4Pt. CU = nominal concentration of carboplatin in the
[CAUTIONGreat care should be taken in handling Carboplatin Sample solution (mg/mL)
because it is a suspected carcinogen.] Acceptance criteria: 90.0%110.0%
IDENTIFICATION PERFORMANCE TESTS
THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 621 UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Standard solution: 10 mg/mL of USP Carboplatin RS in
water IMPURITIES
Sample solution: 10 mg/mL of carboplatin, from contents Organic Impurities
of 1 container in water PROCEDURE: LIMIT OF 1,1-CYCLOBUTANEDICARBOXYLIC ACID
Chromatographic system Solution A: Dissolve 8.5 g of tetrabutylammonium
Mode: TLC hydrogen sulfate in 80 mL of water. Add 3.4 mL of
Adsorbent: 0.25-mm layer of chromatographic silica gel phosphoric acid, and adjust with 10 N sodium hydroxide to
Application volume: 10 L a pH of 7.55.
Developing solvent system: Acetone and water (4:1) Mobile phase: Acetonitrile, Solution A, and water (5:44:1)
Spray reagent: Add 5.6 g of stannous chloride to 10 mL of Standard solution: 0.01 mg/mL of 1,1-
hydrochloric acid, and stir for 5 min. [NOTEIt is not cyclobutanedicarboxylic acid in Mobile phase
necessary that all of the solids dissolve.] Add 90 mL of water System suitability solution: Mix 1.0 mL of the Standard
and 1 g of potassium iodide, and stir. [NOTEPrepare this solution with 1.0 mL of Standard solution from the Assay.
solution fresh daily.] Sample solution: 1 mg/mL carboplatin, from the contents
Analysis of 1 container diluted with Mobile phase
Samples: Standard solution and Sample solution [NOTEComplete the chromatographic analysis of the
Place the plate in a chromatographic chamber lined with solution within 2 h.]
filter paper and equilibrated for 2 h in Developing solvent Chromatographic system
system. Develop the chromatogram until the solvent front (See Chromatography 621, System Suitability.)
has moved 10 cm from the origin. Remove the plate from Mode: LC
the chamber, and air-dry at room temperature for 2 h. Detector: UV 220 nm
Spray with the Spray reagent, and heat at 110 for 10 Column: 4.0-mm 30-cm; packing L1
min. Flow rate: 2 mL/min
Acceptance criteria: The principal spot from the Sample Injection size: 100 L
solution corresponds in appearance and RF value to that of System suitability
the Standard solution. Sample: System suitability solution
[NOTEThe relative retention times for carboplatin and
ASSAY 1,1-cyclobutanedicarboxylic acid are about 0.65 and
PROCEDURE 1.0, respectively.]
Mobile phase: Acetonitrile and water (87:13) Suitability requirements
Standard solution: 1 mg/mL of USP Carboplatin RS Resolution: NLT 2.5 between carboplatin and 1,1-
[NOTEUse this solution within 2 h.] cyclobutanedicarboxylic acid peaks
Sample solution: 1 mg/mL carboplatin, from the contents Column efficiency: NLT 1500 theoretical plates for the
of 1 container diluted with water cyclobutanedicarboxylic acid peak
[NOTEComplete chromatographic analysis of this solution
within 2 h.]
82 Carboplatin / Official Monographs USP 32
Relative standard deviation: NMT 10.0% Carboprost Tromethamine

Analysis (Comment on this Monograph)id=m13140=Carboprost
Sample: Standard solution and Sample solution Tromethamine=Ca-Chl-Monos.pdf)
Calculate the percentage of 1,1-cyclobutanedicarboxylic
acid in the portion of Carboplatin for Injection:
rU = peak response for 1,1-cyclobutanedicarboxylic

acid from the Sample solution
rS = peak response for 1,1-cyclobutanedicarboxylic
acid from the Standard solution C21H36O5 C4H11NO3 489.64
CS = concentration of 1,1-cyclobutanedicarboxylic Prosta-5,13-dien-1-oic acid, 9,11,15-trihydroxy-15-methyl-, (5Z,
acid in the Standard solution (mg/mL) 9,11,13E,15S)-, compound with 2-amino-2-
CU = nominal concentration of carboplatin in the (hydroxymethyl)-1,3-propanediol (1:1);
Sample solution (mg/mL) (Z)-7-[(1R,2R,3R,5S)-3,5-Dihydroxy-2-[(E)-(3S)-3-hydroxy-3-
Acceptance criteria: NMT 1.0% methyl-1-octenyl]cyclopentyl]-5-heptenoic acid compound
SPECIFIC TESTS with 2-amino-2-(hydroxymethyl)-1,3-propanediol (1:1);
BACTERIAL ENDOTOXINS TEST 85: 0.54 USP Endotoxin (15S)-15-Methylprostaglandin F2 tromethamine [58551-69-2].
Unit/mg of carboplatin DEFINITION
STERILITY TESTS 71: Meets the requirements when tested as Carboprost Tromethamine contains NLT 95.0% and NMT
directed in Test for Sterility of the Product to Be Examined, 105.0% of C25H47NO8, calculated on the dried basis.
Membrane Filtration
CONSTITUTED SOLUTION: At the time of use, it meets the [CAUTIONGreat care should be taken to prevent inhaling
requirements for Injections 1, Constituted Solutions. particles of Carboprost Tromethamine and exposing the skin to
PH 791: 5.07.0, in a solution constituted as directed in it.]
the labeling, Sterile Water for Injection being used
WATER DETERMINATION, Method I 921 IDENTIFICATION
Analysis: Use anhydrous formamide as the extraction INFRARED ABSORPTION 197M
solvent. Introduce 50 mL of anhydrous formamide into the
titration vessel, and titrate with the reagent to the ASSAY
electrometric endpoint. Use the formamide thus dried to PROCEDURE
rinse a suitable glass syringe equipped with a 22-gauge Mobile phase: Methylene chloride, 1,3-butanediol, and
needle, 8 cm long. Add the rinse back to the titration vessel, water (992:7:0.5)
and titrate the vessel contents again, if necessary. Via the Internal standard solution: 7 mg/mL of guaifenesin in
syringe, withdraw 5 mL of the formamide thus titrated and, Mobile phase
through the closure of the container, expel the contents into Buffer solution: Dissolve 10.5 g of citric acid in 75 mL of
the container. Shake the container to obtain a solution. With water. Add 5 N sodium hydroxide slowly to adjust to a pH
the same syringe, withdraw all of the contents of the of 4.0, and dilute with water to 100 mL.
container, and transfer to the titration vessel. Titrate to the Standard solution: Transfer 5 mg of USP Carboprost
endpoint, adjusting the feeding speed control to the lowest Tromethamine RS to a stoppered, 50-mL centrifuge tube.
setting to avoid overtitration. Add 20.0 mL of methylene chloride and 2 mL of Buffer
Acceptance criteria: NMT 3.0% solution. Shake the stoppered tube for 10 min, and
centrifuge. Remove and discard the top (aqueous) layer, and
ADDITIONAL REQUIREMENTS transfer a 4.0-mL aliquot of the lower (methylene chloride)
PACKAGING AND STORAGE: Preserve as described in Injections layer to a suitable vial. Evaporate with the aid of a stream of
1, Containers for Sterile Solids, and protect from light. nitrogen to dryness. Add 100 L of a freshly prepared
USP REFERENCE STANDARDS 11 solution of -bromo-2-acetonaphthone in acetonitrile (1 in
USP Carboplatin RS 50). Swirl to wash down the sides of the vial. Add 50 L of
USP Endotoxin RS
USP 32 Official Monographs / Carboprost 83
a freshly prepared solution of diisopropylethylamine in rB = area of any peak at a relative retention time of
acetonitrile (1 in 100), swirl again, and place the vial in a 1.0
suitable heating device maintained at a temperature of 30 rC = area of any peak at a relative retention time of
to 35 for NLT 15 min. Evaporate the acetonitrile from the 1.2
vial with the aid of a stream of nitrogen, add 2.0 mL of Calculate the percentage of the 5-trans isomer (as the
Internal standard solution, mix, and pass the resulting tromethamine salt) in the portion of Carboprost
solution through a fine-porosity filter. Protect the filtered Tromethamine taken:
solution from light prior to injection to prevent degradation
of the naphthacyl ester of carboprost. Result = rC/(rA + rB + rC) 100
Sample solution: Proceed as directed for Standard solution,
except to use Carboprost Tromethamine in place of USP rC = area of any peak at a relative retention time of
Carboprost Tromethamine RS. 1.2
Chromatographic system rA = area of any peak at a relative retention time of
(See Chromatography 621, System Suitability.) 0.7
Mode: LC rB = area of any peak at a relative retention time of
Detector: UV 254 nm 1.0
Column: 3.9-mm 30-cm stainless steel; 10-m packing Acceptance criteria
L3 15R-epimer: NMT 2.0%
Flow rate: 1.8 mL/min 5-trans isomer: NMT 3.0%
System suitability SPECIFIC TESTS
Sample: Standard solution OPTICAL ROTATION, Specific Rotation 781S: +18 to +24
[NOTEThe relative retention times for guaifenesin and 2- Sample solution: 10 mg/mL in alcohol
naphthacyl ester of carboprost are 0.6 and 1.0, LOSS ON DRYING 731: Dry it in a vacuum at a pressure not
respectively.] exceeding 5 mm of mercury at 50 for 16 h: it loses NMT
Suitability requirements 1.0% of its weight.
Resolution: NLT 4.0 between guaifenesin and the 2- ADDITIONAL REQUIREMENTS
naphthacyl ester of carboprost PACKAGING AND STORAGE: Preserve in well-closed containers,
Relative standard deviation: NMT 2.0% and store in a freezer.
Samples: Standard solution and Sample solution USP Carboprost Tromethamine RS
Carboprost Tromethamine taken:
Result = (RU/RS) (CS/CU) 100 Carboprost Tromethamine Injection
RU = peak response ratio of the 2-naphthacyl ester of (Comment on this Monograph)id=m13150=Carboprost
carboprost to the internal standard of the Tromethamine Injection=Ca-Chl-Monos.pdf)
Sample solution DEFINITION
RS = peak response ratio of the 2-naphthacyl ester of Carboprost Tromethamine Injection is a sterile solution of
carboprost to the internal standard of the Carboprost Tromethamine in aqueous solution, which may also
Standard solution contain Benzyl Alcohol, Sodium Chloride, and Tromethamine.
CS = concentration of USP Carboprost Tromethamine It contains NLT 90.0% and NMT 110.0% of the labeled
RS in the Standard solution (mg/mL) amount of carboprost (C21H36O5).
CU = concentration of carboprost tromethamine in the
Acceptance criteria: 95.0%105.0% A. INFRARED ABSORPTION 197K
Sample: Extract the equivalent of 2.5 mg of carboprost
IMPURITIES tromethamine from a volume of Injection, with 1.52 times
Inorganic Impurities its volume of chloroform. Discard the chloroform layer, and
RESIDUE ON IGNITION 281: NMT 0.5% acidify the aqueous layer with 35 drops of hydrochloric
Organic Impurities acid. Extract the acidified solution with an equivalent
PROCEDURE: LIMIT OF 15R-EPIMER AND 5-trans ISOMER volume of chloroform. Filter the chloroform layer through a
Buffer solution, Mobile phase, Internal standard solution, pledget of cotton, and concentrate it to a volume of less
and Sample solution: Proceed as directed in the Assay. than 1 mL. Combine the resulting solution with 150180
Chromatographic system: mg of potassium bromide. Dry the potassium bromide
(See Chromatography 621, System Suitability.) mixture in a vacuum overnight, and prepare a pellet from
Proceed as directed in the Assay, except use the following the dried mixture.
injection size:
System suitability: Proceed as directed in the Assay, using PROCEDURE
the Sample solution in place of the Standard solution. Mobile phase: Methylene chloride, 1,3-butanediol, and
Analysis water (992:7:0.5)
Sample: Sample solution Internal standard solution: 3 mg/mL of guaifenesin in
Calculate the percentage of the 15R-epimer (as the Mobile phase
tromethamine salt) in the portion of Carboprost Buffer solution: Dissolve 10.5 g of citric acid in 75 mL of
Tromethamine taken: water. Add 5 N sodium hydroxide slowly to adjust to a pH
of 4.0, and dilute with water to 100 mL.
Result = rA/(rA + rB + rC) 100 Standard solution: Prepare a solution containing 0.332
mg/mL of USP Carboprost Tromethamine RS and 9 mg/mL
rA = area of any peak at a relative retention time of of benzyl alcohol. Transfer 2.0 mL into a stoppered
0.7
84 Carboprost / Official Monographs USP 32
centrifuge tube. Add 20.0 mL of methylene chloride and 1.0 ADDITIONAL REQUIREMENTS
mL of Buffer solution, shake the stoppered tube for 10 min, PACKAGING AND STORAGE: Preserve in single-dose or
and centrifuge. Remove and discard the top (aqueous) layer, multiple-dose containers, preferably of Type I glass, and store
transfer an 8.0-mL aliquot of the lower (methylene chloride) in a refrigerator.
layer to a suitable vial, and evaporate the solution with the USP REFERENCE STANDARDS 11
aid of a stream of nitrogen. [NOTEThe residue does not USP Carboprost Tromethamine RS
evaporate to dryness because of the presence of benzyl USP Endotoxin RS
alcohol.] Add 100 L of a freshly prepared solution of -
bromo-2-acetonaphthone in acetonitrile (1 in 50), and swirl
to wash down the sides of the vial. Add 50 L of a freshly
prepared solution of diisopropylethylamine in acetonitrile (1 Carboxymethylcellulose Sodium
in 100). Swirl again, and place the vial in a suitable heating (Comment on this
device maintained at a temperature of 30 to 35 for NLT Monograph)id=m13210=Carboxymethylcellulose Sodium=Ca-
15 min. Evaporate the acetonitrile from the vial with the aid Chl-Monos.pdf)
of a stream of nitrogen, add 1.0 mL of Internal standard Cellulose carboxymethyl ether sodium salt [9004-32-4].
solution, mix, and pass the resulting solution through a fine-
porosity filter. Protect the filtered solution from light prior to DEFINITION
injection to prevent degradation of the naphthacyl ester of Carboxymethylcellulose Sodium is the sodium salt of a
carboprost. polycarboxymethyl ether of cellulose. It contains NLT 6.5%
Sample solution: Pipet a volume of Injection, nominally and NMT 9.5% of sodium (Na), calculated on the dried basis.
equivalent to 500 g of carboprost, to a stoppered, 50-mL
centrifuge tube. Proceed as directed for Standard solution, IDENTIFICATION
beginning with Add 20.0 mL of methylene chloride. A. PROCEDURE
Chromatographic system Sample solution: Add about 1 g of powdered
(See Chromatography 621, System Suitability.) Carboxymethylcellulose Sodium to 50 mL of water, while
Mode: LC stirring to produce a uniform dispersion. Continue the
Detector: UV 254 nm stirring until a clear solution is produced.
Column: 3.9-mm 30-cm stainless steel; 10-m packing Analysis: To 1 mL of the Sample solution, diluted with an
L3 equal volume of water in a small test tube, add 5 drops of
Flow rate: 1.8 mL/min 1-naphthol TS. Incline the test tube, and carefully introduce
Injection size: 10 L down the side of the tube 2 mL of sulfuric acid so that it
System suitability forms a lower layer.
Sample: Standard solution Acceptance criteria: A red-purple color develops at the
[NOTEThe relative retention times for guaifenesin and 2- interface.
naphthacyl ester of carboprost are 0.6 and 1.0, B. PROCEDURE
respectively. ] Sample solution: Add 1 g of powdered
Suitability requirements Carboxymethylcellulose Sodium to 50 mL of water, while
Resolution: NLT 4.0 between guaifenesin and the 2- stirring to produce a uniform dispersion. Continue the
naphthacyl ester of carboprost stirring until a clear solution is produced.
Relative standard deviation: NMT 2.0% Analysis: To 5 mL of the Sample solution, add an equal
Analysis volume of barium chloride TS.
Samples: Standard solution and Sample solution Acceptance criteria: A fine, white precipitate is formed.
Calculate the percentage of C21H36O5 in each mL of C. IDENTIFICATON TESTSGENERAL, Sodium 191: A portion
Injection taken: of the Sample solution meets the requirements.
Sample solution: Add 1 g of powdered
Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 Carboxymethylcellulose Sodium to 50 mL of water, while
stirring to produce a uniform dispersion. Continue the
RU = peak response ratios of the 2-naphthacyl ester of stirring until a clear solution is produced.
carboprost to the internal standard of the
Sample solution ASSAY
RS = peak response ratios of the 2-naphthacyl ester of PROCEDURE
carboprost to the internal standard of the Sample solution: Transfer to a beaker 500 mg of
Standard solution Carboxymethylcellulose Sodium, add 80 mL of glacial acetic
CS = concentration of USP Carboprost Tromethamine acid, heat the mixture on a boiling water bath for 2 h, and
RS in the Standard solution (g/mL) cool to room temperature.
CU = nominal concentration of carboprost in the Analysis: Titrate the Sample solution with 0.1 N perchloric
Sample solution (mg/mL) acid VS. Each mL of 0.1 N perchloric acid is equivalent to
Mr1 = molecular weight of carboprost, 368.51 2.299 mg of Na.
Mr2 = molecular weight of carboprost tromethamine, Acceptance criteria: NLT 6.5% and NMT 9.5% of Na,
489.64 calculated on the dried basis
IMPURITIES
BACTERIAL ENDOTOXINS TEST 85: NMT 714.3 USP HEAVY METALS, Method II 231: NMT 20 ppm, adding 1
Endotoxin Units/mg of carboprost tromethamine mL of hydroxylamine hydrochloride solution (1 in 5) to the
PH 791: 7.08.0 solution of the residue
OTHER REQUIREMENTS: Meets the requirements under
Injections 1
USP 32 Official Monographs / Carboxymethylcellulose 85
SPECIFIC TESTS Analysis: Titrate the Sample solution with 0.1 N perchloric
VISCOSITY 911 acid in dioxane VS, determining the endpoint
Analysis: Determine the viscosity in a water solution at the potentiometrically. Each mL of 0.1 N perchloric acid is
concentration stated on the label. Using undried equivalent to 29.67 mg of carboxymethylcellulose sodium.
Carboxymethylcellulose Sodium, weigh the amount that, on Acceptance criteria: 16.0%17.0%
the dried basis, will provide 200 g of solution of the stated
concentration. Add the substance in small amounts to 180 IMPURITIES
mL of stirred water contained in a tared, wide-mouth bottle, Inorganic Impurities
continue stirring rapidly until the powder is well wetted, add HEAVY METALS, Method II 231: NMT 50 ppm, using 400
sufficient water to make the mixture weigh 200 g, and allow mg of Carboxymethylcellulose Sodium Paste and adding 1
to stand, with occasional stirring, until solution is complete. mL of hydroxylamine hydrochloride solution (1 in 5) to the
Adjust the temperature to 25 0.2, and determine the solution of the residue
viscosity, using a rotational type of viscosimeter, making
certain that the system reaches equilibrium before taking the SPECIFIC TESTS
final reading. MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED
Acceptance criteria: The viscosity of solutions of 2% or MICROORGANISMS 62: The total bacterial count does not
higher concentration is NLT 80.0% and NMT 120.0% of exceed 1000 cfu/g, and the tests for Salmonella species and
that stated on the label; the viscosity of solutions of less Escherichia coli are negative.
than 2% concentration is NLT 75.0% and NMT 140.0% of CONSISTENCY
that stated on the label. Apparatus: Determine the consistency of Paste by means of
PH 791: 6.58.5 in a solution (1 in 100) a penetrometer fitted with a polished cone-shaped metal
LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT plunger weighing 150 g, having a detachable steel tip of the
10.0% of its weight. following dimensions: the tip of the cone has an angle of
30, the point being truncated to a diameter of 0.381
ADDITIONAL REQUIREMENTS 0.025 mm, the base of the tip is 8.38 0.05 mm in
PACKAGING AND STORAGE: Preserve in tight containers. diameter, and the length of the tip is 14.94 0.05 mm. The
LABELING: Label it to indicate the viscosity in solutions of remaining portion of the cone has an angle of 90, is 28 mm
stated concentrations. in height, and has a maximum diameter at the base of
about 65 mm. The containers for the test are flat-bottom
metal cylinders that are 100 6 mm in diameter and NLT
65 mm in height. They are constructed of at least 1.6-mm
Carboxymethylcellulose Sodium Paste (16-gauge) metal, and are provided with well-fitting, water-
(Comment on this tight covers.
Monograph)id=m13240=Carboxymethylcellulose Sodium Analysis: Place the required number of containers in an
Paste=Ca-Chl-Monos.pdf) oven, and bring them and a quantity of Paste to a
temperature of 82 2.5, pour the Paste into one or more
DEFINITION of the containers, filling to within 6 mm of the rim. Cool to
Carboxymethylcellulose Sodium Paste contains NLT 16.0% and 25 2.5 over a period of NLT 16 h, protected from drafts.
NMT 17.0% of carboxymethylcellulose sodium. Two h before the test, place the containers in a water bath
at 25 0.5. If the room temperature is below 23.5 or
IDENTIFICATION above 26.5, adjust the temperature of the cone to 25
A. PROCEDURE 0.5 by placing it in the water bath. Without disturbing the
Sample solution: Digest a quantity of Paste equivalent to 1 surface of the substance under test, place the container on
g of carboxymethylcellulose sodium with 50 mL of water the penetrometer table, and lower the cone until the tip just
until the solution is virtually complete, and filter. touches the top surface of the test substance at a spot
Analysis: To 30 mL of the Sample solution, add 3 mL of 2538 mm from the edge of the container. Adjust the zero
hydrochloric acid. setting and quickly release the plunger, then hold it free for
Acceptance criteria: A white precipitate is formed. [NOTE 5 s. Secure the plunger, and read the total penetration from
Filter the solution and save the filtrate for use in Identification the scale. Make three or more trials, each so spaced that
test C.] there is no overlapping of the areas of penetration. Where
B. PROCEDURE the penetration exceeds 20 mm, use a separate container of
Sample solution: Use the remainder of the Sample solution the test substance for each trial. Read the penetration to the
prepared under Identification test A. nearest 0.1 mm. Calculate the average of the three or more
Analysis: To the Sample solution add an equal volume of readings, and conduct further trials to a total of 10 if the
barium chloride TS. individual results differ from the average by more than 3%.
Acceptance criteria: A fine, white precipitate is formed. Acceptance criteria: The final average of the trials is NLT
C. IDENTIFICATION TESTSGENERAL, Sodium 191: The filtrate 30.0 mm and NMT 36.0 mm, indicating a consistency value
from Identification test A responds to the tests. between 300 and 360.
ASSAY 2.0% of its weight.
PROCEDURE
Sample solution: Transfer 2 g of Paste to a glass-stoppered, ADDITIONAL REQUIREMENTS
250-mL conical flask. Add 75 mL of glacial acetic acid, PACKAGING AND STORAGE: Preserve in well-closed containers,
attach a condenser, and reflux for 2 h. Cool, transfer the and avoid prolonged exposure to temperatures exceeding
mixture to a 250-mL beaker with the aid of small volumes of 30.
glacial acetic acid.
86 Carboxymethylcellulose / Official Monographs USP 32
Carboxymethylcellulose Sodium Tablets IDENTIFICATION

(Comment on this A. INFRARED ABSORPTION 197K
Monograph)id=m13250=Carboxymethylcellulose Sodium B. The RF value of the principal spot due to carisoprodol of
Tablets=Ca-Chl-Monos.pdf) the Sample solution obtained in the test for Limit of
Meprobamate corresponds to that obtained in a Standard
DEFINITION solution of USP Carisoprodol RS in chloroform containing
Carboxymethylcellulose Sodium Tablets contain an amount of 100 mg/mL.
sodium (Na) equivalent to NLT 6.5% and NMT 9.5% of the
labeled amount of carboxymethylcellulose sodium. ASSAY
PROCEDURE
IDENTIFICATION Sodium methoxide titrant: Prepare and standardize as
A. PROCEDURE directed for 0.1 N sodium methoxide (in toluene) (see
Sample solution: Digest a quantity of powdered Tablets Reagents, Indicators, and SolutionsVolumetric Solutions),
equivalent to 1 g of carboxymethylcellulose sodium with 50 except to use 200 mL of methanol to dissolve the sodium
mL of water until the solution is virturally complete, and metal, to add 750 mL of toluene, and to dilute with
filter. methanol to 1000 mL.
Analysis: To 30 mL of the Sample solution, add 3 mL of Sample: 400 mg of Carisoprodol
hydrochloric acid. Analysis: Add 10 mL of pyridine to the Sample. Add 1 drop
Acceptance criteria: A white precipitate is formed. [NOTE of phenolphthalein TS, and titrate with Sodium methoxide
Filter the solution and save the filtrate for use in Identification titrant to a permanent pink endpoint. Add 25.0 mL of
test A.] Sodium methoxide titrant, connect the flask to a water-cooled
B. PROCEDURE condenser, and reflux on a hot plate for 30 min. Allow to
Sample solution: Use the remainder of the Sample solution cool, add 40 mL of alcohol and 7 drops of phenolphthalein
prepared under Identification test A. TS. Titrate the excess alkali with 0.1 N hydrochloric acid VS
Analysis: To the Sample solution add an equal volume of until the pink color disappears. Perform a blank
barium chloride TS. determination (see Titrimetry 541, Residual Titrations). Each
Acceptance criteria: A fine, white precipitate is formed. mL of Sodium methoxide titrant is equivalent to 26.03 mg of
C. IDENTIFICATION TESTSGENERAL, Sodium 191: The filtrate C12H24N2O4.
from Identification test A meets the requirements. Acceptance criteria: 98.0%102.0%
ASSAY IMPURITIES
PROCEDURE Inorganic Impurities
Sample solution: Dissolve an amount equivalent to 500 mg HEAVY METALS, Method II 231: NMT 10 ppm
of carboxymethylcellulose sodium from powdered Tablets Organic Impurities
(NLT 20 Tablets) in 80 mL of glacial acetic acid. Heat the PROCEDURE: LIMIT OF MEPROBAMATE
mixture on a steam bath for 2 h and cool to room Standard solution: 1 mg/mL of USP Meprobamate RS in
temperature. chloroform
Analysis: Titrate the Sample solution with 0.1 N perchloric Sample solution: 100 mg/mL of Carisoprodol in
acid VS. Each mL of 0.1 N perchloric acid is equivalent to chloroform
2.299 mg of Na. Chromatographic system
Acceptance criteria: 6.5%9.5% (See Chromatography 621, Thin-Layer Chromatography.)
Mode: TLC
PERFORMANCE TESTS Adsorbent: 0.25-mm layer of chromatographic silica gel
DISINTEGRATION 701 Application volume: 10 L for Sample solution and 5 L
Time: 2 h for Standard solution
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Developing solvent system: Chloroform and acetone
(4:1)
ADDITIONAL REQUIREMENTS Spray reagent 1: Antimony trichloride TS
PACKAGING AND STORAGE: Preserve in tight containers. Spray reagent 2: 3 in 100 solution of furfural in
chloroform
Analysis
Carisoprodol Samples: Standard solution and Sample solution
Proceed as directed in the chapter. Allow the spots to dry
(Comment on this Monograph)id=m13370=Carisoprodol=Ca- in a current of air, and develop the chromatogram in the
Chl-Monos.pdf) Developing solvent system until the solvent front has
moved three-fourths of the length of the plate. Remove
the plate from the developing chamber, mark the solvent
front, allow the solvent to evaporate, and spray the plate
alternately with Spray reagent 1 and Spray reagent 2 until
one or more black spots appear, heat the plate at 110 for
15 min, and examine the plate.
Acceptance criteria: Any spot in the Sample solution
having an RF value corresponding to that of meprobamate
C12H24N2O4 260.33
in the Standard solution is not darker in color than the
()-2-Methyl-2-propyl-1,3-propanediol carbamate
meprobamate spot in the Standard solution (NMT 0.5%).
isopropylcarbamate [78-44-4].
DEFINITION
Carisoprodol contains NLT 98.0% and NMT 102.0% of
C12H24N2O4, calculated on the dried basis.
USP 32 Official Monographs / Carisoprodol 87
SPECIFIC TESTS CU = nominal concentration of carisoprodol in the

MELTING RANGE OR TEMPERATURE, Class I 741: 9194 Sample solution (mg/mL)
LOSS ON DRYING 731: Dry it in vacuum at 60 for 3 h: it Acceptance criteria: 90.0%110.0%
PERFORMANCE TESTS
ADDITIONAL REQUIREMENTS DISSOLUTION 711
PACKAGING AND STORAGE: Preserve in tight containers. Medium: 0.05 M, pH 6.9 phosphate buffer (see Reagents,
USP REFERENCE STANDARDS 11 Indicators, and SolutionsBuffer Solutions) containing 5 units
USP Carisoprodol RS of -amylase per mL; 900 mL
USP Meprobamate RS [NOTEUse only freshly prepared solutions containing -
amylase; and equilibrate the Dissolution Medium at 37 for
NMT one h before beginning the Dissolution test.]
Apparatus 2: 75 rpm
Carisoprodol Tablets Time: 60 min
(Comment on this Monograph)id=m13380=Carisoprodol Determine the amount of C12H24N2O4 dissolved employing
Tablets=Ca-Chl-Monos.pdf) the following method:
Mobile phase, System suitability solution, and System
DEFINITION suitability: Proceed as directed in the Assay.
Carisoprodol Tablets contain NLT 90.0% and NMT 110.0% of Standard solution: 0.4 mg/mL of USP Carisoprodol RS in
the labeled amount of C12H24N2O4. Medium
[NOTEA volume of acetonitrile not exceeding 2% of the
IDENTIFICATION final total volume of solution may be used to aid in
The retention time of the Sample solution corresponds to that dissolving the carisoprodol.]
of the Standard solution, as obtained in the Assay. Sample solution: Sample per Dissolution 711. Filter before
ASSAY analyzing.
PROCEDURE Chromatographic system: Proceed as directed under
Mobile phase: Acetonitrile and water (2:3) Assay, except use the following injection size:
Diluent: Methanol and 0.01 N sulfuric acid (3:2) Injection size: 150 L
System suitability solution: 2.4 mg/mL of 2-methyl-2- Analysis
propyl-1,3-propanediol and 3.4 mg/mL of carisoprodol in Samples: Standard solution and Sample solution
Mobile phase Record the peak responses, and measure the heights for the
Standard solution: 3.5 mg/mL of USP Carisoprodol RS in major peaks. Calculate the amount of C12H24N2O4 dissolved.
Diluent Tolerances: NLT 80% (Q) of the labeled amount of
Sample solution: Transfer an amount equivalent to 350 mg C12H24N2O4 is dissolved.
of carisoprodol from powdered Tablets (NLT 20 Tablets), to UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
a 100-mL volumetric flask and add 50 mL of Diluent, place ADDITIONAL REQUIREMENTS
in an ultrasonic bath for 30 min, and shake by mechanical PACKAGING AND STORAGE: Preserve in well-closed containers.
means for 60 min. Dilute with Diluent to volume. Pass a USP REFERENCE STANDARDS 11
portion of this solution through a membrane filter of USP Carisoprodol RS
0.5m or finer porosity, and use the filtrate as the Sample
solution.
(See Chromatography 621, System Suitability.) Carisoprodol and Aspirin Tablets
Mode: LC (Comment on this Monograph)id=m13383=Carisoprodol and
Detector: Refractive index Aspirin Tablets=Ca-Chl-Monos.pdf)
Temperature: 30 1 for column and detector DEFINITION
Flow rate: 2 mL/min Carisoprodol and Aspirin Tablets contain NLT 90.0% and NMT
Injection size: 35 L 110.0% of the labeled amounts of carisoprodol (C12H24N2O4)
System suitability and aspirin (C9H8O4).
[NOTEThe relative retention times for 2-methyl-2- IDENTIFICATION
propyl-1,3-propanediol and carisoprodol are about 0.5 A. The retention times of the aspirin peak and the
and 1.0, respectively.] carisoprodol peak of the Sample solution correspond to those
Suitability requirements from Standard solution A, as obtained in Assay for Aspirin and
Resolution: NLT 2.0 between the 2-methyl-2-propyl-1,3- carisoprodol.
propanediol and carisoprodol peaks, System suitability
solution ASSAY
Relative standard deviation: NMT 2.0% for three ASPIRIN AND CARISOPRODOL
replicate injections of the Standard solution Mobile phase: Methanol and 0.174 M acetic acid (16:9)
Analysis Diluent: Acetonitrile, glacial acetic acid, and water
Samples: Standard solution and Sample solution (40:1:59)
Calculate the percentage of C12H24N2O4 in the portion of Standard solution A: To 80 mg of USP Aspirin RS and 80J
Tablets taken: mg of USP Carisoprodol RS, in a 25-mL volumetric flask, add
15 mL of Diluent, swirl for 5 min, and sonicate for 2530 s (J
Result = (rU/rS) (CS/CU) 100 being the ratio of the labeled amount, in mg, of
carisoprodol to that of aspirin). Dilute with Diluent to
rU = peak response from the Sample solution volume.
rS = peak response from the Standard solution Standard solution B: 16 g/mL of USP Salicylic Acid RS in
CS = concentration of USP Carisoprodol RS in the Diluent
88 Carisoprodol / Official Monographs USP 32
System suitability solution: 0.5 mg/mL of salicylic acid in Mode: LC

Standard solution A Detector: Refractive index
Sample solution: To an equivalent to 325 mg of aspirin Column: 3.9-mm 30-cm; packing L1
from powdered Tablets (NLT 20), in a 100-mL volumetric Temperature: 30 1 for column and detector
flask, add 50 mL of Diluent, swirl for 5 min, sonicate for Flow rate: 2 mL/min
2530 s, shake by mechanical means for 30 min, dilute with Injection size: 300 L
Diluent to volume, and mix. Pass a portion of this solution System suitability
through a membrane filter of 0.5m or finer porosity, and Samples: Standard solution and System suitability solution
use the filtrate. [NOTEUse within 8 h.] [NOTEThe relative retention times for aspirin and
Chromatographic system carisoprodol are 0.4 and 1.0, respectively.]
Mode: LC Resolution: NLT 1.5 between the aspirin and salicylic acid
Detector: Refractive index peaks, and between the carisoprodol and salicylic acid
Column: 4.6-mm 25-cm; packing L7 peaks, System suitability solution
Temperature: 30 1 column and refractive index Relative standard deviation: NMT 2.0%, Standard
detector solution
System suitability Calculate the percentage of C9H8O4 and C12H24N2O4
Samples: Standard solution A, Standard solution B, and dissolved:
[NOTEThe relative retention times for aspirin, salicylic Result = (rU/rS) (CS/CU) 100
acid, and carisoprodol are about 0.6, 0.7, and 1.0,
respectively.] rU = peak response for aspirin or carisoprodol from
Suitability requirements the Sample solution
Resolution: NLT 1.2 between the solvent and aspirin rS = peak response for aspirin or carisoprodol from
peaks and NLT 1.5 between the aspirin and salicylic acid the Standard solution A
peaks from the System suitability solution CS = concentration of USP Aspirin RS or USP
Relative standard deviation: NMT 2.0% for Standard Carisoprodol RS in the Standard solution A
solution A; NMT 5.0% for Standard solution B (mg/mL)
Analysis CU = nominal concentration of aspirin or carisoprodol
Samples: Standard solution A and Sample solution in the Sample solution (mg/mL)
Analysis: Calculate the percentage of C9H8O4 and Tolerances: NLT 75% (Q) of the labeled amounts of C9H8O4
C12H24N2O4 in the portion of Tablets: and C12H24N2O4 are dissolved.
Result = (rU/rS) (CS/CU) 100 for Content Uniformity with respect to aspirin and to
carisoprodol
rU = peak response for aspirin or carisoprodol from
the Sample solution IMPURITIES
rS = peak response for aspirin or carisoprodol from Organic Impurities
the Standard solution A PROCEDURE: LIMIT OF FREE SALICYLIC ACID
CS = concentration of USP Aspirin RS or USP Mobile phase, Diluent, Standard solution A, Standard
Carisoprodol RS in the Standard solution A solution B, System suitability solution, and Sample
(mg/mL) solution: Proceed as directed in Assay.
CU = nominal concentration of aspirin or carisoprodol Chromatographic system
in the Sample solution (mg/mL) (See Chromatography 621, System Suitability.)
Detector: UV 313 nm
PERFORMANCE TESTS Column: 4.6-mm 25-cm; packing L7
DISSOLUTION 711 Temperature: 30 1
Medium: Water; 900 mL Flow rate: 1 mL/min
Apparatus 2: 75 rpm Injection size: 50 L
Time: 45 min System suitability
Mobile phase: Methanol and glacial acetic acid solution (1 Samples: Standard solution A, Standard solution B and
in 50) (51:49) System suitability solution
Standard solution: To 90 mg of USP Aspirin RS and 90J mg [NOTEThe relative retention times for aspirin, salicylic
of USP Carisoprodol RS in a 250-mL volumetric flask, add 5 acid, and carisoprodol are about 0.6, 0.7, and 1.0,
mL of acetonitrile, previously passed through a membrane respectively.]
filter of 0.5-m or finer porosity, swirl to dissolve, and dilute Suitability requirements
with water to volume (J being the ratio of the labeled Resolution: NLT 1.2 between the solvent and aspirin
amount, in mg, of carisoprodol to that of aspirin). peaks and NLT 1.5 between the aspirin and salicylic acid
System suitability solution: 0.36 mg/mL of salicylic acid in peaks from the System suitability solution
Standard solution Relative standard deviation: NMT 2.0% for Standard
Sample solution: Sample per Dissolution 711. Dilute with solution A; NMT 5.0% for Standard solution B
Medium to a concentration that is similar to the Standard Analysis
solution. Samples: Standard solution A, Standard solution B, and
Chromatographic system Sample solution
USP 32 Official Monographs / Carisoprodol 89
Calculate the percentage of free salicylic acid in the Tablets: Flow rate: 1 mL/min
Result = (rU/rS) (CS/CU) 100 System suitability
rU = peak response for salicylic acid from the Sample System suitability solution
solution [NOTEThe relative retention times for aspirin, salicylic
rS = peak response for salicylic acid from the acid, and carisoprodol are about 0.6, 0.7, and 1.0,
Standard solution B respectively.]
CS = concentration of USP Salicylic Acid RS in the Suitability requirements
Standard solution B (g/mL) Resolution: NLT 1.2 between the solvent and aspirin
CU = nominal concentration of aspirin in the portion peaks and NLT 1.5 between the aspirin and salicylic acid
of Tablets taken in the Sample solution (g/mL) peaks from the System suitability solution
Acceptance criteria: NMT 3.0% is found Relative standard deviation: NMT 2.0% for Standard
solution A; NMT 5.0% for Standard solution B
PACKAGING AND STORAGE: Preserve in well-closed containers. Samples: Standard solution A and Sample solution
USP REFERENCE STANDARDS 11 Calculate the percentage of C9H8O4 and C12H24N2O4 in the
USP Aspirin RS portion of Tablets:
USP Carisoprodol RS
USP Salicylic Acid RS Result = (rU/rS) (CS/CU) 100
rU = peak response for aspirin or carisoprodol from
Carisoprodol, Aspirin, and Codeine the Sample solution
rS = peak response for aspirin or carisoprodol from
Phosphate Tablets the Standard solution A
(Comment on this Monograph)id=m13385=Carisoprodol, CS = concentration of USP Aspirin RS or US
Aspirin, and Codeine Phosphate Tablets=Ca-Chl-Monos.pdf) Carisoprodol RS in the Standard solution A
(mg/mL)
DEFINITION CU = nominal concentration of aspirin or carisoprodol
Carisoprodol, Aspirin, and Codeine Phosphate Tablets contain in the Sample solution (mg/mL)
NLT 90.0% and NMT 110.0% of the labeled amounts of Acceptance criteria: 90.0%110.0% of the labeled
carisoprodol (C12H24N2O4), aspirin (C9H8O4), and codeine amounts of C12H24N2O4 and C9H8O4
phosphate (C18H21NO3 H3PO4 1/2H2O). CODEINE PHOSPHATE
Mobile phase: Dissolve 2.2 g of docusate sodium in 600
IDENTIFICATION mL of methanol. Dissolve 0.8 g of ammonium nitrate in 400
A. The retention times of the aspirin, carisoprodol, and mL of water. Mix these two solutions and adjust with glacial
codeine phosphate peaks from the Sample solutions acetic acid to a pH of 3.3 0.05.
correspond to those of the Standard solutions obtained as Diluent: Methanol and 0.01 N sulfuric acid (1:1)
directed in the Assay for Aspirin and Carisoprodol and the System suitability solution: 0.16 mg/mL of USP Codeine
Assay for Codeine Phosphate Phosphate RS and 0.12 mg/mL of USP Codeine N-Oxide RS
ASSAY in Diluent
[NOTEBoth Aspirin and Carisoprodol and Codeine Phosphate Standard solution: 0.16 mg/mL of USP Codeine Phosphate
must be completed for this test.] RS and 0.16J mg/mL of USP Aspirin RS in Diluent, with the
ASPIRIN AND CARISOPRODOL aid of swirling for 5 min and sonication for 2530 s (J being
Mobile phase: Methanol and 0.174 M acetic acid (16:9) the ratio of the labeled amount, in mg, of aspirin to that of
Diluent: Acetonitrile, glacial acetic acid, and water codeine phosphate)
(40:1:59) Sample solution: To an amount equivalent to 16 mg of
Standard solution A: To 80 mg of USP Aspirin RS and 80J codeine phosphate from powdered Tablets (NLT 20), in a
mg of USP Carisoprodol RS, in a 25-mL volumetric flask, add 100-mL volumetric flask, add 50 mL of Diluent, sonicate for
15 mL of Diluent, swirl for 5 min, and sonicate for 2530 s. 30 min, shake by mechanical means for 30 min, and dilute
Dilute with Diluent to volume (J being the ratio of the with Diluent to volume.
labeled amount, in mg, of carisoprodol to that of aspirin). Chromatographic system
Standard solution B: 16 g/mL of USP Salicylic Acid RS in (See Chromatography 621, System Suitability.)
Diluent Mode: LC
System suitability solution: 0.5 mg/mL of salicylic acid in Detector: UV 254 nm
Standard solution A Column: 3.9-mm 30-cm; packing L1
Sample solution: To an amount equivalent to 325 mg of Flow rate: 1.5 mL/min
aspirin from powdered Tablets (NLT 20), in a 100-mL Injection size: 50 L
volumetric flask, add 50 mL of Diluent, swirl for 5 min, System suitability
sonicate for 2530 s, shake by mechanical means for 30 Samples: System suitability solution and Standard solution
min, dilute with Diluent to volume, and mix. Pass a portion [NOTEThe relative retention times for codeine N-Oxide
of this solution through a membrane filter of 0.5m or and codeine phosphate are 0.9 and 1.0, respectively.]
finer porosity, and use the filtrate. [NOTEUse within 8 h.] Suitability requirements
Chromatographic system Resolution: NLT 1.2 between the codeine phosphate and
(See Chromatography 621, System Suitability.) codeine N-Oxide peaks in the System suitability solution
Mode: LC Relative standard deviation: NMT 2.0% from the
Detector: Refractive index Standard solution
Temperature: 30 1 column and refractive index
detector
90 Carisoprodol / Official Monographs USP 32
Analysis DISSOLUTION 711

Samples: Standard solution and Sample solution Medium: Water; 900 mL
Calculate the percentage of C18H21NO3 H3PO4 1/2H2O in Apparatus 2: 75 rpm
the portion of Tablets: Time: 45 min
Procedure 1: Determination of dissolved aspirin and
Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 carisoprodol
Mobile phase: Methanol and glacial acetic acid solution (1
rU = peak response from the Sample solution in 50) (51:49)
rS = peak response from the Standard solution Standard solution: To 90 mg of USP Aspirin RS and 90J
CS = concentration of USP Codeine Phosphate RS in mg of USP Carisoprodol RS in a 250-mL volumetric flask,
the Standard solution (mg/mL) add 5 mL of acetonitrile, previously passed through a
CU = nominal concentration of the Sample solution membrane filter of 0.5-m or finer porosity, swirl to
(mg/mL) dissolve, and dilute with water to volume (J being the ratio
Mr1 = molecular weight of codeine phosphate of the labeled amount, in mg, of carisoprodol to that of
hemihydrate, 406.37 aspirin).
Mr2 = molecular weight of anhydrous codeine System suitability solution: 0.36 mg/mL of salicylic acid
phosphate, 397.37 in Standard solution
Acceptance criteria: 90.0%110.0% of the labeled amount Sample solution: Sample per Dissolution 711. Dilute with
of C18H21NO3 H3PO4 1/2H2O Medium to a concentration that is similar to the Standard
solution.
IMPURITIES Chromatographic system
Organic Impurities (See Chromatography 621, System Suitability.)
PROCEDURE: LIMIT OF FREE SALICYLIC ACID Mode: LC
Mobile phase, Diluent, Standard solution A, Standard Detector: Refractive index
solution B, System suitability solution, and Sample Column: 3.9-mm 30-cm; packing L1
solution: Proceed as directed in the Assay for Aspirin and Temperature: 30 1 for column and detector
Carisoprodol. Flow rate: 2 mL/min
Chromatographic system Injection size: 300 L
(See Chromatography 621, System Suitability.) System suitability
Mode: LC Samples: Standard solution and System suitability solution
Detector: UV 313 nm [NOTEThe relative retention times for aspirin and
Column: 4.6-mm 25-cm; packing L7 carisoprodol are 0.4 and 1.0, respectively.]
Temperature: 30 1 Suitability requirements
Flow rate: 1 mL/min Resolution: NLT 1.5 between the aspirin and salicylic
Injection size: 50 L acid peaks, and between the carisoprodol and salicylic
System suitability acid peaks, in the System suitability solution
Samples: Standard solution A, Standard solution B and Relative standard deviation: NMT 2.0% of the
System suitability solution Standard solution
[NOTEThe relative retention times for aspirin, salicylic Analysis
acid, and carisoprodol are about 0.6, 0.7, and 1.0, Samples: Standard solution and Sample solution
respectively.] Calculate the percentage of C9H8O4 and C12H24N2O4
Suitability requirements dissolved:
Resolution: NLT 1.2 between the solvent and aspirin
peaks and NLT 1.5 between the aspirin and salicylic acid Result = (rU/rS) (CS/CU) 100
peaks from the System suitability solution
Relative standard deviation: NMT 2.0% for Standard rU = peak response for aspirin or carisoprodol from
solution A; NMT 5.0% for Standard solution B the Sample solution
Analysis rS = peak response for aspirin or carisoprodol from
Samples: Standard solution A, Standard solution B, and the Standard solution A
Sample solution CS = concentration of USP Aspirin RS or USP
Calculate the percentage of free salicylic acid in the Tablets: Carisoprodol RS in the Standard solution A
(mg/mL)
Result = (rU/rS) (CS/CU) 100 CU = nominal concentration of aspirin or carisoprodol
rU = peak response for salicylic acid from the Sample Tolerances: NLT 75% (Q) of the labeled amounts of
solution C9H8O4 and C12H24N2O4 are dissolved.
rS = peak response for salicylic acid from the Standard Procedure 2: Determination of dissolved codeine
solution B phosphate
CS = concentration of USP Salicylic Acid RS in the Mobile phase: Dissolve 2.2 g of docusate sodium and 0.8
Standard solution B (g/mL) g of ammonium nitrate in 550 mL of water, and pass
CU = nominal concentration of aspirin in the portion of through a membrane filter of 0.5-m or finer porosity. Add
Tablets taken in the Sample solution (g/mL) 450 mL of similarly filtered acetonitrile to the filtrate.
Acceptance criteria: NMT 3.0% is found Standard solution: 18 g/mL of USP Codeine Phosphate
PERFORMANCE TESTS RS in water
[NOTEBoth Procedure 1 and Procedure 2 must be completed
for this test.]
USP 32 Official Monographs / Carprofen 91
Sample solution: Sample per Dissolution 711. Dilute with ASSAY

Medium to a concentration that is similar to the Standard PROCEDURE
solution. Mobile phase: Acetonitrile, methanol, glacial acetic acid,
Chromatographic system and water (40:25:0.2:35)
(See Chromatography 621, System Suitability.) System suitability stock solution: 16 g/mL of USP
Mode: LC Carprofen Related Compound A RS in Mobile phase. Sonicate
Detector: UV 254 nm if necessary.
Column: 3.9-mm 30-cm; packing L1 [NOTEUse low-actinic glassware.]
Flow rate: 2 mL/min System suitability solution: Mix 10 mL of System suitability
Injection size: 50 L stock solution and 10 mL of the Standard solution and dilute
System suitability with Mobile phase to 100 mL.
Sample: Standard solution [NOTEUse low-actinic glassware]
Suitability requirements Standard solution: 160 g/mL of USP Carprofen RS in
Relative standard deviation: NMT 2.0% Mobile phase. Sonicate if necessary.
Analysis [NOTEUse low-actinic glassware.]
Samples: Standard solution and Sample solution Sample solution: 160 g/mL of Carprofen in Mobile phase
Calculate the percentage of C18H21NO3 H3PO4 1/2H2O [NOTEUse low-actinic glassware]
dissolved: Chromatographic system
Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 Mode: LC
Detector: UV 239 nm
rU = peak response from the Sample solution Column: 4.6-mm 25-cm; 5-m packing L1
rS = peak response from the Standard solution Flow rate: 1 mL/min
CS = concentration of USP Codeine Phosphate RS in Injection size: 10 L
the Standard solution (mg/mL) System suitability
CU = nominal concentration of the Sample solution Sample: System suitability solution
(mg/mL) Suitability requirements
Mr1 = molecular weight of codeine phosphate Resolution: NLT 2.0 between carprofen and carprofen
hemihydrate, 406.37 related compound A
Mr2 = molecular weight of anhydrous codeine Column efficiency: NLT 5000 theoretical plates
phosphate, 397.37 Tailing factor: NMT 2.0
Tolerances: NLT 75% (Q) of the labeled amounts of Relative standard deviation: NMT 2.0%
C18H21NO3 H3PO4 1/2H2O are dissolved. Analysis
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Samples: Standard solution and Sample solution
for Content Uniformity with respect to aspirin, carisoprodol, Calculate the percentage of C15H12ClNO2 in the portion of
and codeine phosphate Carprofen taken:
USP Aspirin RS rS = peak response from the Standard solution
USP Carisoprodol RS CS = concentration of USP Carprofen RS in the
USP Codeine N-Oxide RS Standard solution (g/mL)
USP Codeine Phosphate RS CU = concentration of carprofen in the Sample solution
USP Salicylic Acid RS (g/mL)
IMPURITIES
Carprofen Inorganic Impurities
(Comment on this Monograph)id=m13600=Carprofen=Ca-Chl- RESIDUE ON IGNITION 281: NMT 0.1%
Monos.pdf) HEAVY METALS, Method II 231: NMT 20 ppm
Organic Impurities
PROCEDURE 1: LIMIT OF ACETONE AND METHYLENE CHLORIDE
Standard solution: 0.5 mg/mL of acetone and 0.06
mg/mL of methylene chloride in N,N-dimethylacetamide
Sample solution: 100 mg/mL of Carprofen in N,N-
dimethylacetamide
Mode: GC
C15H12ClNO2 273.71 Detector: Flame ionization
9H-Carbazole-2-acetic acid, 6-chloro--methyl-, ()-; Column: 0.53-mm 30-m capillary column coated with
()-6-Chloro--methylcarbazole-2-acetic acid [53716-49-7]. 3.0-m G43 stationary phase
DEFINITION
Carprofen contains NLT 98.0% and NMT 102.0% of
C15H12ClNO2, calculated on the dried basis.
IDENTIFICATION
B. The retention time of the Sample solution corresponds to
92 Carprofen / Official Monographs USP 32
Carrier gas: Nitrogen Acceptance criteria

Temperature (See the temperature table below.) Individual impurities (See Impurity Table 1.)
Time Temperature
(min) () Impurity Table 1
Column 0 80 Relative Relative Acceptance
4 80 Retention Response Criteria
Impurity Name Time Factor NMT (%)
7.66 190
Carprofen related 0.9 0.5
10.66 190
compound A
Injection port 210 (carbazole)
Detector 220 2-[1,1-Dimethoxy-2- 1.3 0.5
hydroxypropyl]-6-
Flow rate: 4.9 mL/min chlorocarbazole
Injection size: 1 L 2-[2- 3.3 0.5
Injection type: Split flow ratio, 10:1 Chloropropionyl]-6-
System suitability chloro-9-
Sample: Standard solution acetylcarbazole
[NOTEAcetone elutes before methylene chloride.]
Suitability requirements Any individual 0.1
Resolution: NLT 1.5 between acetone and methylene unknown related
chloride compound
Relative standard deviation: NMT 10.0% for the
acetone peak responses
Samples: Standard solution and Sample solution LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT
Calculate the percentage of each residual solvent in the 0.5% of its weight.
portion of Carprofen taken: ADDITIONAL REQUIREMENTS
Result = (rU/rS) (CS/CU) 100 containers. Store at 25, excursions permitted between 15
rU = peak response of the individual residual solvent and 30.
in the Sample solution LABELING: Label it to indicate that it is intended for
rS = peak response of the individual residual solvent veterinary use only.
in the Standard solution USP REFERENCE STANDARDS 11
CS = concentration of individual residual solvent in USP Carprofen RS
the Standard solution (g/mL) USP Carprofen Related Compound A RS
CU = concentration of carprofen in the Sample
solution (g/mL)
Acceptance criteria Carprofen Tablets
Acetone: NMT 5000 ppm
Methylene chloride: NMT 600 ppm (Comment on this Monograph)id=m13610=Carprofen
Procedure 2 Tablets=Ca-Chl-Monos.pdf)
Mobile phase, Sample solution, System suitability DEFINITION
solution, System suitability, and Chromatographic Carprofen Tablets contain NLT 90.0% and NMT 110.0% of the
system: Proceed as directed in the Assay. labeled amount of carprofen (C15H12ClNO2).
Analysis
Sample: Sample solution IDENTIFICATION
Calculate the percentage of each related compound in A. INFRARED ABSORPTION 197K
the portion of Carprofen taken: Standard: Mix 2 mg of USP Carprofen RS with 200 mg of
potassium bromide, and grind thoroughly for 1015 min.
Result = (rU/rT) 100 Compress the mixture into a clear pellet. Record the IR
spectrum of the pellet immediately after preparation.
rU = peak response of each individual peak (other Sample: Transfer an amount equivalent to 100 mg of
than the major peak of carprofen) carprofen from powdered Tablets (NLT 4 Tablets), to a 125-
rT = sum of the peak responses
USP 32 Official Monographs / Carprofen 93
mL separatory funnel. Add 30 mL of water and 3 drops of rU = peak response from the Sample solution
hydrochloric acid, and shake for 5 min. Add 30 mL of rS = peak response from the Standard solution
methylene chloride, and shake for another 5 min. Allow the Cs = concentration of USP Carprofen RS in the
phases to separate. Carefully drain and collect the lower Standard solution (mg/mL)
methylene chloride layer through anhydrous sodium sulfate Cu = nominal concentration of the Sample solution
that is placed on a cotton pledget into a suitable container. (mg/mL)
Evaporate the methylene chloride on a steam bath with the Acceptance criteria: 90.0%110.0% of the labeled
aid of a stream of nitrogen to dryness. Dry the residue in amount of C15H12ClNO2
vacuum at 60 for about 30 min. Mix 2 mg of the dried
residue with 200 mg of potassium bromide, and grind PERFORMANCE TESTS
thoroughly for 1015 min. Compress the mixture into a DISSOLUTION 711
clear pellet. Record the IR spectrum of the carprofen sample [NOTEUse low-actinic volumetric flasks, dissolution vessels,
pellet immediately after preparation. and evaporation covers.]
B. The retention time of the Sample solution corresponds to Medium: 0.05 M phosphate buffer, pH 7.5 (prepared by
that of the Standard solution, as obtained in the Assay. dissolving 6.8 g of monobasic potassium phosphate in 600
mL of water, mixing, adding 18 mL of 2 N sodium
ASSAY hydroxide, mixing, diluting with water to 1000 mL, and
PROCEDURE adjusting with 0.2 N sodium hydroxide or 0.2 N
Mobile phase: Acetonitrile, phosphoric acid, and water hydrochloric acid to a pH of 7.50 0.05); 900 mL
(500:1:500) Apparatus 2: 50 rpm
Standard solution: 0.05 mg/mL of USP Carprofen RS in Time: 30 min
Mobile phase Determine the amount of C15H12ClNO2 dissolved by
[NOTEUse low-actinic glassware.] employing the following method.
Sample solution: Weigh 20 Tablets, and calculate the Standard solution:
average Tablet weight. Grind the Tablets into a uniform For Tablets labeled to contain 25 mg: 0.028 mg/mL of
powder. Transfer three weighed portions of the powder, USP Carprofen RS in methanol and Medium (1:89)
each equivalent to the weight of one Tablet, into three For Tablets labeled to contain 75 mg: 0.083 mg/mL of
volumetric flasks of a suitable calibrated volume such that an USP Carprofen RS in methanol and Medium (3:87)
interim concentration of 0.5 mg/mL of Mobile phase can be For Tablets labeled to contain 100 mg: 0.111 mg/mL of
prepared. To each flask, add Mobile phase to 80% of the USP Carprofen RS in methanol and Medium (2:43)
calibrated volume, sonicate for 10 min, then stir for 10 min. Sample solutions: Sample per 711 Dissolution. Dilute with
Sonicate again for 10 min, and stir for another 10 min. Cool Medium to a concentration that is similar to the Standard
to room temperature, dilute with Mobile phase to volume to solution. Pass a portion of the solution under test through a
obtain an interim concentration of 0.5 mg of carprofen/mL, suitable 0.45-m filter.
and mix. Quantitatively transfer 5.0 mL of the solution to a System suitability solution: Determine the absorbance of
50.0-mL volumetric flask, and dilute with Mobile phase to the Standard solution, as directed for Analysis, five times: the
volume. Pass the solution through a PVDF filter having a relative standard deviation is NMT 2.0%.
0.45-m or finer porosity, discarding the first 5 mL of the Analysis
filtrate. The final concentration is 0.05 mg/mL of carprofen. Samples: Standard solution, Sample solution, and Blank
[NOTEUse low-actinic glassware.] Determine the amount of C15H12ClNO2 dissolved by
Chromatographic system measuring the absorbance of the Sample solution in
(See Chromatography 621, System Suitability.) comparison with the appropriate Standard solution at the
Mode: LC wavelength of maximum absorbance at 300 nm, using a
Detector: UV 240 nm 0.5-cm cell for Tablets labeled to contain 25 mg, a 0.2-cm
Column: 4.6-mm 15-cm; packing L7 cell for Tablets labeled to contain 75 mg, and a 0.1-cm
Flow rate: 1 mL/min cell for Tablets labeled to contain 100 mg. Use Medium as
Injection size: 20 L the blank.
[NOTEWash the column after each series of analyses with Calculate the percentage of C15H12ClNO2 dissolved:
a mixture of acetonitrile and water (20:80) for 30 min;
gradually change the composition of acetonitrile and Result = (AU/AS) (CS/CU) 100
water to 80:20 over 10 min; continue to wash at 80:20
for 30 min; gradually change the composition to 50:50 AU = absorbance of the Sample solution
over 10 min; and continue to wash at 50:50 for another AS = absorbance of the Standard solution
30 min.] CS = concentration of USP Carprofen RS in the
System suitability Standard solution (mg/mL)
Sample: Standard solution CU = nominal concentration of the Sample solution
Suitability requirements (mg/mL)
Column efficiency: NLT 4000 theoretical plates Tolerances: NLT 80% (Q) of the labeled amount of
Tailing factor: NMT 2.0 C15H12ClNO2 is dissolved.
Relative standard deviation: NMT 2.0% UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
[NOTEInject the Standard solution in duplicate after every for Content Uniformity
12 injections or fewer of any other solution. The ratio of Procedure for content uniformity
the average area of the duplicate injections to that [NOTEUse low-actinic glassware.]
obtained from the initial five replicate injections is Mobile phase, Standard solution, System suitability, and
0.951.05.] Chromatographic system: Prepare as directed in the
Analysis Assay.
Samples: Standard solution and Sample solution Sample solution: Transfer 10 Tablets individually to 10
Calculate the percentage of the labeled content of separate volumetric flasks of a suitable calibrated volume
C15H12ClNO2 in the portion of Tablets taken: such that an interim concentration of 0.5 mg/mL of Mobile
phase can be prepared. To each flask, add Mobile phase to
Result = (rU/rS) (CS/CU) 100 80% of the calibrated volume, sonicate for 10 min, then
94 Carprofen / Official Monographs USP 32
stir for 10 min. Sonicate again for 10 min, and stir for Column efficiency: NLT 4000 theoretical plates,
another 10 min or until the Tablets are completely Standard solution
disintegrated. Cool to room temperature, dilute with Mobile Tailing factor: NMT 2.0, Standard solution
phase to volume to obtain an interim concentration of 0.5 Relative standard deviation: NMT 2.0%, Standard
mg of carprofen/mL, and mix. Quantitatively transfer 5.0 solution
mL of the individual solutions to 10 separate 50.0-mL [NOTEThe carprofen peak should be defined and
volumetric flasks, dilute with Mobile phase to volume, and integratable from injections of the System suitability
mix. Pass the solution through a polyvinylidenefluoride solution.]
(PVDF) filter having a 0.45-m or finer porosity, discarding [NOTEAfter every six injections of any solution, inject a
the first 5 mL of the filtrate. The final concentration is Standard solution in duplicate. The ratio of the average
about 0.05 mg of carprofen/mL. response of the duplicate injections to that obtained
Analysis: Proceed as directed for Analysis in the Assay. from the initial five replicate injections is 0.951.05.]
Calculate the percentage of the labeled content of Analysis
C15H12ClNO2 in the portion of Tablets taken: Samples: Standard solution, Sample solution and Blank
solution
Result = (rU/rS) (CS/CU) 100 Calculate the percentage of carprofen-related compounds
in the portion of Tablets taken:
rU = peak area response from the Sample solution
rS = peak area response rom the Standard solution Result = (rU/rS) (CS/CU) 100
CS = concentration of USP Carprofen RS in the
Standard solution (mg/mL) rU = peak area of any peak other than carprofen
CU = nominal concentration of the Sample solution from the Sample solution
(mg/mL) rS = peak area of carprofen from the Standard
solution
CS = concentration of carprofen in the Standard
IMPURITIES solution (mg/mL)
Organic Impurities CU = nominal concentration of the Sample solution
PROCEDURE (mg/mL)
Mobile phase: Proceed as directed in the Assay. Acceptance criteria
[NOTEUse low-actinic glassware.] Individual impurities: NMT 0.5%
Standard solution: 0.05 g/mL of USP Carprofen RS in Total impurities: NMT 2.0%
Mobile phase [NOTEDisregard any peak also observed in the Blank
System suitability solution: 0.005 g/mL of Carprofen, solution.]
from Standard solution in Mobile phase
Sample solution: Use the Sample solution, prepared as ADDITIONAL REQUIREMENTS
directed in the Assay. PACKAGING AND STORAGE: Preserve in tight containers.
Blank solution: Transfer an amount of Tablet base USP REFERENCE STANDARDS 11
equivalent to the weight of 1 Tablet to a volumetric flask of USP Carprofen RS
the same calibrated volume as that used to prepare the
Sample solution. To each flask, add Mobile phase to 80% of
the calibrated volume. Sonicate for 10 min, then stir for 10
min. Sonicate again for 10 min, and stir for another 10
Carteolol Hydrochloride
min. Cool to room temperature, and dilute with Mobile (Comment on this Monograph)id=m13660=Carteolol
phase to volume. Quantitatively transfer 5.0 mL of the Hydrochloride=Ca-Chl-Monos.pdf)
solution to a 50.0-mL volumetric flask, dilute with Mobile
phase to volume, and mix. Pass the solution through a
PVDF filter having a 0.45-m or finer porosity, discarding
the first 5 mL of the filtrate.
Mode: LC
Detector: UV 240 nm C16H24N2O3 HCl 328.83
Column: 4.6-mm 15-cm; 5-m packing L7 2(1H)-Quinolinone, 5-[3-[(1,1-dimethylethyl)amino]-2-
Flow rate: 1 mL/min hydroxypropoxy]-3,4-dihydro-, monohydrochloride;
Injection size: 50 L 5-[3-(tert-Butylamino)-2-hydroxypropoxy]-3,4-dihydrocarbostyril
[NOTEWash the column after each series of analyses monohydrochloride [51781-21-6].
with a mixture of acetonitrile and water (20:80) for 30
min; gradually change the composition of acetonitrile DEFINITION
and water to 80:20 over 10 min; continue to wash at Carteolol Hydrochloride contains NLT 98.0% and NMT 101.5%
80:20 for 30 min; gradually change the composition to of C16H24N2O3 HCl, calculated on the dried basis.
50:50 over 10 min; and continue to wash at 50:50 for
another 30 min.] IDENTIFICATION
System suitability A. INFRARED ABSORPTION 197K
Samples: Standard solution, Sample solution and System B. ULTRAVIOLET ABSORPTION 197U
suitability solution Solution: 10 g/mL
Suitability requirements C. CHLORIDE AND SULFATE, Chloride 191: A 20 mg/mL
Resolution: NLT 2.0 between carprofen and the nearest solution responds to the tests.
impurity peak, Sample solution
USP 32 Official Monographs / Carteolol 95
ASSAY Standard solution B: Dilute 5.0 mL of Standard solution A

PROCEDURE to 50 mL with methanol.
Solution A: 0.67 mg/mL of dibasic sodium phosphate, Standard solution C: Dilute 5.0 mL of Standard solution B
adjust with 1 M phosphoric acid to a pH of 6.0 0.05 to 10 mL with methanol.
before bringing to volume Sample solution: Transfer 250 mg of Carteolol
Mobile phase: Acetonitrile and Solution A (1:3) Hydrochloride to a 10mL volumetric flask, dissolve in
[NOTEIncreasing the proportion of pH 6.0 buffer increases methanol, using heat or sonication if necessary to achieve
resolution.] dissolution. Dilute with methanol to volume.
Diluent: Methanol and Solution A (1:1) Chromatographic system
Standard stock solution: 1 mg/mL of USP Carteolol (See Chromatography 621, Thin-Layer Chromatography.)
Hydrochloride RS Mode: TLC
Standard solution: Transfer 10.0 mg/mL of Standard stock Adsorbent: 0.25-mm layer of chromatographic silica gel
solution to a 100mL volumetric flask containing 5 mL mixture
acetonitrile, and dilute with water to volume. Application volume: 10 L
System suitability stock solution: Dissolve 50 mg of p- Developing solvent system: Chloroform, methanol, and
acetotoluidide in 50 mL of acetonitrile and dilute with water ammonium hydroxide (50:20:1)
to 100 mL. Analysis
System suitability solution: Combine 10.0 mL of System Samples: Standard solutions A, B, and C; and Sample
suitability stock solution and 10.0 mL of Standard stock solution
solution. Dilute with water to 100 mL (contains 0.05 mg/mL Proceed as directed in the chapter. Allow the spots to dry.
of p-acetotoluidide and 0.1 mg/mL of USP Carteolol Line a chromatographic chamber with filter paper, and
Hydrochloride RS). saturate the paper with the Developing solvent system.
Sample stock solution: 1 mg/mL of Carteolol Place the plate in the chamber, and develop the
Hydrochloride chromatogram until the solvent front has moved three-
Sample solution: Transfer 10.0 mL of Sample stock solution fourths of the length of the plate. Remove the plate from
to a 100mL volumetric flask containing 5 mL of the chamber, and allow to air-dry. Examine the plate
acetonitrile, and dilute with water to volume. under short-wavelength UV light.
Chromatographic system Acceptance criteria: The RF value of the principal spot
(See Chromatography 621, System Suitability.) from the Sample solution corresponds to that from Standard
Mode: LC solution A. Compare the sizes and intensities of any spots
Detector: UV 252 nm other than the principal spot from the Sample solution with
Column: 3.9-mm 30-cm; packing L1 those of the principal spots from the Standard solutions: no
Flow rate: 1 mL/min spot exceeds in size or intensity the principal spot from
Injection size: 20 L Standard solution B (0.2%), and the sum of all the impurity
System suitability spots is NMT 0.5%.
[NOTEThe relative retention times for carteolol and p- ADDITIONAL REQUIREMENTS
acetotoluidide are 0.8 and 1.0, respectively.] PACKAGING AND STORAGE: Preserve in well-closed containers.
Suitability requirements USP REFERENCE STANDARDS 11
Resolution: NLT 3 between the carteolol peak and p- USP Carteolol Hydrochloride RS
acetotoluidide peak, System suitability solution
solution Carteolol Hydrochloride Ophthalmic
Analysis
Samples: Standard solution and Sample solution Solution
Calculate the percentage of C16H24N2O3 HCl in the portion (Comment on this Monograph)id=m13665=Carteolol
of Carteolol Hydrochloride taken: Hydrochloride Ophthalmic Solution=Ca-Chl-Monos.pdf)
Result = (rU/rS) (CS/CU) 100 DEFINITION
Carteolol Hydrochloride Ophthalmic Solution is a sterile,
rU = peak response from the Sample solution aqueous, isotonic solution of Carteolol Hydrochloride. It
rS = peak response from the Standard solution contains a suitable antimicrobial preservative. It contains NLT
CS = concentration of USP carteolol hydrochloride RS 90.0% and NMT 110.0% of the labeled amount of C16H24N2O3
in the Standard solution (mg/mL) HCl.
CU = concentration of carteolol hydrochloride in the
Acceptance criteria: 98.0%101.5% A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
SPECIFIC TESTS mixture
PH 791: 5.06.0, in a solution (1 in 100) Standard solution: 1 mg/mL of USP Carteolol
LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT Hydrochloride RS
0.5% of its weight. Sample solution: 1 mg/mL of carteolol hydrochloride from
a volume of Ophthalmic Solution, diluted if necessary
IMPURITIES Application volume: 10 L
Inorganic Impurities Developing solvent system: Chloroform, methanol, and
RESIDUE ON IGNITION 281: NMT 0.1% ammonium hydroxide (50:20:1)
ARSENIC, Method II 211: NMT 3 ppm Analysis
HEAVY METALS, Method I 231: NMT 20 ppm Samples: Standard solution and Sample solution
Organic Impurities Allow the spots to dry. Line a chromatographic chamber
PROCEDURE with filter paper, and saturate the paper with the
Standard solution A: 0.5 mg/mL of USP Carteolol Developing solvent system. Place the plate in the chamber,
Hydrochloride RS in methanol
96 Carteolol / Official Monographs USP 32
and develop the chromatogram until the solvent front has CS = concentration of USP Carteolol Hydrochloride RS
moved three-fourths of the length of the plate. Remove in the Standard solution (mg/mL)
the plate from the chamber, and allow to air-dry. Examine CU = nominal concentration of carteolol hydrochloride
the plate under short-wavelength UV light. in the Sample solution (mg/mL)
Acceptance criteria: The RF value of the principal spot from Acceptance criteria: 90.0%110.0%
the Sample solution corresponds to that of the Standard
solution. SPECIFIC TESTS
B. The retention time of the Sample solution corresponds to STERILITY TESTS 71: It meets the requirements when tested
that of the Standard solution, as obtained in the Assay. as directed for Test for Sterility of the Product to Be Examined,
ASSAY PH 791: 6.08.0
PROCEDURE
Solution A: 0.67 mg/mL of dibasic sodium phosphate, ADDITIONAL REQUIREMENTS
adjust with 1 M phosphoric acid to a pH of 6.0 0.05, PACKAGING AND STORAGE: Preserve in tight containers.
before bringing to volume USP REFERENCE STANDARDS 11
Mobile phase: Acetonitrile and Solution A (1:3) [NOTE USP Carteolol Hydrochloride RS
Increasing the proportion of pH 6.0 buffer increases
resolution.]
Diluent: Methanol and Solution A (1:1) Carteolol Hydrochloride Tablets
Standard stock solution: 1 mg/mL of USP Carteolol
Hydrochloride RS (Comment on this Monograph)id=m13670=Carteolol
Standard solution: Transfer 10.0 mL of Standard stock Hydrochloride Tablets=Ca-Chl-Monos.pdf)
solution, to a 100mL volumetric flask containing 5 mL of
acetonitrile, and dilute with water to volume. DEFINITION
System suitability stock solution: Dissolve 50 mg of p- Carteolol Hydrochloride Tablets contain NLT 90.0% and NMT
acetotoluidide in 50 mL of acetonitrile and dilute with water 110.0% of the labeled amount of C16H24N2O3 HCl.
to 100 mL. IDENTIFICATION
System suitability solution: Combine 10.0 mL of System The retention time of the Sample solution corresponds to that
suitability stock solution, and 10.0 mL of Standard stock of the Standard solution, as obtained in the Assay.
solution. Dilute with water to 100 mL. (Contains 0.05
mg/mL of p-acetotoluidide and 0.1 mg/mL of USP Carteolol ASSAY
Hydrochloride RS.) PROCEDURE
Sample solution: Transfer an amount of Ophthalmic Solution A: 0.67 mg/mL of dibasic sodium phosphate,
Solution equivalent to 10 mg of carteolol hydrochloride to a adjust with 1 M phosphoric acid to a pH of 6.0 0.05
100mL volumetric flask and dilute with Diluent to volume. before bringing to volume
Pass a portion of this solution through a filter having a Mobile phase: Acetonitrile and Solution A (1:3)
porosity of 0.5 m or finer, discarding the first 2 mL of the [NOTEIncreasing the proportion of pH 6.0 buffer increases
filtrate, and use the filtrate as the Sample solution. resolution.]
Chromatographic system Diluent: Methanol and Solution A (1:1)
(See Chromatography 621, System Suitability.) System suitability stock solution: Dissolve 50 mg of p-
Mode: LC acetotoluidide in 50 mL of acetonitrile and dilute with water
Detector: UV 252 nm to 100 mL.
Column: 3.9-mm 30-cm; packing L1 System suitability solution: Combine 10.0 mL of System
Flow rate: 1 mL/min suitability stock solution, and 10.0 mL of Standard stock
Injection size: 20 L solution. Dilute with water to 100 mL. (Contains 0.05
System suitability mg/mL of p-acetotoluidide and 0.1 mg/mL of USP Carteolol
Sample: System suitability solution and Standard solution Hydrochloride RS.)
[NOTEThe relative retention times for carteolol and p- Standard stock solution: 1 mg/mL of USP Carteolol
acetotoluidide are 0.8 and 1.0, respectively.] Hydrochloride RS
Suitability requirements Standard solution: Transfer 10.0 mL of Standard stock
Resolution: NLT 3 between the carteolol peak and the p- solution to a 100mL volumetric flask containing 5 mL of
acetotoluidide peak, System suitability solution acetonitrile, and dilute with water to volume
Relative standard deviation: NMT 2.0%, Standard Sample solution: Transfer an amount equivalent to 10 mg
solution of carteolol hydrochloride from NLT 20 powdered Tablets to
Analysis a 100mL flask. Add 50 mL of Diluent, and shake by
Samples: Standard solution and Sample solution mechanical means for 1 h. Add 5 mL of acetonitrile, and
Calculate the percentage of C16H24N2O3 HCl in each mL of dilute with Diluent to volume. Pass a portion of this solution
the Ophthalmic Solution taken: through a filter having a 0.5-m or finer porosity, discarding
the first 2 mL of filtrate, and use the clear filtrate as the
Result = (rU/rS) (CS/CU) 100 Sample solution.
rU = peak response from the Sample solution (See Chromatography 621, System Suitability.)
USP 32 Official Monographs / Casanthranol 97
Mode: LC UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements

Detector: UV 252 nm
Column: 3.9-mm 30-cm; packing L1 IMPURITIES
Flow rate: 1 mL/min Organic Impurities
Injection size: 20 L PROCEDURE: LIMIT OF DEHYDROCARTEOLOL HYDROCHLORIDE
System suitability Solution A, Mobile phase, and Diluent: Proceed as
Samples: System suitability solution and Standard solution directed in the Assay.
Suitability requirements Standard solution: 1 g/mL of USP Dehydrocarteolol
Resolution: NLT 3 between the carteolol peak and the p- Hydrochloride RS in Diluent
acetotoluidide peak, System suitability solution Sample solution: Transfer an amount equivalent to 10 mg
Relative standard deviation: NMT 2.0%, Standard of carteolol hydrochloride from NLT 20 powdered Tablets
solution to a 100mL volumetric flask, add 50 mL of Diluent, and
[NOTEThe relative retention times for carteolol and p- shake by mechanical means for 1 h. Dilute with Diluent to
acetotoluidide are about 0.8 and 1.0, respectively.] volume. Pass a portion of this solution through a filter
Analysis having a 0.5-m or finer porosity.
Samples: Standard solution and Sample solution Chromatographic system
[NOTEUse peak areas where peak responses are (See Chromatography 621, System Suitability.)
indicated.] Mode: LC
Calculate the percentage of C16H24N2O3 HCl in the portion Detector: Fluorometric detector, with excitation at 300
of Tablets taken: nm and a 418-nm emission filter
Result = (rU/rS) (CS/CU) 100 Flow rate: 2 mL/min
rU = peak response from the Sample solution System suitability
rS = peak response from the Standard solution Sample: Standard solution
CS = concentration of USP Carteolol Hydrochloride RS Suitability requirements
in the Standard solution (mg/mL) Relative standard deviation: NMT 5.%
CU = nominal concentration of carteolol hydrochloride Analysis
in the Sample solution (mg/mL) Samples: Standard solution and Sample solution
Acceptance criteria: 90.0%110.0% [NOTEUse peak areas where peak responses are
indicated.]
PERFORMANCE TESTS Calculate the percentage of dehydrocarteolol hydrochloride
DISSOLUTION 711 in the portion of Tablets taken:
Medium: Water, 900 mL
Apparatus 2: 50 rpm Result = (rU/rS) (CS/CU) 100
Time: 30 min
Solution A: 2.0 mg/mL of monobasic potassium phosphate rU = peak response of the Sample solution
Mobile phase: Acetonitrile and Solution A (2:3) rS = peak response of the Standard solution
Standard solution: 1.1L g/mL of USP Carteolol CS = concentration of USP Dehydrocarteolol
Hydrochloride RS, L being the labeled amount, in mg, of Hydrochloride RS in the Standard solution
carteolol hydrochloride/Tablet (mg/mL)
Sample solution: Sample per Dissolution 711. Dilute with CU = nominal concentration of carteolol hydrochloride
Medium to a concentration that is similar to Standard in the Sample solution (mg/mL)
solution. [NOTEPass a portion of the Sample solution Acceptance criteria: NMT 1.0%
through a filter having a porosity of 1 m or finer,
discarding the first 2 mL of the filtrate.] ADDITIONAL REQUIREMENTS
Mode: LC USP Carteolol Hydrochloride RS
Detector: UV 252 nm USP Dehydrocarteolol Hydrochloride RS
Flow rate: 1 mL/min
Injection size: 15 L Casanthranol
System suitability
Sample: Standard solution (Comment on this Monograph)id=m13700=Casanthranol=Ca-
Suitability requirements Chl-Monos.pdf)
Relative standard deviation: NMT 2.%
Analysis DEFINITION
Samples: Standard solution and Sample solution Casanthranol is obtained from Cascara Sagrada. It contains in
Calculate the percentage of C16H24N2O3 HCl dissolved: each 100 g NLT 20.0 g of total hydroxyanthracene derivatives
calculated on the dried basis, calculated as cascaroside A. NLT
Result = (rU/rS) (CS/CU) 100 80% of the total hydroxyanthracene derivatives consists of
cascarosides, calculated as cascaroside A.
rS = peak response of the Standard solution ASSAY
CS = concentration of USP Carteolol Hydrochloride RS TOTAL HYDROXYANTHRACENE DERIVATIVES
in the Standard solution (g/mL) [NOTE 1Perform all extractions by shaking vigorously, and
CU = nominal concentration of carteolol hydrochloride allow all phases to separate completely before transferring.
in the Sample solution (g/mL) Entrainment of aglycones into the aqueous phase, as
Acceptance criteria: NLT 80% (Q) of the labeled amount indicated by a value of less than 2.6 for the ratio of the
of C16H24N2O3 HCl
98 Casanthranol / Official Monographs USP 32
absorbance of the final solution at 515 nm to that at 440 absorbance of the final solution at 515 nm to that at 440
nm, may lead to false results.] nm, may lead to false results.]
[NOTE 2Throughout this Assay, use 1 N sodium hydroxide [NOTE 2Throughout this Assay, use 1 N sodium hydroxide
that is prepared without added barium ions as directed that is prepared without added barium ions as directed
under Reagents, Indicators, and SolutionsVolumetric under Reagents, Indicators, and SolutionsVolumetric
Solutions.] Solutions.]
Solution A: 1 g/mL of ferric chloride Solution A: 1 g/mL of ferric chloride
Sample stock solution: Transfer 500 mg of Casanthranol to Sample stock solution: Transfer 500 mg of Casanthranol to
a 100-mL volumetric flask. Add 30 mL of 70% alcohol, swirl a 100-mL volumetric flask. Add 30 mL of 70% alcohol, swirl
to dissolve, and dilute with 70% alcohol to volume. Quickly to dissolve, and dilute with 70% alcohol to volume. Quickly
filter through soft, rapid-flow filter paper, taking precautions filter through soft, rapid-flow filter paper, taking precautions
to minimize loss by evaporation. to minimize loss by evaporation.
Sample solution: Pipet 10 mL of Sample stock solution into Sample solution: Pipet 10 mL of Sample stock solution into
a separatory funnel containing 5 mL of water and 2 drops of a separatory funnel containing 5 mL of water and 2 drops of
1 N hydrochloric acid. Extract with 40 mL of methylene 1 N hydrochloric acid. Extract with 40 mL of methylene
chloride, and transfer the lower layer to a second separatory chloride, and transfer the lower layer to a second separatory
funnel. Add 10 mL of water to the second separatory funnel, funnel. Add 10 mL of water to the second separatory funnel,
and shake. Allow to separate, discard the lower layer, and and shake. Allow to separate, discard the lower layer, and
transfer the water layer to the first separatory funnel. Extract transfer the water layer to the first separatory funnel. Extract
the combined water layers with 40 mL of methylene the combined water layers with 40 mL of methylene
chloride, and transfer the lower layer to the second chloride, and transfer the lower layer to the second
separatory funnel. Add 10 mL of water to the second separatory funnel. Add 10 mL of water to the second
separatory funnel, and shake. Allow to separate, and discard separatory funnel, and shake. Allow to separate, discard the
the lower layer. Transfer the combined water layers, with the lower layer, and transfer the water layer to the first
aid of water, to a 50-mL volumetric flask, filtering through a separatory funnel. Extract the combined aqueous phase with
small pledget of cotton, water-wet, and dilute with water to 30 mL of clear, freshly prepared water-saturated ethyl
volume. acetate, and transfer the water layer to another separatory
Spectrometric conditions funnel. Repeat the extraction with two additional 30-mL
Mode: Visible portions of the freshly prepared water-saturated ethyl
Analytical wavelength: 515 nm acetate. Add 5 mL of water to the combined ethyl acetate
Cell: 1 cm extracts, shake, allow the phases to separate, discard the
Blank: Methanol ethyl acetate extracts, and add 30 mL of the freshly
Analysis prepared water-saturated ethyl acetate to the water wash.
Sample: Sample solution Shake, allow the phases to separate, and discard the ethyl
Pipet 10 mL of Sample solution into a flask containing 2 mL acetate phase. Transfer the combined aqueous phases, with
of Solution A and 12 mL of hydrochloric acid. Attach a the aid of water, to a 50-mL volumetric flask, filtering
condenser arranged for refluxing, and heat for 3 h by through a small pledget of cotton, water-wet, and dilute
keeping the flask immersed in boiling water or with water to volume.
continuously exposed to steam heat. Cool, wash down Spectrometric conditions
the condenser, and transfer to a separatory funnel with Mode: Visible
the aid of 4 mL of 1 N sodium hydroxide and five 6-mL Analytical wavelength: 515 nm
portions of water. Extract with 20 mL of methylene Cell: 1 cm
chloride, and transfer the lower layer to another Blank: Methanol
separatory funnel. Repeat the extraction with three Analysis
additional 20-mL portions of methylene chloride, wash the Sample: Sample solution
combined methylene chloride extracts with two 10-mL Pipet 15 mL of Sample solution into a flask containing 2 mL
portions of water, shaking each time for 2 min, and of Solution A and 12 mL of hydrochloric acid. Attach a
discard the water washings. Transfer the washed condenser arranged for refluxing, and heat for 3 h by
methylene chloride extract to a 100-mL volumetric flask, keeping the flask immersed in boiling water or
and dilute with methylene chloride to volume. Evaporate continuously exposed to steam heat. Cool, wash down
a 20.0-mL portion carefully on a water bath to dryness, the condenser, and transfer to a separatory funnel with
and dissolve the residue in 10.0 mL of a 1in200 solution the aid of 4 mL of 1 N sodium hydroxide and five 6-mL
of magnesium acetate in methanol. portions of water. Extract with 20 mL of methylene
Determine the absorbance and calculate the quantity, in chloride, and transfer the lower layer to another
mg, of total hydroxyanthracene derivatives in the portion separatory funnel. Repeat the extraction with three
of Casanthranol taken: additional 20-mL portions of methylene chloride, wash the
combined methylene chloride extracts with two 10-mL
Result = AU F portions of water, shaking each time for 2 min, and
discard the water washings. Transfer the washed
AU = absorbance of the Sample solution methylene chloride extract to a 100-mL volumetric flask,
F = monograph correction factor, 155 dilute with methylene chloride to volume, and mix.
Acceptance criteria: NLT 20.0 g of total hydroxyanthracene Evaporate a 20.0-mL portion carefully on a water bath to
derivatives/100 g calculated on the dried basis, calculated as dryness, and dissolve the residue in 10.0 mL of a
cascaroside A. 1in200 solution of magnesium acetate in methanol.
CASCAROSIDES Determine the absorbance and calculate the quantity, in
[NOTE 1Perform all extractions by shaking vigorously, and mg, of cascarosides in the portion of Casanthranol taken:
allow all phases to separate completely before transferring.
Entrainment of aglycones into the aqueous phase, as Result = AU F
indicated by a value of less than 2.7 for the ratio of the
USP 32 Official Monographs / Cascara 99
AU = absorbance of the Sample solution 1 N hydrochloric acid. Extract with 40 mL of methylene

F = monograph correction factor, 103.5 mg chloride, and transfer the lower layer to a second separatory
Acceptance criteria: NLT 80% of the Total funnel. Add 10 mL of water to the second separatory funnel,
Hydroxyanthracene Derivatives and shake. Allow to separate, discard the lower layer, and
transfer the water layer to the first separatory funnel. Extract
IMPURITIES the combined water layers with 40 mL of methylene
Inorganic Impurities chloride, and transfer the lower layer to the second
RESIDUE ON IGNITION 281: NMT 4.0% separatory funnel. Add 10 mL of water to the second
HEAVY METALS, Method II 231: NMT 25 ppm separatory funnel, and shake. Allow to separate, discard the
lower layer, and transfer the water layer to the first
SPECIFIC TESTS separatory funnel. Extract the combined aqueous phase with
LOSS ON DRYING 731: Dry it in vacuum at 80 for 16 h: it 30 mL of clear, freshly prepared water-saturated ethyl
loses NMT 10.0% of its weight. acetate, and transfer the water layer to another separatory
ADDITIONAL REQUIREMENTS funnel. Repeat the extraction with two additional 30-mL
PACKAGING AND STORAGE: Preserve in tight, light-resistant portions of the freshly prepared water-saturated ethyl
containers, at a temperature not exceeding 30. acetate. Add 5 mL of water to the combined ethyl acetate
extracts, shake, allow the phases to separate, discard the
ethyl acetate extracts, and add 30 mL of the freshly
prepared water-saturated ethyl acetate to the water wash.
Cascara Sagrada Shake, allow the phases to separate, and discard the ethyl
(Comment on this Monograph)id=m13720=Cascara acetate phase. Transfer the combined aqueous phases, with
Sagrada=Ca-Chl-Monos.pdf) the aid of water, to a 50-mL volumetric flask. Dilute with
water to volume.
DEFINITION Spectrometric system
Cascara Sagrada is the dried bark of Rhamnus purshiana De Mode: Visible
Candolle (Fam. Rhamnaceae). It yields NLT 7.0% of total Analytical wavelength: 515 nm
hydroxyanthracene derivatives, calculated as cascaroside A, Cell: 1 cm
and calculated on the dried basis. NLT 60% of the total Blank: Methanol
hydroxyanthracene derivatives consists of cascarosides, Analysis
calculated as cascaroside A. Sample: Sample solution
[NOTECollect Cascara Sagrada NLT 1 year before use.] Pipet 15 mL of Sample solution into a flask containing 2 mL
of Solution A and 12 mL of hydrochloric acid. Attach a
IDENTIFICATION condenser arranged for refluxing, and heat for 3 h by
A. PROCEDURE keeping the flask immersed in boiling water or
Sample: 100 mg of powdered Cascara Sagrada continuously exposed to steam heat. Cool, wash down
Analysis: Add Sample to 10 mL of hot water, shake the the condenser, and transfer to a separatory funnel with
mixture occasionally until it is cold, filter, dilute the filtrate the aid of 4 mL of 1 N sodium hydroxide and five 6-mL
with water to 10 mL, and add 10 mL of 6 N ammonium portions of water. Extract with 20 mL of methylene
hydroxide. chloride, and transfer the lower layer to another
Acceptance criteria: An orange color is produced. separatory funnel. Repeat the extraction with three
B. It becomes red to reddish brown in color when treated additional 20-mL portions of methylene chloride, wash the
with 6 N ammonium hydroxide. combined methylene chloride extracts with two 10-mL
C. PROCEDURE portions of water, shaking each time for 2 min, and
Sample: 100 mg of powdered Cascara Sagrada discard the water washings. Transfer the washed
Analysis: Macerate Sample with 1 mL of alcohol, add 10 mL methylene chloride extract to a 100-mL volumetric flask,
of water, boil the mixture, then cool, filter, and shake the and dilute with methylene chloride to volume. Evaporate
filtrate with 10 mL of ether: a greenish yellow ether solution a 20.0-mL portion carefully on a water bath to dryness,
separates. Shake 3 mL of this ether solution with 3 mL of 6 and dissolve the residue in 10.0 mL of a solution (1 in
N ammonium hydroxide, and dilute the separated ammonia 200) of magnesium acetate in methanol
solution with 20 mL of water. Determine the absorbance and calculate the amount, in
Acceptance criteria: A distinct orange-pink color remains. mg, of cascarosides in the portion of Cascara Sagrada
taken:
ASSAY
CASCAROSIDES Result = F AU
[NOTE 1Perform all extractions by shaking vigorously, and
allow all phases to separate completely before transferring. F = conversion factor, 103.5
Entrainment of aglycones into the aqueous phase, as AU = absorbance of the Sample solution
indicated by a value of less than 2.7 for the ratio of the Acceptance criteria: NLT 60% of the total
absorbance of the final solution at 515 nm to that at 440 hydroxyanthracene derivatives consists of cascarosides,
nm, may lead to false results.] calculated as cascaroside A.
[NOTE 2Throughout this Assay, use 1 N sodium hydroxide TOTAL HYDROXYANTHRACENE DERIVATIVES
that is prepared without added barium ions as directed [NOTE 1Perform all extractions by shaking vigorously, and
under Reagents, Indicators, and SolutionsVolumetric allow all phases to separate completely before transferring.
Solutions.] Entrainment of aglycones into the aqueous phase, as
Solution A: 1 g/mL of ferric chloride indicated by a value of less than 2.6 for the ratio of the
Sample stock solution: Add 1 g of Cascara Sagrada to 70 absorbance of the final solution at 515 nm to that at 440
mL of boiling water, boil for several min, with stirring, allow nm, may lead to false results.]
to cool, and transfer with the aid of water to a 100-mL [NOTE 2Throughout this Assay, use 1 N sodium hydroxide
volumetric flask. Dilute with water to volume, and filter that is prepared without added barium ions as directed
through suitable filter paper. under Reagents, Indicators, and SolutionsVolumetric
Sample solution: Pipet 10 mL of Sample stock solution into Solutions.]
a separatory funnel containing 5 mL of water and 2 drops of
100 Cascara / Official Monographs USP 32
Solution A and Sample stock solution: Prepare as directed an alkali; starch grains spheroidal, up to 8 m in diameter;
in the Assay for Cascarosides. calcium oxalate in monoclinic prisms or rosette aggregates
Sample solution: Pipet 10 mL of Sample stock solution into from 6 to 20 m in diameter, occasionally up to 45 m in
a separatory funnel containing 5 mL of water and 2 drops of diameter.
1 N hydrochloric acid. Extract with 40 mL of methylene WATER DETERMINATION, Method IIIProcedure for Articles of
chloride, and transfer the lower layer to a second separatory Botanical Origin 921: Dry it at 105 for 5 h: it loses NMT
funnel. Add 10 mL of water to the second separatory funnel, 12.0% of its weight.
and shake. Allow to separate, discard the lower layer, and
transfer the water layer to the first separatory funnel. Extract
the combined water layers with 40 mL of methylene
chloride, and transfer the lower layer to the second Cascara Sagrada Extract
separatory funnel. Add 10 mL of water to the second (Comment on this Monograph)id=m13730=Cascara Sagrada
separatory funnel, and shake. Allow to separate, and discard Extract=Ca-Chl-Monos.pdf)
the lower layer. Transfer the combined water layers, with the
aid of water, to a 50-mL volumetric flask, and dilute with DEFINITION
water to volume. Cascara Sagrada Extract contains, in each 100 g, NLT 10.0 g
Spectrometric system and NMT 12.0 g of hydroxyanthracene derivatives, of which
Mode: Visible NLT 50% consists of cascarosides, both calculated as
Analytical wavelength: 515 nm cascaroside A.
Cell: 1 cm Mix 900 g of Cascara Sagrada, in coarse powder, with 4000 mL
Blank: Methanol of boiling water, and macerate the mixture for 3 h. Then
Analysis transfer it to a percolator, allow it to drain, exhaust it by
Sample: Sample solution percolation, using boiling water as the menstruum, and collect
Proceed as directed for Analysis in the Assay for 5000 mL of percolate. Evaporate the percolate to dryness,
Cascarosides, except to evaporate a 15.0-mL portion of reduce the Extract to a fine powder, and, after assaying, add
the methylene chloride solution instead of 20.0 mL. sufficient starch, dried at 100, or other inert, nontoxic
Calculate the quantity, in mg, of total hydroxyanthracene diluents to make the product contain, in each 100 g, 11 g of
derivatives in the portion of Cascara Sagrada taken: hydroxyanthracene derivatives. Mix the powders, and pass the
Extract through a number 60 sieve.
Result = F AU
ASSAY
F = conversion factor, 138 CASCAROSIDES
AU = absorbance of the Sample solution [NOTE 1Perform all extractions by shaking vigorously, and
Acceptance criteria: NLT 7.0% of the total allow all phases to separate completely before transferring.
hydroxyanthracene derivatives, calculated as cascaroside A, Entrainment of aglycones into the aqueous phase, as
and calculated on the dried basis. indicated by a value of less than 2.7 for the ratio of the
absorbance of the final solution at 515 nm to that at 440
IMPURITIES nm, may lead to false results.]
Organic Impurities [NOTE 2Throughout this assay, use 1 N sodium hydroxide
PROCEDURE: ARTICLES OF BOTANICAL ORIGIN, Foreign Organic that is prepared without added barium ions as directed for
Matter 561: NMT 4.0% Reagents, Indicators, and SolutionsVolumetric Solutions.]
Solution A: 1 g/mL of ferric chloride
SPECIFIC TESTS Sample stock solution: Transfer 1 g of Extract to a 100-mL
BOTANIC CHARACTERISTICS volumetric flask. Add 60 mL of 70% alcohol, swirl or
Cascara Sagrada: Usually in flattened or transversely curved sonicate for 1520 min, several times, allow to stand
pieces, occasionally in quills of variable length and from 1 to overnight, sonicate or swirl for 1015 min, dilute with 70%
5 mm in thickness. The outer surface is brown, purplish alcohol to volume, mix, and filter through suitable filter
brown, or brownish red, longitudinally ridged, with or paper.
without grayish or whitish lichen patches, sometimes with Sample solution: Pipet 10 mL of Sample stock solution into
numerous lenticels and occasionally with moss attached. The a separatory funnel containing 5 mL of water and 2 drops of
inner surface is longitudinally striate, light yellow, weak 1 N hydrochloric acid. Extract with 40 mL of methylene
reddish brown, or moderate yellowish brown. The fracture is chloride, and transfer the lower layer to a second separatory
short with projections of phloem fiber bundles in the inner funnel. Add 10 mL of water to the second separatory funnel,
bark. and shake. Allow to separate, discard the lower layer, and
Histology: It shows a yellowish brown, purple, or reddish transfer the water layer to the first separatory funnel. Extract
brown cork of up to 10 or more rows of small cells; stone the combined water layers with 40 mL of methylene
cells in yellowish, tangentially elongated groups of 2050 chloride, and transfer the lower layer to the second
cells in the cortex, pericycle, and outer phloem regions; separatory funnel. Add 10 mL of water to the second
phloem rays 14 cells wide, 1525 cells deep, frequently separatory funnel, and shake. Allow to separate, discard the
diagonal or curved, forming converging groups; phloem lower layer, and transfer the water layer to the first
fibers in small bundles, more or less surrounded by crystal separatory funnel. Extract the combined aqueous phase with
fibers and located between the phloem rays; parenchyma 30 mL of clear, freshly prepared water-saturated ethyl
with brown walls and containing starch grains and calcium acetate, and transfer the water layer to another separatory
oxalate crystals. funnel. Repeat the extraction with two additional 30-mL
Powdered Cascara Sagrada: Moderate yellowish brown to portions of the freshly prepared water-saturated ethyl
dusky yellowish orange. It shows numerous broken phloem acetate. Add 5 mL of water to the combined ethyl acetate
fiber bundles with accompanying crystal fibers containing extracts, shake, allow the phases to separate, discard the
monoclinic prisms of calcium oxalate; stone cells more or ethyl acetate extracts, and add 30 mL of the freshly
less adherent, in small groups with thick, finely lamellated prepared water-saturated ethyl acetate to the water wash.
and porous walls; fragments of reddish brown to yellow Shake, allow the phases to separate, and discard the ethyl
cork; masses of parenchyma and phloem ray cells colored acetate phase. Transfer the combined aqueous phases, with
reddish brown to orange upon the addition of a solution of
USP 32 Official Monographs / Cascara 101
the aid of water, to a 50-mL volumetric flask. Dilute with Spectrometric conditions
water to volume. Mode: Visible
Spectrometric conditions Analytical wavelength: 515 nm
Mode: Visible Cell: 1 cm
Analytical wavelength: 515 nm Blank: Methanol
Cell: 1 cm Analysis
Blank: Methanol Sample: Sample solution
Analysis Analysis: Pipet 10 mL of Sample solution into a flask
Sample: Sample solution containing 2 mL of Solution A and 12 mL of hydrochloric
Pipet 25 mL of Sample solution into a flask containing 2 mL acid. Attach a condenser arranged for refluxing, and heat
of Solution A and 12 mL of hydrochloric acid. Attach a for 3 h by keeping the flask immersed in boiling water or
condenser arranged for refluxing, and heat for 3 h by continuously exposed to steam heat. Cool, wash down the
keeping the flask immersed in boiling water or condenser, and transfer to a separatory funnel with the aid
continuously exposed to steam heat. Cool, wash down of 4 mL of 1 N sodium hydroxide and five 6-mL portions of
the condenser, and transfer to a separatory funnel with water. Extract with 20 mL of methylene chloride, and
the aid of 4 mL of 1 N sodium hydroxide and five 6-mL transfer the lower layer to another separatory funnel.
portions of water. Extract with 20 mL of methylene Repeat the extraction with three additional 20-mL portions
chloride, and transfer the lower layer to another of methylene chloride, wash the combined methylene
separatory funnel. Repeat the extraction with three chloride extracts with two 10-mL portions of water,
additional 20-mL portions of methylene chloride, wash the shaking each time for 2 min, and discard the water
combined methylene chloride extracts with two 10-mL washings. Transfer the washed methylene chloride extract
portions of water, shaking each time for 2 min, and to a 100-mL volumetric flask, and dilute with methylene
discard the water washings. Transfer the washed chloride to volume. Evaporate a 20.0-mL portion carefully
methylene chloride extract to a 100-mL volumetric flask, on a water bath to dryness, and dissolve the residue in
dilute with methylene chloride to volume, and mix. 10.0 mL of a 1in200 solution of magnesium acetate in
Evaporate a 20.0-mL portion carefully on a water bath to methanol.
dryness, and dissolve the residue in 10.0 mL of a Determine the absorbance and calculate the quantity, in
1in200 solution of magnesium acetate in methanol. mg, of Total Hydroxyanthracene Derivatives in the portion
Determine the absorbance and calculate the quantity, in of Extract taken:
mg, of Cascarosides in the portion of Extract taken:
Result = F AU
Result = F AU
F = correction factor, 155.2
F = correction factor, 62.06 AU = absorbance of the solution from the Sample
AU = absorbance of the solution from the Sample solution
solution Acceptance criteria: 10.0 g12.0 g of hydroxyanthracene
Acceptance criteria: NLT 50% of Total Hydroxyanthracene derivatives/100 g calculated as cascaroside A
Derivatives calculated as cascaroside A.
TOTAL HYDROXYANTHRACENE DERIVATIVES ADDITIONAL REQUIREMENTS
[NOTE 1Perform all extractions by shaking vigorously, and PACKAGING AND STORAGE: Preserve in tight, light-resistant
allow all phases to separate completely before transferring. containers, at a temperature not exceeding 30.
Entrainment of aglycones into the aqueous phase, as
indicated by a value of less than 2.6 for the ratio of the
absorbance of the final solution at 515 nm to that at 440 Cascara Tablets
nm, may lead to false results.]
[NOTE 2Throughout this assay, use 1 N sodium hydroxide (Comment on this Monograph)id=m13740=Cascara Tablets=Ca-
that is prepared without added barium ions as directed for Chl-Monos.pdf)
Reagents, Indicators, and SolutionsVolumetric Solutions..]
Solution A and Sample stock solution: Prepare as directed DEFINITION
in the Assay for Cascarosides. Cascara Tablets are prepared from Cascara Sagrada Extract.
Sample solution: Pipet 10 mL of Sample stock solution into They contain an amount of hydroxyanthracene derivatives,
a separatory funnel containing 5 mL of water and 2 drops of calculated as cascaroside A, NLT 9.35% and NMT 12.65% of
1 N hydrochloric acid. Extract with 40 mL of methylene the labeled amount of Cascara Sagrada Extract. NLT 50% of
chloride, and transfer the lower layer to a second separatory the hydroxyanthracene derivatives are cascarosides, calculated
funnel. Add 10 mL of water to the second separatory funnel, as cascaroside A.
and shake. Allow to separate, discard the lower layer, and ASSAY
transfer the water layer to the first separatory funnel. Extract CASCAROSIDES
the combined water layers with 40 mL of methylene [NOTE 1Perform all extractions by shaking vigorously, and
chloride, and transfer the lower layer to the second allow all phases to separate completely before transferring.
separatory funnel. Add 10 mL of water to the second Entrainment of aglycones into the aqueous phase, as
separatory funnel, and shake. Allow to separate, and discard indicated by a value of less than 2.7 for the ratio of the
the lower layer. Transfer the combined water layers, with the
aid of water, to a 50-mL volumetric flask, and dilute with
water to volume.
102 Cascara / Official Monographs USP 32
absorbance of the final solution at 515 nm to that at 440 F = monograph correction factor 103.5
nm, may lead to false results.] AU = absorbance of the solution from the Sample
[NOTE 2Throughout this assay, use 1 N sodium hydroxide solution
that is prepared without added barium ions as directed for Acceptance criteria: NLT 50% of the Total
Reagents, Indicators, and Solutions, Volumetric Solutions.] Hydroxyanthracene Derivatives, calculated as cascaroside A
Solution A: 1 g/mL of ferric chloride TOTAL HYDROXYANTHRACENE DERIVATIVES
Sample stock solution: Transfer the equivalent to 1 g of [NOTE 1Perform all extractions by shaking vigorously, and
Cascara Sagrada Extract from powdered Tablets (NLT 20), to allow all phases to separate completely before transferring.
a 100-mL volumetric flask. Add 60 mL of 70% alcohol, swirl Entrainment of aglycones into the aqueous phase, as
or sonicate for 1520 min, several times, allow to stand indicated by a value of less than 2.6 for the ratio of the
overnight, sonicate or swirl for 1015 min, dilute with 70% absorbance of the final solution at 515 nm to that at 440
alcohol to volume, mix, and filter through suitable filter nm, may lead to false results.]
paper. [NOTE 2Throughout this assay, use 1 N sodium hydroxide
Sample solution: Pipet 10 mL of Sample stock solution into that is prepared without added barium ions as directed
a separatory funnel containing 5 mL of water and 2 drops of under Reagents, Indicators, and Solutions, Volumetric
1 N hydrochloric acid. Extract with 40 mL of methylene Solutions.]
chloride, and transfer the lower layer to a second separatory Solution A and Sample stock solution: Prepare as directed
funnel. Add 10 mL of water to the second separatory funnel, in the Assay for Cascarosides.
and shake. Allow to separate, discard the lower layer, and Sample solution: Pipet 10 mL of Sample stock solution into
transfer the water layer to the first separatory funnel. Extract a separatory funnel containing 5 mL of water and 2 drops of
the combined water layers with 40 mL of methylene 1 N hydrochloric acid. Extract with 40 mL of methylene
chloride, and transfer the lower layer to the second chloride, and transfer the lower layer to a second separatory
separatory funnel. Add 10 mL of water to the second funnel. Add 10 mL of water to the second separatory funnel,
separatory funnel, and shake. Allow to separate, and discard and shake. Allow to separate, discard the lower layer, and
the lower layer. Transfer the water layer to the first transfer the water layer to the first separatory funnel. Extract
separatory funnel. Extract the combined aqueous phase with the combined water layers with 40 mL of methylene
30 mL of clear, freshly prepared water-saturated ethyl chloride, and transfer the lower layer to the second
acetate, and transfer the water layer to another separatory separatory funnel. Add 10 mL of water to the second
funnel. separatory funnel, and shake. Allow to separate, and discard
Repeat the extraction with two additional 30-mL portions of the lower layer. Transfer the combined water layers, with the
the freshly prepared water-saturated ethyl acetate. Add 5 aid of water, to a 50-mL volumetric flask, and dilute with
mL of water to the combined ethyl acetate extracts, shake, water to volume.
allow the phases to separate, discard the ethyl acetate Spectrometric conditions
extracts, and add 30 mL of the freshly prepared water- Mode: Spectrometry
saturated ethyl acetate to the water wash. Shake, allow the Analytical wavelength: 515 nm
phases to separate, and discard the ethyl acetate phase. Cell: 1 cm
Transfer the combined aqueous phases, with the aid of Blank: Methanol
water, to a 50-mL volumetric flask. Dilute with water to Analysis
volume. Sample: Sample solution
Spectrometric conditions Prepare as directed under Analysis in the Assay for
Mode: Spectrometry Cascarosides, except pipet 10 mL of Sample solution.
Analytical wavelength: 515 nm Continue to and dilute with methylene chloride to
Cell: 1 cm volume.
Blank: Methanol Evaporate a 15.0-mL portion carefully on a water bath to
Analysis dryness, and dissolve the residue in 10.0 mL of a
Sample: Sample solution 1in200 solution of magnesium acetate in methanol.
Pipet 20 mL of Sample solution into a flask containing 2 mL Determine the absorbance and calculate the quantity, in
of Solution A and 12 mL of hydrochloric acid. Attach a mg, of total hydroxyanthracene derivatives in the portion
condenser arranged for refluxing, and heat for 3 h by of Extract:
keeping the flask immersed in boiling water or
continuously exposed to steam heat. Cool, wash down Result = F AU
the condenser, and transfer to a separatory funnel with
the aid of 4 mL of 1 N sodium hydroxide and five 6-mL F = monograph correction factor 206.9
portions of water. Extract with 20 mL of methylene AU = absorbance of the solution from the Sample
chloride, and transfer the lower layer to another solution
separatory funnel. Repeat the extraction with three Acceptance criteria: Contains an amount of
additional 20-mL portions of methylene chloride, wash the hydroxyanthracene derivatives NLT 9.35% and NMT 12.65%
combined methylene chloride extracts with two 10-mL of the labeled amount of Cascara Sagrada Extract calculated
portions of water, shaking each time for 2 min, and as cascaroside A
discard the water washings. Transfer the washed
methylene chloride extract to a 100-mL volumetric flask, PERFORMANCE TESTS
and dilute with methylene chloride to volume. Evaporate DISINTEGRATION 701: 60 min
a 20.0-mL portion carefully on a water bath to dryness, UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
and dissolve the residue in 10.0 mL of a 1in200 solution ADDITIONAL REQUIREMENTS
of magnesium acetate in methanol. PACKAGING AND STORAGE: Preserve in tight containers; if the
Determine the absorbance and calculate the quantity, in Tablets are coated, well-closed containers may be used.
mg, of cascarosides in the portion of Extract:
Result = F AU
USP 32 Official Monographs / Castor 103
Cascara Sagrada Fluidextract Castor Oil

(Comment on this Monograph)id=m13750=Cascara Sagrada (Comment on this Monograph)id=m13790=Castor Oil=Ca-Chl-
Fluidextract=Ca-Chl-Monos.pdf) Monos.pdf)
Prepare Cascara Sagrada Fluidextract as follows. To 1000 g of Castor Oil is the fixed oil obtained from the seed of Ricinus
coarsely ground Cascara Sagrada add 3000 mL of boiling communis Linne (Fam. Euphorbiaceae). It contains no added
water, and allow to macerate in a suitable percolator for 2 h. substances.
Allow the percolation to proceed at a moderate rate, gradually
adding boiling water until the drug is practically exhausted of IMPURITIES
its active principles. Evaporate the percolate on a water bath Inorganic Impurities
or in a vacuum still to NMT 800 mL, cool, add 200 mL of HEAVY METALS, Method II 231: NMT 10 ppm
alcohol and, if necessary, add sufficient water to make the
product measure 1000 mL. SPECIFIC TESTS
OTHER COMPONENTS DISTINCTION FROM MOST OTHER FIXED OILS: It is only partly
ALCOHOL DETERMINATION, Method I 611: 18.0%20.0% of soluble in solvent hexane (distinction from most other fixed
C2H5OH oils), but it yields a clear liquid with an equal volume of
alcohol (foreign fixed oils).
ADDITIONAL REQUIREMENTS FATS AND FIXED OILS, Free Fatty Acids 401: The free fatty
PACKAGING AND STORAGE: Preserve in tight, light-resistant acids in 10 g require NMT 3.5 mL of 0.10 N sodium
containers, and avoid exposure to direct sunlight and to hydroxide for neutralization.
excessive heat FATS AND FIXED OILS, Hydroxyl Value 401
Sample solution: Transfer 2 g to a glass-stoppered, 250-mL
conical flask. Add 5.0 mL of a freshly prepared mixture of
acetic anhydride and pyridine (1:3), and swirl to mix.
Aromatic Cascara Fluidextract Analysis: Connect the flask to a reflux condenser, and heat
(Comment on this Monograph)id=m13760=Aromatic Cascara on a steam bath for 2 h. Add 10 mL of water through the
Fluidextract=Ca-Chl-Monos.pdf) condenser, swirl to mix, heat on a steam bath for an
additional 10 min, and allow to cool to room temperature.
DEFINITION Add through the condenser 15 mL of normal butyl alcohol
Prepare Aromatic Cascara Fluidextract as follows: that previously has been neutralized to phenolphthalein,
remove the condenser, and wash the tip of the condenser
Cascara Sagrada, in very coarse powder 1000 g and the sides of the flask with an additional 10 mL of
neutralized normal butyl alcohol. Add 1 mL of
Magnesium Oxide 120 g
phenolphthalein TS, and titrate with 0.5 N alcoholic
Suitable sweetening agent(s) potassium hydroxide VS to a faint pink endpoint. Perform a
Suitable essential oils(s) blank determination on a 5.0-mL portion of the acetic
anhydridepyridine mixture. To determine the amount of
Suitable flavoring agent(s)
free acid in the Oil, weigh accurately 10 g into a 250-mL
Alcohol 200 mL conical flask, add 10 mL of pyridine that previously has been
Purified Water, a sufficient quantity, neutralized to phenolphthalein, swirl to mix, add 1 mL of
to make 1000 mL phenolphthalein TS, and titrate with 0.5 N alcoholic
potassium hydroxide VS to a faint pink endpoint.
Mix the Cascara Sagrada with the Magnesium Oxide, moisten it Calculate the hydroxyl value in the portion of Oil taken:
uniformly with 2000 mL of boiling water, and set it aside in a
shallow container for 48 h, stirring it occasionally. Pack it in a Result = 56.1N[A + (BW/D) C]/W
percolator, and percolate with boiling water until the drug is
exhausted. Evaporate the percolate, at a temperature not N = normality determined for the alcoholic potassium
exceeding 100, to 750 mL, and at once dissolve in it the hydroxide solution
flavoring agent(s). When the liquid has cooled, add the A = volume of 0.5 N alcoholic potassium hydroxide
Alcohol, in which the sweetening agent(s) and oils have been consumed by the blank (mL)
dissolved, add sufficient water to make the Aromatic B = volume consumed in the free-acid titration (mL)
Fluidextract measure 1000 mL, and mix. W = weight of Oil taken (g)
D = weight of Oil used in the free-acid titration (g)
OTHER COMPONENTS C = volume consumed in the sample titration (mL)
ALCOHOL DETERMINATION 611: Between 18%20% of Acceptance criteria: 160168
C2H5OH, determined by the gasliquid chromatographic FATS AND FIXED OILS, Iodine Value 401: 8388
method, acetone being used as the internal standard FATS AND FIXED OILS, Saponification Value 401: 176182
ADDITIONAL REQUIREMENTS ADDITIONAL REQUIREMENTS

PACKAGING AND STORAGE: Preserve in tight, light-resistant PACKAGING AND STORAGE: Preserve in tight containers, and
containers, and avoid exposure to direct sunlight and to avoid exposure to excessive heat.
excessive heat.
104 Castor / Official Monographs USP 32
Castor Oil Capsules Castor Oil Emulsion

(Comment on this Monograph)id=m13800=Castor Oil (Comment on this Monograph)id=m13810=Castor Oil
Capsules=Ca-Chl-Monos.pdf) Emulsion=Ca-Chl-Monos.pdf)
Castor Oil Capsules contain NLT 90.0% and NMT 110.0% of Castor Oil Emulsion contains NLT 90.0% and NMT 120.0% of
the labeled amount of castor oil, calculated from the tests for the labeled amount of Castor Oil.
Weight variation and Specific gravity.
IDENTIFICATION
INFRARED ABSORPTION 197S Analysis: Shake well, and place 10 mL in a 125-mL
Standard solution: A similar solution of Castor Oil separator. Add 10 mL of 1 N hydrochloric acid and 20 mL
Sample solution: 40 mg/mL of castor oil from Capsules in of solvent hexane. Shake vigorously for 23 min, allow the
chloroform layers to separate, discard the aqueous phase, and filter the
upper layer through anhydrous sodium sulfate into a small
PERFORMANCE TESTS beaker. Evaporate the solvent on a steam bath, and to the
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements residue add 12 drops of sulfuric acid.
Acceptance criteria: A red color indicates the presence of
SPECIFIC TESTS castor oil.
Sample: Capsule contents ASSAY
FATS AND FIXED OILS, Hydroxyl Value 401 PROCEDURE
Sample solution: Transfer 2 g from Capsule contents to a Internal standard solution: 12 mg/mL of di(2-
glass-stoppered, 250-mL conical flask. Add 5.0 mL of a ethylhexyl)phthalate in chloroform
freshly prepared mixture of 1 volume of acetic anhydride Standard solution: Transfer 100 mg of castor oil to a 100-
and 3 volumes of pyridine, and swirl to mix. mL boiling flask equipped with a suitable reflux condenser
Analysis: Connect the flask to a reflux condenser, and heat connected by a ground-glass joint. Add 30 mL of a mixture
on a steam bath for 2 h. Add 10 mL of water through the of 300 mL of methanol and 3.7 mL of sulfuric acid, reflux in
condenser, swirl to mix, heat on a steam bath for an a water bath maintained at 7580 for 2.5 h, cool, and
additional 10 min, and allow to cool to room temperature. rinse down the condenser with 10 mL of water. Transfer the
Add through the condenser 15 mL of normal butyl alcohol contents of the flask to a 125-mL separator with the aid of
that previously has been neutralized to phenolphthalein, 10 mL of water. Rinse the condenser and the flask with 25
remove the condenser, and wash the tip of the condenser mL of solvent hexane, and transfer to the separator. Shake
and the sides of the flask with an additional 10 mL of the separator for 2 min, and draw off the aqueous layer into
neutralized normal butyl alcohol. Add 1 mL of a second 125-mL separator. Add 20 mL of solvent hexane to
phenolphthalein TS, and titrate with 0.5 N alcoholic the second separator, shake for 2 min, discard the aqueous
potassium hydroxide VS to a faint pink endpoint. Perform a layer, and transfer the solvent hexane layer to the first
blank determination on a 5.0 mL portion of the acetic separator with the aid of 10 mL of solvent hexane. Wash the
anhydridepyridine mixture. To determine the amount of combined extracts with three 5-mL portions of water,
free acid in the Oil, weigh accurately 10 g into a 250-mL discarding the washings, and transfer the washed extract to
conical flask, add 10 mL of pyridine that previously has been a 125-mL conical flask, through a funnel containing
neutralized to phenolphthalein, swirl to mix, add 1 mL of anhydrous sodium sulfate, with the aid of 25 mL of solvent
phenolphthalein TS, and titrate with 0.5 N alcoholic hexane. Place the flask in a hot water bath, and evaporate
potassium hydroxide VS to a faint pink endpoint. with the aid of a current of air to dryness. To the residue,
Calculate the hydroxyl value taken: add 10.0 mL of Internal standard solution, and mix until
solution is complete.
Result = 56.1N[A + (BW/D) C]/W Sample solution: Transfer an amount of Emulsion, well-
shaken and nominally equivalent to 100 mg of castor oil, to
N = normality determined for the alcoholic potassium a long-neck, round-bottom, 100-mL boiling flask equipped
hydroxide solution with a suitable reflux condenser connected by a ground-
A = volume of 0.5 N alcoholic potassium hydroxide glass joint. Proceed as directed under Standard solution,
consumed by the blank (mL) beginning with Add 30 mL of a mixture of 300 mL of
B = volume consumed in the free-acid titration (mL) methanol and 3.7 mL of sulfuric acid.
W = weight of Oil taken (g) Chromatographic system
D = weight of Oil used in the free-acid titration (g) (See Chromatography 621, System Suitability.)
C = volume consumed in the sample titration (mL) Mode: GC
Acceptance criteria: 160168 Detector: Flame ionization
FATS AND FIXED OILS, Iodine Value 401: 8388 Column: 1.8-m 4-mm; packed with 4% liquid phase
Sample: Capsule contents G25 on support S1
FATS AND FIXED OILS, Saponification Value 401: 176182 Column conditioning: Flush with helium for 25 min,
Sample: Capsule contents then heat without further flushing at 250 for NLT 30 min,
ADDITIONAL REQUIREMENTS then cool to room temperature, and finally heat while
PACKAGING AND STORAGE: Preserve in tight containers, helium is flowing through it at 250 for NLT 60 min.
preferably at a controlled room temperature.
USP 32 Official Monographs / Cefaclor 105
Temperature solution, beginning with Add 30 mL of a mixture of 300

Column: 245 mL of methanol and 3.7 mL of sulfuric acid
Injection port: 300 Chromatographic system
Detector: 300 (See Chromatography 621, System Suitability.)
Flow rate: Adjust to obtain a peak due to castor oil 5.5 Mode: GC
min after introduction of the specimen and an internal Detector: Flame ionization
standard peak 8 min after introduction of the specimen. Column: 1.8-m 4-mm column packed with 4% liquid
Carrier gas: Helium phase G25 on support S1
Injection size: 5 L Column conditioning: Flush with helium for 25 min,
Analysis then heat without further flushing at 250 for NLT 30 min,
Sample: Standard solution and Sample solution then cool to room temperature, and finally heat while
Measure the heights of the peaks due to castor oil and helium is flowing through it at 250 for NLT 60 min.
internal standard Temperature
Calculate the percentage of castor oil in the portion of Column: 245
Emulsion: Injection port: 300
Detector: 300
Result = (RU/RS) (CS/CU) 100 Flow rate: Adjust to obtain a peak due to castor oil 5.5
min after introduction of the specimen and an internal
RU = ratio of the heights of the peaks due to castor oil standard peak 8 min after introduction of the specimen.
and internal standard from the Sample solution Carrier gas: Helium
RS = ratio of the heights of the peaks due to castor oil Injection size: 5 L
and internal standard from the Standard Analysis
solution Sample: Standard solution and Sample solution
CS = concentration of castor oil in the Standard Measure the heights of the peaks due to castor oil and
solution (mg/mL) Internal standard solution.
CU = nominal concentration of castor oil in the Sample Calculate the percentage, of castor oil in the portion of
solution (mg/mL) Aromatic Castor Oil:
Result = (RU/RS) (Cs/CU) 100
PACKAGING AND STORAGE: Preserve in tight containers. RU = ratio of the heights of the peaks due to castor oil
and internal standard from the Sample solution
RS = ratio of the heights of the peaks due to castor oil
Aromatic Castor Oil and internal standard from the Standard
solution
(Comment on this Monograph)id=m13830=Aromatic Castor CS = concentration of the Standard solution (mg/mL)
Oil=Ca-Chl-Monos.pdf) CU = nominal concentration of castor oil in the Sample
solution (mg/mL)
DEFINITION Acceptance criteria: NLT 95.0%
Aromatic Castor Oil is Castor Oil containing suitable flavoring
agents. It contains NLT 95.0% of castor oil. OTHER COMPONENTS
ALCOHOL DETERMINATION, Method I 611: NMT 4.0% of
ASSAY C2H5OH
PROCEDURE
Internal standard solution: 12 mg/mL of di(2- ADDITIONAL REQUIREMENTS
ethylhexyl)phthalate in chloroform PACKAGING AND STORAGE: Preserve in tight containers.
Standard solution: Transfer 100 mg of castor oil to a 100-
mL boiling flask equipped with a suitable reflux condenser
connected by a ground-glass joint. Add 30 mL of a mixture
of 300 mL of methanol and 3.7 mL of sulfuric acid, reflux in Cefaclor
a water bath maintained at 7580 for 2.5 h, cool, and (Comment on this Monograph)id=m13880=Cefaclor=Ca-Chl-
rinse down the condenser with 10 mL of water. Transfer the Monos.pdf)
contents of the flask to a 125-mL separator with the aid of
10 mL of water. Rinse the condenser and the flask with 25
mL of solvent hexane, and transfer to the separator. Shake
the separator for 2 min, and draw off the aqueous layer into
a second 125-mL separator. Add 20 mL of solvent hexane to
the second separator, shake for 2 min, discard the aqueous
layer, and transfer the solvent hexane layer to the first
separator with the aid of 10 mL of solvent hexane. Wash the
combined extracts with three 5-mL portions of water, C15H14ClN3O4S H2O 385.82
discarding the washings, and transfer the washed extract to 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-
a 125-mL conical flask, through a funnel containing [(aminophenylacetyl)amino]-3-chloro-8-oxo-, monohydrate,
anhydrous sodium sulfate, with the aid of 25 mL of solvent [6R-[6,-7 (R*)]]-;
hexane. Place the flask in a hot water bath, and evaporate (6R,7R)-7-[(R)-2-Amino-2-phenylacetamido]-3-chloro-8-oxo-5-
with the aid of a current of air to dryness. To the residue, thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid
add 10.0 mL of Internal standard solution, and mix until monohydrate;
solution is complete. 3-Chloro-7-D-(2-phenylglycinamido)-3-cephem-4-carboxylic
Sample solution: Transfer 100 mg of Aromatic Castor Oil acid monohydrate [70356-03-5];
to a long-neck, round-bottom, 100-mL boiling flask Anhydrous 367.81
equipped with a suitable reflux condenser connected by a [53994-73-3].
ground-glass joint. Proceed as directed under Standard
106 Cefaclor / Official Monographs USP 32
DEFINITION Mobile Phase: See Gradient Table.

Cefaclor has a potency of NLT 950 g and NMT 1020 g of
C15H14ClN3O4S/mg, calculated on the anhydrous basis. Gradient Table
IDENTIFICATION Time Solution A Solution B
A. INFRARED ABSORPTION 197K (min) (%) (%)
B. The retention time for cefaclor of the Sample solution 0 95 5
the Assay. 30 75 25
45 0 100
ASSAY
55 0 100
PROCEDURE
Mobile phase: Dissolve 1 g of sodium 1-pentanesulfonate 60 95 5
in a mixture of 780 mL of water and 10 mL of triethylamine. 70 95 5
Adjust with phosphoric acid to a pH of 2.5 0.1, and add
220 mL of methanol Standard solution: 0.05 mg/mL of USP Cefaclor RS in
System suitability solution: 0.3 mg/mL of cefaclor and 0.3 Diluent
mg/mL of USP Cefaclor, Delta-3 Isomer RS in Mobile phase [NOTESonicate briefly, if necessary, to dissolve, and
Standard solution: 0.3 mg/mL of USP Cefaclor RS in Mobile avoid heating. Use this solution on the day it is
phase prepared.]
[NOTESonicate briefly, if necessary, to achieve dissolution, System suitability solution: 0.05 mg/mL of USP Cefaclor,
and avoid heating the solution. Use within 8 h if stored at Delta-3 Isomer RS dissolved in Standard solution
room temperature or within 20 h if stored under Sample solutions: Transfer 50 mg of Cefaclor into each of
refrigeration.] two 10-mL volumetric flasks, and dilute each with Diluent
Sample solution: 0.3 mg/mL of Cefaclor in Mobile phase to volume. Sonicate briefly, if necessary, to dissolve, and
[NOTESonicate briefly, if necessary, to achieve dissolution, avoid heating. [NOTEUse these within 2 h when stored at
and avoid heating the solution. Use within 8 h if stored at room temperature or within 20 h when stored under
room temperature, or within 20 h if stored under refrigeration.]
refrigeration.] Chromatographic system
(See Chromatography 621, System Suitability.) [NOTEReducing the acetonitrile content increases the
Mode: LC retention time of cefaclor and increases the resolution
Detector: UV 265 nm between cefaclor, delta-3 isomer and cefaclor.]
Column: 4.6-mm 25-cm; 5-m packing L1 Mode: LC
Flow rate: 1.5 mL/min Detector: UV 220 nm
Injection size: 20 L Column: 4.6-mm 25-cm; packing L1
System suitability Flow rate: 1 mL/min
Sample: System suitability solution Injection size: 20 L
[NOTEThe relative retention times for cefaclor and System suitability
cefaclor, delta-3 isomer are 0.8 and 1.0, respectively.] Samples: System suitability solution and Blank solution
Suitability requirements [NOTEThe retention time for the cefaclor peak is 2329
Resolution: NLT 2.5 Between the cefaclor peak and the min].
cefaclor, delta-3 isomer peak Suitability requirements
Tailing factor: NMT 1.5 Resolution: NLT 2.0 between cefaclor, delta-3 isomer
Relative standard deviation: NMT 2.0% and cefaclor, System suitability solution
Analysis Tailing factor: NMT 1.2 for the cefaclor peak, System
Samples: Standard solution and Sample solution suitability solution
Calculate the potency, in g/mg, of cefaclor C15H14ClN3O4S [NOTEExamine the chromatogram of the Blank solution
in each mg of the Cefaclor taken: for any extraneous peaks, and disregard any
corresponding peaks observed in the chromatogram of
Result = (rU/rS) (CS/CU) P the Sample solutions. Ensure that any extraneous peaks
observed do not represent carryover from previous
rU = peak area of Sample solution injections.]
rS = peak area of the Standard solution Analysis
CS = concentration of USP Cefaclor RS in the Standard Samples: Standard solution and Sample solutions
solution (mg/mL) Calculate the percentage of each cefaclor related
CU = concentration of Cefaclor in the Sample solution compound in the portion of Cefaclor taken:
(mg/mL)
P = designated potency, in g of Result = (ri/rS) (CS/CU) P F 100
(C15H14ClN3O4S)/mg, of USP Cefaclor RS
Acceptance criteria: 950 g/mg1020 g/mg ri = peak area of an individual related compound of
the Sample solution
IMPURITIES rS = peak area of the Standard solution
Organic Impurities CS = concentration of USP Cefaclor RS in the
PROCEDURE Standard solution (mg/mL)
Diluent: 2.4 mg/mL of monobasic sodium phosphate in CU = concentration of Cefaclor in the respective
water, adjust with phosphoric acid to a pH of 2.5 Sample solution (mg/mL)
Blank solution: Use the Diluent. P = designated potency of USP Cefaclor RS (g/mg)
Solution A: 6.9 mg/mL of monobasic sodium phosphate F = conversion factor, 0.001 mg/g
in water; adjust with phosphoric acid to a pH of 4.0
Solution B: Acetonitrile and Solution A (9:11), degassing
for NMT 2 min

Individual Impurities: Determine the mean values for (See Chromatography 621, System Suitability.)
each cefaclor related compound: NMT 0.5% of any Mode: LC
individual cefaclor related compound is found. Detector: UV 265 nm
Total impurities: NMT 2.0% of total cefaclor related Column: 4.6-mm 25-cm; 5-m packing L1
compounds is found. Flow rate: 1.5 mL/min
[NOTEIn an acceptable determination, the difference Injection size: 20 L
between duplicate determinations of total cefaclor System suitability
related compounds is NMT 0.2% absolute, or the Sample: System suitability solution
variation from the mean of the two values is NMT 10%, [NOTEThe relative retention times for cefaclor and
whichever is greater.] cefaclor, delta-3 isomer are 0.8 and 1.0, respectively. ]
SPECIFIC TESTS Resolution: NLT 2.5 between the cefaclor peak and the
CRYSTALLINITY 695: Meets the requirements cefaclor, delta-3 isomer peak
PH 791: 3.04.5, in an aqueous suspension containing 25 Tailing factor: NMT 1.5
mg/mL Relative standard deviation: NMT 2.0%
WATER DETERMINATION, Method I 921: 3.0%6.5% Analysis
ADDITIONAL REQUIREMENTS Calculate the potency, in g/mg, cefaclor C15H14ClN3O4S in
PACKAGING AND STORAGE: Preserve in tight containers. the portion of Cefaclor Capsules taken:
USP Cefaclor RS Result = (rU/rS) (CS /CU) P
USP Cefaclor, Delta-3 Isomer RS
rU = peak area of the Sample solution
rS = peak area of the Standard solution
Cefaclor Capsules CS = concentration of USP Cefaclor RS in the Standard
solution (mg/mL)
(Comment on this Monograph)id=m13890=Cefaclor CU = nominal concentration of Cefaclor in the Sample
Capsules=Ca-Chl-Monos.pdf) solution (mg/mL)
P = designated potency of USP Cefaclor RS (g of
DEFINITION C15H14ClN3O4S/mg )
Cefaclor Capsules contain the equivalent of NLT 90.0% and Acceptance criteria: 90.0%120%
NMT 120.0% of the labeled amount of C15H14ClN3O4S.
PERFORMANCE TESTS
IDENTIFICATION DISSOLUTION 711
PROCEDURE Medium: Water; 900 mL
Sample solution: Mix the contents of 1 capsule with water Apparatus 2: 50 rpm
and filter to obtain a concentration of 2 mg/mL. Time: 30 min
Analysis: Proceed as directed in the Assay. Sample solutions: Sample per Dissolution 711. Dilute with
Acceptance criteria: The retention time of the Sample Medium as needed, and filter.
solution corresponds to that of the Standard solution, as Standard solution: USP Cefaclor RS of a known
obtained in the Assay. concentration in Medium
ASSAY Analysis: Determine the amount of C15H14ClN3O4S dissolved
PROCEDURE from UV absorbances at 264 nm of the Sample solution, in
Mobile phase: Dissolve 1 g of sodium 1-pentanesulfonate comparison with the Standard solution.
in a mixture of 780 mL of water and 10 mL of triethylamine Tolerances: NLT 80% (Q) of the labeled amount of cefaclor
A. Adjust with phosphoric acid to a pH of 2.5 0.1, and add (C15H14ClN3O4S) is dissolved.
220 mL of methanol. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
System suitability solution: 0.3 mg/mL of cefaclor and 0.3 IMPURITIES
mg/mL of USP Cefaclor, Delta-3 Isomer RS in Mobile phase Organic Impurities
Standard solution: 0.3 mg/mL of USP Cefaclor RS in Mobile PROCEDURE
phase Diluent: 2.4 mg/mL of monobasic sodium phosphate in
[NOTESonicate briefly, if necessary, to achieve dissolution, water; adjusted with phosphoric acid to a pH of 2.5
and avoid heating the solution. Use within 8 h if stored at Blank solution: Use the Diluent.
room temperature, or within 20 h if stored under Solution A: Dissolve 6.9 g of monobasic sodium
refrigeration.] phosphate in 1000 mL of water, and adjust with
Sample solution: Remove the contents of NLT 20 Cefaclor phosphoric acid to a pH of 4.0.
Capsules as completely as possible, and weigh. Mix the Solution B: Acetonitrile and Solution A (9:11), degassing
combined contents, and transfer a portion of the powder, for NMT 2 min
nominally equivalent to 75 mg of cefaclor, to a 250-mL
volumetric flask, dilute with Mobile phase to volume, and
mix. Sonicate if necessary to ensure complete dissolution of
the cefaclor. Filter to obtain the clear Sample solution.
Mobile Phase: See Gradient Table. F = conversion factor (0.001 mg/g)

Acceptance criteria
Gradient Table Individual impurities: NMT 0.5% of any individual
cefaclor-related compound is found.
Solution A Solution B Total impurities: NMT 2.0% of all cefaclor-related
Time (%) (%) compounds, not including the contribution of any peak
0 95 5 that gives a result of less than 0.1%
30 75 25 SPECIFIC TESTS
45 0 100 WATER DETERMINATION, Method I 921: NMT 8.0%
55 0 100
60 95 5 PACKAGING AND STORAGE: Preserve in tight containers.
USP Cefaclor RS
Standard solution: 0.05 mg/mL of USP Cefaclor RS in USP Cefaclor, Delta-3 Isomer RS
Diluent
[NOTESonicate briefly, if necessary to dissolve, and avoid
heating. Use this solution on the day it is prepared.] Cefaclor for Oral Suspension
System suitability solution: 0.05 mg/mL of USP Cefaclor,
Delta-3 Isomer RS dissolved in the Standard solution (Comment on this Monograph)id=m13900=Cefaclor for Oral
Sample solution: Remove the contents of NLT 20 Cefaclor Suspension=Ca-Chl-Monos.pdf)
Capsules as completely as possible, and mix. Transfer a
portion of the combined contents, equivalent to about 50 DEFINITION
mg of cefaclor, to a 10-mL volumetric flask. Dissolve in Cefaclor for Oral Suspension is a dry mixture of Cefaclor and
Diluent, using brief sonication, if necessary, to achieve one or more suitable buffers, colors, diluents, and flavors. It
dissolution. Avoid heating. Dilute with Diluent to volume, contains the equivalent of NLT 90.0% and NMT 120.0% of
mix, and filter. Use within 3 h if stored at room the labeled amount of C15H14ClN3O4S.
temperature, or within 20 h when stored under IDENTIFICATION
refrigeration. The retention time of the major peak for cefaclor in the
Chromatographic system Sample solution corresponds to that of the Standard solution,
(See Chromatography 621, System Suitability.) as obtained in the Assay.
[NOTEReducing the acetonitrile content increases the
retention time of cefaclor and increases the resolution ASSAY
between cefaclor, delta-3 isomer and cefaclor.] PROCEDURE
Mode: LC Mobile phase: Dissolve 1 g of sodium 1-pentanesulfonate
Detector: UV 220 nm in a mixture of 780 mL of water and 10 mL of triethylamine.
Column: 4.6-mm 25-cm; packing L1 Adjust with phosphoric acid to a pH of 2.5 0.1, and add
Flow rate: 1 mL/min 220 mL of methanol.
Injection size: 20 L System suitability solution: 0.3 mg/mL of cefaclor and 0.3
System suitability mg/mL of USP Cefaclor, Delta-3 Isomer RS in Mobile phase
Samples: System suitability solution and Blank solution Standard solution: 0.3 mg/mL of USP Cefaclor RS in Mobile
[NOTEThe retention time for the cefaclor peak is 2329 phase
min.] [NOTESonicate briefly, if necessary, to achieve dissolution,
Suitability requirements and avoid heating the solution. Use within 8 h if stored at
Resolution: NLT 2.0 between cefaclor, delta-3 isomer room temperature, or within 20 h if stored under
and cefaclor, System suitability solution refrigeration.]
Tailing factor: NMT 1.2 for the cefaclor peak, System Sample stock solution: Constitute Cefaclor for Oral
suitability solution Suspension as directed in the labeling, freshly mixed and
[NOTEExamine the chromatogram of the Blank solution free from air bubbles.
for any extraneous peaks, and disregard any Sample solution: 0.3 mg/mL of cefaclor from Sample stock
corresponding peaks in the Sample solutions. Ensure solution diluted with Mobile phase. Sonicate if necessary to
that any extraneous peaks observed do not represent ensure complete dissolution of the cefaclor, and filter.
carryover from previous injections.] Chromatographic system
Samples: Standard solution and Sample solution Mode: LC
Calculate the percentage of each related compound in the Detector: UV 265 nm
portion of Cefaclor Capsules taken: Column: 4.6-mm 25-cm; 5-m packing L1
Result = (rU/rS) (CS/CU) P F 100 Injection size: 20 L
System suitability
rU = peak area of the Sample solution Sample: System suitability solution
rS = peak area of the Standard solution [NOTEThe relative retention times for cefaclor and
CS = concentration of USP Cefaclor RS in the cefaclor, delta-3 isomer are about 0.8 and 1.0,
Standard solution (mg/mL) respectively.]
CU = nominal concentration of cefaclor in the Sample Suitability requirements
solution (mg/mL) Resolution: NLT 2.5 between the cefaclor and cefaclor,
P = potency of USP Cefaclor RS (g/mg of delta-3 isomer peaks
C15H14ClN3O4S)
Tailing factor: NMT 1.5 Chromatographic system

Relative standard deviation: NMT 2.0% (See Chromatography 621, System Suitability.)
Analysis [NOTEReducing the acetonitrile content increases the
Samples: Standard solution and Sample solution retention time of cefaclor and increases the resolution
Calculate the potency, in mg, of C15H14ClN3O4S in the between cefaclor, delta-3 isomer and cefaclor.]
portion of the constituted Cefaclor for Oral Suspension Mode: LC
taken: Detector: UV 220 nm
Result = (rU/rS) (CS/CU) P F 100 Flow rate: 1 mL/min
rU = peak area of the cefaclor peak from the Sample System suitability
solution Samples: System suitability solution and Blank solution
rS = peak area of the cefaclor peak from the Standard [NOTEThe retention time for the cefaclor peak is 2329
solution min.]
CS = concentration of USP Cefaclor RS in the Standard Suitability requirements
solution (mg/mL) Resolution: NLT 2.0 between cefaclor, delta-3 isomer
CU = nominal concentration of cefaclor in the Sample and cefaclor, System suitability solution
solution (mg/mL) Tailing factor: NMT 1.2 for the cefaclor peak, System
P = potency of USP Cefaclor RS (mg) suitability solution
F = conversion factor, 0.001 mg/g [NOTEExamine the chromatogram of the Blank solution
Acceptance criteria: 90.0%120% for any extraneous peaks, and disregard any
corresponding peaks observed in the chromatogram of
PERFORMANCE TESTS the Sample solutions. Ensure that any extraneous peaks
DELIVERABLE VOLUME 698: Meets the requirements observed do not represent carryover from previous
UNIFORMITY OF DOSAGE UNITS 905 injections.]
For solid packaged in single-unit containers: Meets the Analysis
requirements Samples: Standard solution and Sample solution
IMPURITIES Calculate the percentage of each related compound in the
Organic Impurities portion of Cefaclor for Oral Suspension taken:
PROCEDURE
Diluent: 2.4 mg/mL of monobasic sodium phosphate in Result = (rU/rS) (CS/CU) P F 100
water, adjust with phosphoric acid to a pH of 2.5 rU = peak response area of an individual related
Blank solution: Use the Diluent. compound from the Sample solution
Solution A: Dissolve 6.9 g of monobasic sodium rS = peak response area from the Standard solution
phosphate in 1000 mL of water, and adjust with CS = concentration of USP Cefaclor RS in the
phosphoric acid to a pH of 4.0. Standard solution (mg/mL)
Solution B: Acetonitrile and Solution A (9:11), degassing CU = nominal concentration of cefaclor in the Sample
for NMT 2 min solution (mg/mL)
Mobile Phase: See Gradient Table. P = designated potency of USP Cefaclor RS (g/mg)
F = conversion factor, 0.001 mg/g
Gradient Table Acceptance criteria
Time Solution A Solution B Individual impurities: NMT 1.0% of any individual
(min) (%) (%) cefaclor-related compound is found.
Total impurities: NMT 3.0% of all cefaclor-related
0 95 5 compounds, not including the contribution of any peak
30 75 25 that gives a result of less than 0.1%.
45 0 100
SPECIFIC TESTS
55 0 100 PH 791: 2.55.0, in the suspension constituted as directed
60 95 5 in the labeling
70 95 5
Standard solution: 0.05 mg/mL of USP Cefaclor RS in PACKAGING AND STORAGE: Preserve in tight containers.
Diluent USP REFERENCE STANDARDS 11
[NOTESonicate briefly, if necessary, to dissolve, and USP Cefaclor RS
avoid heating. Use this solution on the day it is USP Cefaclor, Delta-3 Isomer RS
prepared.]
System suitability solution: 0.05 mg/mL of USP Cefaclor,
Delta-3 Isomer RS dissolved in Standard solution
Sample solution: Constitute Cefaclor for Oral Suspension Cefaclor Chewable Tablets
as directed in the labeling. Transfer a portion of Cefaclor (Comment on this Monograph)id=m662=Cefaclor Chewable
for Oral Suspension, freshly mixed and free from air Tablets=Ca-Chl-Monos.pdf)
bubbles, equivalent to 50 mg of cefaclor, to a 10-mL
volumetric flask. Dissolve in Diluent, using brief sonication, DEFINITION
if necessary, to achieve dissolution. Avoid heating. Dilute Cefaclor Chewable Tablets contain NLT 90.0% and NMT
with Diluent to volume, and filter. Use within 3 h if stored 110.0% of the labeled amount of cefaclor (C15H14ClN3O4 S).
at room temperature, or within 20 h when stored under
refrigeration.
IDENTIFICATION AS = absorbance from the Standard solution

The retention time of the Sample solution corresponds to that CS = concentration of USP Cefaclor RS in the Standard
of the Standard solution, as obtained in the Assay. solution (mg/mL)
CU = nominal concentration of cefaclor in the Sample
ASSAY solution (mg/mL)
PROCEDURE Tolerances: NLT 80% (Q) of the labeled amount of cefaclor
Mobile phase: Dissolve 1 g of sodium 1-pentanesulfonate UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
in a mixture of 780 mL of water and 10 mL of triethylamine.
Adjust with phosphoric acid to a pH of 2.5 0.1, and add IMPURITIES
220 mL of methanol. Organic Impurities
System suitability solution: 0.3 mg/mL of cefaclor and 0.3 PROCEDURE
mg/mL of USP Cefaclor, Delta-3 Isomer RS in Mobile phase Solvent: 2.4 mg/mL of monobasic sodium phosphate in
Standard solution: 0.3 mg/mL of USP Cefaclor RS in the water; adjust with phosphoric acid to a pH of 2.5
Mobile phase. [NOTESonicate briefly, if necessary, to Blank: Use the Solvent.
achieve dissolution, and avoid heating the solution. Use this Solution A: 6.9 mg/mL of monobasic sodium phosphate
Standard solution within 8 h if stored at room temperature, in water; Adjust with phosphoric acid to a pH of 4.0
or within 20 h if stored under refrigeration.] Solution B: Acetonitrile and Solution A (9:11), degassing
Sample solution: Transfer an equivalent to 75 mg of for NMT 2 min
cefaclor, from weighed and finely powdered Chewable Mobile Phase: See Gradient Table.
Tablets (NLT 20), to a 250-mL volumetric flask, and dilute
with Mobile phase to volume. Sonicate, if necessary, to Gradient Table
dissolve the cefaclor. Filter to obtain a clear solution.
Chromatographic system Time Solution A Solution B
(See Chromatography 621, System Suitability.) (min) (%) (%)
Mode: LC 0 95 5
Detector: UV 265 nm 30 75 25
Flow rate: 1.5 mL/min 45 0 100
Injection size: 20 L 55 0 100
System suitability 60 95 5
[NOTEThe relative retention time for cefaclor and cefaclor 70 95 5
delta-3 isomer are 0.8 and 1.0, respectively.]
Suitability requirements System suitability solution: 0.05 mg/mL of USP Cefaclor,
Resolution: NLT 2.5 between the cefaclor peak and the Delta-3 Isomer RS in the Standard solution
cefaclor, delta-3 isomer peak Standard solution: 0.05 mg/mL of USP Cefaclor RS in the
Tailing factor: NMT 1.5 Solvent
Relative standard deviation: NMT 2.0% [NOTESonicate briefly, if necessary, to dissolve, and
Analysis avoid heating. Use this solution on the day it is
Samples: Standard solution and Sample solution prepared.]
Calculate the percentage of cefaclor C15H14ClN3O4S in the Sample solution: Transfer an equivalent to 50 mg of
portion of Chewable Tablets taken: cefaclor, from weighed and finely powdered Chewable
Tablets (NLT 20), to a 10-mL volumetric flask. Dissolve in
Result = (rU/rS) (CS/CU) 100 Solvent, using brief sonication, if necessary, to dissolve.
Avoid heating. Dilute with Solvent to volume and filter.
rU = cefaclor peak area rom the Sample solution [NOTEUse this Sample solution within 3 h if stored at
rS = cefaclor peak area from the Standard solution room temperature, or within 20 h when stored under
CS = concentration of USP Cefaclor RS in the Standard refrigeration.]
solution (mg/mL) Chromatographic system
CU = nominal concentration of cefaclor in the Sample (See Chromatography 621, System Suitability.)
solution (mg/mL) Mode: LC
Acceptance criteria: 90.0%110.0% Detector: UV 220 nm
PERFORMANCE TESTS Flow rate: 1 mL/min
DISSOLUTION 711 Injection size: 20 L
Medium: Water; 900 mL System suitability
Apparatus 2: 50 rpm Sample: System suitability solution
Time: 30 min [NOTEThe retention time for cefaclor is 2329 min.]
Sample solution: Sample per Dissolution 711 Suitability requirements
Analysis: Determine the amount of cefaclor dissolved by Resolution: NLT 2.0 between cefaclor delta-3 isomer and
employing UV absorption at the wavelength of maximum cefaclor
absorption at about 264 nm on filtered portions of the Tailing factor: NMT 1.2
Sample solution, suitably diluted with Medium, if necessary, [NOTEExamine the chromatogram for any extraneous peaks,
in comparison with a Standard solution having a known and disregard any corresponding peaks observed in the
concentration of USP Cefaclor RS in the same Medium. chromatogram of the Sample solution. Ensure that any
Calculate the amount of cefaclor dissolved: extraneous peaks observed do not represent carryover from
previous injections.]
AU = absorbance from the Sample solution
Analysis volumetric flask, and dilute with Mobile phase to volume.

Samples: Standard solution and Sample solution Sonicate, if necessary, to dissolve the cefaclor.
Calculate the percentage of each cefaclor related Chromatographic system
compound in the portion of Cefaclor: (See Chromatography 621, System Suitability.)
Mode: LC
Result = (ri/rs) (CS/CU) 100 Detector: UV 265 nm
ri = peak response of an individual related Flow rate: 1.5 mL/min
compound in the chromatogram from the Injection size: 20 L
Sample solution System suitability
rS = peak response for the cefaclor peak in the Sample: System suitability solution
chromatogram of the Standard solution [NOTEThe relative retention times for cefaclor and
CS = concentration of USP Cefaclor RS in the Standard cefaclor, delta-3 isomer are 0.8 and 1.0, respectively.]
solution (mg/mL) Suitability requirements
CU = nominal concentration of the Sample solution Resolution: NLT 2.5 between the cefaclor peak and the
(mg/mL) cefaclor, delta-3 isomer peak
Acceptance criteria Tailing factor: NMT 1.5
Any individual related compound: NMT 1.0% Relative standard deviation: NMT 2.0%
Total impurities: NMT 3.0%, not including the Analysis
contribution of any peak that gives a result of less than Samples: Standard solution and Sample solution
0.1% Calculate the percentage of label claim of C15H14ClN3O4S in
[NOTEIn an acceptable determination, the difference the portion of Cefaclor Extended Release Tablets taken:
between duplicate determinations of total cefaclor related
compounds NMT 0.2% absolute, or the variation from the Result = (rU/rS) (CS/CU) P F 100
mean of the two values NMT 10%, whichever is greater.]
SPECIFIC TESTS rS = peak area from the Standard solution
WATER DETERMINATION, Method I 921: NMT 5.0% CS = concentration of USP Cefaclor RS in the Standard
solution (mg/mL)
ADDITIONAL REQUIREMENTS CU = nominal concentration of Cefaclor in the Sample
PACKAGING AND STORAGE: Preserve in tight containers. Store solution (mg/mL)
at 25, excursions permitted between 15 and 30. P = designated potency of USP Cefaclor RS [g of
LABELING: The product label and product labeling indicate cefaclor (C15H14ClN3O4S)/mg]
that the Chewable Tablets must be chewed or crushed F = conversion factor, 0.001 mg/g
before administration. Acceptance criteria: 90.0%110%
USP Cefaclor RS PERFORMANCE TESTS
USP Cefaclor Delta-3 Isomer RS DISSOLUTION 711
Apparatus 1 (10-mesh basket): 100 rpm
Cefaclor Extended-Release Tablets Time: 30, 60, and 240 min
Standard solution: 25 g/mL USP Cefaclor RS in Medium
(Comment on this Monograph)id=m13905=Cefaclor Extended- Sample solutions: Dilute filtered portions of the solution
Release Tablets=Ca-Chl-Monos.pdf) under test with Medium to obtain solutions having an
estimated concentration of 25 g/mL cefaclor.
DEFINITION Analysis: Determine the amount of cefaclor (C15H14ClN3O4S)
Cefaclor Extended-Release Tablets contain the equivalent of NLT dissolved in the Sample solution by employing UV absorption
90.0% and NMT 110.0% of the labeled amount of cefaclor at 265 nm, in comparison with the Standard solution.
(C15H14ClN3O4S). Tolerances: The percentages of the labeled amount of
IDENTIFICATION cefaclor (C15H14ClN3O4S) dissolved at the times specified
The retention time for cefaclor from the Sample solution conform to Acceptance Table 2.
the Assay. Time (min) Amount dissolved
30 Between 5% and 30%
ASSAY
PROCEDURE 60 Between 20% and 50%
Mobile phase: Dissolve 1 g of sodium 1-pentanesulfonate 240 NLT 80%
in a mixture of 780 mL of water and 10 mL of triethylamine.
Adjust with phosphoric acid to a pH of 2.5 0.1, add 220 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
mL of methanol.
System suitability solution: 0.3 mg/mL of cefaclor and 0.3 IMPURITIES
mg/mL of USP Cefaclor, Delta-3 Isomer RS in Mobile phase Organic Impurities
Standard solution: 0.3 mg/mL of USP Cefaclor RS in Mobile PROCEDURE
phase Diluent: 2.4 mg/mL of monobasic sodium phosphate in
[NOTESonicate briefly, if necessary, to achieve dissolution, water; adjusted with phosphoric acid to a pH of 2.5
and avoid heating the solution. Use within 8 h if stored at Blank solution: Use the Diluent.
room temperature, or within 20 h if stored under Solution A: Dissolve 6.9 g of monobasic sodium
refrigeration.] phosphate in 1000 mL of water, and adjust with
Sample solution: Weigh and finely powder NLT 20 Cefaclor phosphoric acid to a pH of 4.0.
Extended Release Tablets. Transfer a portion of the powder, Solution B: Acetonitrile and Solution A (9:11), degassing
nominally equivalent to 75 mg of cefaclor, to a 250-mL for NMT 2 min
Mobile Phase: See Gradient Table. Acceptance criteria

Individual impurities: NMT 0.6% of any individual
Gradient Table cefaclor-related compound is found.
Total impurities: NMT 2.0% of all cefaclor-related
Time Solution A Solution B compounds, not including the contribution of any peak
(min) (%) (%) that gives a result of less than 0.1%
0 95 5
SPECIFIC TESTS
30 75 25 WATER DETERMINATION, Method I 921: NMT 7.0%
45 0 100
55 0 100
60 95 5 containers, and store at controlled room temperature.
USP Cefaclor RS
Standard solution: 0.05 mg/mL of USP Cefaclor RS in USP Cefaclor, Delta-3 Isomer RS
Diluent
[NOTESonicate briefly, if necessary, to dissolve, and
avoid heating. Use this solution on the day it is Cefadroxil
prepared.]
System suitability solution: 0.05 mg/mL of USP Cefaclor, (Comment on this Monograph)id=m13930=Cefadroxil=Ca-Chl-
Delta-3 Isomer RS dissolved in Standard solution Monos.pdf)
Sample solution: Weigh and finely powder NLT 20
Tablets. Transfer a portion of the powder, nominally
equivalent to 50 mg of cefaclor, to a 10-mL volumetric
flask. Dissolve in Diluent, using brief sonication, if necessary,
to achieve dissolution. Avoid heating. Dilute with Diluent to
volume, mix, and filter.
[NOTEUse within 3 h if stored at room temperature, or
within 20 h if stored under refrigeration.]
Chromatographic system C16H17N3O5S H2O 381.40
(See Chromatography 621, System Suitability.) 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[
[NOTEReducing the acetonitrile content increases the [amino(4-hydroxyphenyl)acetyl]amino]-3-methyl-8-oxo-,
retention time of cefaclor and increases the resolution monohydrate, [6R-[6,7 (R*)]]-;
between cefaclor, delta-3 isomer and cefaclor.] (6R,7R)-7-[(R)-2-Amino-2-(p-hydroxyphenyl)acetamido]-3-
Mode: LC methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic
Detector: UV 220 nm acid monohydrate [66592-87-8];
Column: 4.6-mm 25-cm; packing L1 Hemihydrate 372.39
Flow rate: 1 mL/min [119922-85-9];
Injection size: 20 L Anhydrous 363.40
System suitability [50370-12-2].
Samples: Blank solution and System suitability solution DEFINITION
[NOTEThe retention time for the cefaclor peak is 2329 Cefadroxil has a potency equivalent to NLT 950 g and NMT
min.] 1050 g of C16H17N3O5S/mg, calculated on the anhydrous
Suitability requirements basis.
Resolution: NLT 2.0 between cefaclor, delta-3 isomer
and cefaclor, System suitability solution IDENTIFICATION
Tailing factor: NMT 1.2 for the cefaclor peak, System A. INFRARED ABSORPTION 197K
suitability solution B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
[NOTEExamine the chromatogram of the Blank solution Standard solution: 2 mg/mL of USP Cefadroxil RS
for any extraneous peaks, and disregard any Sample solution: 2 mg/mL of Cefadroxil
corresponding peaks observed in the chromatogram of Chromatographic system
the Sample solutions. Ensure that any extraneous peaks (See Chromatography 621, Thin-Layer Chromatography.)
observed do not represent carryover from previous Adsorbent: 0.25-mm layer of binder-free silica gel
injections.] Application volume: 20 L
Analysis Pre-developing solvent solution: n-hexane and
Samples: Standard solution and Sample solution tetradecane (95:5)
Calculate the percentage of each related compound in the Developing solvent system: 0.1M citric acid, 0.1M
portion of Cefaclor Extended Release Tablets taken: dibasic sodium phosphate and a 1 in 15 solution of
ninhydrin in acetone (60:40:15)
Result = (rU/rS) (CS/CU) P F 100 Spray reagent: 1in500 solution of ninhydrin in
dehydrated alcohol
rU = peak area from the Sample solution [NOTEProtect Spray reagent from light.]
rS = peak area from the Standard solution Analysis
CS = concentration of USP Cefaclor RS in the Samples: Standard solution and Sample solution
Standard solution (mg/mL) Place the thin-layer chromatographic plate in a chamber
CU = nominal concentration of cefaclor in the Sample containing the Pre-developing solvent solution and allow
solution (mg/mL) the solvent front to move the length of the plate. Remove
P = potency of USP Cefaclor RS [g of cefaclor the plate from the chamber and allow the solvent to
(C15H14ClN3O4S)/mg] evaporate. Apply the Sample solution and Standard solution
F = conversion factor, 0.001 mg/g
USP 32 Official Monographs / Cefadroxil 113
to the plate, allow the spots to dry, and develop the Chromatographic system
chromatogram in the Developing solvent system until the (See Chromatography 621, Thin-Layer Chromatography.)
solvent front has moved three-fourths of the length of the Adsorbent: 0.25-mm layer of chromatographic silica gel
plate. Remove the plate from the developing chamber, mixture
mark the solvent front, and allow to air-dry. Spray the Application volume: 2-L portions of the Sample solution,
plate with Spray reagent, dry for 10 min at 110, and Standard solution A, Standard solution B, and Standard
examine the chromatogram. solution C, and a 4-L portion of the Identification solution
Acceptance criteria: The RF value of the principal spot of Developing solvent system: Ethyl acetate, alcohol,
the Sample solution corresponds to that of the Standard formic acid, and water (14:5:1:5)
solution. Analysis
Samples: Sample solution, Standard solution A, Standard
ASSAY solution B, Standard solution C, and Identification solution
PROCEDURE Proceed as directed for Chromatography 621, Thin-Layer
Buffer solution: Dissolve 13.6 g of monobasic potassium Chromatography. Develop the chromatograms until the
phosphate in water to make 2000 mL of solution. Adjust solvent front has moved three-fourths of the length of
with 10 N potassium hydroxide to a pH of 5.0. the plate. Remove the plate from the developing
Mobile phase: Acetonitrile and Buffer solution (1:24) chamber, mark the solvent front, allow the plate to dry,
Standard solution: 1.06 mg/mL of USP Cefadroxil RS in and examine the chromatograms under short-
Buffer solution wavelength UV light.
[NOTEThis solution contains the equivalent of 1000 g/ [NOTEIn a valid test the chromatogram obtained from
mL of C16H17N3O5S. Use this solution on the day the Identification solution shows three clearly separated
prepared.] spots.]
Sample solution: 1.06 mg/mL of Cefadroxil in Buffer Acceptance criteria: Any secondary spot in the
solution chromatogram of the Sample solution corresponding to 7-
[NOTEStir by mechanical means for 5 min until dissolved. aminodesacetoxycephalosporanic acid or D--4-
Use this solution on the day prepared.] hydroxyphenylglycine is not more intense than the
Chromatographic system corresponding spot in the chromatogram of Standard
(See Chromatography 621, System Suitability.) solution B (1.0%); and any spot, other than the principal
Mode: LC spot and any spot corresponding to 7-
Detector: UV 230 nm aminodesacetoxycephalosporanic acid or D--4-
Column: 4-mm 25-cm; packing L1 hydroxyphenylglycine, is not more intense than the
Flow rate: 1.5 mL/min principal spot in the chromatogram of Standard solution A
Injection size: 10 L (1.0%).
System suitability PROCEDURE 2: DIMETHYLANILINE 223: Meets the requirement
Suitability requirements SPECIFIC TESTS
Capacity factor, k: Between 2.0 and 3.5 OPTICAL ROTATION, Specific Rotation 781S: +165.0 to
Column efficiency: NLT 1800 theoretical plates +178.0
Tailing factor: NMT 2.2 Sample solution: 10 mg/mL
Relative standard deviation: NMT 2.0% CRYSTALLINITY 695: Meets the requirements
Analysis PH 791: 4.06.0, in a suspension containing 50 mg/mL
Samples: Standard solution and Sample solution WATER DETERMINATION, Method I 921: 4.2%6.0%, except
Calculate the quantity, in g, of C16H17N3O5S in each mg of that where it is labeled as being in the hemihydrate form it
Cefadroxil taken: is between 2.4% and 4.5%
Result = (rU/rS) (CS/CU) P ADDITIONAL REQUIREMENTS
rU = peak response of the Sample solution LABELING: The hemihydrate form is so labeled.
rS = peak response of the Standard solution USP REFERENCE STANDARDS 11
CS = concentration of USP Cefadroxil RS in the USP Cefadroxil RS
CU = concentration of Cefadroxil taken to prepare the
P = cefadroxil equivalent (g/mg) of USP Cefadroxil Cefadroxil Capsules
RS (Comment on this Monograph)id=m13940=Cefadroxil
Acceptance criteria: 9501050 g/mg Capsules=Ca-Chl-Monos.pdf)
IMPURITIES DEFINITION
Organic Impurities Cefadroxil Capsules contain the equivalent of NLT 90.0% and
PROCEDURE 1 NMT 120.0% of the labeled amount of cefadroxil
Diluent: Alcohol, 2.4 N hydrochloric acid, and water (C16H17N3O5S).
(75:3:22)
Standard solution A: Dilute 1.0 mL of the Sample solution IDENTIFICATION
to 100 mL with Diluent. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
Standard solution B: 0.25 mg/mL each of 7- Standard solution: 2 mg/mL of USP Cefadroxil RS
aminodesacetoxycephalosporanic acid and D--4- Sample solution: 2 mg/mL of cefadroxil, from the contents
hydroxyphenylglycine in Diluent of 1 Capsule dissolved in water
Standard solution C: 0.25 mg/mL of D--4- Chromatographic system
hydroxyphenylglycine in Solution A (See Chromatography 621, System Suitability.)
Sample solution: 25 mg/mL of Cefadroxil in Diluent
Identification solution: Standard solution B and Sample
solution (1:1)
114 Cefadroxil / Official Monographs USP 32
Adsorbent: 0.25-mm layer of binder-free silica gel CU = nominal concentration of cefadroxil in the
Application volume: 20 L Sample solution (mg/mL)
Pre-developing solvent solution: n-hexane and P = cefadroxil equivalent of USP Cefadroxil RS
tetradecane (95:5) (g/mg)
Developing solvent system: 0.1 M citric acid, 0.1 M F = conversion factor, 0.001 mg/g
dibasic sodium phosphate, and a 1in15 solution of Acceptance criteria: 90.0%120.0%
ninhydrin in acetone (60:40:15)
Spray reagent: 1in500 solution of ninhydrin in PERFORMANCE TESTS
dehydrated alcohol DISSOLUTION 711
[NOTEProtect Spray reagent from light.] Medium: Water; 900 mL
Analysis Apparatus 1: 100 rpm
Samples: Standard solution and Sample solution Time: 30 min
Place the thin-layer chromatographic plate in a chamber Standard solution: USP Cefadroxil RS in Medium of a
containing the Pre-developing solvent solution and allow known concentration
the solvent front to move the length of the plate. Remove Sample solution: Sample per Dissolution 711. Suitably
the plate from the chamber and allow the solvent to dilute with Medium, if necessary, and filter.
evaporate. Apply the Sample solution and Standard solution Analysis: Determine the amount of C16H17N3O5S dissolved
to the plate, allow the spots to dry, and develop the from UV absorbances at 263 nm of the Sample solution in
chromatogram in the Developing solvent system until the comparison to the Standard solution.
solvent front has moved three-fourths of the length of the Tolerances: NLT 80% (Q) of the labeled amount of
plate. Remove the plate from the developing chamber, C16H17N3O5S is dissolved.
mark the solvent front, and allow to air-dry. Spray the UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
plate with the Spray reagent, dry for 10 min at 110, and
examine the chromatogram. SPECIFIC TESTS
Acceptance criteria: The RF value of the principal spot of WATER DETERMINATION, Method I 921: NMT 7.0%
the Sample solution corresponds to that of the Standard ADDITIONAL REQUIREMENTS
solution. PACKAGING AND STORAGE: Preserve in tight containers.
ASSAY LABELING: Capsules prepared using the hemihydrate form of
PROCEDURE Cefadroxil are so labeled.
Buffer solution: Dissolve 13.6 g of monobasic potassium USP REFERENCE STANDARDS 11
phosphate in water to make 2000 mL of solution. Adjust USP Cefadroxil RS
with 10 N potassium hydroxide to a pH of 5.0.
Mobile phase: Acetonitrile and Buffer solution (1:24)
Standard solution: 1.06 mg/mL of USP Cefadroxil RS in Cefadroxil for Oral Suspension
Buffer solution
[NOTEThis solution contains the equivalent of 1000 (Comment on this Monograph)id=m13950=Cefadroxil for Oral
g/mL of C16H17N3O5S. Use this solution on the day Suspension=Ca-Chl-Monos.pdf)
prepared.] DEFINITION
Sample solution: Remove, as completely as possible, the Cefadroxil for Oral Suspension is a dry mixture of Cefadroxil
contents of NLT 10 Capsules, and weigh. Transfer a portion and one or more suitable buffers, colors, diluents, and flavors.
of the powder, nominally equivalent to 200 mg of It contains the equivalent of NLT 90.0% and NMT 120.0% of
cefadroxil, to a 200-mL volumetric flask. Dilute with Buffer the labeled amount of C16H17N3O5S.
solution to volume, and stir by mechanical means for 5 min.
[NOTEUse this solution on the day prepared.] IDENTIFICATION
Chromatographic system THIN-LAYER CHROMATOGRAPHY
(See Chromatography 621, System Suitability.) Standard solution: 2 mg/mL of USP Cefadroxil RS
Mode: LC Sample solution: Constitute 1 container of Cefadroxil for
Detector: UV 230 nm Oral Suspension as directed in the labeling. Dilute a portion
Column: 4-mm 25-cm; packing L1 of the resulting suspension to a concentration of 2 mg/mL.
Flow rate: 1.5 mL/min Chromatographic system
Injection size: 10 L (See Chromatography 621, Thin-Layer Chromatography.)
System suitability Mode: TLC
Sample: Standard solution Adsorbent: 0.25-mm layer of binder-free silica gel
Suitability requirements Application volume: 20 L
Capacity factor, k: 2.03.5 Pre-developing solvent solution: n-hexane and
Column efficiency: NLT 1800 theoretical plates tetradecane (95:5)
Tailing factor: NMT 2.2 Developing solvent system: 0.1M citric acid, 0.1M
Relative standard deviation: NMT 2.0% dibasic sodium phosphate and a 1 in 15 solution of
Analysis ninhydrin in acetone (60:40:15)
Samples: Standard solution and Sample solution Spray reagent: 1in500 solution of ninhydrin in
Calculate the percentage of C16H17N3O5S in the portion of dehydrated alcohol
Capsules taken: [NOTEProtect Spray reagent from light.]
Analysis
Result = (rU/rS) (CS/CU) P F 100 Samples: Standard solution and Sample solution
Place the thin-layer chromatographic plate in a chamber
rU = peak response of cefadroxil from the Sample containing the Pre-Developing solvent solution and allow
solution the solvent front to move the length of the plate. Remove
rS = peak response of cefadroxil from the Standard the plate from the chamber and allow the solvent to
solution evaporate. Apply the Sample solution and Standard solution
CS = concentration of USP Cefadroxil RS in the to the plate, allow the spots to dry, and develop the
USP 32 Official Monographs / Cefadroxil 115
chromatogram in the Developing solvent system until the Analysis Determine the amount of cefadroxil dissolved by
solvent front has moved three-fourths of the length of the employing UV absorption at the wavelength of 263 nm on
plate. Remove the plate from the developing chamber, the Sample solution in comparison with the Standard solution
mark the solvent front, and allow to air-dry. Spray the Calculate the amount of cefadroxil dissolved:
plate with the Spray reagent, dry for 10 min at 110, and
examine the chromatogram. Result = (AU/AS) (Cs/W) (V/D) 100
the Sample solution corresponds to that of the Standard AU = absorbance of the Sample solution
solution. AS = absorbance of the Standard solution
CS = concentration of Standard solution (mg/mL)
ASSAY W = weight of Sample (mg)
PROCEDURE V = volume of Medium (mL), 900
Buffer solution: Dissolve 13.6 g of monobasic potassium D = dilution factor
phosphate in water to make 2000 mL of solution. Adjust Tolerances: NLT 75% (Q) of the labeled amount of
with 10 N potassium hydroxide to a pH of 5.0 cefadroxil is dissolved.
Mobile phase: Acetonitrile and Buffer solution (1:24) UNIFORMITY OF DOSAGE UNITS 905: For solid packaged in
Standard solution: 1.06 mg/mL of USP Cefadroxil RS in single-unit containers: Meets the requirements
Buffer solution DELIVERABLE VOLUME 698: Meets the requirements
[NOTEThis solution contains the equivalent of 1000 g/
mL of cefadroxil (C16H17N3O5S). Use this solution on the SPECIFIC TESTS
day prepared.] PH 791: 4.56.0, in the suspension constituted as directed
Sample solution: Constitute a container of Cefadroxil for in the labeling
Oral Suspension as directed in the labeling. Dilute a portion WATER DETERMINATION, Method I 921: NMT 2.0%, except
of the resulting suspension with Buffer solution to prepare a where it is labeled as containing 100 mg of cefadroxil/mL
solution containing nominally 1.0 mg/mL. Pass through a after constitution, in which case the limit is NMT 3.0%
suitable filter of 0.8m or finer porosity, and use the
filtrate. Use this solution on the day prepared. ADDITIONAL REQUIREMENTS
(See Chromatography 621, System suitability.) USP REFERENCE STANDARDS 11
Mode: LC USP Cefadroxil RS
Detector: UV 230 nm
Flow rate: 1.5 mL/min Cefadroxil Tablets
System suitability (Comment on this Monograph)id=m13955=Cefadroxil
Sample: Standard solution Tablets=Ca-Chl-Monos.pdf)
Capacity factor, k : Between 2.0 and 3.5 Cefadroxil Tablets contain NLT 90.0% and NMT 120.0% of the
Column efficiency: NLT 1800 theoretical plates labeled amount of C16H17N3O5S.
Relative standard deviation: NMT 2.0% IDENTIFICATION
Analysis THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST
Samples: Standard solution and Sample solution Standard solution: 2 mg/mL of USP Cefadroxil RS
Calculate the percentage of C16H17N3O5S of label claim in Sample solution: 2 mg/mL of cefadroxil from the
Cefadroxil for Oral Suspension: powdered Tablets dissolved in water
Result = (rU/rS) (CS/CU) P F 100 (See Chromatography 621, System Suitability.)
Adsorbent: 0.25-mm layer of binder-free silica gel
rU = peak response of the cefadroxil of the Sample Application volume: 20 L
solution Pre-developing solvent solution: n-hexane and
rS = peak response of the cefadroxil of the Standard tetradecane (95:5)
solution Developing solvent system: 0.1 M citric acid, 0.1 M
CS = concentration of USP Cefadroxil RS in the dibasic sodium phosphate and a 1 in 15 solution of
Standard solution (mg/mL) ninhydrin in acetone (60:40:15)
CU = nominal concentration of cefadroxil in the Spray reagent: 1in500 solution of ninhydrin in
Sample solution (mg/mL) dehydrated alcohol
P = cefadroxil equivalent (g/mg) of USP Cefadroxil [NOTEProtect Spray reagent from light.]
RS Analysis
F = conversion factor, 0.001 mg/g Samples: Standard solution and Sample solution
Acceptance criteria: 90.0%120% Place the thin-layer chromatographic plate in a chamber
PERFORMANCE TESTS containing the Pre-developing solvent solution and allow
DISSOLUTION 711 the solvent front to move the length of the plate. Remove
Medium: Water; 900 mL the plate from the chamber and allow the solvent to
Apparatus 2: 25 rpm evaporate. Apply the Sample solution and Standard solution
Time: 30 min to the plate, allow the spots to dry, and develop the
Standard solution: USP Cefadroxil RS in Medium at a chromatogram in the Developing solvent system until the
known concentration solvent front has moved three-fourths of the length of the
Sample solution: Transfer 5.0 mL of the constitued Oral plate. Remove the plate from the developing chamber,
Suspension (weighed) to the dissolution vessel. mark the solvent front, and allow to air-dry. Spray the
116 Cefadroxil / Official Monographs USP 32
plate with the Spray reagent, dry for 10 min at 110, and UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
examine the chromatogram.
Acceptance criteria: The RF value of the principal spot of SPECIFIC TESTS
the Sample solution corresponds to that of the Standard WATER DETERMINATION, Method I 921: NMT 8.0%
solution.
ASSAY PACKAGING AND STORAGE: Preserve in tight containers.
PROCEDURE LABELING: The Tablets prepared using the hemihydrate form
Buffer solution: Dissolve 13.6 g of monobasic potassium of cefadroxil are so labeled.
phosphate in water to make 2000 mL of solution. Adjust USP REFERENCE STANDARDS 11
with 10 N potassium hydroxide to a pH of 5.0. USP Cefadroxil RS
Mobile phase: Acetonitrile and Buffer solution (1:24)
Standard solution: 1.06 mg/mL of USP Cefadroxil RS in
Buffer solution Cefamandole Nafate
[NOTEThis solution contains the equivalent of 1000 g/
mL of cefadroxil (C16H17N3O5S). Use this solution on the (Comment on this Monograph)id=m13957=Cefamandole
day prepared.] Nafate=Ca-Chl-Monos.pdf)
Transfer a portion of the powder, equivalent to 200 mg of
cefadroxil, to a 200-mL volumetric flask, dilute with Buffer
solution to volume, and stir by mechanical means for 5 min.
Use this solution on the day prepared.
(See Chromatography 621, System suitability.)
Mode: LC
Detector: UV 230 nm C19H17N6NaO6S2 512.50
Column: 4-mm 25-cm; packing L1 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[
Flow rate: 1.5 mL/min [(formyloxy)phenylacetyl]amino]-3-[[(1-methyl-1H-tetrazol-5-
Injection size: 10 L yl)thio]methyl]-8-oxo-, monosodium salt, [6R-[6,7 (R*)]]-;
System suitability Sodium (6R,7R)-7-(R)-mandelamido-3-[[(1-methyl-1H-tetrazol-5-
Sample: Standard solution yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-
Suitability requirements carboxylate formate (ester) [42540-40-9].
Capacity factor, k : Between 2.03.5 DEFINITION
Column efficiency: NLT 1800 theoretical plates Cefamandole Nafate has a potency equivalent to NLT 810 g
Tailing factor: NMT 2.2 and NMT 1000 g of cefamandole (C18H18N6O5S2)/mg,
Relative standard deviation: NMT 2.0% calculated on the anhydrous basis.
Analysis
Calculate the percentage of C16H17N3O5S in the portion of THIN-LAYER CHROMATOGRAPHY
Tablets taken: Standard solution: 10 mg/mL of USP Cefamandole Nafate
RS in Developing solvent system
Result = (rU/rS) (CS/CU) P F 100 [NOTEUse the solution promptly after preparation.]
Sample solution: 10 mg/mL of Cefamandole Nafate in
rU = peak response of the Sample solution Developing solvent system
rS = peak response of the Standard solution [NOTEUse the solution promptly after preparation.]
CS = concentration of USP Cefadroxil RS in the Chromatographic system
Standard solution (mg/mL) (See Chromatography 621, Thin-Layer Chromatography.)
CU = nominal concentration of cefadroxil in the Mode: TLC
Sample solution (mg/mL) Adsorbent: 0.25-mm layer of chromatographic silica gel
P = cefadroxil equivalent (g/mg) of USP Cefadroxil mixture
RS Application volume: 10 L
F = conversion factor, 0.001 mg/g Developing solvent system: Ethyl acetate, acetone, glacial
Acceptance criteria: 90.0%120% acetic acid, and water (5:2:1:1)
PERFORMANCE TESTS Analysis
DISSOLUTION 711 Samples: Standard solution and Sample solution
Medium: Water; 900 mL Place the plate in a suitable chromatographic chamber,
Apparatus 2: 50 rpm previously equilibrated with Developing solvent system for
Time: 30 min NLT 30 min, and develop the chromatogram until the
Sample solution: Sample per 711 Dissolution solvent front has moved three-fourths of the length of the
Analysis: Determine the amount of C16H17N3O5S dissolved plate. Remove the plate from the developing chamber,
from UV absorbances at the wavelength of maximum mark the solvent front, and allow to air-dry. Locate the
absorbance at about 263 nm of filtered portions of the spots on the plate by examination under short-wavelength
Sample solution suitably diluted, if necessary, in comparison UV light.
to a Standard solution having a known concentration of USP Acceptance criteria: The RF value of the principal spot of
Cefadroxil RS. the Sample solution corresponds to that of the Standard
Tolerances: NLT 75% (Q) of the labeled amount of solution
C16H17N3O5S
USP 32 Official Monographs / Cefamandole 117
ASSAY Cefamandole Nafate for Injection

PROCEDURE (Comment on this Monograph)id=m13960=Cefamandole
Buffer solution: 3.6 mg/mL of anhydrous dibasic sodium Nafate for Injection=Ca-Chl-Monos.pdf)
phosphate, 39.4 mg/mL of citric acid monohydrate, and
70.8 mg/mL of potassium chloride DEFINITION
Standard solution: Transfer 12 mg of USP Cefamandole Cefamandole Nafate for Injection is a sterile mixture of
Nafate RS to a 50-mL volumetric flask containing 4 mL of Cefamandole Nafate and one or more suitable buffers. It has a
water. Immediately before use, add 30.0 mL of Buffer potency equivalent to NLT 810 g and NMT 1000 g of
solution, and dilute with water to volume. cefamandole (C18H18N6O5S2)/mg, calculated on the anhydrous
Sample solution: Transfer 12 mg of Cefamandole Nafate to and sodium carbonate-free basis. It contains the equivalent of
a 50-mL volumetric flask containing 4 mL of water. NLT 90.0% and NMT 115.0% of the labeled amount of
Immediately before use, add 30.0 mL of Buffer solution, and cefamandole (C18H18N6O5S2).
Analysis: (See Polarography 801.) IDENTIFICATION
Samples: Standard solution and Sample solution THIN-LAYER CHROMATOGRAPHY
Transfer a portion of the Sample solution to a suitable Standard solution: 10 mg/mL of USP Cefamandole Nafate
polarographic cell. Deaerate by bubbling scrubbed RS in Developing solvent system
nitrogen through the solution for 5 min, and redirect the [NOTEUse the solution promptly after preparation.]
nitrogen flow to the surface outlet. Insert the dropping Sample solution: 10 mg/mL of cefamandole nafate, from
mercury electrode of a suitable polarograph capable of Cefamandole Nafate for Injection diluted with Developing
measuring a current of 0.5 microampere or appropriate solvent system.
current to maintain on-scale response, using an average [NOTEUse the solution promptly after preparation.]
capillary, and a drop rate of 1/s. Record the polarogram in Chromatographic system
the differential pulse mode from 0.3 volt to 1.05 volts, (See Chromatography 621, System Suitability.)
using a saturated calomel reference electrode and Mode: TLC
platinum wire counter electrode. Determine the peak Adsorbent: 0.25-mm layer of chromatographic silica gel
height obtained, in microamperes, where the peak height mixture
is defined as the perpendicular distance from the Application volume: 10 L
extrapolated baseline to the highest point of the peak as Developing solvent system: Ethyl acetate, acetone, glacial
compared to the full-scale current range. Similarly, acetic acid, and water (5:2:1:1)
determine the peak current of the Standard solution. Analysis
Calculate the concentration, in g/mg, of C18H18N6O5S2 of Samples: Standard solution and Sample solution
the Cefamandole Nafate: Analysis: Place the plate in a suitable chromatographic
chamber, previously equilibrated with Developing solvent for
Result = (iU/iS) (CS/CU) P NLT 30 min, and develop the chromatogram until the
solvent front has moved three-fourths of the length of the
iU = peak current, in microamperes, from the Sample plate.
solution Acceptance criteria: The RF value of the principal spot of
iS = peak current, in microamperes, from the the Sample solution corresponds to that of the Standard
Standard solution solution.
CS = concentration of USP Cefamandole Nafate RS in
the Standard solution (mg/mL) ASSAY
CU = concentration of Cefamandole Nafate taken to PROCEDURE
prepare the Sample solution (mg/mL) Buffer solution: 3.6 mg/mL of anhydrous dibasic sodium
P = potency of cefamandole of USP Cefamandole phosphate, 39.4 mg/mL of citric acid monohydrate, and
Nafate RS (g/mg) 70.8 mg/mL of potassium chloride
Acceptance criteria: 810 g/mg1000 g/mg Standard solution: Transfer 12 mg of USP Cefamandole
Nafate RS to a 50-mL volumetric flask containing 4 mL of
SPECIFIC TESTS water. Immediately before use, add 30.0 mL of Buffer
PH 791: 3.57.0, in a solution containing 100 mg/mL solution, and dilute with water to volume.
WATER DETERMINATION, Method I 921: NMT 2.0% Sample stock solution A (where the article is represented as
BACTERIAL ENDOTOXINS TEST 85: NMT 0.15 USP Endotoxin being in a single-dose container): Constitute a container of
Unit/mg of cefamandole, when the label states that Cefamondole Nafate for Injection in a volume of water
Cefamandole Nafate is sterile or must be subjected to further corresponding to the volume of solvent specified in the
processing during the preparation of injectable dosage forms labeling. Withdraw all of the withdrawable contents, using a
STERILITY TESTS 71: Meets the requirements for Test for suitable hypodermic needle and syringe, and dilute with
Sterility of the Product to be Examined, Membrane Filtration water to obtain a solution containing 2 mg/mL of
when the label states that Cefamandole Nafate is sterile cefamandole.
Sample solution A: Transfer 5.0 mL of Sample stock solution
ADDITIONAL REQUIREMENTS A to a 50-mL volumetric flask, add 30.0 mL of Buffer
PACKAGING AND STORAGE: Preserve in tight containers. solution, and dilute with water to volume.
LABELING: Where it is intended for use in preparing Sample stock solution B (where the label states the quantity
injectable dosage forms, the label states that it is sterile or of cefamandole in a given volume of constituted solution):
must be subjected to further processing during the Constitute Cefamandole Nafate for Injection in a volume of
preparation of injectable dosage forms. water corresponding to the volume of solvent specified in
USP REFERENCE STANDARDS 11 the labeling. Dilute a volume of the constituted solution
USP Cefamandole Nafate RS
USP Endotoxin RS
118 Cefamandole / Official Monographs USP 32
with water to obtain a solution containing nominally 2 PERFORMANCE TESTS

mg/mL of cefamandole. UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Sample solution B: Transfer 5.0 mL of Sample stock solution Analysis for content uniformity: Perform the Assay on
B to a 50-mL volumetric flask, add 30.0 mL of Buffer individual containers using Sample solution A or Sample
solution, and dilute with water to volume. solution B, or both, as appropriate.
Sample solution C: Transfer a quantity of Cefamondole
Nafate for Injection, equivalent to 12 mg of Cefamandole SPECIFIC TESTS
Nafate, to a 50-mL volumetric flask containing 4 mL of CONSTITUTED SOLUTION: At the time of use, it meets the
water. Immediately before use, add 30.0 mL of Buffer requirements for Constituted Solutions, Injections 1.
solution, and dilute with water to volume. Determine the BACTERIAL ENDOTOXINS TEST 85: NMT 0.15 USP Endotoxin
sodium carbonate content of a separate 1g portion of Unit/mg of cefamandole
Cefamandole for Injection dissolved in 100 mL of water. Add STERILITY TESTS 71: It meets the requirements for Test for
methyl orange TS and titrate with 0.2 N sulfuric acid VS. Sterility of the Product to be Examined, Membrane Filtration.
Each mL of 0.2 N sulfuric acid is equivalent to 10.60 mg of PH 791: 6.08.0, determined after 30 min in a solution
Na2CO3. containing 100 mg/mL
Analysis PARTICULATE MATTER 788: Meets the requirements for
(See Polarography 801.) small-volume injections
Samples: Standard solution and Sample solutions WATER DETERMINATION, Method I 921: NMT 3.0%
Transfer a portion of the Sample solution to a suitable OTHER REQUIREMENTS: It meets the requirements under
polarographic cell. Deaerate by bubbling scrubbed Injections 1.
nitrogen through the solution for 5 min, and redirect the
nitrogen flow to the surface outlet. Insert the dropping ADDITIONAL REQUIREMENTS
mercury electrode of a suitable polarograph capable of PACKAGING AND STORAGE: Preserve in Containers for Sterile
measuring a current of 0.5 microampere or appropriate Solids as described under Injections 1.
current to maintain on-scale response, using an average USP REFERENCE STANDARDS 11
capillary, and a drop rate of 1/s. Record the polarogram in USP Cefamandole Nafate RS
the differential pulse mode from 0.3 volt to 1.05 volts, USP Endotoxin RS
using a saturated calomel reference electrode and
platinum wire counter electrode. Determine the peak
height obtained, in microamperes, where the peak height Cefazolin
is defined as the perpendicular distance from the
extrapolated baseline to the highest point of the peak as (Comment on this Monograph)id=m13970=Cefazolin=Ca-Chl-
compared to the full-scale current range. Similarly, Monos.pdf)
determine the peak current of the Standard solution.
constituted solution taken:
Result = (iU/iS) (CS/CU) P F 100
iU = peak current, in microamperes, from the Sample
solution
iS = peak current, in microamperes, from the C14H14N8O4S3 454.51
Standard solution 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 3-[[(5-
CS = concentration of USP Cefamandole Nafate RS in methyl-1,3,4-thiadiazol-2-yl)thio]methyl]-8-oxo-7-[[1H-
the Standard solution (mg/mL) tetrazol-1-yl)acetyl]amino]-, (6R-trans);
CU = nominal concentration of cefamandole in Sample (6R,7R)-3-[[(5-Methyl-1,3,4-thiadiazol-2-yl)thio]methyl]-8-oxo-7-
solution A or in Sample solution B (mg/mL) [2-(1H-tetrazol-1-yl)acetamido]-5-thia-1-azabicyclo
P = potency of cefamandole of USP Cefamandole [4.2.0]oct-2-ene-2-carboxylic acid [25953-19-9].
Nafate RS (g/mg) DEFINITION
F = conversion factor, 0.001 mg/g Cefazolin contains NLT 95.0% and NMT 103.0% of
Calculate the potency in g of C18H18N6O5S2/mg, of the C14H14N8O4S3, calculated on the anhydrous basis.
Cefamandole Nafate for Injection taken:
IDENTIFICATION
Result = (iU/iS) (CS/CU) P The retention time of the Sample solution corresponds to that
of the Standard solution, as obtained in the Assay.
iU = peak current, in microamperes, from the Sample
solution ASSAY
iS = peak current, in microamperes, from the PROCEDURE
Standard solution Buffer A: 0.9 mg/mL of anhydrous dibasic sodium
CS = concentration of USP Cefamandole Nafate RS in phosphate and 1.298 mg/mL of citric acid monohydrate
the Standard solution (mg/mL) Buffer B: 5.68 mg/mL of anhydrous dibasic sodium
CU = nominal concentration of cefamandole nafate in phosphate and 3.63 mg/mL of monobasic potassium
Sample solution C (mg/mL) phosphate
P = potency of cefamandole of USP Cefamandole Mobile phase: Acetonitrile and Buffer A (1:9)
Nafate RS (g/mg) Internal standard solution: Dissolve 750 mg of salicylic
Acceptance criteria: 8101000 g/mg of C18H18N6O5S2, acid in 10 mL of methanol and dilute to 100 mL with Buffer
calculated on the anhydrous and sodium carbonate-free B.
basis; 90.0%115.0% of the labeled amount of
C18H18N6O5S2
USP 32 Official Monographs / Cefazolin 119
Standard stock solution: 1 mg/mL of USP Cefazolin RS in DEFINITION

Buffer B Cefazolin Sodium has a potency equivalent to NLT 89.1% and
Standard solution: Mix 5.0 mL of Standard stock solution NMT 110.1% of cefazolin sodium (C14H13NaN8O4S3),
and 5.0 mL of Internal standard solution. Dilute with Buffer B calculated on the anhydrous basis.
to 100 mL.
Sample stock solution: 1 mg/mL of Cefazolin in Buffer B IDENTIFICATION
Sample solution: Mix 5.0 mL of Sample stock solution and A. ULTRAVIOLET ABSORPTION 197U
5.0 mL of Internal standard solution. Dilute with Buffer B to Sample solution: 20 g/mL in 0.1 M sodium bicarbonate
100 mL. B. The retention time of the major peak for cefazolin in the
Chromatographic system Sample solution corresponds to that of the Standard solution,
(See Chromatography 621, System Suitability.) as obtained in the Assay.
Mode: LC C. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the
Detector: UV 254 nm requirements
Flow rate: 2 mL/min ASSAY
System suitability Buffer A: 0.9 mg/mL of anhydrous dibasic sodium
Sample: Standard solution phosphate and 1.298 mg/mL of citric acid monohydrate in
[NOTEThe relative retention times for salicylic acid and water
cefazolin are about 0.7 and 1.0, respectively.] Buffer B: 5.68 mg/mL of anhydrous dibasic sodium
Suitability requirements phosphate and 3.63 mg/mL of monobasic potassium
Resolution: NLT 4.0 between the analyte and internal phosphate in water
standard peaks Mobile phase: Acetonitrile and Buffer A (1:9)
Column efficiency: NLT 1500 theoretical plates Pass through a membrane filter having a 10-m or finer
Tailing factor: NMT 1.5 porosity.
Relative standard deviation: NMT 2.0% Internal standard solution: 7.5 mg/mL of salicylic acid in
Analysis methanol and Buffer B (1:9)
Samples: Standard solution and Sample solution Standard stock solution: 1 mg/mL of USP Cefazolin RS in
Calculate the percentage of C14H14N8O4S3 in the portion of Buffer B
Cefazolin taken: Standard solution: Mix 5.0 mL of Standard stock solution
and 5.0 mL of Internal standard solution. Dilute with Buffer B
Result = (RU/RS) (CS/CU) 100 to 100 mL.
Sample stock solution: 1 mg/mL of Cefazolin Sodium in
RU = peak response ratio of Cefazolin to the internal Buffer B
standard from the Sample solution Sample solution: Mix 5.0 mL of Sample stock solution and
RS = peak response ratio of Cefazolin to the internal 5.0 mL of Internal standard solution. Dilute with Buffer B to
standard from the Standard solution 100 mL.
CS = concentration of USP Cefazolin RS, calculated on Chromatographic system
the anhydrous basis, in the Standard solution (See Chromatography 621, System Suitability.)
(mg/mL) Mode: LC
CU = concentration of Cefazolin in the Sample solution Detector: UV 254 nm
(mg/mL) Column: 4.0-mm 30-cm; 10-m packing L1
IMPURITIES System suitability
Inorganic Impurities Sample: Standard solution
HEAVY METALS, Method II 231: NMT 20 ppm [NOTEThe relative retention times for salicylic acid and
Cefazolin are about 0.7 and 1.0, respectively.]
WATER DETERMINATION, Method I 921: NMT 2.0% Resolution: NLT 4.0 between the analyte and the internal
standard peaks
ADDITIONAL REQUIREMENTS Column efficiency: NLT 1500 theoretical plates
PACKAGING AND STORAGE: Preserve in tight containers. Tailing factor: NMT 1.5
USP Cefazolin RS Analysis
Calculate the percentage of C14H13NaN8O4S3 in the portion
Cefazolin Sodium of Cefazolin Sodium taken:
(Comment on this Monograph)id=m13972=Cefazolin Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100
Sodium=Ca-Chl-Monos.pdf)
C14H13N8NaO4S3 476.49 RU = peak response ratio of cefazolin to the internal
5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 3-[[(5- standard from the Sample solution
methyl-1,3,4-thiadiazol-2-yl)thio]methyl]-8-oxo-7-[[(1H- RS = peak response ratio of cefazolin to the internal
tetrazol-1-yl)acetyl]amino]-, monosodium salt (6R-trans); standard from the Standard solution
Monosodium (6R,7R)-3-[[(5-methyl-1,3,4-thiadiazol-2- CS = concentration of USP Cefazolin RS, calculated on
yl)thio]methyl]-8-oxo-7-[2-(1H-tetrazol-1-yl)acetamido]-5- the anhydrous basis, in the Standard solution
thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate (mg/mL)
[27164-46-1].
120 Cefazolin / Official Monographs USP 32
CU = nominal concentration of Cefazolin Sodium in Sample solution: Mix 5.0 mL of Sample stock solution and
the Sample solution (mg/mL) 5.0 mL of Internal standard solution. Dilute to 100 mL with
Mr1 = molecular weight of cefazolin sodium, 476.49 Buffer B.
Mr2 = molecular weight of cefazolin, 454.51 Chromatographic system
Acceptance criteria: 89.1%110.1% (See Chromatography 621, System Suitability.)
Mode: LC
OPTICAL ROTATION, Specific Rotation 781S: 10 to 24 Column: 4.0-mm 30-cm; 10-m packing L1
Sample solution: 55 mg/mL, in 0.1 M sodium bicarbonate Flow rate: 2 mL/min
PH 791: 4.06.0, in a solution containing 100 mg/mL of Injection size: 10 L
cefazolin System suitability
WATER DETERMINATION, Method I 921: NMT 6.0% Sample: Standard solution
STERILITY TESTS 71: Where the label states that Cefazolin [NOTEThe relative retention times for salicylic acid and
Sodium is sterile, it meets the requirements when tested as cefazolin are about 0.7 and 1.0, respectively.]
directed for Test for Sterility of the Product to Be Examined, Suitability requirements
Membrane Filtration. Resolution: NLT 4.0 between the analyte and internal
BACTERIAL ENDOTOXINS TEST 85: Where the label states that standard peaks
Cefazolin Sodium is sterile or must be subjected to further Column efficiency: NLT 1500 theoretical plates
processing during the preparation of injectable dosage Tailing factor: NMT 1.5
forms, it contains NMT 0.15 USP Endotoxin Unit/mg of Relative standard deviation: NMT 2.0%
cefazolin. Analysis
ADDITIONAL REQUIREMENTS Calculate the percentage of C14H14N8O4S3 in each mL of the
PACKAGING AND STORAGE: Preserve in tight containers. Injection taken:
LABELING: Where it is intended for use in preparing
injectable dosage forms, the label states that it is sterile or Result = (RU/RS) (CS/CU) 100
preparation of injectable dosage forms. RU = peak response ratio of cefazolin to the internal
USP REFERENCE STANDARDS 11 standard from the Sample solution
USP Cefazolin RS RS = peak response ratio of cefazolin to the internal
USP Endotoxin RS standard from the Standard solution
CS = concentration of USP Cefazolin RS, calculated on
the anhydrous basis, in the Standard solution
Cefazolin Injection (mg/mL)
CU = nominal concentration of cefazolin in the Sample
(Comment on this Monograph)id=m13974=Cefazolin solution (mg/mL)
DEFINITION
Cefazolin Injection is a sterile solution of Cefazolin and Sodium SPECIFIC TESTS
Bicarbonate in a diluent containing one or more suitable BACTERIAL ENDOTOXINS TEST 85: NMT 0.15 USP Endotoxin
tonicity-adjusting agents. It contains NLT 90.0% and NMT Unit/mg of cefazolin
115.0% of the labeled amount of cefazolin (C14H14N8O4S3). STERILITY TESTS 71: It meets the requirements when tested
as directed for Test for Sterility of the Product to Be Examined,
IDENTIFICATION Membrane Filtration.
The retention time of the major peak of the Sample solution PH 791: 4.57.0
corresponds to that of the Standard solution, as obtained in PARTICULATE MATTER IN INJECTIONS 788: Meets the
the Assay. requirements for small-volume injections

PROCEDURE PACKAGING AND STORAGE: Preserve as described under
Buffer A: 0.9 mg/mL of anhydrous dibasic sodium Injections 1, Containers for Injections. Maintain in the frozen
phosphate and 1.298 mg/mL of citric acid monohydrate in state.
water LABELING: It meets the requirements under Injections 1,
Buffer B: 5.68 mg/mL of anhydrous dibasic sodium Labeling. The label states that it is to be thawed just before
phosphate and 3.63 mg/mL of monobasic potassium use, describes conditions for proper storage of the resultant
phosphate in water solution, and directs that the solution is not to be refrozen.
Mobile phase: Acetonitrile and Buffer A (1:9) USP REFERENCE STANDARDS 11
Pass through a membrane filter having a 10-m or finer USP Cefazolin RS
porosity. USP Endotoxin RS
Internal standard solution: 7.5 mg/mL of salicylic acid in
methanol and Buffer B (1:9)
Standard stock solution: 1 mg/mL of USP Cefazolin RS in Cefazolin for Injection
Buffer B
Standard solution: Mix 5.0 mL of Standard stock solution (Comment on this Monograph)id=m13978=Cefazolin for
and 5.0 mL of Internal standard solution. Dilute to 100 with Injection=Ca-Chl-Monos.pdf)
Buffer B.
Sample stock solution: Allow 1 container of Injection to DEFINITION
thaw, and mix. Transfer a volume of the Injection, nominally Cefazolin for Injection contains an amount of Cefazolin Sodium
equivalent to about 50 mg of cefazolin, to a 50-mL equivalent to NLT 90.0% and NMT 115.0% of the labeled
volumetric flask, dilute with Buffer B to volume, and mix. amount of (C14H14N8O4S3).
USP 32 Official Monographs / Cefazolin 121
IDENTIFICATION RU = peak response ratio of cefazolin to the internal

A. ULTRAVIOLET ABSORPTION 197U standard of the Sample solution
Sample solution: 20 g/mL in 0.1 M sodium bicarbonate RS = peak response ratio of cefazolin to the internal
B. The retention time of the major peak for cefazolin from standard of the Standard solution
the Sample solution corresponds to that of the Standard CS = concentration of USP Cefazolin RS, calculated on
solution, as obtained in the Assay. the anhydrous basis, in the Standard solution
C. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the (mg/mL)
requirements CU = nominal concentration of cefazolin in the Sample
solution (mg/mL)
ASSAY [NOTEWhere the test for Uniformity of Dosage Units has
PROCEDURE been performed using the Analysis for content uniformity,
Buffer A: 0.9 mg/mL of anhydrous dibasic sodium use the average of these determinations as the Assay
phosphate and 1.298 mg/mL of citric acid monohydrate in value.]
water Acceptance criteria: 90.0%115.0%
Buffer B: 5.68 mg/mL of anhydrous dibasic sodium
phosphate and 3.63 mg/mL of monobasic potassium PERFORMANCE TESTS
phosphate in water UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Mobile phase: Acetonitrile and Buffer A (1:9) Analysis for content uniformity: Perform the Assay on
Pass through a membrane filter having a 10-m or finer individual containers using Sample solution A or Sample
porosity. solution B, or both, as appropriate.
Internal standard solution: 7.5 mg/mL of salicylic acid in
methanol and Buffer B (1:9) SPECIFIC TESTS
Standard stock solution: 1 mg/mL of USP Cefazolin RS in CONSTITUTED SOLUTION: At the time of use, it meets the
Buffer B requirements for Injections 1, Constituted Solutions.
Standard solution: Mix 5 mL of Standard stock solution and OPTICAL ROTATION, Specific Rotation 781S: 10 to 24
5 mL of Internal standard solution. Dilute with Buffer B to Sample solution: 55 mg/mL, in 0.1 M sodium bicarbonate
100 mL. BACTERIAL ENDOTOXINS TEST 85: NMT 0.15 USP Endotoxin
Sample stock solution A (where it is packaged for Unit/mg of cefazolin
dispensing and is represented as being in a single-dose STERILITY TESTS 71: It meets the requirements when tested
container): Constitute Cefazolin for Injection in a volume of as directed under Test for Sterility of the Product to be
water corresponding to the volume of solvent specified in Examined, Membrane Filtration.
the labeling. Withdraw all of the withdrawable contents, PH 791: 4.06.0, in a solution containing 100 mg/mL of
using a suitable hypodermic needle and syringe, and dilute cefazolin
with Buffer B to obtain a solution containing nominally 1 WATER DETERMINATION, Method I 921: NMT 6.0%
mg/mL of cefazolin. PARTICULATE MATTER IN INJECTION 788: Meets the
Sample solution A: Mix 5.0 mL of Sample stock solution A requirements for small-volume injections
and 5.0 mL of Internal standard solution. Dilute with Buffer B OTHER REQUIREMENTS: It meets the requirements for
to 100 mL. Injections 1, Labeling.
Sample stock solution B: (where the label states the
quantity of cefazolin in a given volume of constituted ADDITIONAL REQUIREMENTS
solution): Constitute Cefazolin for Injection in a volume of PACKAGING AND STORAGE: Preserve as described under
water corresponding to the volume of solvent specified in Injections 1, Containers for Injections.
the labeling. Dilute a volume of the constituted solution USP REFERENCE STANDARDS 11
with Buffer B to obtain a solution containing nominally 1 USP Cefazolin RS
mg/mL of cefazolin. USP Endotoxin RS
Sample solution B: Mix 5.0 mL of Sample stock solution A
and 5.0 mL of Internal standard solution. Dilute with Buffer B
to 100 mL. Cefazolin Ophthalmic Solution
(See Chromatography 621, System Suitability.) (Comment on this Monograph)id=m13980=Cefazolin
Mode: LC Ophthalmic Solution=Ca-Chl-Monos.pdf)
Column: 4.0-mm 30-cm; 10-m packing L1 Cefazolin Ophthalmic Solution contains an amount of Cefazolin
Flow rate: 2 mL/min Sodium equivalent to NLT 29.7 mg and NMT 36.3 mg of
Injection size: 10 L cefazolin (C14H14N8O4S3) in 10.0 mL of Ophthalmic Solution.
System suitability Use Cefazolin Sodium or Cefazolin for Injection that contains
Sample: Standard solution the designated amount of cefazolin, and prepare the
[NOTEThe relative retention times are about 0.7 for Ophthalmic Solution as follows (see Pharmaceutical
salicylic acid and 1.0 for Cefazolin.] CompoundingNonsterile Preparations 795):
standard peaks Cefazolin Sodium 35 mg
Column efficiency: NLT 1500 theoretical plates Thimerosal 0.2 mg
Tailing factor: NMT 1.5 Sodium Chloride Injection (0.9%) A sufficient quantity
Analysis To make 10.0 mL
Samples: Sample solution A, Sample solution B, and
Standard solution Dissolve quantities of Cefazolin Sodium and Thimerosal in
Calculate the percentage of C14H14N8O4S3 in the container, Sodium Chloride Injection (0.9%), and dilute quantitatively
and in the volume of constituted solution: and stepwise if necessary, with Sodium Chloride Injection
(0.9%) to obtain a solution containing, in each mL, 3.5 mg of
Result = (RU/RS) (CS/CU) 100 Cefazolin Sodium and 0.02 mg of Thimerosal. Filter a 10.0-mL
122 Cefazolin / Official Monographs USP 32
portion of the resulting solution to produce a clear and sterile F = correction factor (to convert mg/mL to mg/10
Ophthalmic Solution. If Cefazolin for Injection is used, prepare mL), 10
the Ophthalmic Solution as follows. Dissolve an accurately Acceptance criteria: 29.736.3 mg
weighed quantity of Thimerosal in Sodium Chloride Injection
(0.9%), and dilute quantitatively and stepwise if necessary, SPECIFIC TESTS
with Sodium Chloride Injection (0.9%) to obtain a solution STERILITY: See Pharmaceutical CompoundingNonsterile
containing 0.3 mg of Thimerosal/mL. Add 9.8 mL of the Preparations 795, Sterility
resulting solution to a vial of Cefazolin for Injection, containing PH 791: 4.56.0
500 mg of cefazolin, and mix to obtain a stock solution.
Transfer 3.3 mL of the stock solution to a 50-mL volumetric ADDITIONAL REQUIREMENTS
flask, dilute with Sodium Chloride Injection (0.9%) to volume, PACKAGING AND STORAGE: Preserve in tight, sterile
and mix. Filter a 10.0-mL portion of the resulting solution to ophthalmic containers. Store in a refrigerator.
produce a clear and sterile Ophthalmic Solution. LABELING: Label it to state that it is intended for use in the
eye, and is not to be used if a precipitate is present.
ASSAY BEYOND-USE DATE: 5 days after the date on which it was
PROCEDURE compounded
Buffer A: 0.9 mg/mL of anhydrous dibasic sodium USP REFERENCE STANDARDS 11
phosphate and 1.298 mg/mL of citric acid monohydrate in USP Cefazolin RS
water
Buffer B: 5.68 mg/mL of anhydrous dibasic sodium
phosphate and 3.63 mg/mL of monobasic potassium
phosphate in water
Solution C: Acetonitrile and Buffer A (1:9) Add the following:
Pass the resulting solution through a filter having a 5-m or
finer porosity.
Solution D: Acetonitrile and Buffer A (4:1)
Cefdinir
Pass the resulting solution through a filter having a 5-m or (Comment on this Monograph)id=m13982=Cefdinir=Ca-Chl-
finer porosity. Monos.pdf)
Time Solution C Solution D

(min) (%) (%)
0 100 0
15 0 100
25 100 0
C14H13N5O5S2 395.41
Standard solution: 0.32 mg/mL of USP Cefazolin RS in 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[(2-
Buffer B amino-4-thiazolyl)(hydroxyimino)acetyl]amino]-3-ethenyl-8-
Maintain at 4 prior to injection. oxo-, [6R-[6, 7(Z)]]-;
Sample solution: Transfer 1.0 mL of Ophthalmic Solution (-)-(6R, 7R)-7-[2-(2-Amino-4-thiazolyl)glyoxylamido]-8-oxo-3-
to a 10-mL low-actinic volumetric flask, dilute with Buffer B vinyl-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid,
to volume, and mix. Maintain at 4 prior to injection. 72-(Z)-oxime [91832-40-5].
(See Chromatography 621, System Suitability.) Cefdinir contains NLT 960 g/mg and NMT 1020 g/mg of
Mode: LC C14H13N5O5S2, calculated on the anhydrous basis.
Detector: UV 273 nm
Column: 3.9-mm 30-cm; 10-m packing L1 IDENTIFICATION
Temperature: 25 A. INFRARED ABSORPTION 197M
Flow rate: 2 mL/min B. The retention time of the major peak of the Sample
Injection size: 10 L solution corresponds to that of the Standard solution, as
System suitability obtained in the Assay.
Column efficiency: NLT 1500 theoretical plates PROCEDURE
Tailing factor: NMT 1.5 Solution A: 14.2 mg/mL anhydrous dibasic sodium
Relative standard deviation: NMT 2.0% phosphate
Analysis Solution B: 13.6 mg/mL monobasic potassium phosphate
Samples: Sample solution and Standard solution Buffer solution: Combine appropriate amounts of Solution
Calculate the quantity, in mg, of C14H14N8O4S3 in the A and Solution B (about 2:1) to obtain a pH 7.0 solution.
portion of Ophthalmic Solution taken: Solution C: Dilute tetramethylammonium hydroxide (10%)
with water to obtain a 1% solution. Adjust with dilute
Result = (rU/rS) CS D F phosphoric acid (1 in 10) to a pH of 5.5.
Solution D: 37.2 mg/mL edetate disodium
rU = peak response from the Sample solution Mobile phase: Acetonitrile, methanol, Solution C, and
rS = peak response from the Standard solution Solution D (60:40:900:0.4)
CS = concentration of USP Cefazolin RS in the System suitability solution: 0.2 mg /mL of USP Cefdinir RS
Standard solution (mg/mL) and 0.5 mg/mL of USP Cefdinir Related Compound A RS,
D = dilution factor used in the preparation of the Buffer solution
Sample solution, 10
USP 32 Official Monographs / Cefdinir 123
Standard solution: 0.2 mg/mL of USP Cefdinir RS, Buffer Time Solution E Solution F
solution (min) (%) (%)
Sample solution: 0.2 mg/mL of Cefdinir, Buffer solution
32 50 50
(See Chromatography 621, System Suitability.) 37 50 50
Mode: LC 38 95 5
Detector: UV 254 nm
58 95 5
Column: 4.6-mm 15-cm; 5 m, packing L1
Temperature: 40
Flow rate: 1 mL/min Chromatographic system
Injection size: 5 L (See Chromatography 621, System Suitability.)
System suitability Mode: LC
Sample: Standard solution and System suitability solution Detector: UV 254 nm
[NOTEUSP Cefdinir Related Compound A RS should Column: 4.6-mm 15-cm; 5 micron packing L1
produce four peaks.] Temperature: 40
Tailing factor: NMT 1.5 for cefdinir, System suitability Flow rate: 1.5 mL/min
solution Injection size: 10 L
Relative standard deviation: NMT 1.0%, Standard System suitability
solution Sample: System suitability solution A, System suitability
Analysis solution B, and System suitability solution C
Sample: Standard solution and Sample solution [NOTEUSP Cefdinir Related Compound A RS should
Calculate the quantity (g/mg) of C14H13N5O5S2 in the produce four peaks.]
portion of Cefdinir taken: [NOTEThe relative retention time of the third peak from
USP Cefdinir Related compound A RS is NLT 1.1,
Result = (rU/rS) (CS/CU) P relative to the cefdinir peak, System suitability solution
C.]
rU = peak response from the Sample solution Suitability requirements
rS = peak response from the Standard solution Linearity: The response of cefdinir in System suitability
CS = concentration of the Standard solution (mg/mL) solution B is between 7% and 13% of that from System
CU = concentration of the Sample solution (mg/mL) suitability solution A.
P = purity of USP Cefdinir RS (g/mg) Column efficiency: NLT 7000 theoretical plates for
Acceptance criteria: 9601020 g/mg cefdinir, System suitability solution C
Tailing factor: NMT 3.0 for cefdinir, System suitability
IMPURITIES solution C
Inorganic Impurities Relative standard deviation: NMT 2.0% for cefdinir,
RESIDUE ON IGNITION 281: NMT 0.1% System suitability solution C
HEAVY METALS, Method II 231: 10 ppm Analysis
Organic Impurities Sample: Sample solution [NOTERecord the
PROCEDURE chromatogram for NLT 40 min.]
Solution A, Solution B, Buffer solution, Solution C, and Calculate the percentage of each impurity in the portion
Solution D: Prepare as directed in the Assay. of Cefdinir taken:
Solution E: To 1000 mL of Solution C, add 0.4 mL of
Solution D. Result = (rU/rS) 100
Solution F: Acetonitrile, methanol, Solution C, and Solution
D (300:200:500:0.4) rU = peak response of each impurity from the Sample
System suitability solution A: 15 g/mL of cefdinir, from solution
the Sample solution, diluted with Solution C r = sum of all the peak responses from the Sample
System suitability solution B: 1.5 g /mL of cefdinir, from solution
System suitability solution A, diluted with Solution C
System suitability solution C: Transfer about 30 mg of
USP Cefdinir RS and 2 mg of USP Cefdinir Related
Compound A RS to a 20-mL volumetric flask, dissolve in 3
mL of Buffer solution, and dilute with Solution C to volume.
Sample stock solution: 10 mg/mL of Cefdinir in Buffer
solution
Sample solution: 1.5 mg/mL of Cefdinir from the Sample
stock solution, in Solution C
[NOTEPrepare fresh immediately before use.]
Time Solution E Solution F

(min) (%) (%)
0 95 5
2 95 5
22 75 25
124 Cefdinir / Official Monographs USP 32
Acceptance criteria IDENTIFICATION

Individual impurities: See Impurity Table 1. A. ULTRAVIOLET ABSORPTION 197U
Total impurities: NMT 3.0% Buffer: Prepare as directed in the Assay.
Blank: Use the Buffer.
Impurity Table 1 Standard solution: 10 g/mL of USP Cefdinir RS in Buffer
Sample solution: Equivalent to 10 g/mL of Cefdinir in
Relative Acceptance Buffer
Retention Criteria, NMT Filter before use.
Name Time (%) Cell size: 1 cm
Impurity Aa 0.10 0.5 Acceptance criteria: Absorptivities of the Sample solution
maxima and minima occur at the same wavelengths as
Impurity Bb 0.12 0.5 those in the Standard solution.
Impurity Cc 0.74 0.7 B. The retention time of the major peak of the Sample
Cefdinir related 0.85, 0.93, 1.11, 0.7 (sum of 4) solution corresponds to that of the Standard solution, as
compound Ad (4 peaks) 1.14 obtained in the Assay.
Impurity Ee 1.22 0.5 ASSAY
Impurity Ff 1.36 0.5 PROCEDURE
Buffer: 10.65 mg/mL of dibasic sodium phosphate and
Impurity Gg 1.51 0.7
3.40 mg/mL of monobasic potassium phosphate
Impurity Hh (2 peaks) 1.61, 1.64 0.5 (sum of 2) Adjust to pH 7.0 0.05 with phosphoric acid or sodium
Any other individual, 0.2 hydroxide before final dilution.
unidentified impurity Solution A: 7.0 mg/mL citric acid monohydrate
Adjust to pH 2.0 0.05 with phosphoric acid.
a1N-[(Z)-2-(2-Aminothiazol-4-yl)-2-(hydroxyimino)acetyl]glycine.
b(Z)-2-(2-Aminothiazol-4-yl)-N-(2,2-dihydroxyethyl)-2-
Mobile phase: Methanol, tetrahydrofuran, and Solution A
(111:28:1000)
(hydroxyimino)acetamide.
c(6R,7R)-7-[(Z)-2-(2-Aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-
System suitability solution: 50 g/mL of cefdinir and 175
g/mL of m-hydroxybenzoic acid in Buffer
methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid.
d2(R)-2-[(Z)-2-(2-Aminothiazol-4-yl)-2-(hydroxyimino)acetamido)]-2-
Standard solution: 50 g/mL of USP Cefdinir RS in Buffer
Sample solution: Equivalent to 50 g/mL of Cefdinir, from
[(2RS,5RS)-5-methyl-7-oxo-2,4,5,7-tetrahydro-1H-furo[3,4-d][1,3]thiazin-2-
Capsule contents in the Buffer
yl]acetic acid.
e(Z)-2-(2-Aminothiazol-4-yl)-2-(hydroxyimino)-N-{(3RS,5aR,6R)-3-
methyl-1,7-dioxo-1,3,4,5a,6,7-hexahydroazeto[2,1-b]furo[3,4-d]
Mode: LC
[1,3]thiazin-6-yl}acetamide.
f(6R,7R)-7-(4-hydroxyisoxazole-3-carboxamido)-8-oxo-3-vinyl-5-thia-1-
Detector: UV 254 nm
Column: 3.9-mm 15-cm; 4 m, packing L1
azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid.
g(6R,7R)-7-[(E)-2-(2-Aminothiazol-4-yl-)-2-(hydroxyimino)acetamido]-8-
oxo-3-vinyl-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid.
h(Z)-2-(2-Aminothiazol-4-yl)-2-(hydroxyimino)-N-{[(2RS,5RS)-5-methyl-7-
System suitability
Sample: Standard solution and System suitability solution
oxo-2,4,5,7-tetrahydro-1H-furo[3,4-d][1,3]thiazin-2-yl]methyl}acetamide.
SPECIFIC TESTS Resolution: NLT 3.0 between cefdinir and m-
OPTICAL ROTATION, Specific Rotation 781S: 61 to 67 at hydroxybenzoic acid, System suitability solution
20 Tailing factor: NMT 2.0 for cefdinir, System suitability
Sample solution: 10 mg/mL in the Buffer Solution as in the solution
Assay Relative standard deviation: NMT 1.0% for cefdinir,
WATER DETERMINATION, Method I 921: NMT 2.0% for Standard solution
anhydrous; 4.0%8.0% for monohydrate, using formamide Analysis
and methanol (2:1) as the solvent Samples: Standard solution and Sample solution
Calculate the percentage of Cefdinir in the portion of
ADDITIONAL REQUIREMENTS Capsules taken:
containers. Result = (rU/rS) (CS/CU) 100
USP Cefdinir RS rU = peak response for cefdinir from the Sample
USP Cefdinir Related Compound A RSUSP32 solution
rS = peak response for cefdinir from the Standard
solution
CS = concentration of the Standard solution (mg/mL)
CU = nominal concentration of cefdinir in the Sample
Add the following: solution (mg/mL)
Cefdinir Capsules
(Comment on this Monograph)id=m2183=Cefdinir
Capsules=Ca-Chl-Monos.pdf)
DEFINITION
Cefdinir Capsules contains NLT 90.0% and NMT 110.0% of the
labeled amount of Cefdinir (C14H13N5O5S2).
Acceptance criteria: 90.0%100.0% flask. Dissolve in 30 mL of Buffer, and dilute with Diluent to
volume to obtain a solution having a known concentration
PERFORMANCE TESTS of about 1.5 mg/mL of cefdinir.
DISSOLUTION 711 Mobile phase: See the gradient table below.
Medium: 50 mM phosphate buffer pH 6.8; 900 mL
Apparatus 2: 50 rpm
Time: 30 min Time Solution E Solution F
Detector: UV 290 nm (min) (%) (%)
Standard solution: 0.33 mg/mL USP Cefdinir RS in Medium 0 95 5
Sample solution: Sample per Dissolution 711 2 95 5
Filter each sample through a suitable 0.45 m filter. Dilute
with Medium to a concentration of about 0.33 mg/mL of 22 75 25
cefdinir. 32 50 50
Blank: Dissolve one empty capsule in 100 mL of Medium, 37 50 50
and dilute to 900 mL; filter if necessary.
Analysis: Determine the percentage of C14H13N5O5S2 38 95 5
dissolved: 58 95 5
Result = (AU/AS) CS D (V/L) 100 Chromatographic system

AU = absorbance of the Sample solution Mode: LC
AS = absorbance of the Standard solution Detector: UV 254 nm
CS = concentration of the Standard solution (mg/mL) Column: Column: 4.6-mm 15-m; packing L1
D = dilution factor of the Sample solution (mL/mL) Column temperature: 40 0.5
V = volume of Medium (mL), 900 Sample solution temperature: 4 3
L = Label claim (mg) Flow rate: 1 mL/min
Tolerances: NLT 80% (Q) of the labeled amount of Injection size: 10 L
C14H13N5O5S2 System suitability
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Sample: Standard solution and System suitability solution
IMPURITIES Suitability requirements
Organic Impurities Resolution: NLT 1.5, between cefdinir and the third
PROCEDURE peak of the USP Cefdinir Related Compound A RS,
Solution A: 14.2 mg/mL anhydrous dibasic sodium System suitability solution
phosphate Tailing factor: NMT 1.5 for cefdinir related compound
Solution B: 13.6 mg/mL monobasic potassium phosphate B, System suitability solution
Buffer: Combine appropriate amounts of Solution A and Relative standard deviation: NMT 2.0%, for cefdinir
Solution B (about 2:1) to obtain a pH 7.0 0.1 solution. peak response, Standard solution
Diluent: Dilute tetramethylammonium hydroxide (10%) Analysis
with water to obtain a 1% solution. Adjust with dilute Sample: Standard solution and Sample solution
phosphoric acid (1 in 10) to a pH of 5.5 0.1. Calculate the percentage of each impurity in the portion
Solution D: 37.2 mg/mL edetate disodium of Capsules taken:
Solution E: To 1000 mL of Diluent, add 0.4 mL of Solution Result = (rU/rS) (CS/CU) 100/RRF
D.
Solution F: Acetonitrile, methanol, Diluent, and Solution D rU = peak response from the Sample solution
(150:100:250:0.2) rS = peak response from the Standard solution
Standard stock solution: 750 mg/mL of USP Cefdinir RS, CS = concentration of the Standard solution (mg/mL)
Buffer CU = concentration of the Sample solution (mg/mL)
Standard solution: 15 g/mL of USP Cefdinir RS, from the RRF = relative response factor (see Impurity Table 1.)
Standard stock solution in Diluent Acceptance criteria
System suitability stock solution 1: 40 g/mL of USP Individual impurities: See Impurity Table 1.
Cefdinir Related Compound A RS in Diluent Total impurities: NMT 5.0%
System suitability stock solution 2: 40 g/mL of USP
Cefdinir Related Compound B RS in Diluent ADDITIONAL REQUIREMENTS
System suitability solution: Transfer 37.5 mg of USP PACKAGING AND STORAGE: Preserve in tight light-resistant
Cefdinir RS to a 25-mL volumetric flask. Add about 10 mL containers, and store at controlled room temperature.
of Buffer. Add 5.0 mL of each of System suitability stock USP REFERENCE STANDARDS 11
solution 1 and System suitability stock solution 2, and dilute USP Cefdinir RS
with Diluent to volume. USP Cefdinir Related Compound A RS
Sample solution: Transfer an equivalent to 300 mg of USP Cefdinir Related Compound B RSUSP32
Cefdinir, from Capsule contents, into a 200-mL volumetric
Add the following: m filter, and transfer 5.0 mL of the filtrate to a 10-mL
volumetric flask, and dilute with methanol to volume.
Cefdinir for Oral Suspension Developing solvent system: Methanol and water (4:1)
(Comment on this Monograph)id=m2184=Cefdinir for Oral Visualization: Shortwave UV
Suspension=Ca-Chl-Monos.pdf) Analysis
Sample: Standard solution and Sample solution
DEFINITION Develop the chromatogram until the solvent front has
Cefdinir for Oral Suspension contains NLT 90.0% and NMT moved about 15 cm. Remove the plate from the
110.0% of the labeled amount of C14H13N5O5S2. It may developing chamber, and allow the solvent to evaporate.
contain one or more suitable buffers, flavors, preservatives, Acceptance criteria: The RF value of the principal spot from
stabilizing agents, sweeteners, and suspending agents. the Sample solution corresponds to that from the Standard
solution.
IDENTIFICATION B. The retention time of the major peak of the Sample
A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 solution corresponds to that of the Standard solution, as
Adsorbent: 0.25-mm layer of chromatographic silica gel, obtained in the Assay.
preconditioned with n-hexane and tetradecane (95:5)
Buffer: Prepare as directed in the Assay. ASSAY
Standard solution: 600 g/mL of USP Cefdinir RS in PROCEDURE
methanol and Buffer (3:1) Buffer: 10.65 mg/mL of anhydrous dibasic sodium
Sample solution: Transfer an equivalent to 125 mg of phosphate and 3.40 mg/mL of monobasic potassium
Cefdinir from reconstituted Suspension, in a 100-mL phosphate in water
volumetric flask, add 50 mL of Buffer and dilute with Adjust to pH 7.0 0.05 with phosphoric acid or sodium
methanol to volume. Pass a portion through a suitable 0.45- hydroxide before final dilution.
Impurity Table 1
Relative Relative Limit of Acceptance
Retention Response Quantification Criteria,
Name Time Factor (% Cefdinir) NMT (%)
Impurity VIII 0.10 1.1 0.1 0.5
Impurity IV 0.13 1.1 0.1 0.5
Impurity XIV 0.36 1.0 0.05 0.2
Impurity V 0.46 1.5 0.05 0.7
Impurity B 0.77 1.0 0.05 0.3
Impurity XI 0.75 1.0 0.05 0.7
Cefdinir Related Compound A (lac- 0.85 1.5 0.1 2.5
tam ring cleavage lactones-a)a
Cefdinir Related Compound A (lac- 0.94 1.5 0.1
tam ring cleavage lactones-b)a
tam ring cleavage lactones-c)a
tam ring cleavage lactones-d)a
Impurity VI 1.18 1.1 0.05 0.2
Impurity I 1.23 1.2 0.05 1.0
Cefdinir Related Compound Ba 1.28 1.1 0.05 0.2
Impurity XIII 1.37 1.4 0.05 0.5
Impurity Ec 1.44 1.0 0.05 0.5
Impurity XV 1.49 1.0 0.05 0.2
Impurity VII 1.51 1.1 0.05 0.7
Impurity IIIab 1.62 1.3 0.05 1.0
Impurity IIIbb 1.64 1.3 0.05
Impurity Dc 1.82 1.0 0.05 0.2
Individual unidentified impurities 1.0 0.2
Total unidentified impuritiesd 1.0
aRS II is a mixture of 4 isomers designated as RS IIa, RS IIb, RS IIc, and RS IId. The sum of all values is reported and the total limit for all 4 isomers combined is
2.5%.
bRS III is a mixture of 2 isomers designated as RS IIIa and RS IIIb. The sum of both values is reported and the total limit for both isomers combined is 1.0%.
cImpurity B, Impurity D, and Impurity E are unidentified impurities.
dThe total unidentified impurities limit includes the % total of unidentified impurities B, D, and E and any other individual unidentified impurities.
Solution A: 7.0 mg/mL citric acid monohydrate UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Adjust to pH 2.0 0.05 with phosphoric acid. DELIVERABLE VOLUME 698 (for Oral Suspension packaged in
Mobile phase: Methanol, tetrahydrofuran, and Solution A multiple-unit containers): Meets the requirements
(111:28:1000)
System suitability solution: 50 g/mL of cefdinir and 175 IMPURITIES
g/mL of m-hydroxybenzoic acid in Buffer Organic Impurities
Standard solution: 50 g/mL of USP Cefdinir RS in Buffer PROCEDURE
Sample solution: Equivalent to 50 g/mL of Cefdinir, from Solution A: 14.2 mg/mL anhydrous dibasic sodium
constituted Suspension in Buffer phosphate
Chromatographic system Solution B: 13.6 mg/mL monobasic potassium phosphate
(See Chromatography 621, System Suitability.) Buffer: Combine appropriate amounts of Solution A and
Mode: LC Solution B (about 2:1) to obtain a pH 7.0 0.1 solution.
Detector: UV 254 nm Diluent: Dilute tetramethylammonium hydroxide (10%)
Column: 3.9-mm 15-cm; 4 m, packing L1 with water to obtain a 1% solution. Adjust with dilute
Flow rate: 1.4 mL/min phosphoric acid (1 in 10) to a pH of 5.5 0.1.
Injection size: 15 L Solution D: 37.2 mg/mL edetate disodium
System suitability Solution E: To 1000 mL of Diluent, add 0.4 mL of Solution
Sample: Standard solution and System suitability solution D.
Suitability requirements Solution F: Acetonitrile, methanol, Diluent, and Solution D
Resolution: NLT 3.0 between cefdinir and m- (150:100:250:0.2)
hydroxybenzoic acid, System suitability solution Standard stock solution: 750 g/mL of USP Cefdinir RS in
Tailing factor: NMT 2.0 for cefdinir, System suitability Buffer
solution Standard solution: 15 g/mL of USP Cefdinir RS, from the
Relative standard deviation: NMT 1.0% for cefdinir, Standard stock solution in Diluent
Standard solution System suitability stock solution 1: 40 g/mL of USP
Analysis Cefdinir Related Compound A RS in Diluent
Sample: Standard solution and Sample solution System suitability stock solution 2: 40 g/mL of USP
Calculate the percentage of Cefdinir in the portion of Oral Cefdinir Related Compound B RS in Buffer
Suspension taken: System suitability solution: Transfer 37.5 mg of USP
Cefdinir RS to a 25-mL volumetric flask. Add about 10 mL
Result = (rU/rS) (CS/CU) 100 of Buffer. Add 5.0 mL of each of System suitability stock
solution 1 and System suitability stock solution 2, and dilute
rU = peak response for cefdinir from the Sample with Diluent to volume.
solution Sample solution: Transfer an equivalent to 150 mg of
rS = peak response for cefdinir from the Standard Cefdinir, from constituted Oral Suspension, into a 100-mL
solution volumetric flask. Dissolve in 30 mL of Buffer, and dilute
CS = concentration of the Standard solution (mg/mL) with Diluent to volume.
CU = nominal concentration of cefdinir in the Sample Mobile phase: See the gradient table below.
solution (mg/mL)
Acceptance criteria: 90.0%110.0% Time Solution E Solution F
(min) (%) (%)
PERFORMANCE TESTS
DISSOLUTION 711 0 95 5
Medium: 50 mM phosphate buffer pH 6.8; 900 mL 2 95 5
Apparatus 2: 50 rpm 22 75 25
Time: 30 min
Detector: UV 290 nm 32 50 50
Sample solution: Dilute a portion of each filtered sample 37 50 50
with Medium as necessary to obtain a solution having a 38 95 5
concentration of about 0.14 mg per mL of cefdinir.
Standard solution: 0.14 mg/mL USP Cefdinir RS in Medium 58 95 5
Blank: Medium
Analysis: Transfer 5 mL, by weight, of the reconstituted Chromatographic system
Oral Suspension into the vessel. After the appropriate time, (See Chromatography 621, System Suitability.)
withdraw a portion of the solution under test and pass Mode: LC
through a suitable 0.45-m filter. Detector: UV 254 nm
Determine the percentage of C14H13N5O5S2 dissolved: Column: 4.6-mm 15-cm; 5 m packing L1
Column temperature: 40 0.5
Result = (AU/AS) ([CS d D V]/W L) 100 Sample solution temperature: 4 3
Flow rate: 1 mL/min
AU = absorbance of the Sample solution Injection size: 10 L
AS = absorbance of the Standard solution System suitability
CS = concentration of the Standard solution (mg/mL) Sample: Standard solution and System suitability solution
d = density of the Oral Suspension (mg/mL) Suitability requirements
D = dilution factor of the Sample solution (mL/mL) Resolution: NLT 1.5, between cefdinir and the third
V = volume of Medium (mL), 900 peak of the USP Cefdinir Related Compound A RS,
W = weight of Oral Suspension taken (mg) System suitability solution
L = Label claim (mg)
Tailing factor: NMT 1.5 for cefdinir related compound Acceptance criteria
B, System suitability solution Individual impurities: See Impurity Table 1.
Relative standard deviation: NMT 2.0% for cefdinir Total impurities: NMT 6.2%
peak response, Standard solution
Sample: Standard solution and Sample solution PH 791: 3.54.5
Calculate the percentage of each impurity in the portion LOSS ON DRYING 731: Dry about 1 g over phosphorous
of Oral Suspension taken: pentoxide in a vacuum not exceeding 5 mm of mercury at
70 for 44.5 h: it loses NMT 1.0% of its weight.
Result = (rU/rS) (CS/CU) 100/RRF
rU = peak response of impurity from the Sample PACKAGING AND STORAGE: Preserve in tight light-resistant
solution containers, and store at controlled room temperature.
rS = peak response from the Standard solution LABELING: The label specifies the directions for the
CS = concentration of the Standard solution (mg/mL) constitution of the powder and states the equivalent amount
CU = nominal concentration of cefdinir in the Sample of C14H13N5O5S2 in a given volume of Oral Suspension after
solution (mg/mL) constitution.
RRF = relative response factor (see Impurity Table 1)
Impurity Table 1
Relative Relative Limit of Acceptance
Retention Response Quantification Criteria,
Name Time Factor (% Cefdinir) NMT (%)
Impurity VIII 0.10 1.1 0.1 0.5
Impurity IV 0.13 1.1 0.1 0.6
Impurity XIV 0.36 1.0 0.05 0.2
Impurity V 0.46 1.5 0.05 0.3
Impurity Bc 0.77 1.0 0.05 0.2
Impurity XI 0.75 1.0 0.05 0.7
Cefdinir Related Compound A 0.85 1.5 0.1 3.3
(lactam ring cleavage lactones-
a)a
Cefdinir Related Compound A 0.94 1.5 0.1
b)a
c)a
d)a
Impurity VI 1.18 1.1 0.05 0.2
Impurity I 1.23 1.2 0.05 0.8
Cefdinir Related Compound B 1.28 1.1 0.05 0.2
Impurity XIII 1.37 1.4 0.05 0.5
Impurity Ec 1.44 1.0 0.05 0.2
Impurity XV 1.49 1.0 0.05 0.2
Impurity VII 1.51 1.1 0.05 1.2
Impurity IIIab 1.62 1.3 0.05 1.1
Impurity IIIbb 1.64 1.3 0.05
Impurity Dc 1.82 1.0 0.05 0.2
Individual unidentified impuri- 1.0 0.2
ties
Total unspecified impuritiesd 0.9
aCefdinir related compound A is a mixture of 4 isomers designated as lactam ring cleavage lactones a, b, c, and d. The sum of all values is reported, and the
total limit for all 4 isomers combined is 3.3%.
bImpurity III is a mixture of 2 isomers designated as Impurity IIIa and Impurity IIIb. The sum of both values is reported, and the total limit for both isomers
combined is 1.5%.
cImpurity B, Impurity D, and Impurity E are unidentified impurities.
dThe total unidentified impurities limit includes the % total of unidentified impurities B, D, and E and any other unidentified impurities detected.
USP 32 Official Monographs / Cefepime 129
USP REFERENCE STANDARDS 11 CS = concentration of USP Cefepime Hydrochloride RS

USP Cefdinir RS in the Standard solution (mg/mL)
USP Cefdinir Related Compound A RS CU = concentration of Cefepime Hydrochloride in the
USP Cefdinir Related Compound B RSUSP32 Sample solution (mg/mL)
P = content of cefepime in USP Cefepime
Hydrochloride RS (g/mg)
Cefepime Hydrochloride
(Comment on this Monograph)id=m13984=Cefepime IMPURITIES
Hydrochloride=Ca-Chl-Monos.pdf) Inorganic Impurities
Organic Impurities
PROCEDURE 1 : LIMIT OF N-METHYLPYRROLIDINE
Mobile phase: Acetonitrile and 0.01 N nitric acid (1:100)
Column rinse solution: Transfer 5.0 mL of nitric acid to a
1-L volumetric flask. Dilute with water to volume, and mix.
Transfer this solution to an appropriate flask, add 1 L of
C19H25ClN6O5S2 HCl H2O 571.50 acetonitrile, and mix.
Pyrrolidinium, 1-[[7-[[(2-amino-4- Standard stock solution: Weigh 0.16 mL of N-
thiazolyl)(methoxyimino)acetyl]amino]-2-carboxy-8-oxo-5- methylpyrrolidine. Transfer to a 100-mL volumetric flask
thia-1-azabicyclo[4.2.0]oct-2-en-3-yl]methyl]-1-methyl-, and dilute with water to volume.
chloride, monohydrochloride, monohydrate, [6R-[6,7(Z)]]-; Standard solution: 4 mL of Standard stock solution diluted
1-[[(6R,7R)-7-[2-(2-Amino-4-thiazolyl)glyoxylamido]-2- to 100 mL with 0.01 N nitric acid
carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3- [NOTEThis solution contains 0.05 mg/mL of N-
yl]methyl]-1-methylpyrrolidinium chloride, 72-(Z)-(O- methylpyrrolidine.]
methyloxime), monohydrochloride, monohydrate Sample solution: 10 mg/mL of Cefepime Hydrochloride in
[123171-59-5]. 0.01 N nitric acid
[NOTEThis solution may be kept up to 6 h if maintained
DEFINITION at 5; otherwise, use this solution within 30 min.]
Cefepime Hydrochloride contains the equivalent of NLT 825 g Chromatographic system
and NMT 911 g of cefepime (C19H24N6O5S2)/mg, calculated (See Chromatography 621, System Suitability.)
on the anhydrous basis. Mode: LC
Detector: Conductivity
IDENTIFICATION [NOTEThe typical background conductance is 3500 S.]
INFRARED ABSORPTION 197M Column: 4.6-mm 5-cm; 5-m packing L52
Sample: Proceed as directed in the chapter, but do not dry. Guard column: 4.4-mm 5-cm; packing L17, placed
between the pump and the injector
ASSAY Flow rate: 1 mL/min
PROCEDURE Injection size: 100 L
Solution A: Dissolve 5.76 g of sodium 1-pentanesulfonate System suitability
in 2000 mL of water, adjust with glacial acetic acid to a pH Sample: Standard solution
of 3.4, and then with potassium hydroxide TS to a pH of Suitability requirements
4.0. Retention time: NLT 8 min for N-methylpyrrolidine
Mobile phase: Acetonitrile and Solution A (3:47) Relative standard deviation: NMT 5.0%
Standard solution: 1.4 mg/mL of USP Cefepime Analysis
Hydrochloride RS in Mobile phase Samples: Standard solution and Sample solution
Sample solution: 1.4 mg/mL of Cefepime Hydrochloride in Calculate the percentage of N-methylpyrrolidine in the
Mobile phase portion of Cefepime Hydrochloride taken:
(See Chromatography 621, System Suitability.) Result = (rU/rS) (CS/CU) 100
Mode: LC
Detector: UV 254 nm rU = peak response of N-methylpyrrolidine from the
Column: 3.9-mm 30-cm; packing L1 Sample solution
Flow rate: 2 mL/min rs = peak response of N-methylpyrrolidine from the
Injection size: 10 L Sample solution
System suitability CS = concentration of N-methylpyrrolidine in the
Sample: Standard solution Standard solution (mg/mL)
Suitability requirements CU = concentration of Cefepime Hydrochloride in the
Column efficiency: NLT 1500 theoretical plates Sample solution (mg/mL)
Tailing factor: NMT 1.7 Acceptance criteria: NMT 0.3%
Relative standard deviation: NMT 2.0% [NOTECefepime from the Sample solution elutes as a
Analysis broad peak at 55 min. To minimize the equilibration time
Samples: Standard solution and Sample solution at the start of the next day, it is recommended that the
Calculate the quantity, in g, of C19H24N6O5S2 in each mg detector be turned on the night before and that the
of Cefepime Hydrochloride taken: Mobile phase be pumped through the system overnight
at a flow rate of 0.2 mL/min. After every Sample solution
Result = (rU/rS) (CS/CU) P injection, it is recommended that the chromatograph be
flushed with Column rinse solution for 30 min at a flow
rU = peak response from the Sample solution rate of 1 mL/min to remove cefepime from the column,
130 Cefepime / Official Monographs USP 32
and that the system then be switched back to Mobile SPECIFIC TESTS
phase at a flow rate of 1 mL/min for reequilibration.] BACTERIAL ENDOTOXINS TESTS 85: Where the label states
PROCEDURE 2 that Cefepime Hydrochloride is sterile or that it must be
Solution A: 0.68mg/mL of monobasic potassium subjected to further processing during the preparation of
phosphate injectable dosage forms, it contains NMT 0.04 USP
Solution B: Acetonitrile and Solution A (1:9) Endotoxin Unit/mg of cefepime hydrochloride.
Adjust with potassium hydroxide or phosphoric acid to a STERILITY TESTS 71: Where the label states that Cefepime
pH of 5.0. Hydrochloride is sterile, it meets the requirements when
Solution C: Acetonitrile and Solution A (1:1) tested as directed for Test for Sterility of the Product to be
Adjust with potassium hydroxide or phosphoric acid to a Examined, Membrane Filtration.
pH of 5.0. CRYSTALLINITY 695: Meets the requirements
Mobile phase: See the gradient table below. WATER DETERMINATION, Method I 921: 3.0%4.5%
Time (min) Solution B (%) Solution C (%)
0 100 0 containers, and store at controlled room temperature.
10 100 0 LABELING: Where it is intended for use in preparing
30 50 50
35 50 50 preparation of injectable dosage forms.
USP Cefepime Hydrochloride RS
System suitability solution: 1.4 mg/mL of USP Cefepime USP Cefepime Hydrochloride System Suitability RS
Hydrochloride System Suitability RS in Solution B USP Endotoxin RS
Sample solution: 1.4 mg/mL of Cefepime Hydrochloride in
Solution B
[NOTEInject this solution immediately, or store in a Cefepime for Injection
refrigerator, and inject within 12 h.]
Chromatographic system (Comment on this Monograph)id=m13987=Cefepime for
(See Chromatography 621, System Suitability.) Injection=Ca-Chl-Monos.pdf)
Mode: LC
Column: 4.6-mm 25-cm; 5-m packing L1 Cefepime for Injection is a sterile mixture of Cefepime
Flow rate: 1 mL/min Hydrochloride and Arginine. It contains the equivalent of NLT
Injection size: 10 L 90.0% and NMT 115.0% of the labeled amount of cefepime
System suitability (C19H24N6O5S2).
Samples: System suitability solution and Sample solution IDENTIFICATION
Suitability requirements A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201
Resolution: NLT 5 between cefepime and cefepime Standard solution: 20 mg/mL of arginine
related compound A, and NLT 10 between cefepime Sample solution: 40 mg/mL of Cefepime for Injection
related compound A and cefepime related compound B, Developing solvent system: n-Propyl alcohol, ammonium
System suitability solution hydroxide, and water (7:4:5)
Capacity factor, k: NLT 0.6, Sample solution Analysis
Column efficiency: NLT 4000 theoretical plates, Sample Samples: Sample solution and Standard solution
solution Proceed as directed in Chromatography 621, Thin-Layer
Tailing factor: NMT 1.5, Sample solution Chromatography, except to spray the plate with ninhydrin
Analysis TS.
Sample: Sample solution Acceptance criteria: Arginine appears as a dark red spot.
Calculate the percentage of each impurity in the portion of The intensity and the RF value of the spot from the Sample
Cefepime Hydrochloride taken: solution correspond to those from the Standard solution.
Result = (rU/rT) 100 solution corresponds to that of the Standard solution, as
rU = peak response for each impurity obtained in the Assay.
rT = sum of all the peak responses ASSAY
Acceptance criteria: See the Impurity Table 1. PROCEDURE
Solution A: 2.88 mg/mL of sodium 1-pentanesulfonate
Impurity Table 1 Adjust with glacial acetic acid to a pH of 3.4, and then with
potassium hydroxide TS to a pH of 4.0.
Impurity Name Time (%) Factor NMT (%)
Standard solution: 1.4 mg/mL of USP Cefepime
Hydrochloride RS in Mobile phase
Cefepime 1.0 Sample solution: 1 mg/mL of cefepime from one container
Cefepime related 2.7 0.3 of Cefepime for Injection with the volume of water specified
compound A in the labeling
Cefepime related 4.3 0.1
[NOTEUsing a suitable hypodermic needle and syringe,
compound B
withdraw the entire contents of the vial, and dilute in
Mobile phase.]
Any other impurity 0.1
USP 32 Official Monographs / Cefepime 131
Chromatographic system rU = peak response of N-methylpyrrolidine from the

(See Chromatography 621, System Suitability.) Sample solution
Mode: LC rS = peak response of N-methylpyrrolidine from the
Detector: UV 254 nm Standard solution
Column: 3.9-mm 30-cm; packing L1 CS = concentration of N-methylpyrrolidine in the
Flow rate: 2 mL/min Standard solution (mg/mL)
Injection size: 10 L CU = nominal concentration of cefepime in the
System suitability Sample solution (mg/mL)
Sample: Standard solution Acceptance criteria: NMT 1.0%
Suitability requirements [NOTECefepime from the Sample solution elutes as a
Column efficiency: NLT 1500 theoretical plates broad peak at 55 min. To minimize equilibration time at
Tailing factor: NMT 1.7 the start of the next day, it is recommended that the
Relative standard deviation: NMT 2.0% detector be turned on the night before and that Mobile
Analysis phase be pumped through the system overnight at a
Samples: Sample solution and Standard solution flow rate of 0.2 mL/min. After every Sample solution
Calculate the percentage of C19H24N6 O5S2 in the container injection, it is recommended that the chromatograph be
of Cefepime for Injection: flushed with Column rinse solution for 30 min at a flow
rate of 1 mL/min to remove cefepime from the column
Result = (rU/rS) (CS/CU) P 100 and that the system then be switched back to Mobile
phase at a flow rate of 1 mL/min for reequilibration.]
rU = peak response from the Sample solution PROCEDURE 2: RELATED COMPOUNDS
rS = peak response from the Standard solution Solution A: 0.68 mg/mL of monobasic potassium
CS = concentration of USP Cefepime Hydrochloride RS phosphate
in the Standard solution (mg/mL) Solution B: Acetonitrile and Solution A (1:9)
CU = nominal concentration of cefepime Adjust with potassium hydroxide or phosphoric acid to a
hydrochloride in the Sample solution (mg/mL) pH of 5.0.
P = content of cefepime in the USP Cefepime Solution C: Acetonitrile and Solution A (1:1)
Hydrochloride RS (g/mg) Adjust with potassium hydroxide or phosphoric acid to a
Acceptance criteria: 90.0%115.0% pH of 5.0.
PERFORMANCE TESTS
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Time Solution B Solution C
IMPURITIES (min) (%) (%)
Organic Impurities 0 100 0
PROCEDURE 1: LIMIT OF N-METHYLPYRROLIDINE
Mobile phase: Acetonitrile and 0.01 N nitric acid (1:100) 10 100 0
Column rinse solution: Transfer 5.0 mL of nitric acid to a 30 50 50
1-L volumetric flask. Dilute with water to volume. Transfer 35 50 50
this solution to an appropriate flask, and add 1 L of
acetonitrile. 36 100 0
Standard stock solution: 1.6 mL of N-
methylpyrrolidine/100 mL System suitability solution: 1.4 mg/mL of USP Cefepime
Standard solution: Standard stock solution and 0.01 N Hydrochloride System Suitability RS in Solution B
nitric acid (1:24) Sample solution: Constitute one container of Cefepime for
[NOTEThis solution contains 0.05 mg/mL of N- Injection with a volume of Solution B equivalent to the
methylpyrrolidine.] volume of solvent specified in the labeling, and shake to
Sample solution: 10 mg/mL of cefepime from one dissolve. Transfer the constituted solution to a volumetric
container of Cefepime for Injection with the volume of water flask, and dilute with Solution B to obtain a solution having
specified in the labeling, in 0.05 N nitric acid a concentration of 2 mg of cefepime/mL.
[NOTEInject this solution immediately.] [NOTEInject this solution immediately, or store in a
Chromatographic system refrigerator and inject within 12 h.]
(See Chromatography 621, System Suitability.) Chromatographic system
Mode: LC (See Chromatography 621, System Suitability.)
Detector: Conductivity Mode: LC
[NOTEThe typical background conductance is 3500 S.] Detector: UV 254 nm
Column: 4.6-mm 5-cm; 5-m packing L52 Column: 4.6-mm 25-cm; 5-m packing L1
Guard column: 4.4-mm 5-cm; packing L17, placed Flow rate: 1 mL/min
between the pump and the injector Injection size: 10 L
Flow rate: 1 mL/min System suitability
Injection size: 100 L Sample: System suitability solution and Sample solution
Sample: Standard solution Resolution: NLT 5 between cefepime and cefepime
Suitability requirements related compound A; NLT 10 between cefepime related
Retention time: NLT 8 min for N-methylpyrrolidine compound A and cefepime related compound B, System
Relative standard deviation: NMT 5.0% suitability solution
Analysis Capacity factor, k: NLT 0.6, Sample solution
Samples: Standard solution and Sample solution Column efficiency: NLT 4000 theoretical plates, Sample
Calculate the percentage of N-methylpyrrolidine in the solution
portion of Cefepime for Injection taken:
132 Cefepime / Official Monographs USP 32
Tailing factor: NMT 1.5, Sample solution DEFINITION

Analysis Cefixime contains the equivalent of NLT 950 g and NMT 1030
Sample: Sample solution g of C16H15N5O7S2/mg, calculated on the anhydrous basis.
Calculate the percentage of each impurity in the portion of
Cefepime for Injection taken: IDENTIFICATION
Result = (rU/rT) 100 Sample: Dissolve 5 mg of it by trituration in 2 mL of
methanol and evaporate with the aid of gentle heat to
rU = peak response for each impurity dryness.
rT = sum of the responses for all the peaks
Acceptance criteria: See Impurity Table 1. ASSAY
PROCEDURE
Solution A: 25 mL of 0.4 M tetrabutylammonium
Impurity Table 1
hydroxide solution diluted to 1000 mL with water, adjusted
Relative Acceptance with 1.5 M phosphoric acid to a pH of 6.5
Retention Criteria, Solution B: Dissolve 6.8 g of monobasic potassium
Name Time NMT (%) phosphate in water to 500 mL.
Cefepime related compound A 2.7 0.5 Solution C: Dissolve 7.1 g of anhydrous dibasic sodium
phosphate in water to 500 mL.
Cefepime related compound B 4.3 0.5 Buffer: Adjust a volume of Solution C with a sufficient
Any other impurity 0.5 volume of Solution B to a pH of 7.0.
Cefepime 1.0 Mobile phase: Acetonitrile and Solution A (1:3)
System suitability solution: 1 mg/mL of USP Cefixime RS
in water
SPECIFIC TESTS Heat this solution at 95 in an oil bath for 45 min, cool, and
INJECTIONS, Constituted Solutions 1: At the time of use, it use promptly.
meets the requirements. Standard solution: 0.2 mg/mL of USP Cefixime RS in Buffer
BACTERIAL ENDOTOXINS TEST 85: NMT 0.06 USP Endotoxin Use this solution promptly.
Unit/mg of cefepime Sample solution: 0.22 mg/mL Cefixime in Buffer
STERILITY TESTS 71: Meets the requirements when tested as Use this solution promptly.
directed for Test for Sterility of the Product to Be Examined, Chromatographic system
Membrane Filtration (See Chromatography 621, System Suitability.)
PH 791: 4.06.0, in a solution containing 100 mg/mL of Mode: LC
cefepime Detector: UV 254 nm
WATER DETERMINATION, Method I 921: NMT 4.0% Column: 4.6-mm 12.5-cm; 4-m packing L1
OTHER REQUIREMENTS: Meets the requirements for Injections Temperature: 40
1, Labels and Labeling Flow rate: Adjusted so that the retention time of cefixime
is 10 min
PACKAGING AND STORAGE: Preserve in tight, light-resistant as System suitability
described under Injections 1, Containers for Sterile Solids, Samples: System suitability solution and Standard solution
and store in a refrigerator or at controlled room [NOTERelative retention times are about 0.9 for cefixime
temperature. Store reconstituted powder in a refrigerator for (E)-isomer and 1.0 for cefixime.]
NMT 7 days. Suitability requirements
LABELING: Label it to indicate that it is to be diluted with a Resolution: NLT 2.0 between cefixime and cefixime (E)-
suitable parenteral vehicle before intravenous infusion. isomer, System suitability solution
USP REFERENCE STANDARDS 11 Column efficiency: NLT 4000 theoretical plates, Standard
USP Cefepime Hydrochloride RS solution, when calculated:
USP Cefepime Hydrochloride System Suitability RS
USP Endotoxin RS 5.545(t/Wh/2)2
Tailing factor: For the analyte peak, between 0.9 and
2.0, Standard solution, when calculated:
Cefixime
(Comment on this Monograph)id=m13990=Cefixime=Ca-Chl- W0.1/2f
Monos.pdf)
W0.1 = width of peak of 10% height
solution
Analysis
Calculate the quantity, in g, of C16H15N5O7S2 in each mg
of Cefixime taken:
C16H15N5O7S2 3H2O 507.50 Result = (rU/rS) (CS/CU) F

5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[(2-
amino-4-thiazolyl)[(carboxymethoxy)imino]acetyl]amino]-3- rU = peak response of cefixime from the Sample
ethenyl-8-oxo-, trihydrate, [6R-[6,7(Z)]]-; solution
(6R,7R)-7-[2-(2-Amino-4-thiazolyl)glyoxylamido]-8-oxo-3-vinyl-5- rS = peak response of cefixime from the Standard
thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 72-(Z)-[O- solution
(carboxymethyl)oxime]trihydrate [79350-37-1]. CS = concentration of cefixime in the Standard
Anhydrous 453.46. solution (mg/mL)
USP 32 Official Monographs / Cefixime 133
CU = concentration of Cefixime in the Sample solution ASSAY

(mg/mL) PROCEDURE
F = conversion factor, 1000 g/mg Solution A: 0.4 M tetrabutylammonium hydroxide solution
Acceptance criteria: 9501030 g/mg and water (1:39)
Adjust with 1.5 M phosphoric acid to a pH of 6.5.
IMPURITIES Solution B: 13.6 mg/mL of monobasic potassium
Organic Impurities phosphate
PROCEDURE 1 Solution C: 14.2 mg/mL of anhydrous dibasic sodium
Solution A, Solution B, Solution C, Buffer, Mobile phase, phosphate
System suitability solution, Standard solution and Adjust a volume of this solution with a sufficient volume of
Sample solution: Proceed as directed in the Assay. Solution B to a pH of 7.0.
Chromatographic system: Proceed as directed in the Mobile phase: Acetonitrile and Solution A (1:3)
Assay. System suitability solution: 1 mg/mL of USP Cefixime RS
System suitability: Proceed as directed in the Assay. [NOTEHeat this solution at 95 in an oil bath for 45 min,
Analysis cool, and use promptly.]
Samples: Standard solution and Sample solution Standard solution: 0.2 mg/mL of USP Cefixime RS in
Calculate the percentage of each impurity in the portion Solution C
of Cefixime taken: [NOTEUse this solution promptly.]
Sample solution: Constitute Cefixime for Oral Suspension
Result = (rU/rS) P F 100 as directed in the labeling. Quantitatively dilute a measured
volume of the suspension thus obtained, freshly mixed and
rU = peak area for each impurity free from air bubbles, with Solution C to obtain a solution
rS = cefixime peak area having a nominal concentration of 0.2 mg of cefixime/mL.
P = potency of cefixime calculated in the Assay Chromatographic system
(g/mg) (See Chromatography 621, System Suitability.)
F = conversion factor, 0.001 mg/g Mode: LC
Acceptance criteria Detector: UV 254 nm
Individual impurities: NMT 1.0% of any individual Column: 4.6-mm 12.5-cm; 4-m packing L1
impurity is found. Temperature: 40
Total impurities: NMT 2.0% Flow rate: 10 min
SPECIFIC ROTATION 781S: 75 to 88 System suitability
Sample solution: 10 mg/mL, in sodium bicarbonate Sample: System suitability solution and Standard solution
solution (2 in 100) [NOTEThe relative retention times for cefixime (E)-isomer
CRYSTALLINITY 695: Meets the requirements and cefixime are about 0.9 and 1.0, respectively.]
PH 791: 2.64.1 Suitability requirements
Sample solution: Contains the equivalent of 0.7 mg/mL of Resolution: NLT 2.0 between cefixime and cefixime (E)-
cefixime isomer, System suitability solution
WATER DETERMINATION, Method I 921: 9.0%12.0% Column efficiency: NLT 4000 theoretical plates when
calculated:
PACKAGING AND STORAGE: Preserve in tight containers. 5.545(t/Wh/2)2, Standard solution
LABELING: Label to indicate that it is the trihydrate form.
Where the quantity of Cefixime is indicated in the labeling of Tailing factor: NLT 0.9 and NMT 2.0 for the analyte peak
any preparation containing Cefixime, this shall be when calculated:
understood to be in terms of anhydrous cefixime
(C16H15N5O7S2). W0.1/2f
USP REFERENCE STANDARDS 11 W0.1 = width of peak of 10% height, Standard solution
USP Cefixime RS Relative standard deviation: NMT 2.0%, Standard
solution
Analysis
Cefixime for Oral Suspension Samples: Standard solution and Sample solution
Calculate the percentage of C16H15N5O7S2 in the constituted
(Comment on this Monograph)id=m13992=Cefixime for Oral suspension prepared from the Cefixime for Oral
Suspension=Ca-Chl-Monos.pdf) Suspension:
DEFINITION
Cefixime for Oral Suspension is a dry mixture of Cefixime and Result = (rU/rS) (CS/CU) 100
one or more suitable diluents, flavors, preservatives, and rU = peak response of cefixime from the Sample
suspending agents. It contains the equivalent of NLT 90.0% solution
and NMT 120.0% of the labeled amount of cefixime rS = peak response of cefixime from the Standard
(C16H15N5O7S2)/mL when constituted as directed in the solution
labeling. CS = concentration of USP Cefixime RS in the
IDENTIFICATION Standard solution (mg/mL)
The retention time of the major peak of the Sample solution CU = nominal concentration of cefixime in the Sample
corresponds to that of the Standard solution, as obtained in solution (mg/mL)
the Assay.
134 Cefixime / Official Monographs USP 32
Acceptance criteria: 90.0%120.0% Injection size: 10 L

System suitability
PERFORMANCE TESTS Sample: System suitability solution and Standard solution
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements [NOTEThe relative retention times for cefixime (E)-isomer
for solids packaged in single-unit containers and cefixime are about 0.9 and 1.0, respectively.]
DELIVERABLE VOLUME 698: Meets the requirements Suitability requirements
Resolution: NLT 2.0 between cefixime and cefixime (E)-
SPECIFIC TESTS isomer, System suitability solution
PH 791: 2.54.5, in the suspension constituted as directed Column efficiency: NLT 4000 theoretical plates for the
in the labeling Standard solution when calculated:
5.545(t/Wh/2)2
PACKAGING AND STORAGE: Preserve in tight containers. Tailing factor: NLT 0.9 and NMT 2.0 for the analyte peak
LABELING: Label it to indicate that the cefixime contained when calculated:
therein is in the trihydrate form.
USP REFERENCE STANDARDS 11 W0.1/2f
USP Cefixime RS
W0.1 = width of peak of 10% height, Standard solution
solution
Cefixime Tablets Analysis
(Comment on this Monograph)id=m13994=Cefixime Samples: Standard solution and Sample solution
Tablets=Ca-Chl-Monos.pdf) Calculate the percentage of C16H15N5O7S2 in the portion of
Cefixime Tablets taken:
DEFINITION
Cefixime Tablets contain the equivalent of NLT 90.0% and NMT Result = (rU/rS) (CS/CU) 100
110.0% of the labeled amount of cefixime (C16H15N5O7S2).
rU = peak response of cefixime from the Sample
A. The retention time of the major peak of the Sample rS = peak response of cefixime from the Standard
solution corresponds to that of the Standard solution, as solution
obtained in the Assay. CS = concentration of USP Cefixime RS in the Standard
solution (mg/mL)
ASSAY CU = nominal concentration of cefixime in the Sample
PROCEDURE solution (mg/mL)
Solution A: 0.4 M tetrabutylammonium hydroxide solution Acceptance criteria: 90.0%110.0%
and water (1:39)
Adjust with 1.5 M phosphoric acid to a pH of 6.5. PERFORMANCE TESTS
Solution B: 13.6 mg/mL of monobasic potassium DISSOLUTION 711
phosphate Medium: 6.8 mg/mL of monobasic potassium phosphate in
Solution C: 14.2 mg/mL of anhydrous dibasic sodium 1000 mL of water.
phosphate Adjust with 1 N sodium hydroxide to a pH of 7.2; 900 mL.
Adjust a volume of this solution with a sufficient volume of Apparatus 1: 100 rpm
Solution B to a pH of 7.0. Time: 45 min
Mobile phase: Acetonitrile and Solution A (1:3) Detector: UV 288 nm
System suitability solution: 1 mg/mL of USP Cefixime RS Standard solution: USP Cefixime RS in Medium
[NOTEHeat this solution at 95 in an oil bath for 45 min, Sample solutions: Sample per Dissolution 711. Dilute with
cool, and use promptly.] Medium to concentration similar to that of the Standard
Standard solution: 0.2 mg/mL of USP Cefixime RS in solution
Solution C [NOTEAn amount of methanol not to exceed 0.1% of the
[NOTEUse this solution promptly.] total volume of the Standard solution may be used to
Sample stock solution: Transfer powdered tablets bring the USP Cefixime RS into solution prior to dilution
equivalent to about 400 mg of cefixime (NLT 20 tablets), to with Medium, and the solution may be sonicated to
a 100-mL volumetric flask, add 75 mL of Solution C and ensure complete dissolution of the USP Cefixime RS.]
sonicate. Dilute with Solution C to volume, mix, and Tolerances: NLT 75% (Q)
centrifuge. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Sample solution: Nominal concentration of 0.2 mg/mL of
Sample stock solution in Solution C SPECIFIC TESTS
Chromatographic system WATER DETERMINATION, Method I 921: NMT 10.0%
Column: 4.6-mm 12.5-cm; 4-m packing L1 LABELING: Label the Tablets to indicate that the cefixime
Temperature: 40 contained therein is in the trihydrate form.
Flow rate: Adjust it so that the retention time of cefixime USP REFERENCE STANDARDS 11
is about 10 min. USP Cefixime RS
USP 32 Official Monographs / Cefmenoxime 135
Cefmenoxime Hydrochloride Mode: LC

(Comment on this Monograph)id=m14000=Cefmenoxime Detector: UV 254 nm
Hydrochloride=Ca-Chl-Monos.pdf) Column: 4-mm 15-cm; packing L1
Flow rate: 1 mL/min
System suitability
Resolution: NLT 2.3 between the phthalimide and
cefmenoxime peaks
(C16H17N9O5S3)2 HCl 1059.58 cefmenoxime peak
5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[(2- Tailing factor: NMT 1.6 for the cefmenoxime peak
amino-4-thiazolyl)(methoxyimino)acetyl]amino]-3-[[(1- Relative standard deviation: NMT 2.0%
methyl-1H-tetrazol-5-yl)thio]methyl]-8-oxo-, hydrochloride Analysis
(2:1), [6R-[6,7(Z)]]-; Samples: Standard solution and Sample solution
(6R,7R)-7-[2-(2-Amino-4-thiazolyl)glyoxylamido]-3-[[(1- Calculate the quantity of C16H17N9O5S3 g in each mg of
methyl-1H-tetrazol-5-yl)-thio]methyl]-8-oxo-5-thia-1- Cefmenoxime Hydrochloride taken:
azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid 72-(Z)-(O-
methyloxime), hydrochloride (2:1) [75738-58-8]. Result = (RU/RS) (CS/CU) P
DEFINITION RU = peak area ratio of cefmenoxime to the internal

Cefmenoxime Hydrochloride contains the equivalent of NLT standard from the Sample solution
869 g and NMT 1015 g of C16H17N9O5S3/mg, calculated on RS = peak area ratio of cefmenoxime to the internal
the anhydrous basis. standard from the Standard solution
CS = concentration of USP Cefmenoxime
IDENTIFICATION Hydrochloride RS in the Standard solution
A. ULTRAVIOLET ABSORPTION 197U (mg/mL)
Solution: 25 g/mL in Solution A prepared as directed in CU = concentration of Cefmenoxime Hydrochloride in
the Assay. the Sample solution (mg/mL)
B. The retention time of the cefmenoxime peak of the P = designated C16H17N9O5S3 content of USP
Sample solution corresponds to that of the Standard solution, Cefmenoxime Hydrochloride RS (g/mg)
both relative to the internal standard, as obtained in the Acceptance criteria: 8691015 g/mg
Assay.
SPECIFIC TESTS
ASSAY CRYSTALLINITY 695: Meets the requirements
PROCEDURE PYROGEN TEST 151: Where the label states that it is sterile
Solution A: 6.4 mg/mL of monobasic potassium phosphate or must be subjected to further processing during the
and 18.9 mg/mL of dibasic sodium phosphate in water solution of injectable dosage forms, it meets the
Adjust with 1 N sodium hydroxide to a pH of 6.8 0.1. requirements, the test dose being 1.0 mL/kg of a solution in
Mobile phase: Acetonitrile, glacial acetic acid, and water pyrogen-free sodium carbonate solution (prepared by
(10:1:50) dissolving 14.0 g of sodium carbonate, previously heated at
Filter through a suitable filter of 0.5 m or finer porosity. 170 for NLT 4 h, in 1000 mL of sterile water for injection)
Internal standard solution: 1.5 mg/mL of phthalimide in containing 60 mg/mL.
methanol STERILITY TESTS 71: Where the label states that it is sterile,
Standard solution: Transfer 50 mg of USP Cefmenoxime it meets the requirements when tested as directed under Test
Hydrochloride RS to a 50-mL volumetric flask, add 10 mL of for Sterility of the Product to Be Examined, Membrane Filtration,
pH 6.8 buffer, and dissolve by swirling. Dilute with Mobile except to add 2.0 g of sodium carbonate (previously
phase to volume. Transfer 4.0 mL of this solution to a sterilized by heating at 180 for 2 h) to each 100 mL of
second 50-mL volumetric flask, add 20.0 mL of Internal Fluid A.
standard solution, and dilute with Mobile phase to volume. WATER DETERMINATION, Method I 921: NMT 1.5%
[NOTEThis solution contains the equivalent of 80 g/mL Sample solution: Prepared as directed for a hygroscopic
of C16H17N9O5S3.] specimen, except use 20 mL of a mixture of formamide
Sample stock solution: Transfer 50 mg of Cefmenoxime (previously dried over anhydrous sodium sulfate for 24 h)
Hydrochloride RS to a 50-mL volumetric flask, add 10 mL of and methanol (2:1), instead of methanol, to dissolve the
pH 6.8 buffer, and dissolve by swirling. Dilute with Mobile specimen, use two 5-mL portions of the same formamide
phase to volume. Transfer 4.0 mL of this solution to a and methanol mixture to rinse the container, and determine
second 50-mL volumetric flask, add 20.0 mL of Internal the water content of the formamide and methanol mixture.
standard solution, and dilute with Mobile phase to volume.
Sample solution: 2.0 mL of Sample stock solution and 10.0 ADDITIONAL REQUIREMENTS
mL of Internal standard solution PACKAGING AND STORAGE: Preserve in tight containers.
Dilute with Mobile phase to 25.0 mL. LABELING: Where it is intended for use in preparing
Chromatographic system injectable dosage forms, the label states that it is sterile or
136 Cefmenoxime / Official Monographs USP 32
must be subjected to further processing during the Chromatographic system

preparation of injectable dosage forms. (See Chromatography 621, System Suitability.)
USP Cefmenoxime Hydrochloride RS Detector: UV 254 nm
Flow rate: 1 mL/min
Cefmenoxime for Injection System suitability
(Comment on this Monograph)id=m14002=Cefmenoxime for Sample: Standard solution
Injection=Ca-Chl-Monos.pdf) Suitability requirements
Resolution: NLT 2.3 between the phthalimide and the
DEFINITION cefmenoxime peaks
Cefmenoxime for Injection contains an amount of Column efficiency: NLT 1200 theoretical plates from the
Cefmenoxime Hydrochloride equivalent to NLT 90.0% and cefmenoxime peak when calculated based on the width of
NMT 115.0% of the labeled amount of cefmenoxime the peak at half height
(C16H17N9O5S3). It may contain sodium carbonate. Tailing factor: NMT 1.6 for the cefmenoxime peak
A. ULTRAVIOLET ABSORPTION 197U Samples: Sample solution A or Sample solution B, and
Solution: 25 g/mL in Solution A (prepared as directed in Standard solution
the Assay) [NOTEUse peak areas where peak responses are
B. The retention time of the major cefmenoxime peak in indicated.]
the chromatogram of the Sample solution corresponds to Calculate the percentage of C16H17N9O5S3 withdrawn from
that of the Standard solution, both relative to the internal the container or in the portion of constituted solution
standard, as obtained in the Assay. taken:
ASSAY
PROCEDURE Result = (RU/RS) (CS/CU) 100
Solution A: 6.4 mg/mL of monobasic potassium phosphate RU = peak response ratio of the cefmenoxime peak to
and 18.9 mg/mL of dibasic sodium phosphate in water the internal standard peak from the Sample
Adjust with 1 N sodium hydroxide to a pH of 6.8 0.1. solution
Mobile phase: Acetonitrile, glacial acetic acid, and water RS = peak response ratio of the cefmenoxime peak to
(10:1:50) the internal standard peak from the Standard
Filter through a suitable filter of 0.5 m or finer porosity. solution
Internal standard solution: 1.5 mg/mL of phthalimide in CS = concentration of USP Cefmenoxime
methanol Hydrochloride RS in the Standard solution
Standard stock solution: 1 mg/mL of USP Cefmenoxime (mg/mL)
Hydrochloride RS in Solution A and Mobile phase (1:4) CU = nominal concentration of cetmenoxime in
Standard solution: 2.0 mL of Standard stock solution and Sample solution A or Sample solution B (mg/mL)
10.0 mL of Internal standard solution Acceptance criteria: 90.0%115.0%
Dilute with Mobile phase to 25.0 mL.
[NOTEThis solution contains the equivalent of 80 g/mL SPECIFIC TESTS
of cefmenoxime.] PARTICULATE MATTER IN INJECTIONS 788: It meets the
Sample stock solution A (where it is represented as being requirements for small-volume injections.
in a single-dose container): 1 mg/mL of cefmenoxime from PYROGEN TEST 151: It meets the requirements, the test
a container of Cefmenoxime for Injection in a volume of dose being 1.0 mL/kg of a solution of Cefmenoxime for
water corresponding to the volume of diluent specified in Injection in Sterile Water for Injection having a concentration
the labeling [NOTEWithdraw all of the withdrawable of 60 mg of cefmenoxime/mL.
contents, using a suitable hypodermic needle and syringe.], STERILITY TESTS 71: It meets the requirements when tested
and quantitatively diluted with water. as directed under Test for Sterility of the Product to be
Sample solution A: 2.0 mL of Sample stock solution A and Examined, Membrane Filtration
10.0 mL of Internal standard solution PH 791: 6.47.9, in a solution containing the equivalent of
Dilute with Mobile phase to 25.0 mL. [ NOTEThis solution 100 mg/mL of cefmenoxime
contains the equivalent of 80 g/mL of cefmenoxime.] LOSS ON DRYING 731: Dry 100 mg in vacuum at a pressure
Sample stock solution B (where the label states the not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT
quantity of cefmenoxime in a given volume of constituted 1.5% of its weight.
solution): 1 mg/mL of cefmenoxime from a container of
Cefmenoxime for Injection in a volume of water equivalent ADDITIONAL REQUIREMENTS
to the volume of diluent specified in the labeling, and PACKAGING AND STORAGE: Preserve as described under
quantitatively diluted with water Injections 1, Containers for Sterile Solids
Sample solution B: 2.0 mL of Sample stock solution B and USP REFERENCE STANDARDS 11
10.0 mL of Internal standard solution USP Cefmenoxime Hydrochloride RS
Dilute with Mobile phase to 25.0 mL. [NOTEThis solution
contains the equivalent of 80 g/mL of cefmenoxime.]
USP 32 Official Monographs / Cefmetazole 137
Cefmetazole Analysis
(Comment on this Monograph)id=m14011=Cefmetazole=Ca- Samples: Sample solution and Standard solution
Chl-Monos.pdf) Calculate the quantity, in g, of C15H17N7O5S3 in each mg
of Cefmetazole taken:
Result = (rU/rS) (CS/CU)
rU = peak response of cefmetazole from the Sample

solution
rS = peak response of cefmetazole from the Standard
solution
C15H17N7O5S3 471.52 CS = concentration of USP Cefmetazole RS in the
5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[ Standard solution (g/mL)
[(cyanomethyl)thio]acetyl]amino]-7-methoxy-3-[[(1- CU = nominal concentration of cefmetazole in the
methyl-1H-tetrazol-5-yl)thio]methyl]-8-oxo-, (6R-cis)-; Sample solution (g/mL)
(6R,7S)-7-[2-[(Cyanomethyl)thio]acetamido]-7-methoxy-3-[[(1- Acceptance criteria: 9701030 g/mg
methyl-1H-tetrazol-5-yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo SPECIFIC TESTS
[4.2.0]oct-2-ene-2-carboxylic acid. [56796-20-4]. WATER DETERMINATION, Method I 921: NMT 0.5%
Cefmetazole contains NLT 970 g and NMT 1030 g of PACKAGING AND STORAGE: Preserve in tight containers.
cefmetazole (C15H17N7O5S3)/mg, calculated on the anhydrous USP REFERENCE STANDARDS 11
basis. USP Cefmetazole RS
IDENTIFICATION
B. The retention time of the major peak of the Sample Cefmetazole Injection
obtained in the Assay. (Comment on this Monograph)id=m14013=Cefmetazole
ASSAY
Mobile phase: To 5.75 g of monobasic ammonium Cefmetazole Injection is a sterile isoosmotic solution of
phosphate in 700 mL of water, add 3.2 mL of a 40% Cefmetazole and Sodium Citrate in Water for Injection. It
solution of tetrabutylammonium hydroxide, 280 mL of contains one or more buffer substances and a tonicity-
methanol, and 25 mL of tetrahydrofuran. Adjust with adjusting agent. It contains NLT 90.0% and NMT 120.0% of
phosphoric acid to a pH of 4.5 0.1, and pass through a the labeled amount of cefmetazole (C15H17N7O5S3).
filter having a 0.5-m or finer porosity. IDENTIFICATION
System suitability stock solution: 1 mg/mL of USP The retention time of the major peak of the Sample solution
Cefmetazole RS in 0.01 N sodium hydroxide corresponds to that of the Standard solution, as obtained in
[NOTEHeat at 95 for 10 min.] the Assay.
System suitability solution: Mix 1 mL of the System
suitabiity stock solution and 2 mL of the Standard solution. ASSAY
Dilute with Mobile phase to 20 mL. PROCEDURE
[NOTEThis solution contains cefmetazole and cefmetazole Mobile phase: To 5.75 g of monobasic ammonium
lactone (resolution compound).] phosphate in 700 mL of water, add 3.2 mL of a 40%
Standard solution: 200 g/mL of cefmetazole by solution of tetrabutylammonium hydroxide, 280 mL of
quantitatively dissolving USP Cefmetazole RS in Mobile phase methanol, and 25 mL of tetrahydrofuran, and mix. Adjust
[NOTEUse this solution within 10 min.] with phosphoric acid to a pH of 4.5 0.1, and pass through
Sample solution: 200 g/mL of Cefmetazole in Mobile a filter having a 0.5-m or finer porosity.
phase System suitability stock solution: 1 mg/mL of USP
[NOTEUse this solution within 10 min.] Cefmetazole RS in 0.01 N sodium hydroxide
Chromatographic system [NOTEHeat at 95 for 10 min.]
(See Chromatography 621, System Suitability.) System suitability solution: System suitability stock solution,
Mode: LC Standard solution, and Mobile phase (1:2:17)
Detector: UV 214 nm [NOTEThis solution contains cefmetazole and cefmetazole
Column: 4.6-mm 25-cm; packing L1 lactone (resolution compound).]
Flow rate: 2 mL/min Standard solution: 200 g/mL of cefmetazole by
Injection size: 10 L quantitatively dissolving USP Cefmetazole RS in Mobile phase
System suitability [NOTEUse this solution within 10 min.]
Samples: System suitability solution and Standard solution Sample solution: Equivalent to 200 g/mL of cefmetazole
Suitability requirements from Ceftemetazole Injection
Resolution: NLT 3.0 between cefmetazole and Allow the contents of a container of Injection to thaw, and
cefmetazole lactone, System suitability solution mix the resultant solution in Mobile phase.
Column efficiency: NLT 1250 theoretical plates, Standard [NOTEUse this solution within 10 min.]
solution Chromatographic system
Tailing factor: 0.941.6, Standard solution (See Chromatography 621, System Suitability.)
solution
138 Cefmetazole / Official Monographs USP 32
Mode: LC USP Endotoxin RS

Detector: UV 214 nm
Flow rate: 2 mL/min
Injection size: 10 L Cefmetazole Sodium
System suitability (Comment on this Monograph)id=m14014=Cefmetazole
Samples: System suitability solution and Standard solution Sodium=Ca-Chl-Monos.pdf)
Resolution: NLT 3.0 between cefmetazole and
cefmetazole lactone, System suitability solution
solution
Tailing factor: 0.941.6, Standard solution
solution
Analysis C15H16N7NaO5S3 493.53
Samples: Sample solution and Standard solution 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[
Calculate the percentage of C15H17N7O5S3 in each mL of [(cyanomethyl)thio]acetyl]amino]-7-methoxy-3-[[(1-
Cefmetazole Injection taken: methyl-1H-tetrazol-5-yl)thio]methyl]-8-oxo-, monosodium salt,
(6R-cis)-;
Result = (rU/rS) (CS/CU) 100 Sodium (6R,7S)-7-[2-[(cyanomethyl)thio]acetamido]-7-
methoxy-3-[[(1-methyl-1 H-tetrazol-5-yl)thio]methyl]-8-oxo-5-
rU = peak response of cefmetazole from the Sample thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate
solution [56796-39-5].
rS = peak response of cefmetazole from the Standard
solution DEFINITION
CS = concentration of USP Cefmetazole RS in the Cefmetazole Sodium contains the equivalent of NLT 860 g
Standard solution (g/mL) and NMT 1003 g of cefmetazole (C15H17N7O5S3)/mg,
CU = nominal concentration of cefmetazole taken to calculated on the anhydrous basis.
prepare the Sample solution (g/mL)
SPECIFIC TESTS B. The retention time of the major peak of the Sample
BACTERIAL ENDOTOXINS TEST 85: NMT 0.2 USP Endotoxin solution corresponds to that of the Standard solution, as
Unit/mg of cefmetazole obtained in the Assay.
as directed under Test for Sterility of the Product to Be ASSAY
Examined, Membrane Filtration, except to use water instead PROCEDURE
of Fluid A. Mobile phase: To 5.75 g of monobasic ammonium
PH 791: 4.26.2 phosphate in 700 mL of water, add 3.2 mL of a 40%
PARTICULATE MATTER IN INJECTIONS 788: Meets the solution of tetrabutylammonium hydroxide, 280 mL of
requirements for small-volume injections methanol, and 25 mL of tetrahydrofuran. Adjust with
phosphoric acid to a pH of 4.5 0.1, and pass through a
ADDITIONAL REQUIREMENTS filter having a 0.5-m or finer porosity.
PACKAGING AND STORAGE: Preserve as described under System suitability stock solution: 1 mg/mL of USP
Injections 1, Containers for Injections. Maintain in the frozen Cefmetazole RS in 0.01 N sodium hydroxide
state. [NOTEHeat at 95 for 10 min.]
LABELING: It meets the requirements under Injections 1, System suitability solution: Mix 1 mL of the System
Labeling. The label states that it is to be thawed just before suitability stock solution and 2 mL of the Standard solution.
use, describes the conditions for proper storage of the Dilute with Mobile phase to volume. [NOTEThis solution
resultant solution, and directs that the solution is not to be contains cefmetazole and cefmetazole lactone (resolution
refrozen. compound).]
USP Cefmetazole RS
USP 32 Official Monographs / Cefmetazole 139
Standard solution: 200 g/mL of cefmetazole by IDENTIFICATION

quantitatively dissolving USP Cefmetazole RS in Mobile phase A. INFRARED ABSORPTION 197M
[NOTEUse this solution within 10 min.] B. The retention time of the major peak of the Sample
Sample solution: 210 g/mL of cefmetazole sodium in solution corresponds to that of the Standard solution, as
Mobile phase obtained in the Assay.
[NOTEUse this solution within 10 min.]
(See Chromatography 621, System Suitability.) PROCEDURE
Mode: LC Mobile phase: 5.75 g of monobasic ammonium phosphate
Detector: UV 214 nm in 700 mL of water.
Column: 4.6-mm 25-cm; packing L1 Add 3.2 mL of a 40% solution of tetrabutylammonium
Flow rate: 2 mL/min hydroxide, 280 mL of methanol, and 25 mL of
Injection size: 10 L tetrahydrofuran, and mix. Adjust with phosphoric acid to a
System suitability pH of 4.5 0.1, and pass through a filter having a 0.5-m
Samples: System suitability solution and Standard solution or finer porosity.
Suitability requirements System suitability stock solution: 1 mg/mL of USP
Resolution: NLT 3.0 between cefmetazole and Cefmetazole RS in 0.01 N sodium hydroxide
cefmetazole lactone, System suitability solution [NOTEHeat at 95 for 10 min.]
Column efficiency: NLT 1250 theoretical plates, Standard System suitability solution: 1.0 mL of System suitability
solution stock solution and 2.0 mL of Standard solution
Tailing factor: 0.941.6, Standard solution Dilute with Mobile phase to 20.0 mL.
Relative standard deviation: NMT 2.0%, Standard [NOTEThis solution contains cefmetazole and cefmetazole
solution lactone (resolution compound).]
Analysis Standard solution: 200 g/mL of cefmetazole by
Samples: Sample solution and Standard solution quantitatively dissolving USP Cefmetazole RS in Mobile phase
Calculate the quantity, in g, of C15H17N7O5S3/mg of [NOTEUse this solution within 10 min.]
Cefmetazole Sodium taken: Sample solution A: Where it is represented as being in a
single-dose container, 200 g/mL of cefmetazole from a
Result = (rU/rS) (CS/CU) container of Cefmetazole for Injection in a volume of water,
corresponding to the volume of diluent specified in the
rU = peak response of cefmetazole from the Sample labeling, and quantitatively diluted with Mobile Phase
solution [NOTEWithdraw all of the withdrawable contents, using a
rS = peak response of cefmetazole from the Standard suitable hypodermic needle and syringe.]
solution [NOTEUse this solution within 10 min.]
CS = concentration of USP Cefmetazole RS in the Sample solution B: Where the label states the quantity of
Standard solution (g/mL) cefmenoxime in a given volume of constituted solution, 200
CU = nominal concentration of cefmetazole sodium in g/mL of cefmetazole from a container of Cefmetazole for
the Sample solution (g/mL) Injection in a volume of water, equivalent to the volume of
Acceptance criteria: 8601003 g/mg diluent specified in the labeling, and quantitatively diluted
with Mobile phase
SPECIFIC TESTS [NOTEUse this solution within 10 min.]
PH 791: 4.26.2, in a solution (1 in 10) Chromatographic system
STERILITY TESTS 71: Where the label states that Mode: LC
Cefmetazole Sodium is sterile, it meets the requirements.Test Detector: UV 214 nm
for Sterility of the Product to Be Examined, Membrane Filtration Column: 4.6-mm 25-cm; packing L1
BACTERIAL ENDOTOXINS TEST 85: Where the label states that Flow rate: 2 mL/min
Cefmetazole Sodium is sterile or must be subjected to Injection size: 10 L
further processing during the preparation of injectable System suitability
dosage forms, it meets the requirements. Samples: System suitability solution and Standard solution
ADDITIONAL REQUIREMENTS Resolution: NLT 3.0 between cefmetazole and
PACKAGING AND STORAGE: Preserve in tight containers. cefmetazole lactone, System suitability solution
LABELING: Where it is intended for use in preparing Column efficiency: NLT 1250 theoretical plates, Standard
injectable dosage forms, the label states that it is sterile or solution
must be subjected to further processing during the Tailing factor: 0.941.6, Standard solution
preparation of injectable dosage forms. Relative standard deviation: NMT 2.0%, Standard
USP Cefmetazole RS Analysis
USP Endotoxin RS Samples: Sample solution A or Sample solution B, and
Standard solution
Calculate the percentage of C15H17N7O5S3 withdrawn from
Cefmetazole for Injection the container, or in the portion of constituted solution
taken:
(Comment on this Monograph)id=m14016=Cefmetazole for
Injection=Ca-Chl-Monos.pdf) Result = (rU/rS) (CS/CU) 100
DEFINITION rU = peak response of cefmetazole from the Sample
Cefmetazole for Injection contains an amount of Cefmetazole solution
Sodium equivalent to NLT 90.0% and NMT 120.0% of the
labeled amount of cefmetazole (C15H17N7O5S3).
140 Cefmetazole / Official Monographs USP 32
rS = peak response of cefmetazole from the Standard Heat on a steam bath for 30 min, and cool. This System
solution suitability solution contains a mixture of cefonicid and
CS = concentration of USP Cefmetazole RS in the desacetyl cefonicid.
Standard solution (g/mL) Standard solution: Equivalent to 200 g/mL of cefonicid
CU = nominal concentration of cefmetazole in Sample from USP Cefonicid Sodium RS idissolved in Mobile phase
solution A or Sample solution B (g/mL) Sample solution: 200 g/mL of Cefonicid Sodium in Mobile
Acceptance criteria: 90.0%120.0% phase
PERFORMANCE TESTS (See Chromatography 621, System Suitability.)
UNIFORMITY OF DOSAGE UNITS 905: It meets the Mode: LC
requirements. Detector: UV 254 nm
PH 791: 4.26.2, in a solution (1 in 10) Injection size: 10 L
BACTERIAL ENDOTOXINS TEST 85: NMT 0.2 USP Endotoxin Samples: System suitability solution and Standard solution
Unit/mg of cefmetazole Suitability requirements
STERILITY TESTS 71: It meets the requirements when tested Resolution: NLT 1.1 between the cefonicid and the
as directed for Test for Sterility of the Product to be Examined, desacetyl cefonicid peaks, System suitability solution
Membrane Filtration. Column efficiency: NLT 1500 from the analyte peak
PARTICULATE MATTER IN INJECTIONS 788: It meets the theoretical plates, Standard solution
requirements for small-volume injections. Tailing factor: NMT 2.0, Standard solution
OTHER REQUIREMENTS: It meets the requirements under Relative standard deviation: NMT 2.0%, Standard
Injections 1, Labeling. solution
PACKAGING AND STORAGE: Preserve as described under Samples: Standard solution and Sample solution
Injections 1, Containers for Sterile Solids. Calculate the quantity, in g, of C18H18N6O8S3 per mg of
USP REFERENCE STANDARDS 11 Cefonicid Sodium taken:
USP Cefmetazole RS
USP Endotoxin RS Result = (rU/rS) (CS/CU) Mr1/Mr2 F

Cefonicid Sodium CS = concentration of USP Cefonicid Sodium RS in the
(Comment on this Monograph)id=m14031=Cefonicid CU = nominal concentration of cefonicid sodium in
Sodium=Ca-Chl-Monos.pdf) the Sample solution (g/mL)
Mr1 = molecular weight of cefonicid
Mr2 = molecular weight of cefonicid sodium
F = conversion factor, 1000 g/mg
SPECIFIC TESTS
OPTICAL ROTATION, Specific Rotation 781S: Between 37
C18H16N6Na2O8S3 586.53 and 47
5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7- Sample solution: 10 mg/mL, in methanol
[(hydroxyphenylacetyl)amino]-8-oxo-3-[[[1-(sulfomethyl)-1H- PH 791: 3.56.5, in a solution (1 in 20)
tetrazol-5-yl]thio]methyl]disodium salt, [6R-[6,7 (R*)]]-; WATER DETERMINATION, Method I 921: NMT 5.0%
(6R,7R)-[7-[(R)-Mandelamido]-8-oxo-3-[[[1-(sulfomethyl)-1H- STERILITY TESTS 71: Where the label states that Cefonicid
tetrazol-5-yl]thio]methyl]-5-thia-1-azabicyclo[4.2.0]oct-2- Sodium is sterile, it meets the requirements when tested as
ene-2-carboxylic acid, disodium salt [61270-78-8]. directed for Test for Sterility of the Product to Be Examined,
DEFINITION BACTERIAL ENDOTOXINS TEST 85: Where the label states that
Cefonicid Sodium contains the equivalent of NLT 832 g and Cefonicid Sodium is sterile or it must be subjected to further
NMT 970 g of cefonicid (C18H18N6O8S3)/mg, calculated on processing during the preparation of injectable dosage
the anhydrous basis. forms, it contains NMT 0.35 USP Endotoxin Unit/mg of
cefonicid.
IDENTIFICATION
A. The retention time of the major peak for cefonicid of the ADDITIONAL REQUIREMENTS
Sample solution corresponds to that of the Standard solution, PACKAGING AND STORAGE: Preserve in tight containers.
as obtained in the Assay. LABELING: Where it is intended for use in preparing
B. IDENTIFICATION TESTSGENERAL, Sodium 191: It meets injectable dosage forms, the label states that it is sterile or
the requirements. must be subjected to further processing during the
PROCEDURE USP Cefonicid Sodium RS
Mobile phase: Methanol, 0.2 M monobasic ammonium USP Endotoxin RS
phosphate, and water (5:2:33)
Pass through a filter having a 0.5-m or finer porosity.
System suitability solution: 200 g/mL of USP Cefonicid
Sodium RS in Mobile phase
USP 32 Official Monographs / Cefoperazone 141
Cefonicid for Injection rS = peak response from the Standard solution

(Comment on this Monograph)id=m14035=Cefonicid for CS = concentration of USP Cefonicid Sodium RS in
Injection=Ca-Chl-Monos.pdf) the Standard solution (g/mL)
CU = nominal concentration of cefonicid in Sample
DEFINITION solution A or Sample solution B (g/mL)
Cefonicid for Injection contains an amount of Cefonicid Sodium Acceptance criteria: 90.0%120.0%
equivalent to NLT 90.0% and NMT 120.0% of the labeled
amount of cefonicid (C18H18N6O8S3). PERFORMANCE TESTS
UNIFORMITY OF DOSAGE UNITS 905: It meets the
IDENTIFICATION requirements.
A. The retention time of the major peak for cefonicid in the
Sample solution corresponds to that of the Standard solution, SPECIFIC TESTS
as obtained in the Assay. OPTICAL ROTATION, Specific Rotation 781S: 37 to 47
B. IDENTIFICATION TESTSGENERAL, Sodium 191 It meets the Sample solution: 10 mg/mL, in methanol
requirements. PH 791: 3.56.5, in a solution (1 in 20)
ASSAY CONSTITUTED SOLUTION: At the time of use, it meets the
PROCEDURE requirements for Injections 1, Constituted Solutions.
Mobile phase: Methanol, 0.2 M monobasic ammonium BACTERIAL ENDOTOXINS TEST 85: NMT 0.35 USP Endotoxin
phosphate, and water (5:3:33) Unit/mg of cefonicid
Pass through a filter having a 0.5-m or finer porosity. STERILITY TESTS 71: It meets the requirements when tested
System suitability solution: 200 g/mL of USP Cefonicid as directed for Test for Sterility of the Product to be Examined,
Sodium RS in Mobile phase Membrane Filtration.
[NOTEHeat on a steam bath for 30 min, and cool. This PARTICULATE MATTER IN INJECTIONS 788: It meets the
System suitability solution contains a mixture of cefonicid requirements for small-volume injections.
and desacetyl cefonicid.] OTHER REQUIREMENTS: It meets the requirements for
Standard solution: 200 g/mL of cefonicid by Injections 1, Labeling.
quantitatively dissolving USP Cefonicid Sodium RS in Mobile
phase ADDITIONAL REQUIREMENTS
Sample solution A (where it is represented as being in a PACKAGING AND STORAGE: Preserve as described under
single-dose container): 200 g/mL of cefonicid in Mobile Injections 1, Containers for Sterile Solids.
phase prepared as follows. Using a suitable hypodermic USP REFERENCE STANDARDS 11
needle and syringe, withdraw all of the withdrawable USP Cefonicid Sodium RS
contents from a container of Cefonicid for Injection in a USP Endotoxin RS
volume of water corresponding to the volume of solvent
specified in the labeling, and dilute quantitatively with
Mobile phase.
Sample solution B (where the label states the quantity of
Cefoperazone Sodium
cefonicid in a given volume of constituted solution): 200 (Comment on this Monograph)id=m14039=Cefoperazone
g/mL of cefonicid in Mobile phase, from a container of Sodium=Ca-Chl-Monos.pdf)
Cefonicid for Injection in a volume of water corresponding
to the volume of solvent specified in the labeling,
quantitatively diluted with Mobile phase
Mode: LC
Detector: UV 254 nm
Column: 4-mm 30-cm; packing L1 C25H26N9NaO8S2 667.65
Flow rate: 2 mL/min 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[[[(4-
Injection size: 10 L ethyl-2,3-dioxo-1-piperazinyl)carbonyl]amino](4-
System suitability hydroxyphenyl)acetyl]amino]-3-[[(1-methyl-1H-tetrazol-5-
Samples: System suitability solution and Standard solution yl)thio]methyl]-8-oxo-, monosodium salt, [6R-[6,7 (R*)]]-;
Suitability requirements Sodium (6R,7R)-7-[(R)-2-(4-ethyl-2,3-dioxo-1-
Resolution: NLT 1.1 between the cefonicid and the piperazinecarboxamido)-2-(p-hydroxyphenyl)acetamido-3-[
desacetyl cefonicid peaks, System suitability solution [(1-methyl-H-tetrazol-5-yl)thio]methyl]-8-oxo-5-thia-1-
Column efficiency: NLT 1500 theoretical plates from the azabicyclo[4.2.0]oct-2-ene-2-carboxylate [62893-20-3].
analyte peak, Standard solution
Tailing factor: NMT 2.0, Standard solution DEFINITION
Relative standard deviation: NMT 2.0%, Standard Cefoperazone Sodium contains the equivalent of NLT 870 g
solution and NMT 1015 g of cefoperazone (C25H27N9O8S2)/mg,
Analysis calculated on the anhydrous basis.
Samples: Sample solution A or Sample solution B, and
Standard solution IDENTIFICATION
Calculate the percentage of C18H18N6O8S3 withdrawn from A. The retention time of the major peak for cefoperazone
the container, or in the portion of constituted solution of the Sample solution corresponds to that of the Standard
taken: solution, as obtained in the Assay.
B. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the
Result = (rU/rS) (CS/CU) 100 requirements
142 Cefoperazone / Official Monographs USP 32
ASSAY Cefoperazone Injection

PROCEDURE (Comment on this Monograph)id=m14041=Cefoperazone
Solution A: Place 14 mL of triethylamine and 5.7 mL of Injection=Ca-Chl-Monos.pdf)
glacial acetic acid in a 100-mL volumetric flask, and dilute
with water to volume. DEFINITION
Mobile phase: Acetonitrile, 1 N acetic acid, Solution A, and Cefoperazone Injection is a sterile solution of Cefoperazone
water (120:2.8:1.2:876) Sodium and a suitable osmolality adjusting substance in Water
Filter through a membrane filter of 1-m or finer porosity. for Injection. It may contain a suitable buffer. It contains the
Standard solution: 160 g/mL of cefoperazone from USP equivalent of NLT 90.0% and NMT 120.0% of the labeled
Cefoperazone Dihydrate RS in Mobile phase amount of cefoperazone (C25H27N9O8S2).
Sample solution: 160 g/mL of Cefoperazone Dihydrate in
Mobile phase IDENTIFICATION
Chromatographic system The retention time of the major peak for cefoperazone from
(See Chromatography 621, System Suitability.) the Sample solution corresponds to that of the Standard
Mode: LC solution, as obtained in the Assay.
Detector: UV 254 nm
Column: 4.0-mm 30-cm; packing L1 ASSAY
Injection size: 10 L Solution A: Triethylamine, glacial acetic acid, and water
System suitability (14:5.7:80.3)
Sample: Standard solution Mobile phase: Acetonitrile, 1 N acetic acid, Solution A, and
Suitability requirements water (120:2.8:1.2:876)
Tailing factor: NMT 1.5 Filter through a membrane filter of 1-m or finer porosity.
Relative standard deviation: NMT 2.0% Standard solution: 160 g/mL of cefoperazone by
Analysis quantitatively dissolving USP Cefoperazone Dihydrate RS in
Sample: Sample solution and Standard solution Mobile phase
Calculate the quantity, in g of cefoperazone/mg, of Sample solution: 160 g/mL of cefoperazone from the
Cefoperazone Sodium: volume of Injection in Mobile phase
Result = (rU/rS) (CS/CU) F (See Chromatography 621, System Suitability.)
Mode: LC
rU = peak response from the Sample solution Detector: UV 254 nm
rS = peak response from the Standard solution Column: 4.0-mm 30-cm; packing L1
CS = concentration of cefoperazone in the Standard Flow rate: 2 mL/min
solution (g/mL) Injection size: 10 L
CU = concentration of Cefoperazone Sodium in the System suitability
Sample solution (g/mL) Sample: Standard solution
F = correction factor, 1000 g/mg Suitability requirements
Acceptance criteria: 8701015 g/mg Tailing factor: NMT 1.5
CRYSTALLINITY 695: Meets the requirements Samples: Sample solution and Standard solution
PH 791: 4.56.5, in a solution (1 in 4) Calculate the percentage of C25H27N9O8S2 in the volume of
WATER DETERMINATION, Method I 921: NMT 5.0% Injection:
STERILITY TESTS 71: Where the label states that
Cefoperazone Sodium is sterile, it meets the requirements Result = (rU/rS) (CS/CU) 100
when tested as directed under Test for Sterility of the Product
to Be Examined, Membrane Filtration. rU = peak response from the Sample solution
BACTERIAL ENDOTOXINS TEST 85: Where the label states that rS = peak response from the Standard solution
Cefoperazone Sodium is sterile or it must be subjected to CS = concentration of USP Cefoperazone Dihydrate
further processing during the preparation of injectable RS in the Standard solution (g/mL)
dosage forms, it contains NMT 0.20 USP Endotoxin Unit/mg CU = nominal concentration of cefoperazone in the
of cefoperazone. Sample solution (g/mL)
PACKAGING AND STORAGE: Preserve in tight containers. SPECIFIC TESTS
LABELING: Where it is intended for use in preparing PARTICULATE MATTER IN INJECTIONS 788: Meets the
injectable dosage forms, the label states that it is sterile or requirements for small-volume injections
must be subjected to further processing during the PH 791: 4.56.5
preparation of injectable dosage forms. BACTERIAL ENDOTOXINS TEST 85: NMT 0.20 USP Endotoxin
USP REFERENCE STANDARDS 11 Unit/mg of cefoperazone
USP Cefoperazone Dihydrate RS
USP Endotoxin RS
USP 32 Official Monographs / Cefoperazone 143
STERILITY TESTS 71: It meets the requirements when tested Chromatographic system
as directed under Test for Sterility of the Product to Be (See Chromatography 621, System Suitability.)
Examined, Membrane Filtration. Mode: LC
Detector: UV 254 nm
ADDITIONAL REQUIREMENTS Column: 4.0-mm 30-cm; packing L1
PACKAGING AND STORAGE: Preserve as described under Flow rate: 2 mL/min
Injections 1, Containers for Injections. Maintain in the frozen Injection size: 10 L
state. System suitability
LABELING: It meets the requirements under Injections 1, Sample: Standard solution
Labeling. The label states that it is to be thawed just prior to Suitability requirements
use, describes conditions for proper storage of the resultant Tailing factor: NMT 1.5
solution, and directs that the solution is not to be refrozen. Relative standard deviation: NMT 2.0%
USP Cefoperazone Dihydrate RS Samples: Sample solution A or Sample solution B, and
USP Endotoxin RS Standard solution
the container, or in the portion of constituted solution
Cefoperazone for Injection taken:
(Comment on this Monograph)id=m14047=Cefoperazone for Result = (rU/rS) (CS/CU) 100
DEFINITION rS = peak response from the Standard solution
Cefoperazone for Injection contains an amount of Cefoperazone CS = concentration of USP Cefoperazone Dihydrate
Sodium equivalent to NLT 90.0% and NMT 120.0% of the RS in the Standard solution (g/mL)
labeled amount of cefoperazone (C25H27N9O8S2). CU = nominal concentration of cefoperazone in
Sample solution A or Sample solution B (g/mL)
A. The retention time of the major peak for cefoperazone
from the Sample solution corresponds to that of the Standard PERFORMANCE TESTS
solution, as obtained in the Assay. UNIFORMITY OF DOSAGE UNITS 905: It meets the
B. IDENTIFICATION TESTSGENERAL, Sodium 191: It meets requirements.
the requirements.
SPECIFIC TESTS
ASSAY CONSTITUTED SOLUTION: At the time of use, it meets the
PROCEDURE requirements for Injections 1, Constituted Solutions.
Solution A: Triethylamine, glacial acetic acid, and water BACTERIAL ENDOTOXINS TEST 85: NMT 0.20 USP Endotoxin
(14:5.7:80.3) Unit/mg of cefoperazone
Mobile phase: Acetonitrile, 1 N acetic acid, Solution A, and STERILITY TESTS 71: It meets the requirements when tested
water (120:2.8:1.2:876) as directed for Test for Sterility of the Product to be Examined,
Pass through a membrane filter having a 1-m or finer Membrane Filtration.
porosity. PARTICULATE MATTER IN INJECTIONS 788: It meets the
Standard solution: 160 g/mL of cefoperazone by requirements for small-volume injections.
quantitatively dissolving USP Cefoperazone Dihydrate RS in PH 791: 4.56.5, in a solution (1 in 4)
Mobile phase WATER DETERMINATION, Method I 921: NMT 5.0%, except
Sample solution A (where it is represented as being in a that where it is in the freeze-dried form, the limit is NMT
single-dose container): 160 g/mL of cefoperazone in 2.0%
Mobile phase prepared as follows. Using a suitable OTHER REQUIREMENTS: It meets the requirements for
hypodermic needle and syringe, withdraw all of the Injections 1, Labeling.
withdrawable contents from a container of Cefoperazone for
Injection in a volume of water corresponding to the volume ADDITIONAL REQUIREMENTS
of solvent specified in the labeling, and dilute quantitatively PACKAGING AND STORAGE: Preserve as described under
with Mobile phase. Injections 1, Containers for Sterile Solids.
Sample solution B (where the label states the quantity of USP REFERENCE STANDARDS 11
cefoperazone in a given volume of constituted solution): USP Cefoperazone Dihydrate RS
160 g/mL of cefoperazone in Mobile phase, from a USP Endotoxin RS
container of Cefoperazone for Injection in a volume of water
corresponding to the volume of solvent specified in the
labeling, quantitatively diluted with Mobile phase.
144 Ceforanide / Official Monographs USP 32
Ceforanide rU = peak response from the Sample solution

(Comment on this Monograph)id=m14072=Ceforanide=Ca-Chl- rS = peak response from the Standard solution
Monos.pdf) CS = concentration of USP Ceforanide RS in the
CU = concentration of Ceforanide taken to prepare
Sample solution A, Sample solution B, or Sample
solution C (mg/mL)
SPECIFIC TESTS
BACTERIAL ENDOTOXINS TEST 85: Where the label states that
C20H21N7O6S2 519.56 Ceforanide is sterile or must be subjected to further
5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[[2- processing during the preparation of injectable dosage
(aminomethyl)phenyl]acetyl]amino]-3-[[[1- forms, it contains NMT 0.25 USP Endotoxin Unit/mg of
(carboxymethyl)-1H-tetrazol-5-yl]thio]methyl]-8-oxo-, (6R- Ceforanide.
trans)-; STERILITY TESTS 71: Where the label states that Ceforanide
(6R,7R)-7-[2-(-Amino-o-tolyl)acetamido]-3-[[[1- is sterile, it meets the requirements when tested as directed
(carboxymethyl)-1H-tetrazol-5-yl]thio]methyl]-8-oxo-5-thia-1- for Test for Sterility of the Product to Be Examined, Membrane
azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid; Filtration, except to dissolve 6 g of Ceforanide in Fluid A to
7-[o-(Aminomethyl)phenylacetamido]-3-[[[1- each 1000 mL of which has been added 10 g of sterile L-
(carboxymethyl)-1H-tetrazol-5-yl]thio]methyl]-3-cephem-4- lysine, and to rinse the membrane with three 100-mL
carboxylic acid [60925-61-3]. portions of Fluid D and one 100-mL portion of Fluid A.
PH 791: 2.54.5, in a suspension containing 50 mg/mL
DEFINITION WATER DETERMINATION, Method I 921: NMT 5.0%
Ceforanide contains NLT 900 g and NMT 1050 g of
ceforanide (C20H21N7O6S2)/mg. ADDITIONAL REQUIREMENTS
IDENTIFICATION LABELING: Where it is intended for use in preparing
A. INFRARED ABSORPTION 197M injectable dosage forms, the label states that it is sterile or
B. The retention time of the major peak for ceforanide of must be subjected to further processing during the
the Sample solution corresponds to that of the Standard preparation of injectable dosage forms.
solution, as obtained in the Assay. USP REFERENCE STANDARDS 11
USP Ceforanide RS
ASSAY USP Endotoxin RS
PROCEDURE
Solution A: 18 mL of tetrabutylammonium hydroxide
solution (1 in 10) and 8.6 mL of 11 N potassium hydroxide
Add the mixture to 700 mL of water. Ceforanide for Injection
Mobile phase: To Solution A, add 200 mL of methanol, (Comment on this Monograph)id=m14073=Ceforanide for
adjust with phosphoric acid to a pH of 7.0, and add water Injection=Ca-Chl-Monos.pdf)
to obtain 1000 mL of solution. Filter, using a filter having a
porosity of 1 m or finer. DEFINITION
Standard solution: 1 mg/mL of USP Ceforanide RS in Ceforanide for Injection is a sterile mixture of Ceforanide and L-
Mobile phase lysine. It contains NLT 900 g and NMT 1050 g of
[NOTEUse this solution within 5 min.] ceforanide (C20H21N7O6S2)/mg on the L-lysinefree basis, and
Sample solution: 1 mg/mL of Ceforanide in Mobile phase NLT 90.0% and NMT 115.0% of the labeled amount of
[NOTEUse this solution within 5 min.] C20H21N7O6S2.
(See Chromatography 621, System Suitability.) IDENTIFICATION
Mode: LC A. The retention time of the major peak for L-lysine in the
Detector: UV 254 nm Sample solution corresponds to that of the Standard solution,
Column: 4-mm 30-cm; 5- to 10-m packing L1 as obtained in the test for Content of L-Lysine.
Flow rate: 1 mL/min B. The retention time of the major peak for ceforanide in
Injection size: 10 L the Sample solution corresponds to that of the Standard
System suitability solution, as obtained in the Assay.
Capacity factor, k: 1.85.0 Solution A: Mix 18 mL of tetrabutylammonium hydroxide
Column efficiency: NLT 1900 theoretical plates from the solution (1 in 10) and 8.6 mL of 11 N potassium hydroxide,
analyte peak and add the mixture to 700 mL of water.
Tailing factor: NMT 1.2 Mobile phase: To Solution A, add 200 mL of methanol,
Relative standard deviation: NMT 1.5% adjust with phosphoric acid to a pH of 7.0, and add water
Analysis to obtain 1000 mL of solution. Filter, using a filter having a
Samples: Standard solution and Sample solution porosity of 1 m or finer.
Calculate the quantity, in g, of C20H21N7O6S2 in each mg
of Ceforanide taken:
Result = (rU/rS) (CS/CU)
USP 32 Official Monographs / Ceforanide 145
Standard solution: 1 mg/mL of USP Ceforanide RS in OTHER COMPONENTS

Mobile phase CONTENT OF L-LYSINE
[NOTEUse this solution within 5 min.] Mobile phase: Methanol and water (31:19)
Sample solution A: 1 mg/mL of Ceforanide for Injection in Adjust with glacial acetic acid to a pH of 3.0.
Mobile phase Standard stock solution: 0.36 mg/mL of L-lysine
[NOTEUse this solution within 5 min.] Standard solution: Transfer 2.0 mL of Standard stock
Sample solution B (where it is represented as being in a solution in a glass-stoppered 10-mL volumetric flask, add 2.0
single-dose container): 1 mg/mL of ceforanide from a mL of a 1.4% solution of tris(hydroxymethyl)aminomethane
container of Ceforanide for Injection in a volume of water and 3.0 mL of a 1.5% solution of 1-fluoro-2,4-
corresponding to the volume of solvent specified in the dinitrobenzene in dehydrated alcohol, and insert the stopper
labeling tightly. Heat at 50 in a water bath for 30 min. Remove the
Withdraw all of the withdrawable contents, using a suitable flask from the water bath, allow to cool, and dilute with
hypodermic needle and syringe, and dilute quantitatively methanol to volume.
and stepwise with Mobile phase. Sample stock solution: 1.5 mg/mL of Ceforanide for
[NOTEUse this solution within 5 min.] Injection
Sample solution C (where the label states the quantity of Sample solution: 2.0 mL of Sample stock solution in a glass-
ceforanide in a given volume of constituted solution): 1 stoppered 10-mL volumetric flask. Add 2.0 mL of a 1.4%
mg/mL of ceforanide from a container of Ceforanide for solution of tris(hydroxymethyl)aminomethane and 3.0 mL of
Injection in a volume of water corresponding to the volume a 1.5% solution of 1-fluoro-2,4-dinitrobenzene in
of solvent specified in the labeling dehydrated alcohol, and insert the stopper tightly. Heat at
Dilute measured volume of the constituted solution 50 in a water bath for 30 min. Remove the flask from the
quantitatively and stepwise with Mobile phase. water bath, allow to cool, and dilute with methanol to
[NOTEUse this solution within 5 min.] volume.
Chromatographic system Chromatographic system
(See Chromatography 621, System Suitability.) (See Chromatography 621, System Suitability.)
Mode: LC Mode: LC
Column: 4-mm 30-cm; 5- to 10-m packing L1 Column: 4.6-mm 25-cm; 5- to 10-m packing L1
Flow rate: 1 mL/min Flow rate: 1.5 mL/min
Injection size: 10 L Injection size: 20 L
System suitability System suitability
Sample: Standard solution Sample: Standard solution
Suitability requirements Suitability requirements
Capacity factor, k: 1.85.0 Resolution: NLT 4.5 between the derivatized L-lysine peak
Column efficiency: NLT 1900 theoretical plates from the and the 1-fluoro-2,4-dinitrobenzene peak.
analyte peak Capacity factor, k: 46
Tailing factor: NMT 1.2 Column efficiency: NLT 1500 theoretical plates from the
Relative standard deviation: NMT 1.5% derivatized L-lysine peak
Analysis 1 Tailing factor: NMT 1.3
Samples: Standard solution and Sample solution A Relative standard deviation: NMT 1.5%
Calculate the g of C20H21N7O6S2 in each mg of Ceforanide Analysis
for Injection taken: Samples: Standard solution and Sample solution
Calculate the percentage of of L-lysine in the Ceforanide for
Result = (rU/rS) (CS/CU) P Injection taken:
rU = peak response from Sample solution A Result = (rU/rS) (CS/CU) 100
CS = concentration of USP Ceforanide RS in the rU = peak response from the Sample solution
Standard solution (mg/mL) rS = peak response from the Standard solution
CU = nominal concentration of ceforanide in Sample CS = concentration of L-lysine in the Standard stock
solution A (mg/mL) solution (g/mL)
P = potency of USP Ceforanide RS (g/mg) CU = nominal concentration of ceforanide in the
Analysis 2 Sample solution (g/mL)
Samples: Standard solution and Sample solution B, or Acceptance criteria: 9001050 g/mg
Sample solution C
Calculate the percentage of label claim of C20H21N7O6S2 in PERFORMANCE TESTS
the Ceforanide for Injection taken: UNIFORMITY OF DOSAGE UNITS 905: It meets the
requirements.
Result = (rU/rS) (CS/CU) P F 100
SPECIFIC TESTS
rU = peak response from the appropriate Sample BACTERIAL ENDOTOXINS TEST 85: NMT 0.25 USP Endotoxin
solution Unit/mg of ceforanide
rS = peak response from the Standard solution STERILITY TESTS 71: It meets the requirements when tested
CS = concentration of USP Ceforanide RS in the as directed for Test for Sterility of the Product to Be Examined,
Standard solution (mg/mL) Membrane Filtration, except to constitute each container with
CU = nominal concentration of ceforanide in the 3 mL of Fluid A for each g of ceforanide contained therein,
appropriate Sample solution (mg/mL) and to rinse the membrane with three 100-mL portions of
P = potency of USP Ceforanide RS (g/mg) Fluid D and one 100-mL portion of Fluid A.
F = unit conversion factor, 0.001 mg/g PH 791: 5.58.5, when constituted as directed in the
Acceptance criteria: 9001050 g/mg of C20H21N7O6S2 (L- labeling
lysinefree basis) and 90.0%115.0% of label claim of
C20H21N7O6S2
146 Ceforanide / Official Monographs USP 32
WATER DETERMINATION, Method I 921: NMT 3.0% Standard solution: 0.8 mg/mL of USP Cefotaxime Sodium
PARTICULATE MATTER IN INJECTIONS 788: It meets the RS in Solution B
requirements for small-volume injections. [NOTEUse this solution promptly. It may be used within
OTHER REQUIREMENTS: It meets the requirements under 24 h if stored in a refrigerator.]
Injections 1, Labels and Labeling. System suitability solution: Mix 1 mL of Standard solution,
7.0 mL of water, and 2.0 mL of methanol. Add 25 mg of
ADDITIONAL REQUIREMENTS sodium carbonate, mix, and allow to stand at room
PACKAGING AND STORAGE: Preserve as described under temperature for 10 min, with occasional swirling. Add 3
Injections 1, Containers for Sterile Solids. drops of glacial acetic acid, and 1 mL of Standard solution.
USP REFERENCE STANDARDS 11 Quantitative limit stock solution: 1.0 mL of Standard
USP Ceforanide RS solution diluted with Solution B to 50.0 mL
USP Endotoxin RS Quantitative limit solution: Quantitative limit stock solution
diluted with Solution B to 10.0 mL
Sample solution: 0.8 mg/mL of Cefotaxime Sodium in
Cefotaxime Sodium Solution B
[NOTEUse this solution promptly. It may be used within
(Comment on this Monograph)id=m14080=Cefotaxime 24 h if stored in a refrigerator.]
Sodium=Ca-Chl-Monos.pdf) Chromatographic system
Mode: LC
Detector: UV 235 nm
Temperature: 30
Flow rate: 1 mL/min
System suitability
C16H16N5NaO7S2 477.45 Sample: Standard solution, System suitability solution, and
5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 3- Quantitative limit solution
(acetyloxy)methyl-7-[[(2-amino-4- [NOTERetention times are 3.5 min for
thiazolyl)(methoxyimino)acetyl]amino]-8-oxo, monosodium desacetylcefotaxime, System suitability solution; 14 min for
salt, [6R-[6,7 (Z)]]-; cefotaxime, System suitability solution; and 1215 min for
Sodium (6R,7R)-7-[2-(2-amino-4-thiazolyl)glyoxylamido]-3- the main cefotaxime peak, Standard solution.]
(hydroxymethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- Suitability requirements
carboxylate 72-(Z)-(O-methyloxime), acetate (ester) [NOTEThe cefotaxime peak response from the
[64485-93-4]. Quantitative limit solution is 0.18%0.22% of the
DEFINITION cefotaxime peak response from the Standard solution.]
Cefotaxime Sodium contains the equivalent of NLT 916 g and Resolution: NLT 20 between the two peaks of
NMT 964 g of cefotaxime (C16H17N5O7S2)/mg, calculated on desacetylcefotaxime and cefotaxime, System suitability
the dried basis. solution
Tailing factor: NMT 2, Standard solution
IDENTIFICATION Relative standard deviation: NMT 1.5%, Standard
A. INFRARED ABSORPTION 197K solution
B. The retention time of the cefotaxime major peak from Analysis
the Sample solution corresponds to that of the Standard Samples: Standard solution and Sample solution
solution, as obtained in the Assay. Calculate the percentage of C16H17N5O7S2 in the portion of
C. IDENTIFICATION TESTSGENERAL, Sodium 191: It meets Cefotaxime Sodium taken:
the requirements.
Result = (rU/rS) (CS/CU) Mr1/Mr2 100
ASSAY
PROCEDURE rU = peak response of cefotaxime of the Sample
Solution A: 7.1 mg/mL of anhydrous dibasic sodium solution
phosphate in water rS = peak response of cefotaxime of the Standard
Adjust with phosphoric acid to a pH of 6.25. solution
Solution B: Methanol and Solution A (7:43) CS = concentration of USP Cefotaxime Sodium RS in
Pass through a filter having a porosity of 0.5 m or less the Standard solution (mg/mL)
before use. CU = nominal concentration of Cefotaxime Sodium in
Solution C: Methanol and Solution A (2:3) the Sample solution (mg/mL)
Pass through a filter having a porosity of 0.5 m or less, Mr1 = molecular weight of cefotaxime, 455.47
before use. Mr2 = molecular weight of cefotaxime sodium, 477.45
Mobile phase: Equilibrate the system with 100% Solution Acceptance criteria: 916964 g/mg
B. Increase the proportion of Solution C linearly from
0%20% at a rate of 10% per min 7 min after injection of IMPURITIES
the solution under test, and maintain at that composition Organic Impurities
for 7 min. Increase the proportion of Solution C linearly at a PROCEDURE
rate of 2.7% /min until the proportion of Solution C is Sample: Sample solution
100%, and hold at that composition for 5 min. Increase the Calculate the percentage of each impurity:
proportion of Solution B linearly to 100% at a rate of
20%/min. Result = rU/(rT + rC) 100
USP 32 Official Monographs / Cefotaxime 147
rU = peak area of a given impurity Solution C: Methanol and Solution A (2:3)

rT = sum of all of the impurity peak areas Pass through a filter having a porosity of 0.5 m or less.
rC = peak area for the main cefotaxime peak Mobile phase: Equilibrate the system with 100% Solution B.
[NOTEDisregard any impurity peak that is less than Increase the proportion of Solution C 7 min after injection of
0.1%.] the solution under test, linearly from 0% to 20% at a rate of
Acceptance criteria 10% per min and maintain at that composition for 7 min.
Individual impurities: NMT 1.0% Increase the proportion of Solution C linearly at a rate of
Total impurities: NMT 3.0% 2.7% per min until the proportion of Solution C is 100%,
and hold at that composition for 5 min. Increase the
SPECIFIC TESTS proportion of Solution B linearly to 100% at a rate of 20%
CLARITY AND COLOR OF SOLUTION: 2.5 g in 25 mL of carbon per min.
dioxidefree water Standard solution: 0.8 mg/mL of USP Cefotaxime Sodium
Mix, and examine immediately: the solution is clear. RS in Solution B
Measure the absorbance of this solution at 430 nm in a 1- [NOTEUse this solution promptly. It may be used within
cm cell, using carbon dioxidefree water as the blank: its 24 h if stored in a refrigerator.]
absorbance is NMT 0.20. Transfer 10 mL of the solution to System suitability solution: Mix 1 mL of Standard solution,
a glass test tube, add 1 mL of glacial acetic acid, mix, and 7.0 mL of water, and 2.0 mL of methanol. Add 25 mg of
examine immediately: the solution is clear. sodium carbonate, and allow to stand at room temperature
OPTICAL ROTATION, Specific Rotation 781S: +58 to +64 for 10 min, with occasional swirling. Add 3 drops of glacial
Sample solution: 10 mg/mL acetic acid and 1 mL of Standard solution.
PH 791: 4.56.5, in a solution (1 in 10) Quantitative limit stock solution: 1.0 mL of Standard
LOSS ON DRYING 731: Dry it at 100105 for 3 h: it loses solution diluted with Solution B to 50.0 mL
NMT 3.0% of its weight. Quantitative limit solution: 1.0 mL of Quantitative limit
STERILITY TESTS 71: Where the label states that Cefotaxime stock solution diluted with Solution B to 10.0 mL
Sodium is sterile, it meets the requirements when tested as Sample solution: Allow one container of Injection to thaw.
directed for Test for Sterility of the Product to Be Examined, Transfer a volume of the Injection, equivalent to 80 mg of
Membrane Filtration. C16H17N5O7S2 to a 100-mL volumetric flask, and dilute with
BACTERIAL ENDOTOXINS TEST 85: Where the label states that Solution B to volume.
Cefotaxime Sodium is sterile or it must be subjected to Chromatographic system
further processing during the preparation of injectable (See Chromatography 621, System Suitability.)
dosage forms, it contains NMT 0.2 Endotoxin Unit/mg of Mode: LC
cefotaxime. Detector: UV 235 nm
ADDITIONAL REQUIREMENTS Temperature: 30
PACKAGING AND STORAGE: Preserve in tight containers. Flow rate: 1 mL/min
LABELING: Where it is intended for use in preparing Injection size: 10 L
injectable dosage forms, the label states that it is sterile or System suitability
must be subjected to further processing during the Samples: System suitability solution, Standard solution and
preparation of injectable dosage forms. Quantitative limit solution [NOTEThe Retention times are:
USP REFERENCE STANDARDS 11 3.5 min for desacetylcefotaxime and 14 min for
USP Cefotaxime Sodium RS cefotaxime, System suitability solution; and 1215 min for
USP Endotoxin RS the main cefotaxime peak, Standard solution]
[NOTEThe response of the cefotaxime peak from the
Quantitative limit solution is between 0.18%0.22% of the
Cefotaxime Injection response of the cefotaxime peak from the Standard
solution.]
(Comment on this Monograph)id=m14084=Cefotaxime Suitability requirements
Injection=Ca-Chl-Monos.pdf) Resolution: NLT 20 between the two peaks of
DEFINITION desacetylcefotaxime and cefotaxime, System suitability
Cefotaxime Injection is a sterile solution of Cefotaxime Sodium solution
in Water for Injection. It contains one or more suitable buffers, Tailing factor: NMT 2.0, Standard solution
and it may contain Dextrose or Sodium Chloride as a tonicity- Relative standard deviation: NMT 1.5%, Standard
adjusting agent. It contains the equivalent of NLT 90.0% and solution
NMT 110.0% of the labeled amount of cefotaxime Analysis
(C16H17N5O7S2). Samples: Standard solution and Sample solution
Calculate the percentage of C16H17N5O7S2 in each mL of the
IDENTIFICATION Injection taken:
The retention time of the major peak for cefotaxime in the
Sample solution corresponds to that of the Standard solution, Result = (rU/rS) (CS/CU) Mr1/Mr2 100
ASSAY rS = peak response of the Standard solution
PROCEDURE CS = concentration of USP Cefotaxime Sodium RS in
Solution A: 7.1 mg/mL of anhydrous dibasic sodium the Standard solution (mg/mL)
phosphate in water CU = nominal concentration of cefotaxime sodium in
Adjust with phosphoric acid to a pH of 6.25 the Sample solution (mg/mL)
Solution B: Methanol and Solution A (7:43) Mr1 = molecular weight of cefotaxime, 455.47
Pass through a filter having a porosity of 0.5 m or less. Mr2 = molecular weight of cefotaxime sodium, 477.45
148 Cefotaxime / Official Monographs USP 32
Acceptance criteria: 90.0%110.0% Solution B: Methanol and Solution A (7:43)

Pass through a filter having a porosity of 0.5 m or less,
IMPURITIES before use.
Organic Impurities Solution C: Methanol and Solution A (2:3)
PROCEDURE Pass through a filter having a porosity of 0.5 m or less,
Analysis: Using the chromatogram of the Sample solution before use.
obtained in the Assay, calculate the percentage of each Mobile phase: Equilibrate the system with 100% Solution
impurity: B. Increase the proportion of Solution C linearly from
0%20% at a rate of 10% per min 7 min after injection of
Result = rU/(rT + rC) 100 the solution under test, and maintain at that composition
for 7 min. Increase the proportion of Solution C linearly at a
rU = peak area of a given impurity rate of 2.7% /min until the proportion of Solution C is
rT = sum of all of the impurity peak areas 100%, and hold at that composition for 5 min. Increase the
rC = peak area for the main cefotaxime peak proportion of Solution B linearly to 100% at a rate of
[NOTEDisregard any impurity peak that is less than 20%/min.
0.1%.] Standard solution: 0.8 mg/mL of USP Cefotaxime Sodium
Acceptance criteria RS in Solution B
Individual impurities: NMT 6.0% of any individual [NOTEUse this solution promptly. It may be used within
impurity is found. 24 h if stored in a refrigerator.]
Total impurities: NMT 10.0% System suitability solution: Mix 1 mL of Standard solution,
SPECIFIC TESTS 7.0 mL of water, and 2.0 mL of methanol. Add 25 mg of
BACTERIAL ENDOTOXINS TEST 85: NMT 0.2 USP Endotoxin sodium carbonate, mix, and allow to stand at room
Unit/mg of cefotaxime temperature for 10 min, with occasional swirling. Add 3
STERILITY TESTS 71: It meets the requirements when tested drops of glacial acetic acid and 1 mL of Standard solution.
as directed for Test for Sterility of the Product to Be Examined, Quantitative limit stock solution: 1.0 mL of Standard
Membrane Filtration. solution diluted with Solution B to 50.0 mL.
PH 791: 5.07.5 Quantitative limit solution: 1.0 mL of Quantitative limit
PARTICULATE MATTER IN INJECTIONS 788: It meets the stock solution diluted with Solution B to 10.0 mL.
requirements for small-volume injections. Sample solution A (for use where the Weight Variation test
is to be performed): 0.8 mg/mL of Cefotaxime for Injection
ADDITIONAL REQUIREMENTS in Solution A
PACKAGING AND STORAGE: Preserve in single-dose containers, [NOTEUse this solution promptly. It may be used within
as described under Injections 1. Maintain in the frozen 24 h if stored in a refrigerator.]
state. Sample solution B (for use in assaying vials and infusion
LABELING: It meets the requirements under Injections 1, bottles packaged for dispensing): Constitute one container
Labeling. The label states that it is to be thawed just prior to of Cefotaxime for Injection with the smallest volume of
use, describes conditions for proper storage of the resultant diluent specified in the labeling. Invert the container, and
solution, and directs that the solution is not to be refrozen. withdraw all of the withdrawable contents of the container
USP REFERENCE STANDARDS 11 with a hypodermic needle and syringe. Transfer the contents
USP Cefotaxime Sodium RS of the syringe to a 100-mL volumetric flask, dilute with
USP Endotoxin RS Solution B to volume, and mix. [NOTEDo not rinse the
syringe or container.] Dilute a measured volume of this
solution quantitatively with Solution B to obtain a solution
having a concentration of 0.8 mg of cefotaxime per mL.
Cefotaxime for Injection [NOTEUse this solution promptly. It may be used within 24
(Comment on this Monograph)id=m14087=Cefotaxime for h if stored in a refrigerator.]
Injection=Ca-Chl-Monos.pdf) Sample solution C (for use in assaying piggyback infusion
bottles): Constitute one container of Cefotaxime for
DEFINITION Injection with the smallest volume of diluent recommended
Cefotaxime for Injection contains an amount of Cefotaxime in the labeling, using the directions specified in the labeling.
Sodium equivalent to NLT 90.0% and NMT 115.0% of the Proceed as directed for Sample solution B beginning with
labeled amount of cefotaxime (C16H17N5O7S2). Invert the container.
Sample solution D (for use in assaying pharmacy bulk
IDENTIFICATION packages where the label states the quantity of cefotaxime
Where the Label Indicates That There Are No Added in a given volume of constituted solution): Constitute one
Substances container of Cefotaxime for Injection with the volume of
A. INFRARED ABSORPTION 197K diluent, specified in the labeling. With a hypodermic needle
B. IDENTIFICATION TESTSGENERAL, Sodium 191: It meets and syringe, withdraw an accurately measured portion of
the requirements. the resultant solution, equivalent to about 1000 mg of
Where the Label Indicates That There Are Added cefotaxime, to a 100-mL volumetric flask, dilute with
Substances Solution B to volume, and mix. [NOTEDo not rinse the
C. The retention time of the major peak of the Sample syringe or container.] Dilute volume of this solution
solution corresponds to that of the Standard solution, as quantitatively with Solution B to obtain a solution having a
obtained in the Assay. concentration of 0.8 mg of cefotaxime per mL. [NOTEUse
ASSAY this solution promptly. It may be used within 24 h if stored
PROCEDURE in a refrigerator.]
Solution A: 7.1 mg/mL of anhydrous dibasic sodium Chromatographic system
phosphate in water (See Chromatography 621, System Suitability.)
Adjust with phosphoric acid to a pH of 6.25.
USP 32 Official Monographs / Cefotetan 149
Mode: LC PARTICULATE MATTER IN INJECTIONS 788: It meets the

Detector: UV 235 nm requirements for small-volume injections.
Column: 3.9-mm 15-cm; 5-m packing L1 PH 791: 4.56.5, in a solution (1 in 10)
Temperature: 30 LOSS ON DRYING 731: Dry it at 100105 for 3 h: it loses
Flow rate: 1 mL/min NMT 3.0% of its weight.
Injection size: 10 L OTHER REQUIREMENTS: It meets the requirements under
System suitability Injections 1, Labeling.
Samples: Standard solution, System suitability solution, and
Quantitative limit solution[NOTEThe Retention times are: ADDITIONAL REQUIREMENTS
3.5 min for desacetylcefotaxime and 14 min for PACKAGING AND STORAGE: Preserve as described under
cefotaxime, System suitability solution; for the main Injections 1, Containers for Sterile Solids.
cefotaxime peak, 1215 min, Standard solution ] USP REFERENCE STANDARDS 11
[NOTEThe response of the cefotaxime peak from USP Cefotaxime Sodium RS
Quantitative limit solution is between 0.18%0.22% of the USP Endotoxin RS
response of the cefotaxime peak of the Standard solution.]
Resolution: NLT 20 between the two peaks of Cefotetan
desacetylcefotaxime and cefotaxime, System suitability
solution (Comment on this Monograph)id=m14093=Cefotetan=Ca-Chl-
Tailing factor: NMT 2, Standard solution Monos.pdf)
solution
Analysis
Samples: Sample solution A, or Sample solution B, or Sample
solution C, or Sample solution D, and Standard solution
Calculate the percentage of C16H17N5O7S2 in each mg of
the Cefotaxime for Injection taken:
Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 C17H17N7O8S4 575.62
5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[[4-(2-
rU = peak response of the Sample solution amino-1-carboxy-2-oxoethylidene)-1,3-dithietan-2-
rS = peak response of the Standard solution yl]carbonyl]amino]-7-methoxy-3-[[(1-methyl-1H-tetrazol-5-yl)-
CS = concentration of USP Cefotaxime Sodium RS in thio]methyl]-8-oxo-, [6R-(6,7)]-;
the Standard solution (mg/mL) (6R,7S)-4-[[2-Carboxy-7-methoxy-3-[[(1-methyl-1H-tetrazol-5-
CU = nominal concentration of cefotaxime sodium in yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-7-
the Sample solution (mg/mL) yl]carbamoyl]-1,3-dithietane-2,-malonamic acid;
Mr1 = molecular weight of cefotaxime, 455.47 (6R,7S)-7-[4-(Carbamoylcarboxymethylene)-1,3-dithiethane-2-
Mr2 = molecular weight of cefotaxime sodium, 477.45 carboxamido]-7-methoxy-3-[[(1-methyl-1H-tetrazol-5-
Acceptance criteria: 90.0%115.0% yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-
carboxylic acid [69712-56-7].
PERFORMANCE TESTS
UNIFORMITY OF DOSAGE UNITS 905 DEFINITION
Analysis: Perform the Assay on individual containers using Cefotetan contains NLT 950 g and NMT 1030 g of cefotetan
Sample solution B, Sample solution C, or Sample solution D, as (C17H17N7O8S4)/mg, calculated on the anhydrous basis.
appropriate.
IDENTIFICATION
IMPURITIES A. INFRARED ABSORPTION 197M
Organic Impurities B. The retention time of the major peak of the Sample
PROCEDURE solution corresponds to that of the Standard solution as
Using the chromatogram of the Sample solution obtained in the directed in the Assay.
Assay, calculate the percentage of each impurity:
ASSAY
Result = rU/(rT + rc) 100 PROCEDURE
[NOTEProtect the Standard solution, the System suitability
rU = peak area of a given impurity solution, and the Sample solution from light, and use within
rT = sum of all of the impurity peak area responses 2 h.]
rc = peak area for the main cefotaxime peak Mobile phase: Methanol, acetonitrile, glacial acetic acid,
[NOTEDisregard any impurity peak that is less than 0.1%.] and 0.1 M phosphoric acid (105:105:100:1700)
Acceptance criteria Standard solution: 20 mg of USP Cefotetan RS in a 100-
Individual impurities: NMT 6.0% of any individual mL volumetric flask, add 5 mL of methanol, swirl for several
impurity is found. min, add 5 mL of acetonitrile, and swirl until dissolved.
Total impurities: NMT 10.0% Dilute with water to volume.
System suitability solution: 10 mL of Standard solution in a
SPECIFIC TESTS glass-stoppered flask containing a few mg of magnesium
CONSTITUTED SOLUTION: At the time of use, it meets the carbonate.
requirements for Injections 1, Constituted Solutions. Sonicate for 10 min. If the solution is not turbid, add a few
BACTERIAL ENDOTOXINS TEST 85: NMT 0.2 USP Endotoxin more mg of magnesium carbonate, and repeat the
Unit/mg of cefotaxime sonication. Filter the turbid solution through a filter of 0.5-
STERILITY TESTS 71: It meets the requirements when tested m or finer porosity. Use the clear filtrate.
as directed for Test for Sterility of the Product to Be Examined,
150 Cefotetan / Official Monographs USP 32
Sample solution: 20 mg of Cefotetan in a 100-mL Cefotetan Injection

volumetric flask (Comment on this Monograph)id=m14094=Cefotetan
Add 5 mL of methanol, swirl for several min, add 5 mL of Injection=Ca-Chl-Monos.pdf)
acetonitrile, and swirl until dissolved. Dilute with water to
volume. DEFINITION
Chromatographic system Cefotetan Injection is a sterile isoosmotic solution of Cefotetan
(See Chromatography 621, System Suitability.) and Sodium Bicarbonate in Water for Injection. It contains one
Mode: LC or more buffer substances and a tonicity-adjusting agent. It
Detector: UV 254 nm contains NLT 90.0% and NMT 120.0% of the labeled amount
Column: 4.6-mm 25-cm; packing L1 of cefotetan (C17H17N7O8S4).
Flow rate: 2 mL/min
Injection size: 20 L IDENTIFICATION
System suitability The retention time of the major peak of the Sample solution
Samples: System suitability solution and Standard solution corresponds to that in the Standard solution obtained as
[NOTEThe relative retention times are 0.75 for cefotetan directed in the Assay.
and 1.0 for cefotetan tautomer, System suitability solution.]
Resolution: NLT 2.0 between the cefotetan peak and the [NOTEProtect the Standard solution, the System suitability
cefotetan tautomer peak, System suitability solution solution, and the Sample solution from light, and use within 2
Column efficiency: NLT 1500 theoretical plates, Standard h.]
solution PROCEDURE
Tailing factor: NMT 1.5, Standard solution Mobile phase: Methanol, acetonitrile, glacial acetic acid,
Relative standard deviation: NMT 2.0%, Standard and 0.1 M phosphoric acid (105:105:100:1700)
solution Standard solution: 20 mg of USP Cefotetan RS in a 100-
Analysis mL volumetric flask
Samples: Standard solution and Sample solution Add 5 mL of methanol, swirl for several min, add 5 mL of
Calculate the quantity, in g, of C17H17N7O8S4 in each mg: acetonitrile, and swirl until dissolved. Dilute with water to
volume.
Result = (rU/rS) (CS/CU) System suitability solution: 10 mL of Standard solution in a
glass-stoppered flask containing a few mg of magnesium
rU = peak response of cefotetan from the Sample carbonate
solution Sonicate for 10 min. If the solution is not turbid, add a few
rS = peak response of cefotetan from the Standard more mg of magnesium carbonate, and repeat the
solution sonication. Filter the turbid solution through a filter of 0.5
CS = concentration of USP Cefotetan RS in the m or finer porosity. Use the clear filtrate.
Standard solution (mg/mL) Sample solution: Allow the contents of a container of
CU = nominal concentration of cefotetan in the Injection to thaw, and mix the resultant solution. Transfer a
Sample solution (mg/mL) volume of this solution, equivalent to about 40 mg of
Acceptance criteria: 9501030 g/mg cefotetan, to a 200-mL volumetric flask, and dilute with
Mobile phase to volume.
SPECIFIC TESTS [NOTEUse the sample solution within 10 min.]
STERILITY TESTS 71: Where the label states that Cefotetan is Chromatographic system
sterile, it meets the requirements when tested as directed for (See Chromatography 621, System Suitability.)
Test for Sterility of the Product to Be Examined, Membrane Mode: LC
Filtration, except to use Fluid A to each 1000 mL of which Detector: UV 254 nm
has been added 10 g of sodium bicarbonate before Column: 4.6-mm 25-cm; packing L1
sterilization. Flow rate: 2 mL/min
BACTERIAL ENDOTOXINS TEST 85: Where the label states that Injection size: 20 L
Cefotetan is sterile or must be subjected to further System suitability
processing during the preparation of injectable dosage Sample: System suitability solution and Standard solution
forms, it contains NMT 0.17 USP Endotoxin Unit/mg of [NOTEThe relative retention times for cefotetan and
cefotetan. cefotetan tautomer, System suitability solution, are about
WATER DETERMINATION, Method I 921: NMT 2.5% 0.75 and 1.0, respectively.]
ADDITIONAL REQUIREMENTS Resolution: NLT 2.0 between the cefotetan peak and the
PACKAGING AND STORAGE: Preserve in tight containers. cefotetan tautomer peak, System suitability solution
LABELING: Where it is intended for use in preparing Column efficiency: NLT 1500 theoretical plates, Standard
must be subjected to further processing during the Tailing factor: NMT 1.5, Standard solution
preparation of injectable dosage forms. Relative standard deviation: NMT 2.0%, Standard
USP Cefotetan RS
USP Endotoxin RS
USP 32 Official Monographs / Cefotetan 151
Analysis Solution A to obtain a solution containing the equivalent of

Sample: Standard solution and Sample solution 200 g of cefotetan/mL.
Calculate the percentage of C17H17N7O8S4 in each mL of the Sample solution B (where the label states the quantity of
Injection taken: cefotetan in a given volume of constituted solution):
Constitute Cefotetan for Injection as directed in the labeling.
Result = (rU/rS) (CS/CU) 100 Dilute a volume of the constituted solution quantitatively
with Solution A to obtain a solution containing the
rU = peak response from the Sample solution equivalent of 200 g of cefotetan/mL.
rS = peak response from the Standard solution Chromatographic system
CS = concentration of USP Cefotetan RS in the (See Chromatography 621, System Suitability.)
Standard solution (mg/mL) Mode: LC
CU = nominal concentration of cefotetan in the Sample Detector: UV 254 nm
solution (mg/mL) Column: 4.6-mm 25-cm; packing L1
BACTERIAL ENDOTOXINS TEST 85: NMT 0.17 USP Endotoxin Samples: System suitability solution and Standard solution
Unit/mg of cefotetan [NOTEThe relative retention times for cefotetan and
STERILITY TESTS 71: Meets the requirements when tested as cefotetan tautomer are 0.75 and 1.0, respectively, in
directed under Test for Sterility of the Product to Be Examined, System suitability solution.]
Membrane Filtration. Suitability requirements
PH 791: 4.06.5 Resolution: NLT 2.0 between the cefotetan peak and the
PARTICULATE MATTER IN INJECTIONS 788: Meets the cefotetan tautomer peak, System suitability solution
requirements for small-volume injections Column efficiency: NLT 1500 theoretical plates, Standard
ADDITIONAL REQUIREMENTS solution
PACKAGING AND STORAGE: Preserve as described under Tailing factor: NMT 1.5, Standard solution
Injections 1, Containers for Injections. Maintain in the frozen Relative standard deviation: NMT 2.0%, Standard
state. solution
LABELING: Meets the requirements for Injections 1, Labeling. Analysis
The label states that it is to be thawed just prior to use, Samples: Standard solution and Sample solution
describes the conditions for proper storage of the resultant Calculate the percentage of C17H17N7O8S4 withdrawn from
solution, and directs that the solution is not to be refrozen. the container, or in the portion of solution taken:
USP Cefotetan RS Result = (rU/rS) (CS/CU) 100
USP Endotoxin RS rU = peak response of the Sample solution
CS = concentration of USP Cefotetan RS in the
Cefotetan for Injection Standard solution (g/mL)
CU = nominal concentration of cefotetan in Sample
(Comment on this Monograph)id=m14095=Cefotetan for solution A or Sample solution B (g/mL)
Injection=Ca-Chl-Monos.pdf) Acceptance criteria: 90.0%120.0%
DEFINITION PERFORMANCE TESTS
Cefotetan for Injection contains an amount of Cefotetan UNIFORMITY OF DOSAGE UNITS 905: Meets the
Disodium equivalent to NLT 90.0% and NMT 120.0% of the requirements
labeled amount of cefotetan (C17H17N7O8S4).
SPECIFIC TESTS
ASSAY INJECTIONS, Constituted Solutions 1: Meets the
PROCEDURE requirements
[NOTEProtect the Standard solution, the System suitability BACTERIAL ENDOTOXINS TEST 85 : NMT 0.17 USP Endotoxin
solution, and the Sample solution from light, and use within Unit/mg of cefotetan
2 h.] STERILITY TESTS 71: Meets the requirements when tested
Solution A: Methanol, acetonitrile, and water (1:1:18) as directed for Test for Sterility of the Product to Be Examined,
Mobile phase: Methanol, acetonitrile, glacial acetic acid, Membrane Filtration
and 0.1 M phosphoric acid (105:105:100:1700) PARTICULATE MATTER IN INJECTIONS 788: Meets the
Standard solution: 20 mg of USP Cefotetan RS in a 100- requirements for small-volume injections
mL volumetric flask PH 791: 4.06.5, in a solution (1 in 10)
Add 5 mL of methanol, swirl for several minutes, add 5 mL
of acetonitrile, and swirl until dissolved. Dilute with water
to volume. Change to read:
System suitability solution: 10 mL of Standard solution in a
carbonate WATER DETERMINATION, Method Ic 921: NMT 2.8%3
Sonicate for 10 min. If the solution is not turbid, add a few OTHER REQUIREMENTS: It meets the requirements of the
more mg of magnesium carbonate, and repeat the Identification tests under Cefotetan Disodium and meets the
sonication. Filter the turbid solution through a filter of 0.5 requirements under Injections 1, Labels and Labeling.
m or finer porosity. Use the clear filtrate.
Sample solution A (where the package is represented as ADDITIONAL REQUIREMENTS
being in a single-dose container): Constitute Cefotetan for PACKAGING AND STORAGE: Preserve as described under
Injection as directed in the labeling. Withdraw all of the Injections 1, Containers for Sterile Solids.
withdrawable contents, and quantitatively dilute with
152 Cefotetan / Official Monographs USP 32

USP Cefotetan RS (See Chromatography 621, System Suitability.)
USP Endotoxin RS Mode: LC
Detector: UV 254 nm
Flow rate: 2 mL/min
Cefotetan Disodium Injection size: 20 L
(Comment on this Monograph)id=m14096=Cefotetan System suitability
Disodium=Ca-Chl-Monos.pdf) Sample: System suitability solution and Standard solution
[NOTEThe relative retention times for cefotetan and
cefotetan tautomer are 0.75 and 1.0, respectively, from
the System suitability solution.]
Resolution: NLT 2.0 between the cefotetan peak and the
cefotetan tautomer peak, System suitability solution
solution
C17H15N7Na2O8S4 619.59 Tailing factor: NMT 1.5, Standard solution
5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[[4-(2- Relative standard deviation: NMT 2.0%, Standard
amino-1-carboxy-2-oxoethylidene)-1,3-dithietan-2-yl]carbonyl] solution
amino]-7-methoxy-3-[[(1-methyl-1H-tetrazol-5- Analysis
yl)thio]methyl]-8-oxo-, disodium salt, [6R-(6,7)]-; Sample: Standard solution and Sample solution
(6R,7S)-4-[[2-Carboxy-7-methoxy-3-[[(1-methyl-1H-tetrazol-5- Calculate the quantity, in g, of C17H17N7O8S4 per mg in
yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-7- the portion of Cefotetan Disodium taken:
yl]carbamoyl]-1,3-dithietane-2,-malonamic acid, disodium
salt; Result = (rU/rS) (CS/CU) (Mr1/Mr2) x P
(6R,7S)-7-[4-(Carbamoylcarboxymethylene)-1,3-dithietane-2-
carboxamido]-7-methoxy-3-[[(1-methyl-1H-tetrazol-5- rU = peak response from the Sample solution
yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- rS = peak response from the Standard solution
carboxylic acid, disodium salt [74356-00-6]. CS = concentration of USP Cefotetan RS in the
DEFINITION CU = nominal concentration of cefotetan disodium in
Cefotetan Disodium contains the equivalent of NLT 830 g and the Sample solution (g/mL)
NMT 970 g of cefotetan (C17H17N7O8S4) per mg, calculated Mr1 = molecular weight of cefotetan, 575.62
on the anhydrous basis. Mr2 = molecular weight of cefotetan disodium, 619.59
P = stated content of cefotetan in USP Cefotetan RS
IDENTIFICATION (mg/mg)
A. The retention time of the major peak from the Sample Acceptance criteria: 830970 g/mg
obtained in the Assay. SPECIFIC TESTS
B. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the PH 791: 4.06.5, in a solution (1 in 10)
requirements
ASSAY Change to read:
PROCEDURE
[NOTEProtect the Standard solution, the System suitability
solution, and the Sample solution from light, and use within WATER DETERMINATION, Method Ic 921: NMT 2.5%3
2 h.] STERILITY TESTS 71: Where the label states that Cefotetan
Mobile phase: Methanol, acetonitrile, glacial acetic acid, Disodium is sterile, it meets the requirements when tested as
and 0.1 M phosphoric acid (105:105:100:1700) directed for Test for Sterility of the Product to Be Examined,
Standard solution: 20 mg of USP Cefotetan RS in a 100- Membrane Filtration.
mL volumetric flask BACTERIAL ENDOTOXINS TEST 85: Where the label states that
Add 5 mL of methanol, swirl for several min, add 5 mL of Cefotetan Disodium is sterile or it must be subjected to
acetonitrile, and swirl until dissolved. Dilute with water to further processing during the preparation of injectable
volume. dosage forms, it contains NMT 0.17 USP Endotoxin Unit/mg
System suitability solution: 10 mL of Standard solution in a of cefotetan.
carbonate ADDITIONAL REQUIREMENTS
Sonicate for 10 min. If the solution is not turbid, add a few PACKAGING AND STORAGE: Preserve in tight containers.
more mg of magnesium carbonate, and repeat the LABELING: Where it is intended for use in preparing
sonication. Filter the turbid solution through a filter of 0.5 injectable dosage forms, the label states that it is sterile or
m or finer porosity. Use the clear filtrate. must be subjected to further processing during the
Sample solution: 40 mg of Cefotetan Disodium in a 200- preparation of injectable dosage forms.
mL volumetric flask USP REFERENCE STANDARDS 11
Add 10 mL of methanol, swirl for several minutes, add 10 USP Cefotetan RS
mL of acetonitrile, and swirl until dissolved. Dilute with USP Endotoxin RS
water to volume.
USP 32 Official Monographs / Cefotiam 153
Cefotiam Hydrochloride Column efficiency: From the cefotiam peak, NLT 1985
(Comment on this Monograph)id=m14098=Cefotiam theoretical plates, Standard solution, when calculated as:
Hydrochloride=Ca-Chl-Monos.pdf)
Result = 5.545(tr/Wh/2)2
Wh/2 = width of peak at half-height
Tailing factor: NMT 1.8, Standard solution
solution
Analysis
C18H23N9O4S3 2HCl 598.56 Calculate the quantity, in g, of C18H23N9O4S3 in each mg
5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7[[-(2- of the Cefotiam Hydrochloride:
amino-4-thiazolyl)acetyl]-amino]-3-[[[1-[2-(dimethylamino)
ethyl]-1H-tetrazol-5-yl]-thio]methyl]-8-oxo, hydrochloride, Result = (rU/rS) (CS/CU)
(6R-trans)-; rU = peak response of the Sample solution
(6R,7R)-7-[2-(2-Amino-4-thiazolyl)acetamido]-3-[[[1-[2- rS = peak response of the Standard solution
(dimethylamino)ethyl]-1H-tetrazol-5-yl]thio]methyl]-8-oxo-5- CS = concentration of cefotiam in the Standard
thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid solution (g/mL)
dihydrochloride; CU = concentration of Cefotiam Hydrochloride in the
7(R)-[2-(2-Amino-4-thiazolyl)acetamido]-3-[[[1-[2- Sample solution (mg/mL)
dimethylamino)ethyl]-1H-tetrazol-5-yl]thio]methyl]-3- Acceptance criteria: 790925 g/mg
cephem-4-carboxylic acid dihydrochloride [66309-69-1].
SPECIFIC TESTS
DEFINITION CRYSTALLINITY 695: Meets the requirements
Cefotiam Hydrochloride contains the equivalent of NLT 790 g PYROGEN TEST 151: Where the label states that it is sterile or
and NMT 925 g of cefotiam (C18H23N9O4S3)/mg, calculated must be subjected to further processing during the
on the anhydrous basis. preparation of injectable dosage forms, it meets the
IDENTIFICATION requirements of the test, the test dose being 1.0 mL/kg of a
A. ULTRAVIOLET ABSORPTION 197U solution in pyrogen-free sodium carbonate solution
Solution: 20 g/mL in water containing 40 mg/mL (prepared by dissolving 25.6 g of
B. The retention time of the cefotiam peak of the Sample sodium carbonate, previously heated at 170 for NLT 4 h, in
solution corresponds to that of the Standard solution, as 1000 mL of Sterile Water for Injection).
obtained in the Assay. STERILITY TESTS 71: Where the label states that it is sterile,
it meets the requirements when tested as directed for Test
ASSAY for Sterility of the Product to Be Examined, Membrane Filtration.
PROCEDURE WATER DETERMINATION, Method I 921: NMT 7.0%, the
Mobile phase: 13.1 g of ammonium sulfate in 850 mL of Sample solution being prepared as directed for a hygroscopic
water specimen, except to use a mixture of 20 mL of formamide
Adjust with 2 N ammonium hydroxide to a pH of 6.5 0.1, (previously dried over anhydrous sodium sulfate for 24 h)
and add 150 mL of acetonitrile. Pass through a suitable and methanol (2:1), instead of methanol, to dissolve the
filter of 0.5 m or finer porosity. specimen, and to determine the water content of the
System suitability stock solution: 1 mg/mL of USP formamide and methanol mixture
Cefotiam Hydrochloride RS
[NOTEHeat this solution at 95 for 3 min, and cool.] ADDITIONAL REQUIREMENTS
System suitability solution: 1 mL of System suitability stock PACKAGING AND STORAGE: Preserve in tight containers.
solution diluted with Mobile phase to 100 mL LABELING: Where it is intended for use in preparing
Standard stock solution: 1 mg/mL of cefotiam from USP injectable dosage forms, the label states that it is sterile or
Cefotiam Hydrochloride RS must be subjected to further processing during the
Standard solution: 50 g/mL cefotiam from Standard stock preparation of injectable dosage forms.
solution diluted with Mobile phase USP REFERENCE STANDARDS 11
[NOTEUse this solution without delay.] USP Cefotiam Hydrochloride RS
Sample stock solution: 1.2 mg/mL of Cefotiam
Hydrochloride
Sample solution: 60 g/mL from Sample stock solution Cefotiam for Injection
diluted with Mobile phase
[NOTEUse this solution without delay.] (Comment on this Monograph)id=m14099=Cefotiam for
Chromatographic system Injection=Ca-Chl-Monos.pdf)
Mode: LC Cefotiam for Injection contains an amount of Cefotiam
Detector: UV 254 nm Hydrochloride equivalent to NLT 90.0% and NMT 120.0% of
Column: 4-mm 25-cm column; packing L1 the labeled amount of cefotiam (C18H23N9O4S3). It may contain
Flow rate: 1.5 mL/min Sodium Carbonate.
Samples: System suitability solution and Standard solution A. ULTRAVIOLET ABSORPTION 197U
[NOTEThe relative retention times for de-tetrazol-cefotiam Solution: 20 g/mL
and cefotiam are 0.6 and 1.0, respectively.] B. The retention time of the major peak for cefotiam in the
Suitability requirements Sample solution corresponds to that in the Standard solution
Resolution: NLT 4.0 between the de-tetrazol-cefotiam as obtained in the Assay.
peak and the cefotiam peak, System suitability solution
154 Cefotiam / Official Monographs USP 32
ASSAY rU = peak response from the Sample solution

PROCEDURE rS = peak response from the Standard solution
Mobile phase: 13.1 g of ammonium sulfate in 850 mL of CS = concentration of cefotiam in the Standard
water solution (mg/mL)
Adjust with 2 N ammonium hydroxide to a pH of 6.5 0.1, CU = nominal concentration of cefotiam in Sample
and add 150 mL of acetonitrile. Pass through a suitable solution A or Sample solution B (g/mL)
filter of 0.5 m or finer porosity. Acceptance criteria: 90.0%120.0%
System suitability stock solution: 1 mg/mL of USP
Cefotiam Hydrochloride RS SPECIFIC TESTS
[NOTEHeat this solution at 95 for 3 min and cool.] PYROGEN TEST 151: Meets the requirements of the test; the
System suitability solution: Mobile phase and System dose being 1.0 mL/kg of a solution prepared by diluting
suitability stock solution (99:1) Cefotiam for Injection with Sterile Water for Injection to a
Standard stock solution: 1 mg/mL of USP Cefotiam concentration of 40 mg/mL of cefotiam
Hydrochloride RS STERILITY TESTS 71: It meets the requirements when tested
Standard solution: 50 g/mL from the Standard stock as directed under Test for Sterility of the Product to Be
solution in Mobile phase Examined, Membrane Filtration.
[NOTEUse this solution without delay.] PH 791: 5.77.2, in a solution of 100 mg of cefotiam/mL
Sample stock solution A (where it is represented as being LOSS ON DRYING 731: Dry 100 mg in vacuum at a pressure
in a single-dose container): Constitute a container of not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT
Cefotiam for Injection in a volume of water, measured 6.0% of its weight.
corresponding to the volume of diluent specified in the PARTICULATE MATTER IN INJECTIONS 788: Meets the
labeling. Withdraw all of the withdrawable contents, using a requirements for small-volume injections
suitable hypodermic needle and syringe, and dilute
quantitatively with water to obtain a solution containing the ADDITIONAL REQUIREMENTS
equivalent of 1 mg of cefotiam/mL. PACKAGING AND STORAGE: Preserve as described under
Sample solution A: 50 g/mL from Sample stock solution A Injections 1, Containers for Sterile Solids.
in Mobile phase USP REFERENCE STANDARDS 11
[NOTEUse this solution without delay.] USP Cefotiam Hydrochloride RS
Sample stock solution B (where the label states the
quantity of cefotiam in a given volume of constituted
solution): Constitute a container of Cefotiam for Injection in Cefoxitin Sodium
a volume of water, measured and equivalent to the volume
of diluent specified in the labeling. Dilute a measured (Comment on this Monograph)id=m14104=Cefoxitin
volume of the constituted solution quantitatively with water Sodium=Ca-Chl-Monos.pdf)
to obtain a solution containing 1 mg of cefotiam/mL.
Sample solution B: 50 g/mL from Sample stock solution B
in Mobile phase
[NOTEUse this solution without delay.]
Mode: LC
Detector: UV 254 nm C16H16N3NaO7S2 449.44
Column: 4-mm 25-cm column; packing L1 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid,
Flow rate: 1.5 mL/min 3-[[(aminocarbonyl)oxy]methyl]-7-methoxy-8-oxo-7-
Injection size: 10 L [(2-thienylacetyl)-amino]-, sodium salt (6R-cis)-;
System suitability Sodium (6R,7S)-3-(hydroxymethyl)-7-methoxy-8-oxo-7-[2-(2-
Samples: System suitability solution and Standard solution thienyl)acetamido]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-
[NOTEThe relative retention times for de-tetrazol-cefotiam carboxylate carbamate (ester) [33564-30-6; 35607-66-0].
and cefotiam are about 0.6 and 1.0, respectively, System
suitability solution.] DEFINITION
Suitability requirements Cefoxitin Sodium contains the equivalent of NLT 927 g/mg
Resolution: NLT 4.0 between the de-tetrazol-cefotiam and NMT 970 g/mg of cefoxitin (C16H17N3O7S2),
peak and the cefotiam peak, System suitability solution corresponding to NLT 97.5% and NMT 102.0% of cefoxitin
Column efficiency: NLT 1985 theoretical plates from the sodium (C16H16N3NaO7S2), calculated on the anhydrous and
cefotiam peak of the Standard solution when calculated by acetone- and methanol-free basis.
the following formula:
IDENTIFICATION
5.545(TR/WH/2)2 A. The retention time of the major peak for cefoxitin in the
Sample solution corresponds to that of the Standard solution,
Tailing factor: NMT 1.8, Standard solution obtained as directed in the Assay.
Relative standard deviation: NMT 1.0%, Standard B. ULTRAVIOLET ABSORPTION 197U
solution Buffer solution: 1.0 mg/mL of monobasic potassium
Analysis phosphate and 1.8 mg/mL of anhydrous dibasic sodium
Sample: Sample solution A or Sample solution B, and phosphate
Standard solution
taken:
USP 32 Official Monographs / Cefoxitin 155
Sample solution: 20 g/mL in Buffer solution Mode: GC

C. IDENTIFICATION TESTSGENERAL, Sodium 191 Detector: Flame ionization
Sample solution: 50 mg/mL Column: 1.8-m 6.3-mm glass column containing
Acceptance criteria: Meets the requirements support S2, and a precolumn packed with 60- to 80-mesh
silane-treated glass beads
ASSAY Temperature
PROCEDURE Injection port: 100
Mobile phase: Acetonitrile, water, and glacial acetic acid Columns: 110
(16:84:1) Detector: 200
Filter through a membrane filter of 1-m or finer porosity. Carrier gas: Nitrogen
Buffer solution: 1.0 g of monobasic potassium phosphate Flow rate: 50 mL/min
and 1.8 g of dibasic sodium phosphate in 900 mL of water Injection size: 2 L
Adjust with phosphoric acid or 10 N sodium hydroxide to a System suitability
pH of 7.1 0.1, and dilute with water to make 1000 mL. Sample: Standard solution
Pass through a membrane filter of 1-m or finer porosity. Suitability requirements
Standard solution: 0.3 mg/mL of USP Cefoxitin RS in Buffer Column efficiency: NLT 160 and NLT 200 theoretical
solution plates, respectively, for the acetone and methanol peaks
[NOTESonicate, if necessary, to dissolve the specimen and Tailing factor: NMT 1.3 and NMT 2.3, respectively, for
use this solution within 5 h.] the acetone and methanol peaks
Sample solution: 0.3 mg/mL of Cefoxitin Sodium in Buffer Relative standard deviation: NMT 5.0%
solution Analysis
[NOTEUse this solution within 5 h.] Samples: Sample solution and Standard solution
Chromatographic system Calculate the percentages of acetone and methanol in the
(See Chromatography 621, System Suitability.) portion of Cefoxitin Sodium taken:
Mode: LC
Detector: UV 254 nm Result = (rU/rS) CS/CU) D
Column: 3.9-mm 30-cm; 5- to 10-m packing L1
Flow rate: 1 mL/min rU = peak response of acetone or methanol in the
Injection size: 10 L Sample solution
System suitability rS = peak response of acetone or methanol in the
Sample: Standard solution Standard solution
Suitability requirements CS = concentration of acetone or methanol in the
Column efficiency: NLT 2800 theoretical plates Standard solution (%)
Tailing factor: NMT 1.5 CU = nominal concentration of Cefoxitin Sodium in
Relative standard deviation: NMT 1.0% the Sample solution (g/mL)
Analysis D = density of acetone and methanol at 20 (g/mL),
Samples: Sample solution and Standard solution 0.79
Calculate the concentration, in g/mg, of C16H17N3O7S2 in Acceptance criteria: NMT 0.7% of acetone and NMT
the portion of Cefoxitin Sodium taken: 0.1% of methanol
Result = (rU/rS) CS/CU) P SPECIFIC TESTS
OPTICAL ROTATION, Specific Rotation 781S: +206 to +214,
rU = peak response from the Sample solution calculated on the anhydrous and acetone- and methanol-free
rS = peak response from the Standard solution basis
CS = concentration of cefoxitin in the Standard Sample solution: 10 mg/mL, in methanol
solution (mg/mL) CRYSTALLINITY 695: Meets the requirements
CU = nominal concentration of Cefoxitin Sodium PH 791: 4.27.0, in a solution containing 100 mg/mL
taken to prepare the Sample solution (mg/mL) WATER DETERMINATION, Method I 921: NMT 1.0%, a
P = potency of USP Cefoxitin RS (g/mg) mixture of ethylene glycol and pyridine (3:1) being used in
Acceptance criteria: 927970 g/mg place of methanol in the titration vessel
BACTERIAL ENDOTOXINS TEST 85: Where the label states that
IMPURITIES Cefoxitin Sodium is sterile or must be subjected to further
Inorganic Impurities processing during the preparation of injectable dosage
HEAVY METALS, Method II 231: NMT 20 ppm forms, it contains NMT 0.13 USP Endotoxin Unit/mg of
Organic Impurities cefoxitin.
LIMIT OF ACETONE AND METHANOL STERILITY TESTS 71: Where the label states that Cefoxitin
Standard stock solution A: 0.5% acetone in water Sodium is sterile, it meets the requirements when tested as
Standard stock solution B: 0.5% methanol in water directed for Test for Sterility of the Product to Be Examined,
Standard solution: 0.050% acetone and 0.005%
methanol from Standard stock solution A and Standard stock Membrane Filtration
solution B diluted with water ADDITIONAL REQUIREMENTS
Sample solution: To a 50-mL volumetric flask, add 5.0 g PACKAGING AND STORAGE: Preserve in tight containers, and
of Cefoxitin Sodium, and dissolve in and dilute with water store in a cold place.
to volume. Transfer 3.0 mL of the resulting solution to a LABELING: Where it is intended for use in preparing
15-mL centrifuge tube, cool in an ice-water bath for 2 min, injectable dosage forms, the label states that it is sterile or
and add 3.0 mL of 0.24 N hydrochloric acid while swirling must be subjected to further processing during the
vigorously. Centrifuge to obtain a clear solution, and use preparation of injectable dosage forms.
the supernatant. USP REFERENCE STANDARDS 11
Chromatographic system USP Cefoxitin RS
156 Cefoxitin / Official Monographs USP 32
USP Endotoxin RS STERILITY TESTS 71: It meets the requirements when tested
as directed under Test for Sterility of the Product to be
Examined, Membrane Filtration.
PH 791: 4.58.0
Cefoxitin Injection PARTICULATE MATTER IN INJECTIONS 788: It meets the
(Comment on this Monograph)id=m14106=Cefoxitin requirements for small-volume injections.
DEFINITION PACKAGING AND STORAGE: Preserve as described under
Cefoxitin Injection is a sterile solution of Cefoxitin Sodium and Injections 1, Containers for Injections. Maintain in the frozen
one or more suitable buffer substances in Water for Injection. state.
It contains Dextrose or Sodium Chloride as a tonicity-adjusting LABELING: It meets the requirements for Injections 1,
agent. It contains the equivalent of NLT 90.0% and NMT Labeling. The label states that it is to be thawed just prior to
120.0% of the labeled amount of cefoxitin (C16H17N3O7S2). use, describes conditions for proper storage of the resultant
solution, and directs that the solution is not to be refrozen.
IDENTIFICATION USP REFERENCE STANDARDS 11
The retention time of the major peak for cefoxitin in the USP Cefoxitin RS
Sample solution corresponds to that in the Standard solution, USP Endotoxin RS
ASSAY
PROCEDURE Cefoxitin for Injection
Mobile phase: Acetonitrile, glacial acetic acid, and water (Comment on this Monograph)id=m14107=Cefoxitin for
(16:1:84) Injection=Ca-Chl-Monos.pdf)
Filter through a membrane filter of 1 m or finer porosity.
Solution A: Dissolve 1.0 mg/mL of monobasic potassium DEFINITION
phosphate and 1.8 mg/mL of dibasic sodium phosphate in Cefoxitin for Injection contains Cefoxitin Sodium equivalent to
water, and adjust with phosphoric acid or 10 N sodium NLT 90.0% and NMT 120.0% of the labeled amount of
hydroxide to a pH of 7.1 0.1 cefoxitin (C16H17N3O7S2).
[NOTEFilter through a membrane filter of 1 m or finer
porosity.] IDENTIFICATION
Standard solution: 0.3 mg/mL of USP Cefoxitin RS in A. The retention time of the major peak of the Sample
Solution A solution corresponds to that of the Standard solution, as
[NOTESonicate, if necessary, to dissolve the specimen, obtained in the Assay.
and use this solution within 5 h.] B. ULTRAVIOLET ABSORPTION 197U
Sample solution: Allow one container of Injection to thaw, Solution: 20 g/mL
and mix. Dilute a volume of Injection quantitatively with Medium: 1 mg/mL of monobasic potassium phosphate and
Solution A to obtain a solution containing 0.3 mg of 1.8 mg/mL of anhydrous dibasic sodium phosphate in water
cefoxitin/mL. [NOTEUse this solution within 5 h.] C. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the
Chromatographic system requirements
Mode: LC ASSAY
Detector: UV 254 nm PROCEDURE
Column: 3.9-mm 30-cm; 5 to 10-m packing L1 Mobile phase: Acetonitrile, glacial acetic acid, and water
Flow rate: 1 mL/min (16:1:84)
Injection size: 10 L Filter through a membrane filter of 1-m or finer porosity.
System suitability Solution A: 1.0 mg/mL of monobasic potassium phosphate
Sample: Standard solution and 1.8 mg/mL of dibasic sodium phosphate in water
Suitability requirements Adjust with phosphoric acid or 10 N sodium hydroxide to a
Column efficiency: NLT 2800 theoretical plates pH of 7.1 0.1. Filter through a membrane filter of 1-m
Tailing factor: NMT 1.5 or finer porosity.
Relative standard deviation: NMT 1.0% Standard solution: 0.3 mg/mL of USP Cefoxitin RS in
Analysis Solution A
Samples: Standard solution and Sample solution [NOTESonicate, if necessary, to dissolve the specimen,
Calculate the percentage of C16H17N3O7S2 in each mL of the and use this solution within 5 h.]
Injection taken: Sample solution A (where it is represented as being in a
single-dose container): Constitute Cefoxitin for Injection in
Result = (rU/rS) (CS/CU) 100 a volume of water, corresponding to the volume of solvent
specified in the labeling. Withdraw all of the withdrawable
rU = peak response of the Sample solution contents, using a suitable hypodermic needle and syringe,
rS = peak response of the Standard solution and dilute quantitatively with water to obtain a solution
CS = concentration of cefoxitin in the Standard having a nominal concentration of 0.3 mg of cefoxitin/mL.
solution (mg/mL) [NOTEUse this solution within 5 h.]
CU = nominal concentration of cefoxitin in the Sample Sample solution B (where the label states the quantity of
solution (mg/mL) cefoxitin in a given volume of constituted solution):
Acceptance criteria: 90.0%120.0% Constitute Cefoxitin for Injection in a volume of water,
corresponding to the volume of solvent specified in the
SPECIFIC TESTS labeling. Dilute a measured volume of the constituted
BACTERIAL ENDOTOXINS TEST 85: NMT 0.13 USP Endotoxin solution quantitatively with water to obtain a solution
Unit/mg of cefoxitin
USP 32 Official Monographs / Cefpiramide 157
having a nominal concentration of 0.3 mg of cefoxitin/mL. USP Endotoxin RS

[NOTEUse this solution within 5 h.]
Mode: LC Cefpiramide
Detector: UV 254 nm (Comment on this Monograph)id=m14109=Cefpiramide=Ca-
Column: 3.9-mm 30-cm; 5 to 10-m packing L1 Chl-Monos.pdf)
Flow rate: 1 mL/min
System suitability
Analysis C25H24N8O7S2 612.64
Samples: Sample solution A or Sample solution B and 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid,
Standard Solution 7-[[[[(4-hydroxy-6-methyl-3-pyridinyl)carbonyl]amino](4-
Calculate the percentage of C16H17N3O7S2 in the portion of hydroxyphenyl)acetyl]amino]-3-[[(1-methyl-1H-tetrazol-5-
constituted solution taken: yl)thio]methyl]-8-oxo, [6R-[6,7 (R*)]]-;
(6R,7R)-7-[(R)-2-(4-Hydroxy-6-methylnicotinamido)-2-(p-
Result = (rU/rS) (CS/CU) 100 hydroxyphenyl)acetamido]-3-[[(1-methyl-1H-tetrazol-5-
yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-
rU = peak response from the Sample solution carboxylic acid [70797-11-4].
CS = concentration of cefoxitin in the Standard DEFINITION
solution (mg/mL) Cefpiramide contains NLT 974 g and NMT 1026 g of
CU = nominal concentration of cefoxitin in Sample C25H24N8O7S2/mg, calculated on the anhydrous basis.
solution A or Sample solution B (mg/mL)
PERFORMANCE TESTS B. The retention time of the major peak of the Sample
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements solution corresponds to that of the Standard solution, as
SPECIFIC TESTS
CONSTITUTED SOLUTION: At the time of use, it meets the ASSAY
requirements for Injections 1, Constituted Solutions. PROCEDURE
BACTERIAL ENDOTOXINS TEST 85: NMT 0.13 USP Endotoxin Solution A: 1.36 mg/mL of monobasic potassium
Unit/mg of cefoxitin phosphate in water
STERILITY TESTS 71: It meets the requirements when tested Adjust with 1 N sodium hydroxide to a pH of 6.8 0.1
as directed for Test for Sterility of the Product to Be Examined, Mobile phase: Tetrahydrofuran, acetonitrile, methanol, and
Membrane Filtration. Solution A (3:1:1:45)
PARTICULATE MATTER IN INJECTIONS 788: It meets the System suitability solution: 1 mg/mL of USP Cefpiramide
requirements for small-volume injections. RS in 0.01 N sodium hydroxide
PH 791: 4.27.0, in a solution containing 100 mg/mL Heat this solution at 95 for 10 min. Mix 1 mL of this
WATER DETERMINATION, Method I 921: NMT 1.0%, a solution with 19 mL of Mobile phase. This solution contains
mixture of ethylene glycol and pyridine (3:1) being used in a mixture of cefpiramide and cefpiramide lactone.
place of methanol in the titration vessel Standard solution: 0.25 mg/mL of USP Cefpiramide RS in
OTHER REQUIREMENTS: It meets the requirements for Mobile phase
Injections 1, Labeling. Sample solution: 0.25 mg/mL of Cefpiramide in Mobile
phase
PACKAGING AND STORAGE: Preserve as described under (See Chromatography 621, System Suitability.)
Injections 1, Containers for Sterile Solids.
USP Cefoxitin RS
158 Cefpiramide / Official Monographs USP 32
Mode: LC System suitability: Proceed as directed in the Assay.

Column: 4.6-mm 25-cm; packing L1 Samples: Standard solution and Sample solution 2
Flow rate: 1.5 mL/min Calculate the percentage of 5-mercapto-1-methyltetrazole
Injection size: 20 L in the portion of Cefpiramide taken:
System suitability
Samples: System suitability solution and Standard solution Result = (rU/rS) (Mr1/Mr2) [5CS (100-w)/100W] 100
[NOTEThe relative retention times for cefpiramide and
cefpiramide lactone are about 0.7and 1.0, respectively.] rU = peak area of 5-mercapto-1-methyltetrazole from
Suitability requirements Sample solution 2
Resolution: NLT 5 between the cefpiramide lactone peak rS = peak area of 5-mercapto-1-methyltetrazole from
and the cefpiramide peak, System suitability solution the Standard solution
Capacity factor, k: Between 2 and 5, Standard solution Mr1 = molecular weight of 5-mercapto-1-
Column efficiency: NLT 1700 theoretical plates, from the methyltetrazole, 115.14
Standard solution when calculated: Mr2 = molecular weight of anhydrous sodium 5-
mercapto-1-methyltetrazole, 138.13
Result = 5.545(tr/Wh/2)2 CS = concentration of sodium 5-mercapto-1-
methyltetrazole in the Standard solution
Tailing factor: 0.951.4 for the cefpiramide peak, (g/mL)
Standard solution w = percentage of water in the sodium 5-
Relative standard deviation: NMT 2.0%, Standard mercapto-1-methyltetrazole used to prepare
solution the Standard solution.
Analysis W = weight of Cefpiramide taken to prepare the
Samples: Standard solution and Sample solution Sample solution 2 (mg)
Calculate the quantity, in g, of C25H24N8O7S2 in each mg Calculate the percentage of each other impurity in the
of the Cefpiramide taken: portion of Cefpiramide taken:
Result = (rU/rS) (CS/CU) P Result = (rU/rS) (CS/CU) P F 100
rU = peak response from the Sample solution rU = peak response of each other impurity from
rS = peak response from the Standard solution Sample solution 2
CS = concentration of USP Cefpiramide RS in the rS = peak response of cefpiramide from the Standard
Standard solution (mg/mL) solution
CU = concentration of Cefpiramide taken to prepare CS = concentration of USP Cefpiramide RS in the
the Sample solution (mg/mL) Standard solution (g/mL)
P = designated cefpiramide content of USP CU = concentration of Cefpiramide in Sample solution
Cefpiramide RS (g/mg) 2 (g/mL)
Acceptance criteria: 9741026 g/mg P = designated cefpiramide content of USP
Cefpiramide RS (g/mg)
IMPURITIES F = correction factor, 0.001 mg/g
Organic Impurities Acceptance criteria
PROCEDURE Individual impurities: NMT 0.7% of 5-mercapto-1-
Solution: 4.08 mg/mL of monobasic potassium phosphate methyltetrazole; NMT 0.7% of any other individual
in water, adjust with 1 N sodium hydroxide to a pH of 7.5 impurity
0.1 Total impurities: The sum of 5-mercapto-1-
Mobile phase: Methanol and Solution A (1:3) methyltetrazole and all other impurities is NMT 2.0%.
System suitability solution: Proceed as directed in the
Assay. SPECIFIC TESTS
Sodium 5-mercapto-1-methyltetrazole standard: OPTICAL ROTATION, Specific Rotation 781S: 100 to 112
Determine the water content of the Sodium 5-mercapto-1- Sample solution: 10 mg/mL, in dimethylformamide
methyltetrazole standard (see Water Determination 921). CRYSTALLINITY 695: Meets the requirements
Sample solution 1: Transfer 100 mg of Sodium 5- PH 791: 3.05.0, in a suspension (1 in 200)
mercapto-1-methyltetrazole standard to a stopper WATER DETERMINATION, Method I 921: NMT 9.0%
centrifuge tube. Add 10.0 mL of a solution of N- PYROGEN TEST 151: Where the label states that
ethylmaleimide in methanol (4 in 100), and sonicate for Cefpiramide is sterile, or it must be subjected to further
15 min. Titrate 5.0 mL of this solution as directed in Water processing during the preparation of injectable dosage
Determination 921. forms, it meets the requirements, the test dose being 1.0
Standard stock solution: 0.15 mg/mL of sodium 5- mL/kg of a solution prepared by diluting Cefpiramide with
mercapto-1-methyltetrazole and 0.25 mg/mL of USP Sterile Water for Injection to a concentration of 50 mg/mL of
Cefpiramide RS in Solution A cefpiramide.
Standard solution: 3 g/mL of sodium 5-mercapto-1- STERILITY TESTS 71: Where the label states that
methyltetrazole and 5 g/mL of USP Cefpiramide RS from Cefpiramide is sterile, it meets the requirements when tested
the Standard stock solution diluted with Mobile phase as directed for Test for Sterility of the Product to Be Examined,
Sample solution 2: 0.5 mg/mL of Cefpiramide in Mobile Membrane Filtration.
phase
Assay.
USP 32 Official Monographs / Cefpodoxime 159
ADDITIONAL REQUIREMENTS Tailing factor: 0.951.4 for the cefpiramide peak,

PACKAGING AND STORAGE: Preserve in tight containers. Standard solution
LABELING: Where it is intended for use in preparing Relative standard deviation: NMT 2.0%, Standard
must be subjected to further processing during the Analysis
preparation of injectable dosage forms. Samples: Sample Solution A or Sample solution B, and
USP REFERENCE STANDARDS 11 Standard Solution
USP Cefpiramide RS Calculate the percentage of C25H24N8O7S2 withdrawn from
taken:
Cefpiramide for Injection Result = (rU/rS) (CS/CU) 100
(Comment on this Monograph)id=m14110=Cefpiramide for
Injection=Ca-Chl-Monos.pdf) rU = peak area of the Sample solution
rS = peak area of the Standard solution
DEFINITION CS = concentration of USP Cefpiramide RS in the
Cefpiramide for Injection contains NLT 90.0% and NMT Standard solution (mg/mL)
120.0% of the labeled amount of cefpiramide (C25H24N8O7S2). CU = nominal concentration of cefpiramide in Sample
solution A or Sample solution B (mg/mL)
IDENTIFICATION
The retention time for the cefpiramide peak of the Sample Acceptance criteria: 90.0%120.0%
solution corresponds to that of the Standard solution, as SPECIFIC TESTS
obtained in the Assay. PYROGEN TEST 151: It meets the requirements, the test
ASSAY dose being 1.0 mL/kg of a solution prepared by diluting
PROCEDURE Cefpiramide for Injection with Sterile Water for Injection to a
Solution A: 1.36 mg/mL of monobasic potassium concentration of 50 mg/mL of cefpiramide.
phosphate in water STERILITY TESTS 71: It meets the requirements when tested
Adjust with 1 N sodium hydroxide to a pH of 6.8 0.1. as directed for Test for Sterility of the Product to Be Examined,
Mobile phase: Tetrahydrofuran, acetonitrile, methanol, and Membrane Filtration.
Solution A (3:1:1:45) PH 791: 6.08.0
System suitability solution: 1 mg/mL of USP Cefpiramide Sample solution: Containing the equivalent of 100 mg/mL
RS in 0.01 N sodium hydroxide of cefpiramide
Heat this solution at 95 for 10 min. Mix 1 mL of this WATER DETERMINATION, Method I 921: NMT 3.0%
solution with 19 mL of Mobile phase. This solution contains PARTICULATE MATTER IN INJECTIONS 788: It meets the
a mixture of cefpiramide and cefpiramide lactone. requirements for small-volume injections
Standard solution: 0.25 mg/mL of USP Cefpiramide RS in ADDITIONAL REQUIREMENTS
Mobile phase PACKAGING AND STORAGE: Preserve as described in Injections
Sample solution A (where it is represented as being in a 1, Containers for Sterile Solids.
single-dose container): Constitute a container of USP REFERENCE STANDARDS 11
Cefpiramide for Injection in a volume of water USP Cefpiramide RS
corresponding to the volume of diluent specified in the
labeling. Withdraw all of the withdrawable contents, using a
suitable hypodermic needle and syringe, and dilute with
Mobile phase to obtain a solution containing the nominal Cefpodoxime Proxetil
equivalent of 0.25 mg/mL of cefpiramide. (Comment on this Monograph)id=m14111=Cefpodoxime
Sample solution B (where the label states the quantity of Proxetil=Ca-Chl-Monos.pdf)
cefpiramide in a given volume of constituted solution):
Constitute a container of Cefpiramide for Injection in a
volume of water equivalent to the volume of diluent
specified in the labeling. Dilute the constituted solution with
water to obtain a solution nominally containing 0.25 mg/mL
of cefpiramide.
Mode: LC C21H27N5O9S2 557.60
Detector: UV 254 nm 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid,
Column: 4.6-mm 25-cm; packing L1 7-[[(2-amino-4-thiazolyl)(methoxyimino)acetyl]amino]-3-
Flow rate: 1.5 mL/min (methoxymethyl)-8-oxo-,1-[[(1-methylethoxy)carbonyl]oxy]
Injection size: 20 L ethyl ester, [6R-[6,7(Z)]]-;
System suitability ()-1-Hydroxyethyl(+)-(6R,7R)-7-[2-(2-amino-4-thiazolyl)
Samples: System suitability solution and Standard solution glyoxylamido]-3-methoxymethyl)-8-oxo-5-thia-1-azabicyclo
[NOTEThe relative retention times for cefpiramide and [4.2.0]oct-2-ene-2-carboxylate, 72-(Z)-(O-methyloxime),
cefpiramide lactone are 0.7 and 1.0, respectively.] isopropyl carbonate (ester) [87239-81-4].
Resolution: NLT 5 between the cefpiramide lactone peak DEFINITION
and the cefpiramide peak, System suitability solution Cefpodoxime Proxetil contains the equivalent of NLT 690 g/
Capacity factor, k: 25, Standard solution mg and NMT 804 g/mg of cefpodoxime (C15H17N5O6S2),
Column efficiency: NLT 1700 theoretical plates, Standard calculated on the anhydrous basis.
solution, when calculated:
Result = 5.545 (tr/Wh/2)2
160 Cefpodoxime / Official Monographs USP 32
IDENTIFICATION Acceptance criteria: 690804 g/mg

B. ULTRAVIOLET ABSORPTION 197U IMPURITIES
Solution: 15 g/mL in acetonitrile Inorganic Impurities
C. PROCEDURE RESIDUE ON IGNITION 281: NMT 0.2%
Sample: 1 mg HEAVY METALS, Method II 231: NMT 20 ppm
Analysis: Dissolve in 4 mL of water. Add 1 mL of 1 N Organic Impurities
sulfuric acid while cooling in an ice bath, add 1 mL of a PROCEDURE
freshly prepared solution of sodium nitrite (1 in 100), allow Solution A: 0.02 M ammonium acetate
to stand for 2 min, then add 1 mL of ammonium sulfamate Solution B: Acetonitrile
solution (1 in 100). Allow to stand for 1 min, and add 1 mL Mobile Phase: See the gradient table below.
of N-(1-naphthyl)ethylenediamine dihydrochloride TS.
Acceptance criteria: A red-purple color develops. Time Solution A Solution B
(min) (%) (%)
ASSAY
PROCEDURE 0 90 10
Mobile phase: Acetonitrile and 0.02 M ammonium acetate 10 68 32
(2:3) 40 68 32
Diluent: Acetonitrile and water (2:3)
Standard stock solution: 5 mg/mL of USP Cefpodoxime 80 50 50
Proxetil RS in methanol 85 50 50
Standard solution: 0.025 mg/mL from the Standard stock 90 25 75
solution diluted with Diluent
[NOTEPass through a filter having a 0.45-m or finer 95 25 75
porosity.] 100 90 10
Sample stock solution: 5 mg/mL of Cefpodoxime Proxetil
in methanol Diluent: Acetonitrile and water (1:2)
Sample solution: 0.025 mg/mL from the Sample stock System suitability solution: 10 g/mL of USP
solution diluted with Diluent Cefpodoxime Proxetil RS in Diluent
[NOTEPass through a filter having a 0.45-m or finer [NOTEA volume of methanol not exceeding 10% of the
porosity.] total volume in the final solution may be used to
Chromatographic system facilitate dissolution.]
(See Chromatography 621, System Suitability.) Sample stock solution: 10 mg/mL of Cefpodoxime
Mode: LC Proxetil in methanol
Detector: UV 235 nm [NOTESonicate if necessary.]
Column: 4.6-mm 25-cm; 5-m packing L1 Sample solution: 1.0 mg/mL of Cefpodoxime Proxetil
Temperature: 30 from the Sample stock solution diluted with Diluent
Flow rate: 2 mL/min [NOTEThis solution should be injected promptly, but
Injection size: 20 L may be analyzed within 24 h when stored at 8.]
[NOTERelative retention times are about 0.9 for Mode: LC
cefpodoxime proxetil S-epimer and 1.0 for cefpodoxime Detector: UV 260 nm
proxetil R-epimer.] Column: 4.6-mm 25-cm; 5-m packing L1
Suitability requirements Temperature: 30
Resolution: NLT 2.5 between cefpodoxime proxetil S- Flow rate: 2 mL/min
epimer and cefpodoxime proxetil R-epimer Injection size: 20 L
Tailing factor: NMT 1.5 for cefpodoxime proxetil R- System suitability
epimer Sample: System suitability solution
Relative standard deviation: NMT 1.0% from the sum [NOTEThe relative retention times are about 0.9 for
of the areas of the cefpodoxime proxetil S-epimer and cefpodoxime proxetil S-epimer and 1.0 for cefpodoxime
cefpodoxime proxetil R-epimer peaks proxetil R-epimer. The retention time for the
Analysis cefpodoxime proxetil R-epimer is between 37 and 42
Samples: Standard solution and Sample solution min.]
Calculate the quantity, in g, of C15H17N5O6S2 in each mg Suitability requirements
of Cefpodoxime Proxetil: Resolution: NLT 4.0 between cefpodoxime proxetil S-
epimer and cefpodoxime proxetil R-epimer
Result = (rU/rT) (CS/CU) F Column efficiency: NLT 19,000 theoretical plates
determined from the cefpodoxime proxetil R-epimer
rU = sum of the peak responses for cefpodoxime peak
proxetil S-epimer and cefpodoxime proxetil R- Relative standard deviation: NMT 2.0% from the sum
epimer of the Sample solution of the areas of the cefpodoxime proxetil S-epimer and
rT = sum of the peak responses for cefpodoxime cefpodoxime proxetil R-epimer peaks
proxetil S-epimer and cefpodoxime proxetil R- Analysis
epimer of the Standard solution Samples: Sample solution
CS = concentration of USP Cefpodoxime Proxetil RS in Calculate the percentage of each impurity in the portion
the Standard solution (g/mL) of Cefpodoxime Proxetil taken:
CU = concentration of Cefpodoxime Proxetil in the
Sample solution (g/mL) Result = (rU/rT) 100
F = correction factor, 1000 g/mg
USP 32 Official Monographs / Cefpodoxime 161
rU = peak area for each impurity labeling. Shake the resulting suspension thoroughly, and
rT = sum of the areas of all the peaks determine its density. Transfer a weighed quantity of the
Acceptance criteria suspension, nominally equivalent to 50 mg of cefpodoxime,
Individual impurities: See Impurity Table 1. to a 100-mL volumetric flask. Add 10 mL of water, and
Total impurities: NMT 6.0%, impurity peaks of less than shake to disperse. Add 20 mL of acetonitrile, and sonicate
0.05% being disregarded for 15 min. Cool to room temperature, and dilute with
Diluent to volume.
Impurity Table 1 Sample solution: 0.025 mg/mL from the Sample stock
solution diluted with Diluent
Relative Acceptance [NOTEPass through a filter having a 0.45-m or finer
Retention Criteria, porosity.]
Impurity Name Time NMT(%) Chromatographic system
Individual impurities 0.86 3.0 (See Chromatography 621, System Suitability.)
Individual impurities 1.271.39 1.0
Mode: LC
Detector: UV 235 nm
Individual impurities >2.0 1.0 Column: 4.6-mm 25-cm; 5-m packing L1
All other individual impurities 0.5 Temperature: 30
Flow rate: 2 mL/min
SPECIFIC ROTATION 781S: +35.0 to +48.0 Sample: Standard solution
Sample solution: 10 mg/mL, in methanol [NOTEThe relative retention times for cefpodoxime
WATER DETERMINATION, Method I 921: NMT 3.0% proxetil S-epimer and cefpodoxime proxetil R-epimer are
ISOMER RATIO 0.9 and 1.0, respectively.]
Analysis: Using the chromatogram of the Sample solution Suitability requirements
obtained in the Assay, calculate the ratio of the cefpodoxime Resolution: NLT 2.5 between cefpodoxime proxetil S-
proxetil R-epimer peak response to the sum of the peak epimer and cefpodoxime proxetil R-epimer
responses of the cefpodoxime proxetil S-epimer peak and Tailing factor: NMT 1.5 for cefpodoxime proxetil R-
the cefpodoxime proxetil R-epimer peak. epimer
Acceptance criteria: The ratio is between 0.5 and 0.6. Relative standard deviation: NMT 1.0% from the sum
of the areas of the cefpodoxime proxetil S-epimer and
ADDITIONAL REQUIREMENTS cefpodoxime proxetil R-epimer peaks for replicate
PACKAGING AND STORAGE: Preserve in tight containers, at a injections
temperature not exceeding 25. Analysis
USP Cefpodoxime Proxetil RS Calculate the percentage of C15H17N5O6S2 in the portion of
Oral Suspension taken:
Cefpodoxime Proxetil for Oral Result = (rU/rS) (CS/CU) P F 100

Suspension rU = sum of the peak responses for cefpodoxime
(Comment on this Monograph)id=m14112=Cefpodoxime proxetil S-epimer and cefpodoxime proxetil R-
Proxetil for Oral Suspension=Ca-Chl-Monos.pdf) epimer from the Sample solution
rS = sum of the peak responses for cefpodoxime
DEFINITION proxetil S-epimer and cefpodoxime proxetil R-
Cefpodoxime Proxetil for Oral Suspension contains epimer from the Standard solution
Cefpodoxime Proxetil and one or more buffers, suspending CS = concentration of USP Cefpodoxime Proxetil RS in
agents, sweeteners, flavorings, and preservatives. When the Standard solution (mg/mL)
constituted as directed in the labeling, it contains the CU = nominal concentration of cefpodoxime proxetil
equivalent of NLT 90.0% and NMT 110.0% of the labeled in the Sample solution (mg/mL)
amount of cefpodoxime (C15H17N5O6S2). P = stated content of cefpodoxime in USP
Cefpodoxime Proxetil RS (g/mg)
IDENTIFICATION F = conversion factor, 0.001 mg/g
The retention times of the cefpodoxime proxetil R-epimer Acceptance criteria: 90.0%110.0%
peak and the cefpodoxime proxetil S-epimer peak of the
Sample solution correspond to those in the Standard solution, PERFORMANCE TESTS
as obtained in the Assay. DELIVERABLE VOLUME 698: Meets the requirements
UNIFORMITY OF DOSAGE UNITS 905: It meets the
ASSAY requirements for solids packaged in single-unit containers.
PROCEDURE
Mobile phase: Acetonitrile and 0.02 M ammonium acetate SPECIFIC TESTS
(2:3) PH 791: 4.05.5, in the suspension constituted as directed
Diluent: Acetonitrile and water (2:3) in the labeling
Standard stock solution: 6 mg/mL USP Cefpodoxime WATER DETERMINATION 921: NMT 1.5%
Proxetil RS in methanol
Standard solution: 0.03 mg/mL USP Cefpodoxime Proxetil ADDITIONAL REQUIREMENTS
RS from Standard stock solution diluted with Diluent PACKAGING AND STORAGE: Preserve in tight containers, at a
[NOTEPass through a filter having a 0.45-m or finer temperature not exceeding 30. Store the constituted Oral
porosity.] Suspension in a refrigerator.
Sample stock solution: Constitute a container of USP REFERENCE STANDARDS 11
Cefpodoxime Proxetil for Oral Suspension as directed in the USP Cefpodoxime Proxetil RS
162 Cefpodoxime / Official Monographs USP 32
Cefpodoxime Proxetil Tablets Analysis

(Comment on this Monograph)id=m14113=Cefpodoxime Samples: Standard solution and Sample solution
Proxetil Tablets=Ca-Chl-Monos.pdf) Calculate the percentage of C15H17N5O6S2 in the portion of
Tablets taken:
DEFINITION
Cefpodoxime Proxetil Tablets contain an amount of Result = (rU/rS) (CS/CU) P F 100
Cefpodoxime Proxetil equivalent to NLT 90.0% and NMT
110.0% of the labeled amount of cefpodoxime rU = sum of the peak responses for cefpodoxime
(C15H17N5O6S2). proxetil S-epimer and cefpodoxime proxetil R-
epimer from the Sample solution
IDENTIFICATION rS = sum of the peak responses for cefpodoxime
The retention times of the cefpodoxime proxetil R-epimer proxetil S-epimer and cefpodoxime proxetil R-
peak and the cefpodoxime proxetil S-epimer peak of the epimer from the Standard solution
Sample solution correspond to those in the Standard solution, CS = concentration of USP Cefpodoxime Proxetil RS in
as obtained in the Assay. the Standard solution (mg/mL)
CU = nominal concentration of cefpodoxime proxetil
ASSAY in the Sample solution (mg/mL)
PROCEDURE P = potency of cefpodoxime in USP Cefpodoxime
Mobile phase: Acetonitrile and 0.02 M ammonium acetate Proxetil RS (g/mg)
(2:3) F = conversion factor, 0.001 mg/g
Diluent: Acetonitrile and water (2:3) Acceptance criteria: 90.0%110.0%
Standard stock solution: Transfer about 30mg of USP
Cefpodoxime Proxetil RS to a 50-mL volumetric flask, PERFORMANCE TESTS
dissolve in 5 mL of methanol, dilute with Diluent to volume, DISSOLUTION 711
and mix. Solution A: 54.5 g of glycine and 42.6 g of sodium chloride
Standard solution: 0.03 mg/mL USP Cefpodoxime Proxetil in about 500 mL of water in a 1000-mL volumetric flask
RS from the Standard stock solution diluted with Diluent Cautiously add, with swirling, 14.2 mL of hydrochloric acid,
[NOTEPass through a filter having a 0.45-m or finer and allow to cool. Dilute with water to volume, and mix.
porosity.] Transfer 50 mL of this stock solution to a flask, and dilute
Sample stock solution: Weigh and finely powder NLT 20 with water to 900 mL to obtain a solution having a pH of
Tablets. Transfer a portion of the powder, nominally 3.0 0.1.
equivalent to 50 mg of cefpodoxime, to a 100-mL [NOTEIf necessary, adjust the pH of the stock solution
volumetric flask. Dissolve in 40 mL of Diluent, sonicating for with 10 N sodium hydroxide so that when 50 mL is
5 min. Cool to room temperature, and dilute with Diluent to diluted with water to 900 mL, the pH of the Dissolution
volume. Medium is 3.0 0.1.]
Sample solution: 0.025 mg/mL from the Sample stock Medium: Solution A, 900 mL
solution, diluted with Diluent Apparatus 2: 75 rpm
[NOTEPass through a filter having a 0.45-m or finer Time: 30 min
porosity.] Sample solutions: Sample per Dissolution 711, filtered
Chromatographic system Standard solution: USP Cefpodoxime Proxetil RS dissolved
(See Chromatography 621, System Suitability.) in a small volume of methanol, and diluted with Medium to
Mode: LC a known concentration
Column: 4.6-mm 25-cm; 5-m packing L1 Sample: Standard solution and Sample solution
Temperature: 30 Determine the amount of C15H17N5O6S2 dissolved by
Flow rate: 2 mL/min employing UV absorption at 259 nm on the Sample
Injection size: 20 L solution in comparison with the Standard solution.
System suitability Tolerances: NLT 70% (Q) of the labeled amount of
Sample: Standard solution C15H17N5O6S2
[NOTEThe relative retention times for cefpodoxime proxetil S- UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
epimer and cefpodoxime proxetil R-epimer are 0.9 and 1.0,
respectively.] SPECIFIC TESTS
Suitability requirements WATER DETERMINATION 921: NMT 5.0%
Resolution: NLT 2.5 between cefpodoxime proxetil S- ADDITIONAL REQUIREMENTS
epimer and cefpodoxime proxetil R-epimer PACKAGING AND STORAGE: Preserve in tight containers, at
Tailing factor: NMT 1.5 for cefpodoxime proxetil R- controlled room temperature.
epimer USP REFERENCE STANDARDS 11
Relative standard deviation: NMT 1.0% from the sum USP Cefpodoxime Proxetil RS
of the areas of the cefpodoxime proxetil S-epimer and
cefpodoxime proxetil R-epimer peaks for replicate
injections
USP 32 Official Monographs / Cefprozil 163
Cefprozil Suitability requirements

(Comment on this Monograph)id=m14115=Cefprozil=Ca-Chl- Resolution: NLT 2.5 between the cefprozil (Z)-isomer
Monos.pdf) peak and the cefprozil (E)-isomer peak, System suitability
solution
Column efficiency: NLT 2500 theoretical plates in
Standard solution A from the cefprozil (Z)-isomer peak,
when calculated:
Result = 5.545(tr/Wh/2)2
Tailing factor: 0.91.1, in Standard solution A from the

C18H19N3O5S H2O 407.44 cefprozil (Z)-isomer peak, when calculated:
5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[ami-
no(4-hydroxyphenyl)acetyl]amino]-8-oxo-3-(1-propenyl)- Result = W0.1/2f
monohydrate, [6R-[6, 7(R*)]]-; W0.1 = width of the peak at 10% height
(6R,7R)-7-[(R)-2-Amino-2-(p-hydroxyphenyl)acetamido]-8-oxo-3- Relative standard deviation: NMT 2.0% in Standard
propenyl-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic solution A
acid [121123-17-9]; Analysis
Anhydrous 389.43 Samples: Standard solution A, Standard solution B, and
[92665-29-7]. Sample solution B,
DEFINITION Calculate the quantity (g) of cefprozil (Z)-isomer and
Cefprozil contains NLT 900 g and NMT 1050 g of cefprozil cefprozil (E)-isomer in each mg of Cefprozil taken:
(C18H19N3O5S)/mg, calculated on the anhydrous basis.
IDENTIFICATION
A. INFRARED ABSORPTION 197K rU = peak response of the cefprozil (Z)-isomer or the
Standard solution: USP Cefprozil (Z)-Isomer RS cefprozil (E)-isomer, as appropriate, of the
B. The retention times of the cefprozil (Z)-isomer and Sample solution
cefprozil (E)-isomer peaks of the Sample solution correspond rS = peak response of the cefprozil (Z)-isomer or the
to those of the Standard solutions, as obtained in the Assay. cefprozil (E)-isomer, as appropriate, of the
Standard solution
ASSAY CS = concentration of USP Cefprozil (Z)-isomer RS in
PROCEDURE Standard solution A or of the USP Cefprozil (E)-
Solution A: 11.5 mg/mL of monobasic ammonium isomer RS in Standard solution B, as appropriate
phosphate in water (mg/mL)
Adjust, if necessary, with phosphoric acid to a pH of 4.4. CU = nominal concentration of Cefprozil taken to
Mobile phase: Acetonitrile and Solution A (1:9) prepare the Sample solution (mg/mL)
Pass this solution through a filter having a porosity of 0.5 P = the assigned potency (g/mg) of the appropriate
m or finer. USP Reference Standard
[NOTEDecreasing the proportion of acetonitrile increases Calculate the quantity, in g, of C18H19N3O5S in each mg of
retention times and improves the separation of the the Cefprozil taken by adding the values, in g/mg, of the
cefprozil isomer peaks.] cefprozil (Z)-isomer and the cefprozil (E)-isomer.
Standard solution A: 0.25 mg/mL of USP Cefprozil (Z)- Acceptance criteria: 9001050 g/mg
Isomer RS
[NOTEUse this solution within 6 h.] SPECIFIC TESTS
Standard stock solution B: 0.25 mg/mL of USP Cefprozil CRYSTALLINITY 695: Meets the requirements
(E)-Isomer RS PH 791: 3.56.5, in a solution containing 5 mg/mL
[NOTEUse this solution within 6 h.] WATER DETERMINATION, Method I 921: 3.5%6.5%
Standard solution B: 0.025 mg/mL from Standard stock CEFPROZIL (E)-ISOMER RATIO
solution B diluted with water Analysis: Calculate the ratio of the cefprozil (E)-isomer to
System suitability solution: Standard solution A and the total cefprozil taken:
Standard stock solution B (1:1)
[NOTEUse this solution within 6 h.] Result = E/(E + Z)
Sample solution: 0.3 mg/mL of Cefprozil in water E = content of cefprozil (E)-isomer (g/mg) as
Shake to assure dissolution. determined in the Assay
[NOTEUse this solution within 6 h.] Z = content of cefprozil (Z)-isome (g/mg) as
Chromatographic system determined in the Assay
(See Chromatography 621, System Suitability.) Acceptance criteria: The ratio is 0.060.11.
Mode: LC
Column: 3.9-mm 25-cm; 5-m packing L1 PACKAGING AND STORAGE: Preserve in tight containers.
Injection size: 10 L USP Cefprozil (E)-Isomer RS
System suitability USP Cefprozil (Z)-Isomer RS
Samples: System suitability solution and Standard solution A
[NOTEThe relative retention times for cefprozil (Z)-isomer
and cefprozil (E)-isomer are about 0.7 and 1.0,
respectively.]
164 Cefprozil / Official Monographs USP 32
Cefprozil for Oral Suspension Sample solution: Transfer 15.0 mL of the Sample stock
(Comment on this Monograph)id=m14117=Cefprozil for Oral solution to a 50-mL volumetric flask, dilute with water to
Suspension=Ca-Chl-Monos.pdf) volume, and mix. Pass a portion of this solution through a
filter having a 0.5-m or finer porosity.
DEFINITION [NOTEUse this solution within 6 h.]
Cefprozil for Oral Suspension is a dry mixture of Cefprozil and Chromatographic system
one or more suitable buffers, flavors, preservatives, suspending (See Chromatography 621, System Suitability.)
agents, and sweeteners. It contains NLT 90.0% and NMT Mode: LC
120.0% of the labeled amount of cefprozil (C18H19N3O5S). Detector: UV 280 nm
IDENTIFICATION Flow rate: 1 mL/min
A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Injection size: 10 L
Standard solution: 5 mg/mL of USP Cefprozil (Z)-Isomer System suitability
RS in a mixture of acetone and 0.1 N hydrochloric acid (4:1) Samples: System suitability solution and Standard solution A
Sample solution: Equivalent to 50 mg of cefprozil from [NOTERelative retention times are about 0.7 for the
Cefprozil for Oral Suspension powder, in a 20-mL glass- cefprozil (Z)-isomer and 1.0 for the cefprozil (E)-isomer.]
stoppered test tube Suitability requirements
Add 10 mL of a mixture of acetone and 0.1 N hydrochloric Resolution: NLT 2.5, between the cefprozil (Z)-isomer
acid (4:1), shake for 5 min, and allow to settle. Use the peak and the cefprozil (E)-isomer peak, System suitability
supernatant. solution
Chromatographic system Column efficiency: NLT 2500 theoretical plates from the
(See Chromatography 621, Thin-Layer Chromatography.) cefprozil (Z)-isomer peak, Standard solution A, when
Mode: TLC calculated:
mixture Result = 5.545(tr/Wh/2)2
Developing solvent system: Butyl alcohol, glacial acetic Tailing factor: 0.91.1 from the cefprozil (Z)-isomer
acid, and water (60:20:20) peak, Standard solution A when calculated:
Analysis
Samples: Sample solution and Standard solution Result = W0.1/2f
Allow the spots to dry, and develop the chromatogram in
an equilibrated chromatographic chamber with the W0.1 = width of the peak at 10% height
solvent system, until the solvent front has moved three- Relative standard deviation: NMT 2.0%, Standard
fourths of the length of the plate. Remove the plate from solution A
the chamber, and allow the plate to air-dry in a hood. Analysis
Place the dry plate in a chamber containing iodine vapors. Samples: Sample solution and Standard solution
Examine the plate, and locate the spots. Calculate the concentration of the cefprozil (Z)-isomer and
Acceptance criteria: The RF value of the principal spot of the cefprozil (E)-isomer (mg/mL) in the Sample solution
the Sample solution corresponds to that of the Standard taken:
solution.
B. The retention times of the cefprozil (Z)-isomer and Result = (rU/rS) CS P F
cefprozil (E)-isomer peaks of the Sample solution correspond rU = peak response of the cefprozil (Z)-isomer or the
to those of the Standard solutions, as obtained in the Assay. cefprozil (E)-isomer, as appropriate, of the
ASSAY Sample solution
PROCEDURE rS = peak response of the cefprozil (Z)-isomer or the
Solution A: 11.5 mg/mL of monobasic ammonium cefprozil (E)-isomer, as appropriate, of the
phosphate in water Standard solution
Adjust, if necessary, with phosphoric acid to a pH of 4.4. CS = concentration of USP Cefprozil (Z)-Isomer RS in
Mobile phase: Acetonitrile and Solution A (1:9) Standard solution A or of the USP Cefprozil (E)-
Pass this solution through a filter having a porosity of 0.5 Isomer RS in Standard solution B, as appropriate
m or finer. [NOTEDecreasing the proportion of (mg/mL)
acetonitrile increases retention times and improves the P = the assigned potency of the appropriate USP
separation of the cefprozil isomer peaks.] Reference Standard (g/mg)
Standard solution A: 0.25 mg/mL of USP Cefprozil (Z)- F = correction factor, 0.001 mg/g
Isomer RS Calculate the percentage of C18H19N3O5S in the portion of
[NOTEUse this solution within 6 h.] Cefprozil for Oral Suspension taken:
Standard stock solution B: 0.25 mg/mL of USP Cefprozil
(E)-Isomer RS Result = 100 (CZ + CE)/CU
Standard solution B: 0.025 mg/mL from Standard stock CZ = concentration of the cefprozil (Z)-isomer in the
solution B diluted with water Sample solution (mg/mL)
[NOTEUse this solution within 6 h.] CE = concentration of the cefprozil (E)-isomer in the
System suitability solution: Standard solution A and Sample solution (mg/mL)
Standard stock solution B (1:1) CU = nominal concentration of cefprozil in the Sample
[NOTEUse this solution within 6 h.] solution (mg/mL)
Sample stock solution: Constitute one container of Acceptance criteria: 90%120.0%
Cefprozil for Oral Suspension as directed in the labeling.
Transfer a volume of Cefprozil for Oral Suspension, freshly PERFORMANCE TESTS
mixed and free from air bubbles, nominally equivalent to UNIFORMITY OF DOSAGE UNITS 905: It meets the
250 mg of cefprozil, to a 250-mL volumetric flask, dilute requirements for solids packaged in single-unit containers.
with water to volume, and mix, sonicating briefly.
USP 32 Official Monographs / Cefprozil 165
DELIVERABLE VOLUME 698: It meets the requirements for Standard solution B: 0.025 mg/mL from Standard stock
powder packaged in single-unit containters. solution B, diluted with water
SPECIFIC TESTS System suitability solution: Standard solution A and
PH 791: 4.06.0, in the Cefprozil for Oral Suspension Standard stock solution B (1:1)
constituted as directed in the labeling [NOTEUse this solution within 6 h.]
WATER DETERMINATION, Method I 921: NMT 3.0% Sample stock solution: Transfer a number of Tablets,
nominally equivalent to 1500 mg of cefprozil, to a 250-mL
ADDITIONAL REQUIREMENTS volumetric flask containing 180 mL of water. Allow the
PACKAGING AND STORAGE: Preserve in tight containers. Tablets to disintegrate with the aid of swirling and
USP REFERENCE STANDARDS 11 sonication. Dilute with water to volume.
USP Cefprozil (E)-Isomer RS Sample solution: Nominally 0.3 mg/mL of cefprozil from
USP Cefprozil (Z)-Isomer RS the Sample stock solution diluted with water
[NOTEPass a portion of this solution through a filter
having a 0.5-m or finer porosity, and use the filtrate as
Cefprozil Tablets the Sample solution. Use this solution within 6 h.]
(Comment on this Monograph)id=m14118=Cefprozil (See Chromatography 621, System Suitability.)
Tablets=Ca-Chl-Monos.pdf) Mode: LC
Cefprozil Tablets contain NLT 90.0% and NMT 120.0% of the Column: 3.9-mm 25-cm; 5-m packing L1
labeled amount of cefprozil (C18H19N3O5S). Flow rate: 1 mL/min
IDENTIFICATION System suitability
A. THIN-LAYER CHROMATOGRAPHY Samples: System suitability solution and Standard solution A
Standard solution: 5 mg/mL of USP Cefprozil (Z)-Isomer [NOTEThe relative retention times for cefprozil (Z)-isomer
RS in a mixture of acetone and 0.1 N hydrochloric acid (4:1) and cefprozil (E)-isomer are 0.7 and 1.0, respectively.]
Sample solution: Equivalent to 2.5 mg/mL of cefprozil from Suitability requirements
powdered Tablets in a mixture of acetone and 0.1 N Resolution: NLT 2.5 between the cefprozil (Z)-isomer
hydrochloric acid (4:1) peak and the cefprozil (E)-isomer peak, System suitability
Shake for 5 min, and allow the mixture to settle. Use the solution
supernatant as the Sample solution. Column efficiency: NLT 2500 theoretical plates in
Chromatographic system Standard solution A from the cefprozil (Z)-isomer peak,
(See Chromatography 621, Thin-layer Chromatography.) when calculated:
Mode: TLC
Application volume: 10 L Result = 5.545(tr/Wh/2)2
Developing solvent system: Butyl alcohol, glacial acetic
acid, and water (60:20:20) Tailing factor: 0.91.1 in Standard solution A from the
Analysis cefprozil (Z)-isomer peak, when calculated:
Allow the spots to dry, and develop the chromatogram in Result = W0.1/2f
an equilibrated chromatographic chamber with the W0.1 = width of the peak at 10% height
solvent system, until the solvent front has moved three- Relative standard deviation: NMT 2.0% in Standard
fourths of the length of the plate. Remove the plate from solution A
the chamber, and allow the plate to air-dry in a hood. Analysis
Place the dry plate in a chamber containing iodine vapors. Samples: Sample solution and Standard solution
Examine the plate, and locate the spots. Calculate the concentration of the cefprozil (Z)-isomer and
Acceptance criteria: The RF value of the principal spot from cefprozil (E)-isomer, in mg/mL, of the Sample solution
the Sample solution corresponds to that from the Standard taken:
solution.
B. The retention times of the cefprozil (Z)-isomer and Result = (rU/rS) CS P F
cefprozil (E)-isomer peaks of the Sample solution correspond
to those of the Standard solutions, as obtained in the Assay. rU = peak response of the cefprozil (Z)-isomer or the
cefprozil (E)-isomer, as appropriate, from the
ASSAY Sample solution
PROCEDURE rS = peak response of the cefprozil (Z)-isomer or the
Solution A: 11.5 mg/mL of monobasic ammonium cefprozil (E)-isomer, as appropriate, from the
phosphate in water Standard solution
Adjust, if necessary, with phosphoric acid to a pH of 4.4. CS = concentration of USP Cefprozil (Z)-Isomer RS in
Mobile phase: Acetonitrile and Solution A (1:9) Standard solution A or of the USP Cefprozil (E)-
Filter this solution through a filter having a porosity of 0.5 Isomer RS in Standard solution B, as appropriate
m or finer. (mg/mL)
[NOTEDecreasing the proportion of acetonitrile increases P = assigned potency of the appropriate USP
retention times and improves the separation of the Reference Standard (g/mg)
cefprozil isomer peaks.] F = correction factor, 0.001 mg/g
Standard solution A: 0.25 mg/mL of USP Cefprozil (Z)- Calculate the percentage of label claim of C18H19N3O5S in
Isomer RS the portion of Tablets taken:
Standard stock solution B: 0.25 mg/mL of USP Cefprozil Result = (CZ + CE)/CU 100
(E)-Isomer RS
166 Cefprozil / Official Monographs USP 32
CZ = concentration of the cefprozil (Z)-isomer in the Ceftazidime

Sample solution (mg/mL) (Comment on this Monograph)id=m14120=Ceftazidime=Ca-
CE = concentration of the cefprozil (E)-isomer in the Chl-Monos.pdf)
CU = nominal concentration of the cefprozil in the
Acceptance criteria: 90%120.0%
PERFORMANCE TESTS
DISSOLUTION 711
Apparatus 1: 100 rpm C22H22N6O7S2 5H2O 636.65
Time: 45 min Pyridinium, 1-[[7-[[(2-amino-4-thiazolyl)[(1-carboxy-1-
Analysis: Determine the amount of C18H19N3O5S dissolved methylethoxy)imino]acetyl]amino]-2-carboxy-8-oxo-5-thia-1-
in the Dissolution Medium, as directed in the Assay, using azabicyclo[4.2.0]oct-2-en-3-yl]methyl]-, hydroxide, inner salt,
instead of the Assay Sample solution, a filtered portion of the pentahydrate, [6R[6,7 (Z)]]-;
Dissolution Medium, diluted if necessary, to obtain a Sample 1-[[(6R,7R)-7-[2-(2-Amino-4-thiazolyl)glyoxylamido]-2-
solution containing 0.3 mg of cefprozil/mL. carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en3-
Calculate the quantity, in mg, of cefprozil (Z)-isomer and yl]methyl]pyridinium hydroxide, inner salt, 72-(Z)-[O-(1-
cefprozil (E)-isomer dissolved: carboxy-1-methylethyl)oxime], pentahydrate [78439-06-2].
Anhydrous 546.59
Result = V (rU/rS) CS P F D
DEFINITION
V = volume of Medium in mL, 900 Ceftazidime contains NLT 95.0% and NMT 102.0% of
rU = peak response of the cefprozil (Z)-isomer or the C22H22N6O7S2, calculated on the dried basis.
cefprozil (E)-isomer, as appropriate, from the
Sample solution IDENTIFICATION
rS = peak response of the cefprozil (Z)-isomer or the The retention time of the Standard solution corresponds to
cefprozil (E)-isomer, as appropriate, from the that of the major peak for ceftazidime from the Sample
Standard solution solution as obtained in the Assay.
CS = concentration of USP Cefprozil (Z)-Isomer RS in
Standard solution A or of USP Cefprozil (E)- ASSAY
Isomer RS in Standard solution B, as appropriate PROCEDURE
(mg/mL) Buffer solution: 42.59 mg/mL of anhydrous dibasic sodium
F = correction factor, 0.001 mg/g phosphate and 27.22 mg/mL of monobasic potassium
P = assigned potency of the appropriate USP phosphate in water
Reference Standard (g/mg) Mobile phase: Mix 40 mL of acetonitrile and 200 mL of
D = 1 or, where the filtered Dissolution Medium was Buffer solution, and dilute with water to obtain 2000 mL of
diluted to prepare the Sample solution, the solution. Filter using a filter having a porosity of 1 m or
appropriate dilution factor finer, and degas.
Calculate the percentage of label claim of C18H19N3O5S Standard stock solution: Transfer 29 mg of USP
dissolved: Ceftazidime Pentahydrate RS to a 25-mL volumetric flask
containing 2.5 mL of Buffer solution, and shake until
Result = (MZ + ME) 100/L dissolved. Dilute with water to volume.
[NOTEProtect this solution from light.]
MZ = quantity of cefprozil (Z)-isomer dissolved (mg) Standard solution: 100 g/mL from the Standard stock
ME = quantity of cefprozil (E)-isomer dissolved (mg) solution diluted with water.
L = label claim of cefprozil in Tablet (mg) [NOTEPrepare the solution immediately prior to
Tolerances: NLT 75% (Q) of the labeled amount of chromatography.]
cefprozil (C18H19N3O5S) System suitability stock solution: 0.1 mg/mL of USP
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Ceftazidime, Delta-3-Isomer RS in Buffer solution
System suitability solution: Mix 1 mL of System suitability
SPECIFIC TESTS stock solution with 8 mL of water and 1 mL of the Standard
WATER DETERMINATION, Method I 921: NMT 7.0% stock solution.
ADDITIONAL REQUIREMENTS [NOTEPrepare immediately prior to chromatography.]
PACKAGING AND STORAGE: Preserve in tight containers. Sample stock solution: Transfer 115 mg of Ceftazidime to
USP REFERENCE STANDARDS 11 a 100-mL volumetric flask containing 10.0 mL of Buffer
USP Cefprozil (E)-Isomer RS
USP Cefprozil (Z)-Isomer RS
USP 32 Official Monographs / Ceftazidime 167
solution, and shake until dissolved. Dilute with water to Ceftazidime Injection
volume. (Comment on this Monograph)id=m14122=Ceftazidime
[NOTEProtect this solution from light.] Injection=Ca-Chl-Monos.pdf)
Sample solution: Dilute 5.0 mL of Sample stock solution to
50 mL with water. [NOTEPrepare immediately prior to DEFINITION
chromatography.] Ceftazidime Injection is a sterile isoosmotic solution of
Chromatographic system Ceftazidime in Water for Injection. It contains one or more
(See Chromatography 621, System Suitability.) suitable buffers and a tonicity-adjusting agent. It contains NLT
Mode: LC 90.0% and NMT 120.0% of the labeled amount of
Detector: UV 254 nm C22H22N6O7S2.
Injection size: 20 L The Sample solution exhibits a major peak for ceftazidime, the
System suitability retention time of which corresponds to that exhibited in the
Samples: Standard solution and System suitability solution Standard solution obtained as directed in the Assay.
Resolution: NLT 2.0 between ceftazidime and ASSAY
ceftazidime, delta-3-isomer, System suitability solution PROCEDURE
Tailing factor: 0.751.5, Standard solution Buffer solution: 42.59 mg/mL of anhydrous dibasic sodium
Relative standard deviation: NMT 1.0%, Standard phosphate and 27.22 mg/mL of monobasic potassium
solution phosphate in water
Analysis Mobile phase: Mix 40 mL of acetonitrile and 200 mL of
Samples: Standard solution and Sample solution Buffer solution, and dilute with water to obtain 2000 mL of
Calculate the percentage of C22H22N6O7S2 in the portion solution. Filter, using a filter having a porosity of 1 m or
taken: finer, and degas.
Standard stock solution: Transfer 29 mg of USP
Result = (rU/rS) (CS/CU) 100 Ceftazidime Pentahydrate RS to a 25-mL volumetric flask
containing 2.5 mL of Buffer solution, and shake until
rU = peak response of the Sample solution dissolved. Dilute with water to volume.
rS = peak response of the Standard solution [NOTEProtect this solution from light.]
CS = concentration of Ceftazidime in the Standard Standard solution: 100 g/mL from the Standard stock
solution (g/mL) solution diluted with water
CU = concentration of Ceftazidime in the Sample [NOTEPrepare the solution immediately before
solution (g/mL) chromatography.]
Acceptance criteria: 95.0%102.0% System suitability stock solution: 0.1 mg/mL of USP
Ceftazidime, Delta-3-Isomer RS in Buffer solution
SPECIFIC TESTS System suitability solution: Mix 1 mL of System suitability
CRYSTALLINITY 695: Meets the requirements stock solution with 8 mL of water and 1 mL of the Standard
STERILITY TESTS 71: Where the label states that it is sterile, stock solution.
it meets the requirements when tested as directed for Test [NOTEPrepare immediately before chromatography.]
for Sterility of the Product to Be Examined, Membrane Filtration, Sample stock solution: Allow a container of the Injection
except to use Fluid A. [NOTETo each 1000 mL of Fluid A, to thaw, and mix the solution. Transfer a volume of the
10 g of sodium bicarbonate have been added before Injection, nominally equivalent to 50 mg of ceftazidime, to a
sterilization.] 50-mL volumetric flask, and dilute with Buffer solution to
PH 791: 3.04.0, in a solution containing 5 mg/mL volume.
LOSS ON DRYING 731 Sample solution: Dilute 5.0 mL of Sample stock solution to
Sample: 300 mg 50 mL with water.
Analysis: Dry in vacuum at a pressure not exceeding 5 mm [NOTEPrepare immediately before chromatography.]
of mercury at 60 for 3 h. Chromatographic system
Acceptance criteria: It loses 13.0%15.0% of its weight. (See Chromatography 621, System Suitability.)
BACTERIAL ENDOTOXINS TEST 85: Where the label states that Mode: LC
Ceftazidime is sterile or that it must be subjected to further Detector: UV 254 nm
processing during the preparation of injectable or other Column: 4.6-mm 15-cm; 5-m packing L1
sterile dosage forms, it contains NMT 0.1 USP Endotoxin Flow rate: 2 mL/min
Unit/mg of ceftazidime. Injection size: 20 L
System suitability
ADDITIONAL REQUIREMENTS Samples: System suitability solution and Standard solution
PACKAGING AND STORAGE: Preserve in tight containers. Suitability requirements
LABELING: Where it is intended for use in preparing Resolution: NLT 2.0 between ceftazidime and
injectable dosage forms, the label states that it is sterile or ceftazidime, delta-3-isomer, System suitability solution
must be subjected to further processing during the Tailing factor: 0.751.5, Standard solution
preparation of injectable or other sterile dosage forms. Relative standard deviation: NMT 1.0%, Standard
USP Ceftazidime Delta-3-Isomer RS
USP Ceftazidime Pentahydrate RS
USP Endotoxin RS
168 Ceftazidime / Official Monographs USP 32
Analysis containing 2.5 mL of Buffer solution, and shake until

Samples: Sample solution and Standard solution dissolved. Dilute with water to volume.
Calculate the percentage of C22H22N6O7S2 in the Injection [NOTEProtect this solution from light.]
taken: Standard solution: 100 g/mL from the Standard stock
solution diluted with water
Result = (rU/rS) (CS/CU) 100 [NOTEPrepare the solution immediately prior to use.]
rU = peak response of the Sample solution Ceftazidime, Delta-3-Isomer RS in the Buffer solution
rS = peak response of the Standard solution System suitability solution: Mix 1 mL of System suitability
CS = concentration of ceftazidime in the Standard stock solution with 8 mL of water and 1 mL of the Standard
solution (g/mL) stock solution.
CU = nominal concentration of ceftazidime in the [NOTEPrepare immediately prior to use.]
Sample solution (g/mL) Sample stock solution A: Transfer a quantity of Ceftazidime
Acceptance criteria: 90.0%120.0% for Injection, nominally equivalent to 250 mg of ceftazidime,
to a 250-mL volumetric flask, and dilute with water to
SPECIFIC TESTS volume.
PYROGEN TEST 151: Meets the requirements, the test dose [NOTEProtect this solution from light.]
being a volume of undiluted Injection providing 80 mg of Sample solution A: Dilute 5.0 mL of Sample stock solution A
ceftazidime/kg with water to 50 mL.
STERILITY TESTS 71: Meets the requirements when tested as [NOTEPrepare immediately prior to use.]
directed under Test for Sterility of the Product to Be Examined, Sample stock solution B (where it is represented as being in
Membrane Filtration a single-dose container): Constitute a container of
PH 791: 5.07.5 Ceftazidime for Injection in a volume of water
PARTICULATE MATTER IN INJECTIONS 788: Meets the corresponding to the volume of solvent specified in the
requirements for small-volume injections labeling. Withdraw all of the withdrawable contents, using a
ADDITIONAL REQUIREMENTS suitable hypodermic needle and syringe, and dilute with
PACKAGING AND STORAGE: Preserve as described under water to obtain a solution containing nominally 1 mg of
Injections 1, Containers for Injections. Maintain in the frozen ceftazidime/mL.
state. [NOTEProtect this solution from light.]
LABELING: It meets the requirements for Injections 1, Sample solution B: Dilute 5.0 mL of Sample stock solution B
Labeling. The label states that it is to be thawed just before with water to 50 mL.
use, describes conditions for proper storage of the resultant [NOTEPrepare immediately prior to use.]
solution, and directs that the solution is not to be refrozen. Sample stock solution C (where the label states the quantity
USP REFERENCE STANDARDS 11 of ceftazidime in a given volume of constituted solution):
USP Ceftazidime Delta-3-Isomer RS Constitute a container of Ceftazidime for Injection in a
USP Ceftazidime Pentahydrate RS volume of water corresponding to the volume of solvent
specified in the labeling. Dilute a volume of the constituted
solution with water to obtain a solution containing
nominally 1 mg of ceftazidime/mL.
Ceftazidime for Injection [NOTEProtect this solution from light.]
(Comment on this Monograph)id=m14124=Ceftazidime for Sample solution C: Dilute 5.0 mL of Sample stock solution C
Injection=Ca-Chl-Monos.pdf) with water to 50 mL.
[NOTEPrepare immediately prior to use.]
DEFINITION Chromatographic system:
Ceftazidime for Injection is a sterile mixture of Sterile (See Chromatography 621, System Suitability.)
Ceftazidime and Sodium Carbonate or Arginine. It contains Mode: LC
NLT 90.0% and NMT 105.0% of ceftazidime (C22H22N6O7S2) Detector: UV 254 nm
on the dried and sodium carbonate- or arginine-free basis, and Column: 4.6-mm 15-cm; 5-m packing L1
NLT 90.0% and NMT 120.0% of the labeled amount of Flow rate: 2 mL/min
ceftazidime (C22H22N6O7S2). Injection size: 20 L
System suitability
IDENTIFICATION Samples: System suitability solution and Standard solution
A. The retention time of the major peak for ceftazidime Suitability requirements
from the Sample solutions corresponds to that of the Resolution: NLT 2.0 between ceftazidime and
Standard solution, as obtained in the Assay. ceftazidime, delta-3-isomer, System suitability solution
B. It dissolves in 1 N hydrochloric acid with effervescence, Tailing factor: 0.751.5, Standard solution
evolving a colorless gas, which when passed into calcium Relative standard deviation: NMT 1.0%, Standard
hydroxide TS produces a white precipitate immediately. solution
Analysis
ASSAY I. Samples: Standard solution and Sample solution A
PROCEDURE Calculate the percentage of C22H22N6O7S2 on the dried and
Buffer solution: 42.59 mg/mL of anhydrous dibasic sodium sodium carbonate-free or arginine-free basis in the portion
phosphate and 27.22 mg/mL of monobasic potassium of Ceftazidime for Injection taken:
phosphate in water
Mobile phase: Mix 40 mL of acetonitrile and 200 mL of Result = (rU/rS) {C/[W (100-m-s)]} F V D 100
Buffer solution, and dilute with water to obtain 2000 mL of
solution. Filter, using a filter having a porosity of 1 m or rU = peak response from the Sample solution
finer, and degas. rS = peak response from the Standard solution
Standard stock solution: Transfer 29 mg of USP
Ceftazidime Pentahydrate RS to a 25-mL volumetric flask
USP 32 Official Monographs / Ceftazidime 169
CS = concentration of C22H22N6O7S2 in the Standard CONTENT OF ARGININE (WHERE PRESENT)

solution (mg/mL) Solution A: 1.15 mg/mL of monobasic ammonium
W = mg of Ceftazidime for Injection taken to prepare phosphate in water, adjusted with phosphoric acid to a pH
Sample stock solution A of 2.0 0.1
m = total percentage of loss on drying Mobile phase: Acetonitrile and Solution A (3:1)
s = percentage of sodium carbonate or arginine in Standard solution: 0.2 mg/mL each of USP Ceftazidime
the Ceftazidime for Injection taken Pentahydrate RS and of USP L-Arginine RS
F = correction factor 100 Sample solution: 0.2 mg/mL from Ceftazidime for Injection
V = volume of Sample stock solution A, 250 mL diluted with water
D = dilution factor to prepare Sample solution A from Chromatographic system:
Sample stock solution A, 10 (See Chromatography 621, System Suitability.)
II. Samples: Sample solution B or Sample solution C, and Detector: UV 206 nm
Standard solution Column: 4-mm 25-cm; packing L20
Calculate the percentage of C22H22N6O7S2 withdrawn from Saturator pre-column: 4.6-mm 50-cm; packing L27
the container or in the portion of constituted solution Flow rate: 1 mL/min
taken: Injection size: 20 L
System suitability
Result = (rU/rS) (CS/CU) 100 Sample: Standard solution
rU = peak response from the Sample solution Resolution: NLT 6.0 between the ceftazidime and the
rS = peak response from the Standard solution arginine peaks
CS = concentration of USP Ceftazidime Pentahydrate Tailing factor: NMT 4.0 for the arginine peak
RS in the Standard solution (g/mL) Analysis
CU = nominal concentration of ceftazidime in Sample Samples: Standard solution and Sample solution
solution B or Sample solution C (g/mL) Calculate the percentage of C6H14N4O2 in the Ceftazidime
Acceptance criteria: 90.0%120.0% for Injection taken:
OTHER COMPONENTS Result = (rU/rS) (CS/CU) 100
SODIUM CARBONATE (WHERE PRESENT)
Solution A: 19.07 mg/mL of potassium chloride rU = peak response of arginine from the Sample
Standard stock solution: 14 g/mL of sodium chloride, solution
previously dried at 105 for 2 h rS = peak response of arginine from the Standard
Standard solution: Transfer 10.0 mL of Standard stock solution
solution to a 100-mL volumetric flask, add 10.0 mL of CS = concentration of USP L-Arginine RS in the
Solution A, and dilute with water to volume. Standard solution (mg/mL)
Blank: Transfer 10.0 mL of Solution A to a 100-mL CU = concentration of Ceftazidime for Injection in the
volumetric flask, and dilute with water to volume. Sample solution (mg/mL)
Sample stock solution: Use Sample stock solution A, and [NOTEUse this percentage to calculate the assay result
dilute an aliquot with water to obtain a solution containing from Sample solution A.]
12.5 g of sodium carbonate per mL.
Sample solution: Transfer 10.0 mL of Sample stock solution PERFORMANCE TESTS
to a 100-mL volumetric flask, add 10.0 mL of Solution A and Uniformity of Dosage Units 905: Meets the
dilute with water to volume. requirements
Spectrophotometric conditions
(See Spectrophotometry and Light-Scattering 851.) IMPURITIES
Mode: Atomic absorption spectrophotometer equipped Organic Impurities
with a sodium hollow-cathode lamp and an airacetylene LIMIT OF PYRIDINE
flame Solution A: 5.68 mg/mL of anhydrous dibasic sodium
Analytical wavelength: 589.0 nm (the sodium emission phosphate and 3.63 mg/mL of monobasic potassium
line) phosphate in water
Analysis Mobile phase: Acetonitrile, water, and 0.25 M monobasic
Samples: Standard solution and Sample solution ammonium phosphate (3:6:1), adjusted with ammonium
Calculate the percentage of Na2CO3 in the portion of hydroxide to a pH of 7.0 0.1
Ceftazidime for Injection taken: Pass this solution through a filter having a 1-m or finer
porosity. (See Chromatography 621, System Suitability.)
Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 Standard stock solution: 2.5 mg/mL of pyridine in water
Standard solution: 25 g/mL from the Standard stock
AU = absorbance of the Sample solution solution in Solution A.
AS = absorbance of the Standard solution [NOTEPrepare immediately prior to use.]
CS = concentration of sodium chloride in the Standard Sample solution: To 660 mg of Ceftazidime for Injection,
solution (g/mL) just removed from its container, in a 100-mL volumetric
CU = concentration of Ceftazidime for Injection in the flask, promptly add Solution A to volume, and mix. Store
Sample solution (g/mL) this solution in a cool place, and use it within 1 h.
Mr1 = molecular weight of sodium carbonate, 105.99 Chromatographic system
Mr2 = twice the molecular weight of sodium chloride, (See Chromatography 621, System Suitability.)
116.88
[NOTEUse this percentage to calculate the result from
Sample solution A.]
170 Ceftazidime / Official Monographs USP 32
Mode: LC USP Ceftazidime Pentahydrate RS

System suitability Ceftizoxime Sodium
Sample: Standard solution (Comment on this Monograph)id=m14139=Ceftizoxime
Suitability requirements Sodium=Ca-Chl-Monos.pdf)
Tailing factor: NMT 2.5 in the Standard solution
Relative standard deviation: NMT 3.0% in the
Standard solution
Analysis
Calculate the percentage of pyridine in the portion of
Ceftazidime for Injection taken:
Result = (rU/rS) (CS/CU) 100 C13H12N5NaO5S2 405.39
5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[(2,3-
RU = peak response of pyridine from the Sample dihydro-2-imino-4-thiazoly)(methoxyimino)acetyl]amino]-8-
solution oxomonosodium salt, [6R-[6,7(Z)]]-;
RS = peak response of pyridine from the Standard Sodium (6R,7R)-7-[2-(2-imino-4-thiazolin-4-yl)glyoxylamido]-8-
solution oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylate 72-(Z)-
CS = concentration of pyridine in the Standard (O-methyloxime) [68401-82-1].
solution (g/mL)
CU = concentration of the Sample solution (g/mL) DEFINITION
Impurity Acceptance criteria: NMT 0.4% of pyridine is Ceftizoxime Sodium contains the equivalent of NLT 850 g and
found where it contains sodium carbonate; and NMT NMT 995 g of ceftizoxime (C13H13N5O5S2)/mg, calculated on
0.3% where it contains arginine. the anhydrous basis.
SPECIFIC TESTS IDENTIFICATION

BACTERIAL ENDOTOXINS TEST 85: NMT 0.1 USP Endotoxin A. The chromatogram of the Sample solution exhibits a
Unit/mg of ceftazidime major peak for ceftizoxime, the retention time of which
STERILITY TESTS 71: Meets the requirements for Test for corresponds to that exhibited by the Standard solution,
Sterility of the Product to be Examined, Membrane Filtration obtained as directed in the Assay.
PH 791: 5.07.5, in a solution constituted in the sealed B. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the
container (taking care to relieve the pressure inside the requirements of the tests
container during constitution) containing 100 mg of
ceftazidime/mL ASSAY
LOSS ON DRYING 731: Dry 300 mg in vacuum at a pressure PROCEDURE
not exceeding 5 mm of mercury at 25 for 4 h: where it Buffer A: 1.42 mg/mL of citric acid monohydrate and 1.73
contains arginine, it loses NMT 12.5% of its weight. Where it mg/mL of dibasic sodium phosphate in water
contains sodium carbonate, it loses NMT 13.5% of its Buffer B: 3.63 mg/mL of monobasic potassium phosphate
weight. Where it contains arginine, use the percentage loss and 10.73 mg/mL of dibasic sodium phosphate in water
obtained, m, to calculate the result from Sample solution A. Mobile phase: Acetonitrile and Buffer A (1:9). Filter through
Where it contains sodium carbonate, heat the residue in a filter of 1 m or finer porosity.
vacuum at a pressure not exceeding 5 mm of mercury at [NOTEAdjust the composition, if necessary, to meet the
100 an additional 3 h, and calculate the total percentage of performance requirements under Chromatographic system.]
weight loss. Use this percentage, m, to calculate the result Internal standard solution: Dissolve 1.2 g of salicylic acid
from Sample solution A. in 10 mL of methanol, and dilute with Buffer B to 200 mL.
PARTICULATE MATTER IN INJECTIONS 788: Meets the Standard stock solution: 1 mg/mL of USP Ceftizoxime RS
requirements for small-volume injections in Buffer B
OTHER REQUIREMENTS: Meets the requirements for Injections Standard solution: Add 2.0 mL of Standard stock solution
1, Labeling. and 5.0 mL of Internal standard solution. Dilute with Buffer B
to 100 mL.
ADDITIONAL REQUIREMENTS Sample stock solution: 1 mg/mL of Ceftizoxime Sodium in
PACKAGING AND STORAGE: Preserve as described under Buffer B
Injections 1, Containers for Sterile Solids, protected from Sample solution: Add 2.0 mL of Sample stock solution and
light. 5.0 mL of Internal standard solution. Dilute with Buffer B to
USP REFERENCE STANDARDS 11 100 mL.
USP L-Arginine RS Chromatographic system
USP Ceftazidime Delta-3-Isomer RS (See Chromatography 621, System Suitability.)
USP 32 Official Monographs / Ceftizoxime 171
Detector: UV 254 nm The retention time of the major peak for ceftizoxime from the
Column: 4.0-mm 30-cm; 5- to 10-m packing L1 Sample solution corresponds to that of the Standard solution
Flow rate: 2 mL/min as obtained in the Assay.
Sample: Standard solution PROCEDURE
[NOTEThe relative retention times for ceftizoxime and Buffer A: 1.42 mg/mL of citric acid monohydrate and 1.73
salicylic acid are about 0.6 and 1.0, respectively.] mg/mL of dibasic sodium phosphate
Suitability requirements Buffer B: 3.63 mg/mL of monobasic potassium phosphate
Resolution: NLT 4 between the analyte and internal and 10.73 mg/mL of dibasic sodium phosphate
standard peaks Mobile phase: Acetonitrile and Buffer A (1:9)
Column efficiency: NLT 2000 theoretical plates Pass through a filter of 1-m or finer porosity.
Tailing factor: NMT 2.0 [NOTEAdjust the composition, if necessary, to meet the
Relative standard deviation: NMT 2.0% performance requirements under Chromatographic system.]
Analysis Internal standard solution: Dissolve 1.2 g of salicylic acid
Sample: Standard solution and Sample solution in 10 mL of methanol, and dilute with Buffer B to 200 mL.
Calculate the quantity, in g, of ceftizoxime/mg of the Standard stock solution: 1 mg/mL of USP Ceftizoxime RS
Ceftizoxime Sodium: in Buffer B
Standard solution: Add 2.0 mL of Standard stock solution
Result = (RU/RS) (CS/CU) F and 5.0 mL of Internal standard solution. Dilute with Buffer B
to 100 mL.
RU = peak response ratio of ceftizoxime to the internal Sample stock solution: Allow 1 container of Injection to
standard peak from the Sample solution thaw, and mix. Transfer a volume of the Injection, nominally
RS = peak response ratio of ceftizoxime to the internal equivalent to about 40 mg of ceftizoxime, to a 100-mL
standard peak from the Standard solution volumetric flask, and dilute with Buffer B to volume.
CS = concentration of USP Ceftizoxime RS in the Sample solution: To 5.0 mL of Sample stock solution, add
Standard solution (g/mL) 5.0 mL Internal standard solution. Dilute with Buffer B to 100
CU = concentration of Ceftizoxime Sodium in the mL.
Sample solution (g/mL) Chromatographic system:
F = conversion factor, 1000 g/mg (See Chromatography 621, System Suitability.)
Acceptance criteria: 850995 g/mg Mode: LC
Detector: UV 254 nm
SPECIFIC TESTS Column: 4.0-mm 30-cm; 5- to 10-m packing L1
CRYSTALLINITY 695: Meets the requirements Flow rate: 2 mL/min
PH 791: 6.08.0, in a solution (1 in 10) Injection size: 10 L
STERILITY TESTS 71: Where the label states that Ceftizoxime Sample: Standard solution
Sodium is sterile, it meets the requirements when tested as [NOTEThe relative retention times are about 0.6 for
directed under Test for Sterility of the Product to Be Examined, ceftizoxime and 1.0 for salicylic acid.]
Membrane Filtration. Suitability requirements
BACTERIAL ENDOTOXINS TEST 85: Where the label states that Resolution: NLT 4 between the analyte and internal
Ceftizoxime Sodium is sterile, or it must be subjected to standard peaks.
further processing during the preparation of injectable Column efficiency: NLT 2000 theoretical plates
dosage forms, it contains NMT 0.10 USP Endotoxin Unit/mg Tailing factor: NMT 2.0
of ceftizoxime. Relative standard deviation: NMT 2.0%
Analysis
ADDITIONAL REQUIREMENTS Samples: Sample solution and Standard solution
PACKAGING AND STORAGE: Preserve in tight containers. Calculate the percentage of label claim of C13H13N5O5S2 in
LABELING: Where it is intended for use in preparing the Injection
must be subjected to further processing during the Result = (RU/RS) (CS/CU) 100
USP REFERENCE STANDARDS 11 RU = peak response ratio of the ceftizoxime peak to
USP Ceftizoxime RS the internal standard peak from the Sample
USP Endotoxin RS solution
RS = peak response ratio of the ceftizoxime peak to
the internal standard peak from the Standard
Ceftizoxime Injection solution
CS = concentration of USP Ceftizoxime RS in the
(Comment on this Monograph)id=m14141=Ceftizoxime Standard solution (g/mL)
Injection=Ca-Chl-Monos.pdf) CU = nominal concentration of ceftizoxime sodium in
DEFINITION
Ceftizoxime Injection is a sterile solution of Ceftizoxime Sodium Acceptance criteria: 90.0%115.0%
in a diluent containing one or more tonicity-adjusting agents
in Water for Injection. It contains the equivalent of NLT 90.0% SPECIFIC TESTS
and NMT 115.0% of the labeled amount of ceftizoxime BACTERIAL ENDOTOXINS TEST 85: NMT 0.10 USP Endotoxin
(C13H13N5O5S2). Unit/mg of ceftizoxime
172 Ceftizoxime / Official Monographs USP 32
as directed for Test for Sterility of the Product to be Examined, (See Chromatography 621, System Suitability.)
Membrane Filtration. Mode: LC
PARTICULATE MATTER 788: It meets the requirements for Column: 4.0-mm 30-cm; 5- to 10-m packing L1
small-volume injections. Flow rate: 2 mL/min
PACKAGING AND STORAGE: Preserve as described under Sample: Standard solution
Injections 1, Containers for Injections. Maintain in the frozen [NOTEThe relative retention times are about 0.6 for
state. ceftizoxime and 1.0 for salicylic acid.]
LABELING: It meets the requirements for Injections 1, Suitability requirements
Labeling. The label states that it is to be thawed just before Resolution: NLT 4 between the analyte and internal
use, describes conditions for proper storage of the resultant standard peaks
solution, and directs that the solution is not to be refrozen. Column efficiency: NLT 2000 theoretical plates for the
USP REFERENCE STANDARDS 11 analyte peak
USP Ceftizoxime RS Tailing factor: NMT 2.0 for the analyte peak
USP Endotoxin RS Relative standard deviation: NMT 2%
Analysis
Samples: Sample solution A or Sample solution B, and
Ceftizoxime for Injection Standard solution
(Comment on this Monograph)id=m14143=Ceftizoxime for the container, or in the portion of constituted solution
Injection=Ca-Chl-Monos.pdf) taken:
DEFINITION Result = (RU/RS) (CS/CU) 100
Ceftizoxime for Injection contains an amount of Ceftizoxime
Sodium equivalent to NLT 90.0% and NMT 115.0% of the RU = peak response ratio of ceftizoxime to the internal
labeled amount of ceftizoxime (C13H13N5O5S2). standard peak from Sample solution A or
Sample solution B
ASSAY RS = peak response ratio of ceftizoxime to the internal
PROCEDURE standard peak from the Standard solution
Solution A: 1.42 mg/mL of citric acid monohydrate and CS = concentration of ceftizoxime in the Standard
1.73 mg/mL of dibasic sodium phosphate in water solution (mg/mL)
Solution B: 3.63 mg/mL of monobasic potassium CU = nominal concentration of ceftizoxime in Sample
phosphate and 10.73 mg/mL of dibasic sodium phosphate solution A or Sample solution B (mg/mL)
in water Acceptance criteria: 90.0%115.0%
Mobile phase: Acetonitrile and Solution A (about 1:9). Filter
through a filter (1 m or finer porosity) and adjust the PERFORMANCE TESTS
composition, if necessary, to meet the performance INJECTIONS, Constituted Solutions 1: At the time of use, it
requirements under Chromatographic system. meets the requirements.
Internal standard solution: Dissolve 1.2 g of salicylic acid UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
in 10 mL of methanol and dilute to 200 mL with Solution B.
Standard stock solution: 1 mg/mL of USP Ceftizoxime RS SPECIFIC TESTS
in Solution B BACTERIAL ENDOTOXINS TEST 85: NMT 0.10 USP Endotoxin
Standard solution: Transfer 2.0 mL of the Standard stock Unit/mg of ceftizoxime
solution and 5.0 mL of the Internal standard solution to a STERILITY TESTS, Test for Sterility of the Product to be Examined
100-mL volumetric flask, and dilute with Solution B (0.02 71: It meets the requirements when tested as directed for
mg/mL of ceftizoxime) to volume. Membrane Filtration.
Sample stock solution A: [NOTEPrepare where it is PARTICULATE MATTER IN INJECTIONS 788: Meets the
represented as being in a single-dose container.] Constitute requirements for small-volume injections
Ceftizoxime for Injection in a volume of water corresponding INJECTIONS, Labeling 1: Meets the requirements
to the volume of solvent specified in the labeling. Withdraw OTHER REQUIREMENTS: It meets the requirements for the
all of the withdrawable contents, using a suitable Identification tests under Ceftizoxime Sodium.
hypodermic needle and syringe, and dilute with Solution B CRYSTALLINITY 695: Meets the requirements
to obtain a solution containing 1 mg/mL of ceftizoxime. PH 791: 6.08.0, in a solution (1 in 10)
Sample solution A: Transfer 2.0 mL of Sample stock solution WATER DETERMINATION, Method I 921: NMT 8.5%
A and 5.0 mL of Internal standard solution to a 100-mL
volumetric flask, and dilute with Solution B to volume. ADDITIONAL REQUIREMENTS
Sample stock solution B: [NOTEPrepare where the label PACKAGING AND STORAGE: Preserve as described under
states the quantity of ceftizoxime in a given volume of Injections 1, Containers for Sterile Solids.
constituted solution.] Constitute Ceftizoxime for Injection in USP REFERENCE STANDARDS 11
a volume of water corresponding to the volume of solvent USP Ceftizoxime RS
specified in the labeling. Dilute a volume of the constituted USP Endotoxin RS
solution with Solution B to obtain a solution containing 1
mg/mL of ceftizoxime.
Sample solution B: Transfer 2.0 mL of Sample stock solution
B and 5.0 mL of Internal standard solution to a 100-mL
volumetric flask, and dilute with Solution B to volume.
USP 32 Official Monographs / Ceftriaxone 173
Ceftriaxone Sodium Column efficiency: NLT 1500 theoretical plates, Standard

(Comment on this Monograph)id=m14145=Ceftriaxone solution
Sodium=Ca-Chl-Monos.pdf) Tailing factor: NMT 2.0, Standard solution
Analysis
Calculate the quantity, in g, of C18H18N8O7S3/mg of the
Ceftriaxone Sodium taken:
C18H16N8Na2O7S3 31/2H2O 661.60 rU = peak response of ceftriaxone from the Sample

5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[(2- solution
amino-4-thiazolyl)(methoxyimino)acetyl]amino]-8-oxo-3-[ rS = peak response of ceftriaxone from the Standard
[(1,2,5,6-tetrahydro-2-methyl-5-,6-dioxo-1,2,4-triazin-3- solution
yl)thio]methyl]-, disodium salt, [6R-[6,7(Z)]]-, hydrate, CS = concentration of USP Ceftriaxone Sodium RS in
(2:7); the Standard solution (mg/mL)
(6R,7R)-7-[2-(2-Amino-4-thiazolyl)glyoxylamido]-8-oxo-3-[ CU = concentration of Ceftriaxone Sodium in the
[(1,2,5,6-tetrahydro-2-methyl-5,6-dioxo-as-triazin-3- Sample solution (mg/mL)
yl)thio]methyl]-5-thia-1-azabicyclo[4.2.0]-oct-2-ene-2- P = potency of USP Ceftriaxone Sodium RS in g of
carboxylic acid, 72-(Z)-(O-methyloxime), disodium salt, C18H18N8O7S3/mg
sesquaterhydrate [104376-79-6]. Acceptance criteria: NLT 795 g/mg
Anhydrous 598.56 SPECIFIC TESTS
DEFINITION CRYSTALLINITY 695: Meets the requirements
Ceftriaxone Sodium contains the equivalent of NLT 795 g of PH 791: 6.08.0, in a solution (1 in 10)
ceftriaxone (C18H18N8O7S3)/mg, calculated on the anhydrous WATER DETERMINATION, Method I 921: 8.0%11.0%
basis. STERILITY TESTS 71: It meets the requirements when tested
as directed for Test for Sterility of the Product to be Examined,
IDENTIFICATION Membrane Filtration.
A. INFRARED ABSORPTION 197K BACTERIAL ENDOTOXINS TEST 85: Where the label states that
B. The retention time of the major peak for ceftriaxone it is sterile or must be subjected to further processing during
from the Sample solution corresponds to that of the Standard the preparation of injectable dosage forms, it contains NMT
solution, as obtained in the Assay. 0.20 USP Endotoxin Unit/mg of ceftriaxone.
C. IDENTIFICATION TESTSGENERAL, Sodium 191 ADDITIONAL REQUIREMENTS
ASSAY PACKAGING AND STORAGE: Preserve in tight containers.
PROCEDURE LABELING: Where it is intended for use in preparing
Solution A: 13.6 g/L of dibasic potassium phosphate and injectable dosage forms, the label states that it is sterile or
4.0 g/L of monobasic potassium phosphate in water must be subjected to further processing during the
Adjust with phosphoric acid or 10 N potassium hydroxide preparation of injectable dosage forms.
to a pH of 7.0 0.1. USP REFERENCE STANDARDS 11
Solution B: 25.8 mg/mL of sodium citrate in water USP Ceftriaxone Sodium E-Isomer RS
Adjust with citric acid solution (1 in 5) to a pH of 5.0 0.1. USP Ceftriaxone Sodium RS
Mobile phase: Dissolve 3.2 g of tetraheptylammonium USP Endotoxin RS
bromide in 400 mL of acetonitrile, add 44 mL of Solution A
and 4 mL of Solution B, and add water to make 1000 mL.
Filter through a membrane filter of 0.5-m or finer porosity, Ceftriaxone Injection
and degas.
System suitability solution: 160 g/mL of USP Ceftriaxone (Comment on this Monograph)id=m14147=Ceftriaxone
Sodium E-Isomer RS and 160 g/mL of USP Ceftriaxone Injection=Ca-Chl-Monos.pdf)
Sodium RS in Mobile phase DEFINITION
[NOTEUse this solution promptly after preparation.] Ceftriaxone Injection is a sterile solution of Ceftriaxone Sodium
Standard solution: 0.2 mg/mL of USP Ceftriaxone Sodium in a diluent containing one or more tonicity-adjusting agents
RS in Mobile phase in Water for Injection. It contains the equivalent of NLT 90.0%
[NOTEUse this solution promptly after preparation.] and NMT 115.0% of the labeled amount of ceftriaxone
Sample solution: 0.2 mg/mL of Ceftriaxone Sodium in (C18H18N8O7S3).
Mobile phase
[NOTEUse this solution promptly after preparation.] IDENTIFICATION
Chromatographic system The retention time of the major peak for ceftriaxone from the
(See Chromatography 621, System Suitability.) Sample solution corresponds to that of the Standard solution,
Mode: LC as obtained in the Assay.
Detector: UV 270 nm
Column: 4.0-mm 15-cm; 5-m packing L1 ASSAY
Injection size: 20 L Solvent A: 13.6 mg/mL of dibasic potassium phosphate
System suitability and 4.0 mg/mL of monobasic potassium phosphate in water
Samples: System suitability solution and Standard solution Adjust this solution with phosphoric acid or 10 N potassium
Suitability requirements hydroxide to a pH of 7.0 0.1.
Resolution: NLT 3 between the ceftriaxone E-isomer and Solvent B: 25.8 mg/mL of sodium citrate in water
ceftriaxone peaks, System suitability solution Adjust with citric acid solution (1 in 5) to a pH of 5.0 0.1.
174 Ceftriaxone / Official Monographs USP 32
Mobile phase: Dissolve 3.2 g of tetraheptylammonium USP Ceftriaxone Sodium E-Isomer RS

bromide in 400 mL of acetonitrile, add 44 mL of Solvent A, USP Endotoxin RS
4 mL of Solvent B, and water to 1000 mL. Filter through a
membrane filter of 0.5 m or finer porosity. Make
adjustments if necessary.
System suitability solution: 160 g/mL of USP Ceftriaxone Ceftriaxone for Injection
Sodium E-Isomer RS and 160 g/mL of USP Ceftriaxone (Comment on this Monograph)id=m14149=Ceftriaxone for
Sodium RS in Mobile phase. Injection=Ca-Chl-Monos.pdf)
[NOTEUse this solution promptly after preparation.]
Standard solution: 0.2 mg/mL of USP Ceftriaxone Sodium DEFINITION
RS in Mobile phase Ceftriaxone for Injection contains an amount of Ceftriaxone
[NOTEUse this solution promptly after preparation.] Sodium equivalent to NLT 776 g of ceftriaxone
Sample solution: Allow one container of Injection to thaw (C18H18N8O7S3) per mg, calculated on the anhydrous basis, and
and mix. Transfer a volume of Injection equivalent to 40 mg the equivalent of NLT 90.0% and NMT 115.0% of the labeled
of ceftriaxone to a 200-mL volumetric flask, and dilute to amount of ceftriaxone (C18H18N8O7S3).
volume with Mobile phase.
[NOTEUse this solution promptly after preparation.] ASSAY
(See Chromatography 621, System Suitability.) Solvent A: 13.6 mg/mL of dibasic potassium phosphate
Mode: LC and 4.0 mg/mL of monobasic potassium phosphate in water
Detector: UV 270 nm Adjust this solution with phosphoric acid or 10 N potassium
Column: 4.0-mm 15-cm; 5-m packing L1 hydroxide to a pH of 7.0 0.1.
Flow rate: 2 mL/min Solvent B: 25.8 mg/mL of sodium citrate in water
Injection size: 20 L Adjust with citric acid solution (1 in 5) to a pH of 5.0 0.1.
System suitability Mobile phase: Dissolve 3.2 g of tetraheptylammonium
Samples: Standard solution and System suitability solution bromide in 400 mL of acetonitrile, add 44 mL of Solvent A,
Suitability requirements 4 mL of Solvent B, and water to 1000 mL. Filter through a
Resolution: NLT 3 between the ceftriaxone E-isomer and membrane filter of 0.5 m or finer porosity. Make
ceftriaxone peaks, System suitability solution adjustments if necessary.
Column efficiency: NLT 1500 theoretical plates, Standard System suitability solution: 160 g/mL of USP Ceftriaxone
solution Sodium E-Isomer RS and 160 g/mL of USP Ceftriaxone
Tailing factor: NMT 2, Standard solution Sodium RS in Mobile phase.
Relative standard deviation: NMT 2%, Standard solution [NOTEUse this solution promptly after preparation.]
Analysis Standard solution: 0.2 mg/mL of USP Ceftriaxone Sodium
Samples: Standard solution and Sample solution RS in Mobile phase
Calculate the percentage of ceftriaxone (C18H18N8O7S3) in [NOTEUse this solution promptly after preparation.]
the Injection taken: Sample solution A: 0.2 mg/mL of Ceftriaxone for Injection
in Mobile phase
Result = (rU/rS) (CS/CU) P 100 [NOTEUse this solution promptly after preparation.]
Sample solution B: [NOTEPrepare where it is represented
rU = peak response of ceftriaxone from the Sample as being in a single-dose container.] Constitute Ceftriaxone
solution for Injection in a volume of water corresponding to the
rS = peak response of ceftriaxone from the Standard volume of solvent specified in the labeling. Withdraw all of
solution the withdrawable contents using a suitable hypodermic
CS = concentration of USP Ceftriaxone Sodium RS in needle and syringe, and dilute with Mobile phase to obtain a
the Standard solution (mg/mL) solution containing 180 g/mL of ceftriaxone. [NOTEUse
CU = nominal concentration of ceftriaxone in the this solution promptly after preparation.]
Sample solution (g/mL) Sample solution C: [NOTEPrepare where the label states
P = designated potency of ceftriaxone in the USP the quantity of ceftriaxone in a given volume of constituted
Ceftriaxone Sodium RS (g/mg) solution.] Constitute Ceftriaxone for Injection in a volume of
water corresponding to the volume of solvent specified in
Acceptance criteria: 90.0%115.0% the labeling. Dilute a measured volume of the constituted
SPECIFIC TESTS solution with Mobile phase to obtain a solution containing
BACTERIAL ENDOTOXINS TEST 85: NMT 0.20 USP Endotoxin 180 g/mL of ceftriaxone. [NOTEUse this solution promptly
Unit/mg of ceftriaxone after preparation.]
as directed under Test for Sterility of the Product to be (See Chromatography 621, System Suitability.)
Examined, Membrane Filtration. Mode: LC
PARTICULATE MATTER IN INJECTIONS 788: Meets the Column: 4.0-mm 15-cm; 5-m packing L1
requirements for small-volume injections Flow rate: 2 mL/min
PACKAGING AND STORAGE: Preserve as described under Samples: Standard solution and System suitability solution
Injections 1, Containers for Injections. Maintain in the frozen Suitability requirements
state. Resolution: NLT 3 between the ceftriaxone E-isomer and
LABELING: It meets the requirements under Injections 1, ceftriaxone peaks, System suitability solution
Labeling . The label states that it is to be thawed just prior to Column efficiency: NLT 1500 theoretical plates, Standard
use, describes conditions for proper storage of the resultant solution
solution, and directs that the solution is not to be refrozen.
USP Ceftriaxone Sodium RS
USP 32 Official Monographs / Cefuroxime 175
Tailing factor: NMT 2, Standard solution Cefuroxime Axetil

Relative standard deviation: NMT 2%, Standard solution (Comment on this Monograph)id=m14152=Cefuroxime
Analysis Axetil=Ca-Chl-Monos.pdf)
Samples: Standard solution and Sample solution A, Sample
solution B, or Sample solution C
Calculate the quantity, in g/mg, of C18H18N8O7S3 in the
portion of Ceftriaxone for Injection taken:
Result = (rU/rS) [(CS P)/CU]
rU = peak response of ceftriaxone from Sample
solution A C20H22N4O10S 510.47
rS = peak response of ceftriaxone from the Standard 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid,
solution 3-[[(aminocarbonyl)oxy]methyl]-7-[[2-
CS = concentration of USP Ceftriaxone Sodium RS in furanyl(methoxyimino)acetyl]amino]-8-oxo-, 1-
the Standard solution (mg/mL) (acetyloxy)ethyl ester, [6R-[67(Z)]]-;
P = designated potency of ceftriaxone in USP (RS)-1-Hydroxyethyl (6R,7R)-7-[2-(2-furyl)glyoxylamido]-3-
Ceftriaxone Sodium RS (g/mg) (hydroxymethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene 2-
CU = concentration of Ceftriaxone for Injection in carboxylate, 72-(Z)-(O-methyloxime), 1-acetate 3-carbamate
Sample solution A (mg/mL) [64544-07-6].
the container, or in the portion of constituted solution DEFINITION
taken: Cefuroxime Axetil is a mixture of the diastereoisomers of
C20H22N4O10S. It contains the equivalent of NLT 745 g and
Result = (rU/rS) (CS/CU) P 100 NMT 875 g of C16H16N4O8S/mg, calculated on the anhydrous
basis.
rU = peak response of ceftriaxone from the Sample
solution IDENTIFICATION
rS = peak response of ceftriaxone from the Standard INFRARED ABSORPTION 197K: Meets the requirements
solution
CS = concentration of USP Ceftriaxone Sodium RS in ASSAY
the Standard solution (mg/mL) PROCEDURE
CU = nominal concentration of ceftriaxone in Sample Solvent A: 0.2M monobasic ammonium phosphate
solution B or Sample solution C based on the Mobile phase: Methanol and Solvent A (19:31), filtered
labeled quantity in the container or in the Internal standard solution: 5.4 mg/mL of acetanilide in
portion of constituted solution taken, and the methanol
extent of dilution (g/mL) System suitability stock solution: 0.16 mg/mL of USP
P = stated content of ceftriaxone in USP Ceftriaxone Cefuroxime Axetil Delta-3 Isomers RS in methanol
Sodium RS (g/mg) Standard stock solution: 1.2 mg/mL of USP Cefuroxime
Acceptance criteria: Contains an amount of Ceftriaxone Axetil RS in methanol
Sodium equivalent to NLT 776 g of ceftriaxone per mg, System suitability solution: 10.0 mL of Standard stock
calculated on the anhydrous basis, and 90.0%115.0% of solution, 5.0 mL of Internal standard solution, and 3.8 mL of
the labeled amount of C18H18N8O7S3. System suitability stock solution made to 50.0 mL with Solvent
A
SPECIFIC TESTS Standard solution: 10.0 mL of Standard stock solution, 5.0
CONSTITUTED SOLUTION: At the time of use, it meets the mL of Internal standard solution, and 3.8 mL of methanol
requirements for Constituted Solutions under Injections 1, made to 50.0 mL with Solvent A
Labeling. [NOTEUse this Standard solution promptly, or refrigerate
BACTERIAL ENDOTOXINS 85: NMT 0.20 USP Endotoxin and use on the day prepared.]
Unit/mg of ceftriaxone Sample stock solution: 1.2 mg/mL of Cefuroxime Axetil in
STERILITY 71: It meets the requirements when tested as methanol
directed under Test for Sterility of the Product to be Examined, Sample solution: 10.0 mL of Sample stock solution, 5.0 mL
Membrane Filtration. of Internal standard solution, and 3.8 mL of methanol made
PARTICULATE MATTER IN INJECTIONS 788: Meets the to 50.0 mL with Solvent A
requirements for small-volume injections [NOTEUse this Sample solution promptly, or refrigerate
CRYSTALLINITY 695: Meets the requirements and use on the day prepared.]
PH 791: 6.08.0, in a solution (1 in 10) Chromatographic system
WATER DETERMINATION, Method I 921: 8.0%11.0% (See Chromatography 621, System Suitability.)
OTHER REQUIREMENTS: It responds to the Identification tests Mode: LC
under Ceftriaxone Sodium. It also meets the requirements Detector: UV 278 nm
for Uniformity of Dosage Units 905 and for Injections 1, Column: 4.6-mm 25-cm; 5-m packing L13
Labeling. Flow rate: 1.5 mL/min
PACKAGING AND STORAGE: Preserve as described under Samples: System suitability solution and Standard solution
Injections 1, Containers for Sterile Solids. [NOTEThe relative retention times for acetanilide,
USP REFERENCE STANDARDS 11 cefuroxime axetil diastereoisomer B, cefuroxime axetil
USP Ceftriaxone Sodium RS diastereoisomer A, and cefuroxime axetil delta-3 isomers
USP Ceftriaxone Sodium E-Isomer RS are 0.4, 0.8, 0.9, and 1.0, respectively.]
USP Endotoxin RS
176 Cefuroxime / Official Monographs USP 32
Suitability requirements Cefuroxime Axetil for Oral Suspension

Resolution: NLT 1.5 between cefuroxime axetil (Comment on this Monograph)id=m14153=Cefuroxime Axetil
diastereoisomer A and B; NLT 1.5 between cefuroxime for Oral Suspension=Ca-Chl-Monos.pdf)
axetil diastereoisomer A and cefuroxime axetil delta-3
isomers, System suitability solution DEFINITION
Column efficiency: NLT 3000 theoretical plates when Cefuroxime Axetil for Oral Suspension contains NLT 90.0% and
measured using the cefuroxime axetil diastereoisomer A NMT 110.0% of the labeled amount of cefuroxime
peak, Standard solution (C16H16N4O8S).
Relative standard deviation: NMT 2.0%, Sample solution
Samples: Standard solution and Sample solution The retention times of the major peaks for cefuroxime axetil
Calculate the quantity, in g, of C16H16N4O8S in each mg of diastereoisomers A and B of the Sample solution correspond
Cefuroxime Axetil taken: to those of the Standard solution, both relative to the
internal standard, as obtained in the Assay.
Result = (RU/RS) (CS/CU) (PS/100) (100 K)
ASSAY
RU = ratio of the sum of the peak responses of PROCEDURE
cefuroxime axetil diastereoisomers A and B to Solution A: 23.0 mg/mL of monobasic ammonium
the peak response of the internal standard phosphate in water
from the Sample solution Mobile phase: Methanol and Solution A (19:31)
RS = ratio of the sum of the peak responses of System suitability stock solution A: 1.2 mg/mL of USP
cefuroxime axetil diastereoisomers A and B to Cefuroxime Axetil RS in methanol
the peak response of the internal standard System suitability stock solution B: 0.16 mg/mL of USP
from the Standard solution Cefuroxime Axetil Delta-3 Isomers RS in methanol
CS = concentration of USP Cefuroxime Axetil RS in System suitability solution: Transfer 10.0 mL of System
the Standard solution (mg/mL) suitability stock solution A to a 50-mL volumetric flask. Add
CU = concentration of Cefuroxime Axetil in the Sample 5.0 mL of methanol and 3.8 mL of System suitability stock
solution (mg/mL) solution B. Dilute with Solution A to volume.
PS = designated C16H16N4O8S content of anhydrous Standard stock solution: 1.2 mg/mL of USP Cefuroxime
USP Cefuroxime Axetil RS (g/mg) Axetil RS in methanol
K = percentage water content of USP Cefuroxime Standard solution: Transfer 10.0 mL of Standard stock
Axetil RS solution to a 50-mL volumetric flask, add 8.8 mL of
Acceptance criteria: 745875 g/mg methanol, and dilute with Solution A to volume.
[NOTEUse this Standard solution promptly, or refrigerate
SPECIFIC TESTS and use on the day prepared.]
CRYSTALLINITY 695: Particles that do not show Sample stock solution: Transfer to a 100-mL volumetric
birefringence or exhibit extinction positions are amorphous, flask a portion equivalent to 250 mg of cefuroxime from
and particles that show birefringence and exhibit extinction Cefuroxime Axetil for Oral Suspension, freshly mixed and
positions are crystalline. free from air bubbles, constituted as directed in the labeling
WATER DETERMINATION, Method I 921: NMT 1.5% in 50 mL of methanol, and shake by mechanical means for
DIASTEREOISOMER RATIO: 10 min. Dilute with methanol to volume, and mix. Filter a
Solvent A, Mobile phase, Internal standard solution, portion of this stock solution.
System suitability solution, System suitability stock Sample solution: Transfer 5.0 mL of the filtered Sample
solution, Standard stock solution, Standard solution, stock solution to a 50-mL volumetric flask. Add 13.8 mL of
Sample stock solution, Sample solution, and methanol, dilute to volume with Solution A.
Chromatographic system: Prepare as directed in the [NOTEProtect this Sample solution from light and use
Assay. promptly, or refrigerate and use on the day prepared.]
Analysis: Proceed as directed in the Assay. Chromatographic system
Calculate the ratio of cefuroxime axetil diastereoisomer A to (See Chromatography 621, System Suitability.)
the sum of the cefuroxime axetil diastereoisomers A and B. Mode: LC
Detector: UV 278 nm
Result = rA/(rA + rB) Column: 4.6-mm 25-cm; 5-m packing L13
rA = peak response of cefuroxime axetil Injection size: 10 L
diastereoisomers A System suitability
rB = peak response of cefuroxime axetil Samples: System suitability solution and Standard solution
diastereoisomers B [NOTEThe relative retention times for acetanilide,
Acceptance criteria: 0.480.55 cefuroxime axetil diastereoisomer B, cefuroxime axetil
ADDITIONAL REQUIREMENTS diastereoisomer A, and cefuroxime axetil delta-3 isomers
PACKAGING AND STORAGE: Preserve in tight containers. are 0.4, 0.8, 0.9, and 1.0, respectively.]
LABELING: Label it to indicate whether it is amorphous or Suitability requirements
crystalline. Resolution: NLT 1.5 between cefuroxime axetil
USP REFERENCE STANDARDS 11 diastereoisomer A and B; NLT 1.5 between cefuroxime
USP Cefuroxime Axetil RS axetil diastereoisomer A and cefuroxime axetil delta-3
USP Cefuroxime Axetil Delta-3 Isomers RS isomers, System suitability solution
Column efficiency: NLT 3000 theoretical plates when Cefuroxime Axetil Tablets
measured using the cefuroxime axetil diastereoisomer A (Comment on this Monograph)id=m14154=Cefuroxime Axetil
peak, Standard solution Tablets=Ca-Chl-Monos.pdf)
solution DEFINITION
Analysis Cefuroxime Axetil Tablets contain the equivalent of NLT 90.0%
Samples: Standard solution and Sample solution and NMT 110.0% of the labeled amount of C16H16N4O8S.
Calculate the percentage of C16H16N4O8S in each mL taken:
IDENTIFICATION
Result = (RU/RS) (CS/CU) PS F (100 K) The retention times of the major peaks for cefuroxime axetil
diastereoisomers A and B of the Sample solution correspond
RU = sum of the peak responses of the cefuroxime to those in the Standard solution, both relative to the
axetil diastereoisomers A and B from the internal standard, as obtained in the Assay.
Sample solution
RS = sum of the of the peak responses of the ASSAY
cefuroxime axetil diastereoisomers A and B PROCEDURE
from the Standard solution Solution A: 23.0 mg/mL of monobasic ammonium
CS = concentration of the Standard solution (mg/mL) phosphate in water
CU = nominal concentration of cefuroxime axetil in Mobile phase: Methanol and Solution A (19:31), filtered
the Sample solution (mg/mL) Internal standard solution: 5.4 mg/mL of acetanilide in
PS = designated cefuroxime content of anhydrous methanol
USP Cefuroxime Axetil RS (g/mg) System suitability stock solution A: 1.2 mg/mL of USP
F = conversion correction factor (0.001 mg/g) Cefuroxime Axetil RS in methanol
K = percentage of water content of USP Cefuroxime System suitability stock solution B: 0.16 mg/mL of USP
Axetil RS Cefuroxime Axetil Delta-3 Isomers RS in methanol
Acceptance criteria: 90.0%110.0% System suitability solution: In a 50-mL volumetric flask,
mix 10.0 mL of System suitability stock solution A, 5.0 mL of
PERFORMANCE TESTS Internal standard solution, and 3.8 mL of System suitability
DISSOLUTION 711 stock solution B. Dilute with Solution A to volume.
Medium: 0.07 M pH 7.0 phosphate buffer, prepared by Standard stock solution: 1.2 mg/mL of USP Cefuroxime
dissolving 3.7 mg/mL of monobasic sodium phosphate and Axetil RS in methanol
5.7 mg/mL of anhydrous dibasic sodium phosphate in Standard solution: Transfer 10.0 mL of Standard stock
water; 900 mL solution to a 50-mL volumetric flask, add 5.0 mL of Internal
Apparatus 2: 50 rpm standard solution and 3.8 mL of methanol, and dilute with
Time: 30 min Solution A to volume.
Analysis: Test 5.0 mL of constituted Cefuroxime Axetil for [NOTEUse this Standard solution promptly, or refrigerate
Oral Suspension equivalent to 125 or 250 mg of cefuroxime. and use on the day prepared.]
Determine the amount of cefuroxime equivalent dissolved Sample stock solution: Finely powder NLT 10 Tablets.
by using UV absorption at the wavelength of maximum Transfer the powder with the aid of methanol to a
absorbance at 280 nm on filtered portions of the solution volumetric flask of such capacity that when filled to volume,
under test, suitably diluted with Dissolution Medium, if the solution will contain the equivalent of about 2 mg/mL of
necessary, in comparison with a Standard solution having a C16H16N4O8S. Add methanol to fill the volumetric flask to
known concentration of USP Cefuroxime Axetil RS in the about half its capacity, and shake by mechanical means for
same Medium. about 10 min. Dilute with methanol to volume. Filter a
Tolerances: NLT 60% (Q) of the labeled amount of portion of this stock mixture.
C16H16N4O8S Sample solution: Transfer 5.0 mL of the filtrate from the
UNIFORMITY OF DOSAGE UNITS 905 Sample stock soluton to a 50-mL volumetric flask. Add 5.0
Constitute Cefuroxime Axetil for Oral Suspension as directed mL of Internal standard solution and 8.8 mL of methanol,
in the labeling for a solid packaged in single-unit and dilute with Solution A to volume.
containers. Mix, and allow the contents of the container to [NOTEUse this Sample solution promptly, or refrigerate
drain into a beaker for 5 s. Withdraw and assay 5.0 mL of and use on the day prepared.]
Cefuroxime Axetil for Oral Suspension from the beaker, or Chromatographic system
the total amount if it is less than 5 mL. It meets the (See Chromatography 621, System Suitability.)
requirements. Mode: LC
Detector: UV 278 nm
SPECIFIC TESTS Column: 4.6-mm 25-cm; 5-m packing L13
PH 791: 3.57.0 Flow rate: 1.5 mL/min
DELIVERABLE VOLUME 698 Injection size: 10 L
For multiple-unit containers: Constitute Cefuroxime Axetil System suitability
for Oral Suspension as directed in the labeling for a solid Samples: System suitability solution and Standard solution
packaged in multiple-unit containers. It meets the [NOTEThe relative tretention times for acetanilide,
requirements. cefuroxime axetil diastereoisomer B, cefuroxime axetil
WATER DETERMINATION, Method I 921: NMT 6.0% diastereoisomer A, and cefuroxime axetil delta-3 isomers
ADDITIONAL REQUIREMENTS are 0.4, 0.8, 0.9, and 1.0, respectively.]
PACKAGING AND STORAGE: Preserve in well-closed containers, Suitability requirements
and store at a controlled room temperature. Resolution: NLT 1.5 between cefuroxime axetil
USP REFERENCE STANDARDS 11 diastereoisomer A and B; NLT 1.5 between cefuroxime
USP Cefuroxime Axetil RS axetil diastereoisomer A and cefuroxime axetil delta-3
USP Cefuroxime Axetil Delta-3 Isomers RS isomers, System suitability solution
Column efficiency: NLT 3000 theoretical plates when USP Cefuroxime Axetil Delta-3 Isomers RS
measured using the cefuroxime axetil diastereoisomer A
peak, Standard solution
solution Cefuroxime Sodium
Analysis (Comment on this Monograph)id=m14155=Cefuroxime
Samples: Standard solution and Sample solution Sodium=Ca-Chl-Monos.pdf)
Calculate the percentage of C16H16N4O8S in each Tablet:
Result = (RU/RS) (CS/CU) (PS) (100 K)

RU = ratios of the sum of the peak responses of the
cefuroxime axetil diastereoisomers A and B to
the peak responses of the Internal standard of
the Sample solution
RS = ratios of the sum of the peak responses of the C16H15N4NaO8S 446.37
cefuroxime axetil diastereoisomers A and B to 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 3-[
the peak responses of the Internal standard of [(aminocarbonyl)oxy]methyl]-7-[[2-
the Standard solution furanyl(methoxyimino)acetyl]amino]-8-oxo-, monosodium salt
CS = concentration of USP Cefuroxime Axetil RS in [6R-[6,7 (Z)]]-;
the Standard solution (mg/mL) Sodium (6R, 7R)-7-[2-(2-furyl)glyoxylamido]-3-
CU = nominal concentration of cefuroxime axetil in (hydroxymethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-
the Sample solution (mg/mL) carboxylate, 72-(Z)-(O-methyloxime), carbamate (ester)
PS = designated C16H16N4O8S content of anhydrous [56238-63-2]
USP Cefuroxime Axetil RS (g/mg)
K = percentage of water content of USP Cefuroxime DEFINITION
Axetil RS Cefuroxime Sodium contains the equivalent of NLT 855 g and
Acceptance criteria: 90.0%110.0% NMT 1000 g of cefuroxime (C16H16N4O8S), calculated on the
anhydrous basis.
PERFORMANCE TESTS
DISSOLUTION 711 IDENTIFICATION
Test 1 A. The retention time of the major peak for cefuroxime
Medium: 0.07 N hydrochloric acid; 900 mL from the Sample solution corresponds to that of the Standard
Apparatus 2: 55 rpm solution, as obtained in the Assay.
Time: 15 and 45 min B. IDENTIFICATION TESTSGENERAL, Sodium 191: It meets
Analysis: Determine the amount of C16H16N4O8S dissolved the requirements.
by employing UV absorption at the wavelength of maximum
absorbance at 278 nm on filtered portions of the solution ASSAY
under test, suitably diluted with Dissolution Medium, if PROCEDURE
necessary, in comparison with a Standard solution having a Solution A: pH 3.4 acetate buffer
known concentration of USP Cefuroxime Axetil RS, Transfer 50 mL of 0.1 M sodium acetate to a 1000-mL
equivalent to 0.01 to 0.02 mg of C16H16N4O8S/mL in the volumetric flask, and dilute with 0.1 N acetic acid to
same Medium. volume.
Acceptance criteria: NLT 60% (Q) of the labeled amount Mobile phase: Acetonitrile and Solution A (1:10)
of C16H16N4O8S is dissolved in 15 min, and NLT 75% (Q) is Filter through a membrane filter of 1-m or finer porosity.
dissolved in 45 min; except that where Tablets are labeled Internal standard solution: 1.5 mg/mL of orcinol in water
to contain the equivalent of 500 mg of cefuroxime, NLT Standard solution: Prepare a 1 mg/mL of C16H16N4O8S
50% (Q) of the labeled amount of C16H16N4O8S is dissolved from USP Cefuroxime Sodium RS in water. Immediately
in 15 min, and NLT 70% (Q) is dissolved in 45 min. transfer 5.0 mL of the resulting solution to a 100-mL
Test 2: If the product complies with this test, the labeling volumetric flask, add 20.0 mL of Internal standard solution,
indicates that it meets USP Dissolution Test 2. and dilute with water to volume. This Standard solution
Apparatus 2: 100 rpm contains 0.05 mg of cefuroxime/mL.
Medium, Times, and Analysis: Proceed as directed under Sample solution: Prepare 1 mg/mL of Cefuroxime Sodium
Test 1. in water. Immediately transfer 5.0 mL of the resulting
Acceptance criteria: NLT 60% (Q) of the labeled amount solution to a 100-mL volumetric flask, add 20.0 mL of
of C16H16N4O8S is dissolved in 15 min, and NLT 75% (Q) of Internal standard solution, and dilute with water to volume.
the labeled amount of C16H16N4O8S is dissolved in 45 min. Chromatographic system
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements (See Chromatography 621, System Suitability.)
Mode: LC
WATER DETERMINATION, Method I 921: NMT 6.0% Column: 4.6-mm 15-cm; 5-m packing L15
Flow rate: 2 mL/min
PACKAGING AND STORAGE: Preserve in well-closed containers. System suitability
LABELING: The labeling indicates whether the Tablets contain Sample: Standard solution
amorphous or crystalline Cefuroxime Axetil. If Tablets contain [NOTEThe relative retention times for cefuroxime and
a mixture of amorphous and crystalline Cefuroxime Axetil, orcinol are about 0.5 and 1.0, respectively.]
label to indicate the percentage of each contained therein. Suitability requirements
When more than one Dissolution test is given, the labeling Resolution: NLT 3.5 between the analyte and internal
states the Dissolution test used only if Test 1 is not used. standard peaks in the Standard solution
USP Cefuroxime Axetil RS
Column efficiency: NLT 1300 theoretical plates in the Internal standard solution: 1.5 mg/mL of orcinol in water
Standard solution Standard solution: 1 mg/mL of C16H16N4O8S from USP
Tailing factor: NMT 2.0 in the Standard solution Cefuroxime Sodium RS in water
Relative standard deviation: NMT 2.0% in the Standard Immediately transfer 5.0 mL of the resulting solution to a
solution 100-mL volumetric flask, add 20.0 mL of Internal standard
Analysis solution, and dilute with water to volume.
Samples: Standard solution and Sample solution Sample solution: Allow a container of Injection to thaw,
Calculate the quantity, in g, of cefuroxime per mg in the and mix the solution. Transfer a measured volume of
portion of Cefuroxime Sodium taken: Injection, equivalent to about 50 mg of cefuroxime, to a 50-
mL volumetric flask, and dilute with water to volume.
Result = (RU/RS) (CS/CU) F Immediately transfer 5.0 mL of the resulting solution to a
100-mL volumetric flask, add 20.0 mL of Internal standard
RU = peak response ratio of cefuroxime to the internal solution, and dilute with water to volume.
standard peak from the Sample solution Chromatographic system
RS = peak response ratio of cefuroxime to the internal (See Chromatography 621, System Suitability.)
standard peak from the Standard solution Mode: LC
CS = concentration of USP Cefuroxime Sodium RS in Detector: UV 254 nm
the Standard solution (mg/mL) Column: 4.6-mm 15-cm; 5-m packing L15
CU = concentration of Cefuroxime Sodium in the Flow rate: 2 mL/min
Sample solution (mg/mL) Injection size: 10 L
F = conversion factor, 1000 g/mg System suitability
Acceptance criteria: 8551000 g/mg Sample: Standard solution
[NOTEThe relative retention times for cefuroxime and
SPECIFIC TESTS orcinol are about 0.5 and 1.0, respectively.]
PH 791: 6.08.5, in a solution (1 in 10) Suitability requirements
WATER DETERMINATION, Method I 921: NMT 3.5% Resolution: NLT 3.5 between the analyte and internal
STERILITY TESTS 71: Where the label states that Cefuroxime standard peaks, Standard solution
Sodium is sterile, it meets the requirements when tested as Column efficiency: NLT 1300 theoretical plates, Standard
directed for Test for Sterility of the Product to Be Examined , solution
Membrane Filtration. Tailing factor: NMT 2.0, Standard solution
BACTERIAL ENDOTOXINS TEST 85 Where the label states that Relative standard deviation: NMT 2.0%, Standard
Cefuroxime Sodium is sterile or must be subjected to further solution
processing during the preparation of injectable dosage Analysis
forms, it contains NMT 0.10 USP Endotoxin Unit/mg of Sample: Sample solution and Standard solution
cefuroxime. Calculate the percentage of C16H16N4O8S in each mL of the
ADDITIONAL REQUIREMENTS Injection:
LABELING: Where it is intended for use in preparing Result = (RU/RS) (CS/CU) 100
injectable dosage forms, the label states that it is sterile, or RU = peak response ratio of cefuroxime to the internal
must be subjected to further processing during the standard peak from the Sample solution
preparation of injectable dosage forms. RS = peak response ratio of cefuroxime to the internal
USP REFERENCE STANDARDS 11 standard peak from the Standard solution
USP Cefuroxime Sodium RS CS = concentration of USP Cefuroxime Sodium RS in
USP Endotoxin RS the Standard solution (mg/mL)
CU = nominal concentration of cefuroxime taken to
prepare the Sample solution (mg/mL)
Cefuroxime Injection Acceptance criteria: 90.0%120.0%
(Comment on this Monograph)id=m14156=Cefuroxime PERFORMANCE TESTS
Injection=Ca-Chl-Monos.pdf) UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Cefuroxime Injection is a sterile isoosmotic solution of BACTERIAL ENDOTOXINS TEST 85: NMT 0.10 USP Endotoxin
Cefuroxime Sodium in Water for Injection. It contains one or Unit/mg of cefuroxime
more suitable buffers and a tonicity-adjusting agent. It STERILITY TESTS 71: Meets the requirements when tested as
contains NLT 90.0% and NMT 120.0% of the labeled amount directed for Test for Sterility of the Product to Be Examined,
of cefuroxime (C16H16N4O8S). Membrane Filtration
PH 791: 5.07.5
IDENTIFICATION PARTICULATE MATTER IN INJECTIONS 788: Meets the
The retention time of the major peak for cefuroxime from the requirements for small-volume injections
Sample solution corresponds to that in the Standard solution,
as obtained in the Assay. ADDITIONAL REQUIREMENTS
PACKAGING AND STORAGE: Preserve as described under
ASSAY Injections 1, Packaging, Containers for Injections. Maintain in
PROCEDURE the frozen state.
Solution A: Transfer 50 mL of 0.1 M sodium acetate to a LABELING: It meets the requirements under Injections 1,
1000-mL volumetric flask, and dilute with 0.1 N acetic acid Labels and Labeling. The label states that it is to be thawed
to volume. just before use, describes conditions for proper storage of
Filter through a membrane filter of 1 m or finer porosity.
the resultant solution, and directs that the solution is not to Calculate the percentage of C16H16N4O8S withdrawn from
be refrozen. the container, or in the portion of constituted solution or
USP REFERENCE STANDARDS 11 suspension taken:
USP Cefuroxime Sodium RS
USP Endotoxin RS Result = (RU/RS) (CS/CU) 100
RU = peak response ratio of the cefuroxime peak to
the internal standard from the Sample solution
Cefuroxime for Injection RS = peak response ratio of the cefuroxime peak to
(Comment on this Monograph)id=m14158=Cefuroxime for the internal standard from the Standard solution
Injection=Ca-Chl-Monos.pdf) CS = concentration of cefuroxime in the Standard
solution (mg/mL)
DEFINITION CU = nominal concentration of cefuroxime, of Sample
Cefuroxime for Injection contains an amount of Cefuroxime solution A or Sample solution B, based on the
Sodium equivalent to NLT 90.0% and NMT 120.0% of the labeled quantity in the container or in the
labeled amount of cefuroxime (C16H16N4O8S). portion of constituted solution or suspension
taken, and the extent of dilution (mg/mL)
ASSAY [NOTEWhere the test for Uniformity of Dosage Units has
PROCEDURE been performed using the Analysis for content uniformity,
Solution A: Transfer 50 mL of 0.1 M sodium acetate to a use the average of these determinations as the Assay
1000-mL volumetric flask, and dilute with 0.1 N acetic acid value.]
to volume. Acceptance criteria: 90.0%120.0%
Pass through a membrane filter of 1 m or finer porosity. PERFORMANCE TESTS
Internal standard solution: 1.5 mg/mL of orcinol in water UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Standard stock solution: Prepare a solution of USP Procedure for content uniformity: Perform the Assay on
Cefuroxime Sodium RS equivalent to 1 mg/mL of individual containers using Sample solution A or Sample
cefuroxime. Immediately transfer 5.0 mL of the resulting solution B, or both, as appropriate.
solution to a 100-mL volumetric flask, add 20.0 mL of
Internal standard solution, and dilute with water to volume. SPECIFIC TESTS
Sample solution A (where it is represented as being in a CONSTITUTED SOLUTION: At the time of use, the constituted
single-dose container): Constitute Cefuroxime for Injection in solution for intravenous administration prepared from
a volume of water corresponding to the volume of solvent Cefuroxime for Injection meets the requirements for
specified in the labeling. Withdraw all the withdrawable Constituted Solutions under Injections 1, Labeling.
contents, using a suitable hypodermic needle and syringe, BACTERIAL ENDOTOXINS TEST 85: NMT 0.10 USP Endotoxin
and dilute quantitatively with water to obtain a solution Unit/mg of cefuroxime
containing about 1 mg/mL of cefuroxime. Immediately STERILITY TESTS 71: It meets the requirements when tested
transfer 5.0 mL of the resulting solution to a 100-mL as directed for Test for Sterility of the Product to be Examined,
volumetric flask, add 20.0 mL of Internal standard solution, Membrane Filtration.
and dilute with water to volume. PARTICULATE MATTER IN INJECTIONS 788: Meets the
Sample solution B (where the label states the quantity of requirements for small-volume injections
cefuroxime in a given volume of constituted solution or PH 791: 6.08.5, in a solution (1 in 10)
suspension): Constitute Cefuroxime for Injection in a volume WATER DETERMINATION, Method I 921: NMT 3.5%
of water, corresponding to the volume of solvent specified OTHER REQUIREMENTS: Meets the requirements for Injections
in the labeling. Dilute a measured volume of the constituted 1, Labeling and meets the requirements of the Identification
solution or suspension quantitatively with water to obtain a tests under Cefuroxime Sodium.
solution containing about 1 mg/mL of cefuroxime.
Immediately transfer 5.0 mL of the resulting solution to a ADDITIONAL REQUIREMENTS
100-mL volumetric flask, add 20.0 mL of Internal standard PACKAGING AND STORAGE: Preserve as described under
solution, and dilute with water to volume. Injections 1, Containers for Sterile Solids.
Chromatographic system USP REFERENCE STANDARDS 11
(See Chromatography 621, System Suitability.) USP Cefuroxime Sodium RS
Mode: LC USP Endotoxin RS
Detector: UV 254 nm
Flow rate: 2 mL/min Oxidized Cellulose
System suitability (Comment on this Monograph)id=m14200=Oxidized
Sample: Standard solution Cellulose=Ca-Chl-Monos.pdf)
[NOTEThe relative retention times are 0.5 for cefuroxime DEFINITION
and 1.0 for orcinol.] Oxidized Cellulose contains NLT 16.0% and NMT 24.0% of
Suitability requirements carboxyl groups (COOH), calculated on the dried basis. It is
Resolution: NLT 3.5 between the analyte and internal sterile.
standard peaks in the Standard solution
Column efficiency: NLT 1300 theoretical plates, Standard IDENTIFICATION
solution PROCEDURE
Tailing factor: NMT 2.0, Standard solution Sample solution: 200 mg in 10 mL of 0.25 N sodium
Relative standard deviation: NMT 2.0%, Standard hydroxide
solution Analysis: Shake the Sample solution for 1 min. Add 10 mL
Analysis of water, and shake: the solution so obtained shows no
Samples: Sample solution A, or Sample solution B, and more than a slight haze, and is substantially free from fibers
Standard solution
USP 32 Official Monographs / Cellulose 181
and foreign particles. Allow to stand for 10 min: any swollen ADDITIONAL REQUIREMENTS
fibers initially present are no longer visible. Acidify with 3 N PACKAGING AND STORAGE: Preserve as described under
hydrochloric acid. Injections 1, Containers for Sterile Solids, protected from
Acceptance criteria: A flocculent white precipitate is direct sunlight. Store in a cold place.
formed. LABELING: The package bears a statement to the effect that
the sterility of Oxidized Cellulose cannot be guaranteed if
ASSAY the package bears evidence of damage, or if the package
PROCEDURE has been previously opened. Oxidized Cellulose meets the
Sample solution: 500 mg of Oxidized Cellulose, previously requirements for Injections 1, Labeling.
dried over phosphorus pentoxide in vacuum for 18 h, in a
125-mL conical flask
Add 50.0 mL of calcium acetate solution (1 in 50), swirl
until the sample is completely covered, allow the mixture Oxidized Regenerated Cellulose
to stand for 30 min, then add phenolphthalein TS. (Comment on this Monograph)id=m14210=Oxidized
Analysis: Titrate the solution with 0.1 N sodium hydroxide Regenerated Cellulose=Ca-Chl-Monos.pdf)
VS. Perform a blank determination by titrating 50.0 mL of
the calcium acetate solution, and make any necessary DEFINITION
correction (see Titrimetry 541). Each mL of 0.1 N sodium Oxidized Regenerated Cellulose contains NLT 18.0% and NMT
hydroxide is equivalent to 4.502 mg of COOH. 24.0% of carboxyl groups (COOH), calculated on the dried
Acceptance criteria: 16.0%24.0% basis. It is sterile.
IMPURITIES IDENTIFICATION
Inorganic Impurities PROCEDURE
LIMIT OF NITROGEN Sample solution: 200 mg in 10 mL of 0.25 N sodium
Sample: 1 g hydroxide
Analysis: Previously dried sample in vacuum over Analysis: Shake the Sample solution for 1 min. Add 10 mL
phosphorus pentoxide for 18 h, in a 500-mL Kjeldahl flask of water, and shake: the solution so obtained shows no
Arrange a 125-mL conical flask, containing 30 mL of boric more than a slight haze and is substantially free from fibers
acid solution (1 in 25) and 6 drops of mixed indicator (1 and from foreign particles. Allow to stand for 10 min: any
part of methyl red TS and 4 parts of bromocresol green swollen fibers initially present are no longer visible. Acidify
TS), beneath the condenser of the distillation apparatus so with 3 N hydrochloric acid.
that the tip of the condenser is well below the surface of Acceptance criteria: A flocculent white precipitate is
the boric acid solution. To the Kjeldahl flask containing the formed.
Sample, add 1 g of Devardas alloy, 100 mL of recently
boiled water, a small lump of paraffin, and 100 mL of 1 N ASSAY
sodium hydroxide. Connect the Kjeldahl flask to the PROCEDURE
condenser by a suitable trap bulb. Heat the mixture in the Sample solution: 1 g of Oxidized Regenerated Cellulose,
flask until 4550 mL of distillate has collected in the previously dried at 90 for 2 h, in a 250-mL conical flask.
receiver. Rinse the condenser, and titrate the boric acid Pipet 10 mL of 0.5 N sodium hydroxide VS into the flask,
solution with 0.02 N sulfuric acid VS to a pale pink swirl to dissolve, and add 100 mL of water.
endpoint. Perform a blank determination, and make any Analysis: Immediately titrate with 0.1 N hydrochloric acid
necessary correction. Each mL of 0.02 N sulfuric acid is VS to a phenolphthalein endpoint. Perform a blank
equivalent to 0.2801 mg of nitrogen. determination, and note the difference in volumes required.
Acceptance criteria: NMT 0.5% Each mL of the difference in volumes of 0.1 N hydrochloric
RESIDUE ON IGNITION 281: NMT 0.15% acid consumed is equivalent to 4.50 mg of COOH.
Organic Impurities Acceptance criteria: 18.0%24.0%
LIMIT OF FORMALDEHYDE
Sample: 500 mg OTHER COMPONENTS
Analysis: Transfer the Sample to a 500-mL iodine flask. NITROGEN CONTENT
Add 250 mL of water, and allow to stand for NLT 2 h with Sample: 1 g
intermittent shaking. Pipet 0.50 mL of the supernatant into Analysis: Transfer a previously dried Sample in a 500-mL
a glass-stoppered test tube, and add 10 mL of Kjeldahl flask. Arrange a 125-mL conical flask, containing 30
chromotropic acid TS. Stopper the tube loosely, and heat mL of boric acid solution (1 in 25) and 6 drops of mixed
in a boiling water bath for 30 min. Cool, and determine indicator (1 part of methyl red TS and 4 parts of
the absorbance of the solution at 570 nm with a suitable bromocresol green TS), beneath the condenser of the
spectrophotometer, using a mixture of 0.5 mL of water distillation apparatus so that the tip of the condenser is well
and 10 mL of chromotropic acid TS as the blank. below the surface of the boric acid solution. To the Kjeldahl
Acceptance criteria: The absorbance does not exceed that flask containing the Sample, add 1 g of Devardas alloy, 100
produced when 0.50 mL of dilute formaldehyde solution (1 mL of recently boiled water, a small lump of paraffin, and
in 40,000) is treated in the same manner (0.5%). 100 mL of 1 N sodium hydroxide. Connect the Kjeldahl flask
to the condenser by a suitable trap bulb. Heat the mixture
SPECIFIC TESTS in the flask until 4550 mL of distillate has collected in the
STERILITY TESTS 71: Meets the requirements, the test receiver. Rinse the condenser, and titrate the boric acid
specimen weighing approximately 250 mg, and 0.5 mL of solution with 0.02 N sulfuric acid VS to a pale pink endpoint
0.1 N sodium hydroxide being added to the portions of that persists for 30 s. Perform a blank determination, and
media used make any necessary correction. Each mL of 0.02 N sulfuric
LOSS ON DRYING 731: Dry in vacuum over phosphorus acid is equivalent to 0.2801 mg of nitrogen.
pentoxide for 18 h: it loses NMT 15.0% of its weight.
182 Cellulose / Official Monographs USP 32
Acceptance criteria: NMT 0.5% follows. Add 10 mL of 5 N nitric acid, 10.0 mL of

ammonium vanadate TS, and about 60 mL of water.
IMPURITIES Swirl, and add 10.0 mL of a freshly prepared solution of
Inorganic Impurities 2.5 g of ammonium molybdate in 50 mL of warm water.
RESIDUE ON IGNITION 281: NMT 0.15% Dilute with water to volume. Concomitantly determine
Organic Impurities the absorbances, AU and AS, of the solutions from the
LIMIT OF FORMALDEHYDE Standard solution and the Sample solution, respectively, at
Sample: 500 mg 400 nm with a suitable spectrophotometer, using the
Analysis: Transfer the Sample to a 500-mL iodine flask. reagent blank to set the instrument.
Add 250 mL of water, and allow to stand for NLT 2 h with Calculate the percentage of total phosphate taken:
intermittent shaking. Pipet 0.5 mL of the supernatant into a
glass-stoppered test tube, and add 10 mL of chromotropic (AU/AS) (10,000/W)
acid TS. Stopper the tube loosely, and heat in a boiling
water bath for 30 min. Cool, and determine the AU = absorbance of the solution from the Sample
absorbance of the solution at 570 nm, with a suitable solution
spectrophotometer, using a mixture of 0.5 mL of water AS = absorbance of the solution from the Standard
and 10 mL of chromotropic acid TS as the blank. solution
Acceptance criteria: The absorbance does not exceed that W = weight of Cellulose Sodium Phosphate taken
produced when 0.5 mL of dilute formaldehyde solution (1 (mg)
in 40,000) is treated in the same manner (0.5% CH2O). Calculate the percentage of inorganic bound phosphate in
the undried Cellulose Sodium Phosphate by subtracting
SPECIFIC TESTS from this result the percentage of Free phosphate.
STERILITY TESTS 71: Meets the requirements; the test
specimen weighing approximately 250 mg and 0.5 mL of Acceptance criteria: 31.0%36.0%
0.1 N sodium hydroxide being added to the portions of OTHER COMPONENTS
media used. NITROGEN DETERMINATION, Method I 461: NMT 1.0%
LOSS ON DRYING 731: Dry about 150 mg at 90 for 2 h: it FREE PHOSPHATE
loses NMT 15% of its weight. Standard solution: Prepare as directed under Inorganic
ADDITIONAL REQUIREMENTS Bound Phosphate.
PACKAGING AND STORAGE: Preserve as described under Sample solution: 2 g of Cellulose Sodium Phosphate in 100
Injections 1, Containers for Sterile Solids, protected from mL of water, stir, allow to stand for 5 min, stir again, and
direct sunlight. Store at controlled room temperature. filter through moderately retentive filter paper, collecting the
LABELING: The package bears a statement to the effect that filtrate in a dry flask.
the sterility of Oxidized Regenerated Cellulose cannot be Analysis: Transfer 2.0 mL of the Standard solution and 5.0
guaranteed if the package bears evidence of damage, or if mL of the Sample solution to separate 100-mL volumetric
the package has been previously opened. Oxidized flasks. Proceed as directed in the Analysis under Inorganic
Regenerated Cellulose meets the requirements for Injections Bound Phosphate, beginning with Treat each of these.
1, Labeling. Calculate the percentage of free phosphate taken:
(AU/AS) (4,000/W)
Cellulose Sodium Phosphate AU = absorbance of the solution from the Sample

solution
(Comment on this Monograph)id=m14260=Cellulose Sodium AS = absorbance of the solution from the Standard
Phosphate=Ca-Chl-Monos.pdf) solution
DEFINITION W = weight of undried Cellulose Sodium Phosphate
Cellulose Sodium Phosphate is prepared by phosphorylation of taken (mg)
alpha cellulose. It has an inorganic bound phosphate content Acceptance Criteria: NMT 3.5% of free phospate,
of NLT 31.0% and NMT 36.0%, calculated on the dried basis. calculated on the dried basis
SODIUM CONTENT
ASSAY Standard stock solution: 508.5 mg of sodium chloride,
INORGANIC BOUND PHOSPHATE previously dried at 105 for 2 h, in 100 mL of water
Standard solution: Transfer 358.2 mg of monobasic Transfer to a 1000-mL volumetric flask, and dilute with
potassium phosphate, primary standard grade, to a 250-mL water to volume.
volumetric flask, and dissolve in and dilute with water to [NOTEEach mL of this solution contains 200 g of sodium]
volume. Standard solutions: Transfer 5.0, 10.0, 15.0, and 20.0 mL
[NOTEEach mL of this solution contains 1.0 mg of of the Standard stock solution to separate 100-mL volumetric
phosphate.] flasks, and dilute each with water to volume.
Sample solution: Transfer 250 mg of Cellulose Sodium Sample solution: 250 mg of Cellulose Sodium Phosphate in
Phosphate to a 250-mL conical flask. Rinse to the bottom 10 mL of a mixture of 20 mL of perchloric acid and 15 mL
with a few mL of water. Add 10 mL of a mixture of 20 mL of nitric acid. Heat cautiously to the production of dense,
of perchloric acid and 15 mL of nitric acid. Heat cautiously white fumes, cool, transfer to a 1000-mL volumetric flask,
to the production of dense, white fumes, cool the clear, and dilute with water to volume.
almost colorless, solution, and transfer to a 100-mL Analysis: Concomitantly determine the emission of the
volumetric flask with the aid of water. Dilute with water to Sample solution and of each Standard solution at the sodium
volume. emission line of 589 nm, with a suitable flame photometer.
Analysis Plot the emissions of the Standard solutions versus their
Samples: Standard solution and Sample solution concentration of sodium, and draw a straight line best
Transfer 2.0-mL portions of the Standard solution and the fitting the four plotted points. From the graph so obtained,
Sample solution to separate 100-mL volumetric flasks. Treat determine the concentration of sodium in the Sample
each of these and a third flask, providing the blank, as
USP 32 Official Monographs / Cellulose 183
solution. Calculate the percentage of sodium in the undried ADDITIONAL REQUIREMENTS

Cellulose Sodium Phosphate. PACKAGING AND STORAGE: Preserve in well-closed containers.
Acceptance criteria: 9.5%13.0%, calculated on the dried
basis
IMPURITIES Cellulose Sodium Phosphate for Oral
Inorganic Impurities Suspension
HEAVY METALS, Method III 231: NMT 40 ppm (Comment on this Monograph)id=m14265=Cellulose Sodium
SPECIFIC TESTS Phosphate for Oral Suspension=Ca-Chl-Monos.pdf)
PH 791: Place 3 g of it in a 100-mL beaker, add 60 mL of DEFINITION
water, and stir occasionally for 5 min. Filter through a Cellulose Sodium Phosphate for Oral Suspension contains
sintered-glass crucible. The pH of the filtrate is 6.09.0. Cellulose Sodium Phosphate. It has an inorganic bound
LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT phosphate content of NLT 28.0% and NMT 36.0%, calculated
10.0% of its weight. on the dried basis.
CALCIUM BINDING CAPACITY
Standard solution: Transfer 0.33 g of dried calcium ASSAY
carbonate, primary standard grade, in a 250-mL beaker with INORGANIC BOUND PHOSPHATE
the aid of a few mL of water, and dilute with water to about Standard solution: Transfer 358.2 mg of monobasic
50 mL. Carefully and dropwise, add 2 N hydrochloric acid potassium phosphate, primary standard grade, to a 250-mL
until all of the solid dissolves, and add 2 drops in excess. volumetric flask, and dissolve in and dilute with water to
Heat the solution to boiling, and boil for 5 min. Cool the volume.
solution, and transfer to a 1000-mL volumetric flask. Dilute [NOTEEach mL of this solution contains 1.0 mg of
with water to volume. phosphate.]
Calculate the molarity, M, of the solution taken: Sample solution: Transfer the equivalent to 250 mg of
cellulose sodium phosphate from Cellulose Sodium
Result = g/100.09 Phosphate for Oral Suspension to a 250-mL conical flask.
Rinse to the bottom with a few mL of water. Add 10 mL of
g = weight of calcium carbonate taken (g) a mixture of 20 mL of perchloric acid and 15 mL of nitric
Standard edetate disodium titrant: Dissolve 10 g of acid. Heat cautiously to the production of dense, white
edetate disodium in 100 mL of water. Slowly add alcohol fumes, cool the clear, almost colorless, solution, and transfer
until the first permanent precipitate is formed. Filter, and to a 100-mL volumetric flask with the aid of water. Dilute
discard the solid. Add an equal volume of alcohol to the with water to volume.
filtrate. Filter the resulting precipitate, discard the filtrate, Analysis
and wash the residue on the filter, first with acetone, then Samples: Standard solution and Sample solution
with ethyl ether. Dry at 80 for 4 days at about 50% relative Transfer 2.0-mL portions of the Standard solution and the
humidity. Transfer 3.72 g of this purified edetate disodium Sample solution to separate 100-mL volumetric flasks. Treat
to a 1000-mL volumetric flask, and dissolve with water. each of these and a third flask, providing the blank, as
Dilute with water to volume. follows. Add 10 mL of 5 N nitric acid, 10.0 mL of
Calculate the molarity, MS, of the solution taken: ammonium vanadate TS, and about 60 mL of water.
Result = w/372.24 Swirl, and add 10.0 mL of a freshly prepared solution of
2.5 g of ammonium molybdate in 50 mL of warm water.
w = weight of the purified edetate disodium taken (g) Dilute with water to volume. Concomitantly determine
Analysis: Transfer 0.15 0.02 g of Cellulose Sodium the absorbances, AU and AS, of the solutions from the
Phosphate to a 250-mL beaker. Add 150.0 mL of the Standard solution and the Sample solution, respectively, at
Standard solution, and stir the mixture for 5 min on a 400 nm with a suitable spectrophotometer, using the
magnetic stirrer. Filter, discarding the first few mL of the reagent blank to set the instrument.
filtrate. To 50.0 mL of the filtrate, add about 50 mL of Calculate the percentage of total phosphate in the portion
water, 15 mL of 1 N sodium hydroxide, and 300 mg of of Cellulose Sodium Phosphate for Oral Suspension:
hydroxy naphthol blue. Titrate with Standard edetate
disodium titrant to a permanent deep blue color, and (AU/AS) (10,000/W)
designate the number of mL consumed as VS. AU = absorbance of the solution from the Sample
Calculate the calcium binding capacity of the undried solution
Cellulose Sodium Phosphate, in mmol/g: AS = absorbance of the solution from the Standard
Result = (150M 3VS MS)/W solution
W = weight of Cellulose Sodium Phosphate for Oral
W = weight of Cellulose Sodium Phosphate taken (g) Suspension taken (mg)
Acceptance criteria: The calcium binding capacity is NLT Calculate the percentage of inorganic bound phosphate in
1.8 mmol/g, calculated on the dried basis. the portion of undried Cellulose Sodium Phosphate for Oral
184 Cellulose / Official Monographs USP 32
Suspension by subtracting from this result the percentage edetate disodium titrant to a permanent deep blue color, and
of free phosphate. designate the number of mL consumed as VS.
Acceptance criteria: 28.0%36.0% of inorganic bound Calculate the calcium binding capacity, in mmol/g, of the
phosphate, calculated on the dried basis portion of undried Cellulose Sodium Phosphate for Oral
Suspension:
OTHER COMPONENTS
FREE PHOSPHATE Result = (150M 3VS MS)/W
Standard solution: Prepare as directed under Inorganic
Bound Phosphate. W = weight of Cellulose Sodium Phosphate for Oral
Sample solution: Equivalent to 2 g of cellulose sodium Suspension taken, and the other terms are as
phosphate from Cellulose Sodium Phosphate for Oral defined above (g)
Suspension in 100 mL of water Acceptance criteria: The calcium binding capacity is NLT
Stir, allow to stand for 5 min, stir again, and filter through 1.8 mmol/g, calculated on the dried basis
moderately retentive filter paper, collecting the filtrate in a
dry flask. ADDITIONAL REQUIREMENTS
Analysis: Transfer 2.0 mL of the Standard solution and 5.0 PACKAGING AND STORAGE: Preserve in tight containers. Store
mL of the Sample solution to separate 100-mL volumetric in a refrigerator.
flasks. Proceed as directed in the Analysis under Inorganic
Bound Phosphate, beginning with Treat each of these.
Calculate the percentage of free phosphate in the portion of Cephalexin
Cellulose Sodium Phosphate for Oral Suspension:
(Comment on this Monograph)id=m14280=Cephalexin=Ca-Chl-
(AU/AS) (4000/W) Monos.pdf)
AU = absorbance of the solution from the Sample

solution
As = absorbance of the solution from the Standard
solution
W = weight of undried Cellulose Sodium Phosphate
for Oral Suspension taken (mg)
Acceptance criteria: NMT 6.0% free phosphate, calculated
on the dried basis. C16H17N3O4S H2O 365.40
5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-
SPECIFIC TESTS [(aminophenylacetyl)amino]-3-methyl-8-oxo-, monohydrate,
LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT [6R-[6,7 (R*)]]-;
10.0% of its weight. (6R,7R)-7-[(R)-2-Amino-2-phenylacetamido]-3-methyl-8-oxo-5-
CALCIUM BINDING CAPACITY thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid
Standard solution: Transfer 0.33 g of dried calcium monohydrate [23325-78-2]
carbonate, primary standard grade, in a 250-mL beaker with Anhydrous [15686-71-2].
the aid of a few mL of water, and dilute with water to about 347.40
50 mL. Carefully and dropwise add 2 N hydrochloric acid
until all of the solid dissolves, and add 2 drops in excess. DEFINITION
Heat the solution to boiling, and boil for 5 min. Cool the Cephalexin has a potency of NLT 950 g and NMT 1030 g of
solution, and transfer to a 1000-mL volumetric flask. Dilute C16H17N3O4S/mg, calculated on the anhydrous basis.
Calculate the molarity, M, of the solution taken: IDENTIFICATION
A. The IR absorption spectrum of a potassium bromide
Result = g/100.09 dispersion of it exhibits maxima only at the same
wavelengths as that of a similar solution of USP Cephalexin
g = weight of calcium carbonate taken (g) RS.
Standard edetate disodium titrant: Dissolve 10 g of B. The UV absorption spectrum of a solution (1 in 50,000)
edetate disodium in 100 mL of water. Slowly add alcohol exhibits maxima and minima at the same wavelengths as
until the first permanent precipitate is formed. Filter, and that of a similar solution of USP Cephalexin RS,
discard the solid. Add an equal volume of alcohol to the concomitantly measured, and the absorptivity, calculated on
filtrate. Filter the resulting precipitate, discard the filtrate, the anhydrous basis, at the wavelength of maximum
and wash the residue on the filter, first with acetone, then absorbance at 262 nm is NLT 95.0% and NMT 104.0% of
with ethyl ether. Dry at 80 for 4 days at about 50% relative that of USP Cephalexin RS, the potency of the Reference
humidity. Transfer about 3.72 g of this purified edetate Standard being taken into account.
disodium to a 1000-mL volumetric flask, and dissolve with C. THIN-LAYER CHROMATOGRAPHY
water. Dilute with water to volume. Standard solution: 25 mg/mL of USP Cephalexin RS in
Calculate the molarity, MS, of the solution taken: water, with 0.1 N hydrochloric acid
Sample solution: 25 mg/mL in water, with 0.1 N
Result = w/372.24 hydrochloric acid
w = weight of the purified edetate disodium taken (g) See (Chromatography 621, Thin-Layer Chromatography.)
Analysis: Transfer an equivalent to 0.15 g of cellulose Mode: TLC
sodium phosphate from Oral Suspension, to a 250-mL Adsorbent: 0.25-mm layer of chromatographic silica gel
beaker. Add 150.0 mL of the Standard solution, and stir the mixture
mixture for 5 min on a magnetic stirrer. Filter, discarding Application volume: 5 L
the first few mL of the filtrate. To 50.0 mL of the filtrate add Developing solvent system: Ethyl acetate, acetonitrile,
about 50 mL of water, 15 mL of 1 N sodium hydroxide, and glacial acetic acid, and water (21:7:7:9)
300 mg of hydroxy naphthol blue. Titrate with Standard
USP 32 Official Monographs / Cephalexin 185
Analysis Acceptance criteria: 9501030 g/mg

Allow the spots to dry, place the plate in a saturated IMPURITIES
chamber containing the solvent system and lined with Organic Impurities
filter paper. Develop the chromatogram until the solvent PROCEDURE 1
front has moved three-fourths of the length of the plate. Solution A: Dissolve 1 g of sodium 1-pentanesulfonate in a
Remove the plate from the developing chamber, mark the mixture of 1000 mL of water and 15 mL of triethylamine.
solvent front, allow the plate to air-dry, and examine Adjust with phosphoric acid to a pH of 2.5 0.1.
under short-wavelength UV light. Solution B: Dissolve 1 g of sodium 1-pentanesulfonate in a
Acceptance criteria: The RF value of the principal spot of mixture of 300 mL of water and 15 mL of triethylamine.
the Sample solution corresponds to that of the Standard Adjust with phosphoric acid to a pH of 2.5 0.1, and add
solution. 350 mL of acetonitrile and 350 mL of methanol.
Mobile Phase: See the gradient table below.
ASSAY
PROCEDURE Time Solution A Solution B
Mobile phase: Prepare 1015 mL of a suitable mixture of (min) (%) (%)
acetonitrile, methanol, triethylamine, and water
(100:50:15:850). Dissolve 1.0 g sodium-1-pentanesulfonate 0 100 0
in this mixture and adjust with phosphoric acid to a pH of 1 100 0
3.0 0.1. 33.3 0 100
Internal standard stock solution: Dissolve 300 mg of 1-
hydroxyphenzotriazole in 10 mL of methanol. 34.3 0 100
Internal standard solution: 0.3 mg/mL of 1-
hydroxyphenzotriazole from Internal standard stock solution Diluent: 18 mg/mL of monobasic potassium phosphate in
diluted with Mobile phase water
Standard stock solution: 1 mg/mL of USP Cephalexin RS Standard solutions: 0.08 mg/mL and 0.16 mg/mL of
in water C16H17N3O4S from USP Cephalexin RS in Diluent, taking into
Standard solution: Mix 10 mL of Standard stock solution account the stated potency of the USP Cephalexin RS
with 15 mL of Internal standard solution. Sample solution: 5 mg/mL of Cephalexin in Diluent
Sample stock solution: 1 mg/mL of Cephalexin in water Chromatographic system
Sample solution: Mix 10 mL of Standard stock solution with (See Chromatography 621, System Suitability.)
15 mL of Internal standard solution. Mode: LC
Chromatographic system Detector: UV 254 nm
(See Chromatography 621, System Suitability.) Column: 4.6-mm 25-cm; packing L1 of low acidity
Mode: LC Flow rate: 1 mL/min
Column: 4.6-mm 25-cm; packing L1 of low acidity Analysis
Flow rate: 1.5 mL/min Samples: Standard solutions and Sample solution
Injection size: 20 L Plot the responses of the cephalexin peaks from the
System suitability Standard solutions versus their concentrations, calculated
Sample: Standard solution on the anhydrous basis, in mg/mL, and draw a straight
[NOTEThe relative retention times for 1- line through the two points and zero. From the line so
hydroxybenzotriazole and cephalexin are about 0.35 and obtained and the peak responses of the Sample solution,
1.0, respectively.] determine the concentration, I, in mg/mL, of each
Suitability requirements cephalexin-related substance of the Sample solution other
Resolution: NLT 5 between the internal standard and the than the cephalexin peak.
analyte peaks, Standard solution Calculate the percentage of each cephalexin-related
Relative standard deviation: NMT 2.0%, Standard substance:
solution
Analysis Result = I/C 100
Samples: Standard solution and Sample solution C = concentration, calculated on the anhydrous basis
Calculate the quantity, in g, of C16H17N3O4S per mg of the of Cephalexin taken to prepare the Sample
Cephalexin taken: solution (mg/mL)
Result = (RU/RS) (CS/CU) P Acceptance criteria
Individual impurities: NMT 1.0% of any individual
RU = ratio of the responses of the cephalexin peak to cephalexin-related substance
the 1-hydroxybenzotriazole peak from the Total impurities: NMT 5.0 %
Sample solution PROCEDURE 2: DIMETHYLANILINE 223: Meets the
RS = ratio of the responses of the cephalexin peak to requirement
the 1-hydroxybenzotriazole peak from the SPECIFIC TESTS
Standard solution OPTICAL ROTATION, Specific Rotation 781S: +149 to +158
CS = concentration of USP Cephalexin RS in the Sample solution: 5 mg/mL, in pH 4.4 neutralized phthalate
Standard stock solution (mg/mL) buffer (See Reagents, Indicators, and SolutionsBuffer
CU = concentration of cephalexin in the Sample stock Solutions)
solution (mg/mL)
P = designated content of cephalexin in USP
Cephalexin RS (g/mg)
186 Cephalexin / Official Monographs USP 32
CRYSTALLINITY 695: Meets the requirements Standard stock solution: 1 mg/mL of USP Cephalexin RS
PH 791: 3.05.5, in an aqueous suspension containing 50 in water
mg/mL Standard solution: Mix 10 mL of Standard stock solution
WATER DETERMINATION, Method I 921: 4.0%8.0% and 15 mL of Internal standard solution.
Sample stock solution: 1.15 mg/mL of Cephalexin
ADDITIONAL REQUIREMENTS Hydrochloride in water
PACKAGING AND STORAGE: Preserve in tight containers. Sample solution: Mix 10 mL of Sample stock solution with
USP REFERENCE STANDARDS 11 15 mL of Internal Standard solution.
USP Cephalexin RS Chromatographic system
Mode: LC
Cephalexin Hydrochloride Detector: UV 254 nm
Column: 4.6-mm 25-cm; packing L1 of low acidity
(Comment on this Monograph)id=m14282=Cephalexin Flow rate: 1.5 mL/min
Hydrochloride=Ca-Chl-Monos.pdf) Injection size: 20 L
C16H17N3O4S HCl H2O 401.87 System suitability
5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7- Sample: Standard solution
[(aminophenylacetyl)amino]-3-methyl-8-oxo-, [NOTEThe relative retention times for 1-
monohydrochloride, monohydrate, [6R-[6,7 (R*)]]-; hydroxybenzotriazole and cephalexin are 0.35 and 1.0,
(6R,7R)-7-[(2R)-2-Amino-2-phenylacetamido]-3-methyl-8-oxo-5- respectively.]
thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, Suitability requirements
monohydrochloride, monohydrate; Resolution: NLT 5 between the internal standard and the
7-(D-2-Amino-2-phenylacetamido)-3-methyl-3-cephem-4- analyte peaks, Standard solution
carboxylic acid hydrochloride monohydrate [105879-42-3]. Relative standard deviation: NMT 2.0%, Standard
solution
DEFINITION Analysis
Cephalexin Hydrochloride contains the equivalent of NLT 800 Samples: Standard solution and Sample solution
g and NMT 880 g of cephalexin (C16H17N3O4S) per mg. Calculate the quantity, in g, of C16H17N3O4S in each mg of
Cephalexin Hydrochloride taken:
IDENTIFICATION
A. THIN-LAYER CHROMATOGRAPHY Result = (RU/RS) (CS/CU) P
Standard solution: 25 mg/mL of USP Cephalexin RS in
water with 0.1 N hydrochloric acid RU = ratio of the responses of the cephalexin peak to
Sample solution: 25 mg/mL in water with 0.1 N the 1-hydroxybenzotriazole peak from the
hydrochloric acid Sample solution
Chromatographic system RS = ratio of the responses of the cephalexin peak to
(See Chromatography 621, Thin-Layer Chromatography.) the 1-hydroxybenzotriazole peak from the
Mode: TLC Standard solution
Adsorbent: 0.25-mm layer of chromatographic silica gel CS = concentration of USP Cephalexin RS, mg/mL in
mixture the Standard stock solution
Application volume: 5 L CU = concentration of Cephalexin Hydrochloride from
Developing solvent system: Ethyl acetate, acetonitrile, the Sample stock solution (mg/mL)
glacial acetic acid, and water (21:7:7:9) P = stated content of cephalexin in USP Cephalexin
Analysis RS (g/mg)
Samples: Standard solution and Sample solution Acceptance criteria: 800880 g/mg
Allow the spots to dry, place the plate in a saturated
chamber containing the solvent system and lined with IMPURITIES
filter paper. Develop the chromatogram until the solvent Organic Impurities
front has moved three-fourths of the length of the plate. PROCEDURE 1
Remove the plate from the developing chamber, mark the Solution A: 1 g of sodium 1-pentanesulfonate in a mixture
solvent front, allow the plate to air-dry, and examine of 1000 mL of water and 15 mL of triethylamine. Adjust
under short-wavelength UV light. with phosphoric acid to a pH of 2.5 0.1.
Acceptance criteria: The RF value of the principal spot of Solution B: 1 g of sodium 1-pentanesulfonate in a mixture
the Sample solution corresponds to that of the Standard of 300 mL of water and 15 mL of triethylamine. Adjust
solution. with phosphoric acid to a pH of 2.5 0.1, and add 350 mL
B. The UV absorption spectrum of a solution (1 in 50,000) of acetonitrile and 350 mL of methanol.
exhibits maxima and minima at the same wavelengths as Mobile phase: See the gradient table below.
that of a similar solution of USP Cephalexin RS,
concomitantly measured. Time Solution A Solution B
C. IDENTIFICATION TESTSGENERAL, Chloride 191: 10 mg/mL (min) (%) (%)
meets the requirements for Chloride. 0 100 0
ASSAY 1 100 0
PROCEDURE 33.3 0 100
Mobile phase: 1.0 g of sodium 1-pentanesulfonate in a
mixture of acetonitrile, methanol, triethylamine, and water 34.3 0 100
(100:50:15:850), adjusted with phosphoric acid to a pH of
3.0 0.1 Diluent: 18 mg/mL of monobasic potassium phosphate in
Internal standard solution: 0.3 mg/mL of 1- water
hydroxybenzotriazole dissolved in methanol and diluted with
Mobile phase (1:99) to volume.
Standard solutions: 0.08 mg/mL and 0.16 mg/mL of Mode: TLC

C16H17N3O4S from USP Cephalexin RS in Diluent, taking into Adsorbent: 0.25-mm layer of binder-free silica gel
account the stated potency of the USP Cephalexin RS Application volume: 10 L
Sample solution: 6 mg/mL of Cephalexin Hydrochloride Pre-developing solvent system: n-Hexane and
in Diluent tetradecane (95:5)
Chromatographic system Developing solvent system: 0.1 M citric acid, 0.1 M
(See Chromatography 621, System Suitability.) dibasic sodium phosphate, and a solution (1 in 15) of
Mode: LC ninhydrin in acetone (60:40:1.5)
Column: 4.6-mm 25-cm; packing L1 of low acidity Samples: Standard solution and Sample solution
Flow rate: 1 mL/min Allow the solvent front to move the length of the plate in
Injection size: 20 L the Pre-developing solvent system, remove the plate from
Analysis the chamber, and allow the solvent to evaporate. On this
Samples: Standard solutions and Sample solution plate apply 10 L each of the Sample solution and
Plot the responses of the cephalexin peaks of the Standard Standard solution. Allow the spots to dry, and develop the
solutions versus their concentrations, calculated on the chromatogram in the Developing solvent system until the
anhydrous basis, in mg/mL, and draw a straight line solvent front has moved three-fourths of the length of the
through the two points and zero. From the line so plate. Remove the plate from the developing chamber,
obtained and the peak responses of the Sample solution, mark the solvent front, dry the plate for 10 min at 110,
determine the concentration, I, in mg/mL, of each and examine the chromatogram.
cephalexin-related substance from the Sample solution Acceptance criteria: The RF value of the principal spot of
other than the cephalexin peak. the Sample solution corresponds to that of the Standard
Calculate the percentage of each cephalexin-related solution.
substance represented by each peak of the Sample
solution, other than the cephalexin peak. ASSAY
PROCEDURE
Result = (I/C) 100 Mobile phase: 1.0 g of sodium 1-pentanesulfonate in a
mixture of acetonitrile, methanol, triethylamine, and water
I = concentration of each cephalexin-related (100:50:15:850), adjusted with phosphoric acid to a pH of
substance other than cephalexin in the Sample 3.0 0.1
solution (mg/mL) Internal standard solution: 0.3 mg/mL of 1-
C = concentration mg/mL of cephalexin from the hydroxybenzotriazole dissolved in methanol and diluted with
Sample solution Mobile phase (1:99)
Acceptance criteria Standard stock solution: 1 mg/mL of USP Cephalexin RS
Individual impurities: NMT 1.0% of any individual in water
cephalexin-related substance is found. Standard solution: Mix 10 mL of Standard stock solution
Total impurities: NMT 5.0 % with 15 mL of Internal standard solution.
PROCEDURE 2: DIMETHYLANILINE 223: Meets the requirement Sample stock solution: Equivalent to 1 mg/mL of
cephalexin from combined contents of NLT 20 Capsules in
SPECIFIC TESTS water. Sonicate, if necessary, to dissolve the cephalexin.
CRYSTALLINITY 695: Meets the requirements Filter, if necessary, to obtain a clear solution.
PH 791: 1.53.0, in a solution containing 10 mg/mL Sample solution: Mix 10 mL of Sample stock solution with
WATER DETERMINATION, Method I 921: 3.0%6.5% 15 mL of Internal standard solution.
ADDITIONAL REQUIREMENTS (See Chromatography 621, System Suitability.)
PACKAGING AND STORAGE: Preserve in tight containers. Mode: LC
USP Cephalexin RS Column: 4.6-mm 25-cm; packing L1 of low acidity
Cephalexin Capsules System suitability
(Comment on this Monograph)id=m14290=Cephalexin [NOTEThe relative retention times for 1-
Capsules=Ca-Chl-Monos.pdf) hydroxybenzotriazole and cephalexin are 0.35 and 1.0,
DEFINITION respectively.]
Cephalexin Capsules contain the equivalent of NLT 90.0% and Suitability requirements
NMT 120.0% of the labeled amount of cephalexin Resolution: NLT 5 between the internal standard and the
(C16H17N3O4S). analyte peaks, Standard solution
A. THIN-LAYER CHROMATOGRAPHY Analysis
Standard solution: 3 mg/mL of USP Cephalexin RS in Samples: Standard solution and Sample solution
water Calculate the percentage of C16H17N3O4S in the portion of
Sample solution: 3 mg/mL of cephalexin from Capsule in Capsules taken:
water and filter
Chromatographic system Result = (RU/RS) (CS/CU) P F 100
RU = ratio of the response of the cephalexin peak to Analysis

the 1-hydroxybenzotriazole peak from the Samples: Standard solution and Sample solution
Sample solution Allow the solvent front to move the length of the plate in
RS = ratio of the response of the cephalexin peak to the Pre-developing solvent system, remove the plate from
the 1-hydroxybenzotriazole peak from the the chamber, and allow the solvent to evaporate. On this
Standard solution plate apply 10 L each of the Sample solution and the
CS = concentration of USP Cephalexin RS in the Standard solution. Allow the spots to dry, and develop the
Standard stock solution (mg/mL) chromatogram in the Developing solvent system until the
CU = nominal concentration of cephalexin from the solvent front has moved three-fourths of the length of the
Sample stock solution (mg/mL) plate. Remove the plate from the developing chamber,
P = designated potency of USP Cephalexin RS mark the solvent front, dry the plate for 10 min at 110,
(g/mg) and examine the chromatogram.
F = unit conversion factor, 0.001 mg/g Acceptance criteria: The RF value of the principal spot of
Acceptance criteria: 90.0%120.0% the Sample solution corresponds to that of the Standard
solution
PERFORMANCE TESTS
DISSOLUTION 711 ASSAY
Medium: Water; 900 mL PROCEDURE
Apparatus 1: 100 rpm Mobile phase: 1.0 g of sodium 1-pentanesulfonate in a
Time: 30 min mixture of acetonitrile, methanol, triethylamine, and water
Sample solution: Sample per Dissolution 711. Dilute with (100:50:15:850), adjusted with phosphoric acid to a pH of
Medium as needed. 3.0 0.1.
Standard solution: 20 g/mL of USP Cephalexin RS at a Standard stock solution: 1 mg/mL of USP Cephalexin RS
known concentration in the Medium in water
Spectrometric conditions Standard solution: Mix 10 mL of Standard stock solution
Mode: UV with 15 mL of Internal standard solution.
Analytical wavelength: 262 nm System suitability stock solution: 0.3 mg/mL of 1-
Analysis hydroxybenzotriazole dissolved in methanol, and diluted
Samples: Standard solution and Sample solution with Mobile phase (1:99)
Tolerances: NLT 80% (Q) of the labeled amount of System suitability solution: 10 mL of System suitability
C16H17N3O4S is dissolved. stock solution and 15 mL of Internal standard solution
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Sample stock solution: Nominally equivalent to 1 mg/mL
of cephalexin from Oral Suspension, constituted as directed
SPECIFIC TESTS in the labeling, freshly mixed and free from air bubbles.
WATER DETERMINATION, Method I 921: NMT 10.0% Sonicate, if necessary, to assure complete dissolution of the
cephalexin. Filter, if necessary, to obtain a clear solution.
ADDITIONAL REQUIREMENTS Sample solution: Mix 10 mL of Sample stock solution and
PACKAGING AND STORAGE: Preserve in tight containers. 15 mL of Internal standard solution.
USP Cephalexin RS (See Chromatography 621, System Suitability.)
Mode: LC
Detector: UV 254 nm
Cephalexin for Oral Suspension Column: 4.6-mm 25-cm; packing L1 of low acidity
Flow rate: 1 mL/min
(Comment on this Monograph)id=m14300=Cephalexin for Oral Injection size: 20 L
Suspension=Ca-Chl-Monos.pdf) System suitability
DEFINITION Sample: Standard solution and System suitability solution
Cephalexin for Oral Suspension is a dry mixture of Cephalexin [NOTEThe relative retention times for 1-
and one or more suitable buffers, colors, diluents, and flavors. hydroxybenzotriazole and cephalexin are about 0.35 and
It contains the equivalent of NLT 90.0% and NMT 120.0% of 1.0, respectively.]
the labeled amount of C16H17N3O4S per mL when constituted Suitability requirements
as directed in the labeling. Resolution: NLT 5 between the 1-hydroxybenzotriazole
peak and the cephalexin peak, System suitability solution
IDENTIFICATION Relative standard deviation: NMT 2.0%, Standard
A. THIN-LAYER CHROMATOGRAPHY solution
Standard solution: 3 mg/mL of USP Cephalexin RS in Analysis
water Samples: Standard solution and Sample solution
Sample solution: 3 mg/mL of cephalexin from Oral Calculate the percentage of C16H17N3O4S in each mL of
Suspension, constituted as directed in the labeling and Oral Suspension taken:
filtered
Chromatographic system Result = (rU/rS) (CS/CU) P F 100
Mode: TLC
Adsorbent: 0.25-mm layer of binder-free silica gel rU = cephalexin peak response from the Sample
Application volume: 10 L solution
Pre-developing solvent system: n-Hexane and rS = cephalexin peak response from the Standard
tetradecane (95:5) solution
Developing solvent system: 0.1 M citric acid, 0.1 M CS = concentration of USP Cephalexin RS in the
dibasic sodium phosphate, and a solution (1 in 15) of Standard stock solution (mg/mL)
ninhydrin in acetone (60:40:1.5)
CU = nominal concentration of cephalexin from the Standard stock solution: 1 mg/mL of USP Cephalexin RS
Sample solution (mg/mL) in water
P = designated potency of USP Cephalexin RS Standard solution: Standard stock solution and Internal
(g/mg) standard solution (2:3)
F = unit conversion factor, 0.001 mg/g Sample stock solution: Equivalent to 1 mg/mL of
Acceptance criteria: 90.0%120.0% cephalexin from combined contents of powdered Tablets
(NLT 20) in water. Sonicate, if necessary, to assure complete
SPECIFIC TESTS dissolution of the cephalexin. Filter, if necessary, to obtain a
WATER DETERMINATION, Method I 921: NMT 2.0% clear solution.
DELIVERABLE VOLUME 698: Meets the requirements Sample solution: Sample stock solution and Internal
PH 791: 3.06.0 standard solution (2:3)
PERFORMANCE TESTS (See Chromatography 621, System Suitability.)
UNIFORMITY OF DOSAGE UNITS 905 For solid packaged in Mode: LC
single-unit containers: meets the requirements Detector: UV 254 nm
ADDITIONAL REQUIREMENTS Column: 4.6-mm 25-cm; packing L1 of low acidity
PACKAGING AND STORAGE: Preserve in tight containers. Flow rate: 1.5 mL/min
USP REFERENCE STANDARDS 11 Injection size: 20 L
USP Cephalexin RS System suitability
[NOTEThe relative retention times for 1-
hydroxybenzotriazole and cephalexin are about 0.35 and
Cephalexin Tablets 1.0, respectively.]
(Comment on this Monograph)id=m14310=Cephalexin Suitability requirements
Tablets=Ca-Chl-Monos.pdf) Resolution: NLT 5 between the internal standard and the
analyte peaks, Standard solution
DEFINITION Relative standard deviation: NMT 2.0%, Standard
Cephalexin Tablets are prepared from Cephalexin or Cephalexin solution
Hydrochloride. They contain the equivalent of NLT 90.0% and Analysis
NMT 120.0% of the labeled amount of cephalexin Samples: Standard solution and Sample solution
(C16H17N3O4S). Calculate the percentage of C16H17N3O4S in the portion of
Tablets taken:
IDENTIFICATION
THIN-LAYER CHROMATOGRAPHY Result = (RU/RS) (CS/CU) P F 100
Standard solution: 3 mg/mL of USP Cephalexin RS in
water RU = peak response ratio of cephalexin to the 1-
Sample solution: 3 mg/mL of cephalexin from powdered hydroxybenzotriazole peak from the Sample
Tablets in water and filter solution
Chromatographic system RS = peak response ratio of cephalexin to the 1-
(See Chromatography 621, Thin-Layer Chromatography.) hydroxybenzotriazole peak from the Standard
Mode: TLC solution
Adsorbent: 0.25-mm layer of binder-free silica gel CS = concentration of USP Cephalexin RS in the
Application volume: 10 L Standard stock solution used to prepare the
Pre-developing solvent system: n-Hexane and Standard solution (mg/mL)
tetradecane (95:5) CU = nominal concentration of cephalexin taken to
Developing solvent system: 0.1 M citric acid, 0.1 M prepare the Sample solution (mg/mL)
dibasic sodium phosphate, and a solution (1 in 15) of P = designated potency of USP Cephalexin RS
ninhydrin in acetone (60:40:1.5) (g/mg)
Analysis F = unit conversion factor, 0.001 mg/g
Allow the solvent front to move the length of the plate in
the Pre-developing solvent system. On this plate apply 10 PERFORMANCE TESTS
L each of the Sample solution and Standard solution. DISSOLUTION 711
Allow the spots to dry, and develop the chromatogram in Cephalexin
the Developing solvent system until the solvent front has Medium: Water; 900 mL
moved three-fourths of the length of the plate. Remove Apparatus 1: Use 40-mesh cloth and 100 rpm
the plate from the developing chamber, mark the solvent Time: 30 min
front, dry the plate for 10 min at 110, and examine the Sample solution: Sample per Dissolution 711. Dilute with
chromatogram. Medium to a concentration that is similar to the Standard
Acceptance criteria: The RF value of the principal spot of solution.
the Sample solution corresponds to that of the Standard Standard solution: 20 g/mL of USP Cephalexin RS in the
solution. Medium
ASSAY Mode: UV
PROCEDURE Analytical wavelength: 262 nm
Mobile phase: 1.0 g of sodium 1-pentanesulfonate in a Analysis
mixture of acetonitrile, methanol, triethylamine, and water Samples: Standard solution and Sample solution
(100:50:15:850). Adjust with phosphoric acid to a pH of 3.0 Tolerances: NLT 80% (Q) of the labeled amount of
0.1. C16H17N3O4S is dissolved.
Internal standard solution: 0.3 mg/mL of 1-
hydroxybenzotriazole dissolved in methanol and diluted with
Mobile phase (1:99)
Cephalexin hydrochloride Internal standard solution: 0.3 mg/mL of 1-

Medium, Sample solution, Spectrometric conditions, hydroxybenzotriazole dissolved in methanol and diluted with
Standard solution, and Analysis: Proceed as directed Mobile phase (1:99)
under Cephalexin. Standard stock solution: 1 mg/mL of USP Cephalexin RS
Apparatus 1: Use 10-mesh cloth and 150 rpm in water
Time: 45 min Standard solution: Mix 10 mL of Standard stock solution
Tolerances: NLT 75% (Q) of the labeled amount of and 15 mL of Internal Standard solution.
C16H17N3O4S is dissolved. Sample stock solution: Nominally equivalent to 1 mg/mL
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements of cephalexin from combined contents of NLT 20 powdered
Tablets for Oral Suspension in water. Pass a portion of the
SPECIFIC TESTS solution through a filter having a 1-m or finer porosity.
WATER DETERMINATION, Method I 921: NMT 9.0% where Sample solution: Mix 10 mL of Sample stock solution with
Tablets contain Cephalexin; NMT 8.0% where Tablets 15 mL of Internal Standard solution.
contain Cephalexin Hydrochloride Chromatographic system
(See Chromatography 621, Syste

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USP 32 MONOGRAPH LIST / 1

Acarbose Acetaminophen and Pseudoephedrine Hydrochloride Tablets

Oral Powder Containing at Least Three of the Following Adenine

Dehydrated Alcohol Injection Aluminum Chloride

Alumina, Magnesia, and Simethicone Tablets Aluminum Zirconium Trichlorohydrate

Alumina, Magnesium Carbonate, and Magnesium Oxide Amantadine Hydrochloride

Amcinonide Cream Amitriptyline Hydrochloride

Ampicillin for Oral Suspension Arginine Hydrochloride

Aurothioglucose Injectable Suspension Azathioprine Tablets

rU = peak response from the Sample solution

Impurity Table (continued) USP Acarbose System Suitability Mixture RS

trihydroxy-3-(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl]- C18H28N2O4 HCl 372.89

Mode: LC ADDITIONAL REQUIREMENTS

Acceptance criteria: 90.0%110.0% rS = peak response from the Standard solution

Acepromazine Maleate Acceptance criteria: 98.0%101.0%

Result = (rU/rS) (CS/CU) 100 IDENTIFICATION

CU = nominal concentration of the Sample solution Acceptance criteria: 98.0%101.0%

Chromatographic system having a 0.5-m or finer porosity, discarding the first 10 mL

Acceptance criteria: Meets the requirements Acetaminophen for Effervescent Oral

CS = concentration of USP Acetaminophen RS in the Mode: LC

Apparatus 2: 75 rpm rS = peak response from the Standard solution

Acetaminophen, Aspirin, and Caffeine Result = (RU/RS) (CS/CU) 100

Acceptance criteria: 90.0%110.0% Oral Solution Containing at Least Three

Sample solution: Equivalent to 0.05 mg/mL of Acceptance criteria: 90.0%110.0%

rU = acetaminophen peak response from the Sample SPECIFIC TESTS

PH 791: 2.67.5 Standard stock solution: 2.5 mg/mL of USP

Mode: LC Injection size: 20 L

Dextromethorphan, and Pseudoephedrine contain NLT 90.0% Mode: LC

Acceptance criteria: 90.0%110.0% CU = nominal concentration of the Sample solution

Chlorpheniramine, Dextromethorphan, and Mode: LC

USP Dextromethorphan Hydrobromide RS CS = concentration of USP Acetaminophen RS in the

Sample solution: Transfer an equivalent to 6 mg of Acetaminophen and Codeine Phosphate

Mode: LC rU = peak response from the Sample solution

ASSAY CS = concentration of USP Codeine Phosphate RS in

Application volume: 10 L rS = peak response from the Standard solution

Acetaminophen, Dextromethorphan Acceptance criteria: 90.0%110.0%

CU = concentration of doxylamine succinate in the ASSAY

65 mL of Solution A, and shake by mechanical means for 15 DEFINITION

rU = peak response from the Sample solution ASSAY

rU = peak response for the corresponding analyte of

ASSAY Acetazolamide for Injection

Acetazolamide Oral Suspension Result = (rU/rS) (CS/CU) 100

white precipitate. Collect the precipitate on a medium- Acceptance criteria: 95.0%105.0%

CHLORIDE AND SULFATE, Chloride 221 Acetic Acid Otic Solution

IDENTIFICATION Sample solution: Quantitatively dilute Sample stock solution

Acetohexamide Tablets ADDITIONAL REQUIREMENTS

ASSAY the best-fit straight line from the fluorescence intensities

dichlorofluorescein TS. Titrate with 0.1 N silver nitrate VS ASSAY

ADDITIONAL REQUIREMENTS RU = internal standard ratio of the Sample solution

ASSAY Acetylcysteine and Isoproterenol

Mode: LC RU = ratio of the peak responses of isoproterenol

IDENTIFICATION Dilute quantitatively with alcohol.

USP Tretinoin RS Mode: LC

Analysis Dissolve both in a volume of 0.1 N sodium hydroxide of

rU = peak response from the Sample solution

IDENTIFICATION System suitability solution: 0.5 g/mL each of purine and

SPECIFIC TESTS rU = peak response from the Sample solution

DEFINITION Result = (rU/rS) (CS/CU) 100

Suitability requirements Adenine

ASSAY the Assay with Acidified methanol, prepared as directed for

Acceptance criteria: 90.0%110.0% ADDITIONAL REQUIREMENTS

Albuterol SPECIFIC TESTS

obtain a Standard solution having a known concentration of Alclometasone Dipropionate