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Triamterene
Molecular formula: C12H11N7
Molecular weight: 253.3
CAS Registry No.: 396-01 -0

SAMPLE
Matrix: bile, blood, urine
Sample preparation: 1 mL Plasma, whole blood, bile, or urine + 100 |xL MeOH + 1
(plasma, blood, urine) or 0.5 (bile) mL 100 mM sodium bicarbonate + 6 mL diethyl ether,
shake horizontally for 10 min, centrifuge at 2500 g at 10 for 10 (plasma, blood, urine) or
20 (bile) min, freeze in dry ice/acetone. Remove the organic layer and evaporate it to
dryness, reconstitute the residue in 500 |JLL mobile phase, inject a 20 jxL aliquot. (Hydro-
lyze conjugates as follows. 500 jxL Plasma, whole blood, bile, or urine + 500 |xL pH 5
buffer + 50 |JLL glusulase, heat at 50 for 24 h, add 50 |JLL 1 M NaOH, add 100 |xL 1
(xg/mL triamterene in MeOH, add 1 mL 100 mM sodium bicarbonate, proceed as above.
In order to determine only the sulfate add 60 |JLL neutralized 100 mM 1,4-saccharolactone
solution to inhibit p-glucuronidase.)

HPLC VARIABLES
Guard column: 25 X 2.3 5 |xm PRP-I (Hamilton)
Column: 150 X 4.1 5 |xm PRP-I (Hamilton)
Mobile phase: MeOH:buffer 65:35 (Buffer was 1 g sodium carbonate and 2.902 g sodium
bicarbonate in 1 L water, pH 9.8.)
Flow rate: 0.5
Injection volume: 20
Detector: F ex 330 em 420

CHROMATOGRAM
Retention time: 10
Internal standard: triamterene

OTHER SUBSTANCES
Extracted: dilevalol
Noninterfering: acebutolol, brefanolol, carteolol, carvedilol, enalapril, furosemide, hy-
drochlorothiazide, isosorbide dinitrate, metoprolol, nifedipine, piretanide, propranolol,
verapamil, xamoterol, xipamide

KEYWORDS
plasma; whole blood; triamterene is IS; pharmacokinetics

REFERENCE
Neubeck, M.; Becker, C; Henke, D.; Rosch, W.; Spahn-Langguth, H.; Mutschler, E. Pharmacokinetics of
dilevalol and its conjugates in man. Assay method for plasma, blood, urine and bile samples and
preliminary pharmacokinetic studies. Arzneimittelforschung, 1993, 43, 953-957

SAMPLE
Matrix: blood
Sample preparation: 100 jxL Plasma or whole blood + 400 fxL 3.5 |xg/mL furosemide in
MeCN, vortex for 1 min, centrifuge at 3200 rpm for 10 min. Remove the supernatant and
evaporate it to 200 jxL under a stream of nitrogen, inject an aliquot.

HPLCVARIABLES
Column: 300 X 4 10 jjim Micro Pak MCH reverse phase
Mobile phase: MeCN: 0.02% phosphoric acid 30:70 adjusted to pH 4.0 with 500 mM NaOH
Flow rate: 2
Detector: F ex 365 em 440

CHROMATOGRAM
Retention time: 5.7
Internal standard: furosemide (8.1)
Limit of quantitation: 1 ng/mL

KEYWORDS
whole blood; plasma; pharmacokinetics

REFERENCE
Sorgel, R; Lin, E.T.; Hasegawa, J.; Benet, L.Z. Liquid chromatographic analysis of triamterene and its
major metabolite, hydroxytriamterene sulfate, in blood, plasma, and urine. J.Pharm.Sci., 1984, 73,
831-833

SAMPLE
Matrix: blood, urine
Sample preparation: Plasma. Dilute 200 [xL plasma with 600 (xL water, inject a 20 |xL
aliquot. Urine. Dilute 10 |JLL urine with 2 mL water, inject a 20 JJIL aliquot.

HPLCVARIABLES
Column: 125 X 4 5 |xm Spherisorb-amino
Mobile phase: Buffer containing 10 mM perchloric acid, 2 mM triethylamine, and 100 mM
ammonium acetate
Flow rate: 3
Injection volume: 20
Detector: F ex 360 em 436

CHROMATOGRAM
Retention time: 0.5
Limit of detection: 1 ng/mL

OTHER SUBSTANCES
Simultaneous: hydroxytriamterene sulfate
Noninterfering: bendroflumethiazide, butizide, chlorthalidone, furosemide, hydrochloro-
thiazide, hydroxytriamterene, nifedipine

KEYWORDS
plasma

REFERENCE
Mascher, H.; Wasilewski, M. Simple and fast HPLC determination of triamterene and hydroxytriam-
terenesulphate in plasma and urine. J.Liq.Chromatogr., 1994, 17, 1577-1585

SAMPLE
Matrix: blood, urine
Sample preparation: Plasma. 150 |JLL Plasma + 10 JLJLL 70% perchloric acid, shake for 15
min, centrifuge at 900 g for 10 min, inject 30 |xL of the supernatant. Urine. Dilute urine
with 2 volumes of MeOH, centrifuge at 900 g for 10 min, inject a 5 JJLL aliquot.

HPLCVARIABLES
Column: 150 X 4.6 5 jxm Nova-Pak C18
Mobile phase: MeCN:MeOH-.buffer 14:8:70 (Buffer obtained from 6.75 mL 89% phos-
phoric acid in 900 mL water, pH adjusted to 2.8 with triethylamine, volume made up to
IL.)
Flow rate: 0.8
Injection volume: 5-30
Detector: F ex 340-380 em 400-600

CHROMATOGRAM
Retention time: 3.3
Limit of quantitation: 1 ng/mL

OTHER SUBSTANCES
Simultaneous: metabolites

KEYWORDS
plasma

REFERENCE
Swart, K. J.; Botha, H. Rapid method for the determination of the diuretic triamterene and its metab-
olites in plasma and urine by high-performance liquid chromatography. J.Chromatogr., 1987, 413,
315-319

SAMPLE
Matrix: blood, urine
Sample preparation: Dilute 100 JULL urine with 1 mL water. 1 mL Plasma or diluted urine
+ 40 |xL 1 |xg/mL p-methoxytriamterene in MeOH + 200 mg sodium bicarbonate + 5 mL
ethyl acetate, shake for 1 min, centrifuge for 10 min. Remove the organic layer and evap-
orate it to dryness under a stream of nitrogen at 37, wash sides of tube with ethyl acetate
and again evaporate to dryness, dissolve residue in 20-50 |xL MeOH, inject a 5-12 JULL
aliquot.

HPLCVARIABLES
Column: 300 X 4 10 |xm uJBondapak C18
Mobile phase: MeOH: 0.1% KH2PO4 45:55, final pH adjusted to 3.8 with phosphoric acid
Flow rate: 2
Injection volume: 5-12
Detector: F ex 365 em 440; UV 230

CHROMATOGRAM
Retention time: 2.6
Internal standard: p-methoxytriamterene (3.1)
Limit of detection: 1 ng/mL (F); 10 ng/mL (UV)

OTHER SUBSTANCES
Noninterfering: metabolites

KEYWORDS
plasma

REFERENCE
Brodie, R.R.; Chasseaud, L.F.; Taylor, T.; Walmseley, L.M. Determination of the diuretic triamterene in
the plasma and urine of humans by high-performance liquid chromatography. J.Chromatogr, 1979,
164, 527-533

SAMPLE
Matrix: formulations, urine
Sample preparation: Tablets. Pulverize tablets, add MeOH, shake for 30 min, sonicate for
5 min, filter (Albet 242 paper), wash solid with MeOH, make up nitrate to 50 mL with
MeOH, inject a 20 \xL aliquot. Urine. Adjust pH of 2 mL urine to 10.0 with 2 M KOH,
add 1.5 mg NaCl, add 4 mL ethyl acetate, shake for 10 min, centrifuge at 2500 rpm for
5 min. Remove the organic layer and evaporate it to dryness under a stream of nitrogen
at 40, reconstitute the residue in 2 mL mobile phase, sonicate, inject a 20 jxL aliquot.

HPLCVARIABLES
Guard column: ixBondapak C18
Column: 300 X 3.9 10 |xm ixBondapak C18
Mobile phase: MeCN: water 30:70 containing 5 mM KH2PO4ZK2HPO4, pH adjusted to 5.5
Flow rate: 1
Injection volume: 20
Detector: E, EG&G Princeton Applied Research PAR Model 400, glassy carbon working elec-
trode + 1300 mV, Ag^AgCl reference electrode (At the end of each day clean electrode with
mobile phase of MeOH at 1.5 mL/min, -800 mV for 2 min then +1600 mV for 5 min.)

CHROMATOGRAM
Retention time: 5.01
Limit of detection: 0.1 ng/mL

OTHER SUBSTANCES
Extracted: furosemide

KEYWORDS
tablets; pharmacokinetics

REFERENCE
Barroso, M.B.; Alonso, R.M.; Jimenez, R.M. Simultaneous determination of the diuretics triamterene
and furosemide in pharmaceutical formulations and urine by HPLC-EC. J.Liq.Chrom.Rel.TechnoL,
1996, 19, 231-246

SAMPLE
Matrix: solutions

HPLCVARIABLES
Column: 250 X 4.6 5 jxm Supelcosil LC-DP (A) or 250 X 4 5 yna LiChrospher 100 RP-8 (B)
Mobile phase: MeCN:0.025% phosphoric acid:buffer 25:10:5 (A) or 60:25:15 (B) (Buffer
was 9 mL concentrated phosphoric acid and 10 mL triethylamine in 900 mL water, adjust
pH to 3.4 with dilute phosphoric acid, make up to 1 L.)
Flow rate: 0.6
Injection volume: 25
Detector: UV 229

CHROMATOGRAM
Retention time: 6.73 (A), 3.90 (B)

OTHER SUBSTANCES
Also analyzed: acebutolol, acepromazine, acetaminophen, acetazolamide, acetophenazine,
albuterol, alprazolam, amitriptyline, amobarbital, amoxapine, antipyrine, atenolol, atro-
pine, azatadine, baclofen, benzocaine, bromocriptine, brompheniramine, brotizolam,
bupivacaine, buspirone, butabarbital, butalbital, caffeine, carbamazepine, cetirizine,
ehlorcyclizine, chlordiazepoxide, chlormezanone, chloroquine, chlorpheniramine,
chlorpromazine, chlorpropamide, chlorprothixene, chlorthalidone, chlorzoxazone, cimeti-
dine, cisapride, clomipramine, clonazepam, clonidine, clozapine, cocaine, codeine, colchi-
cine, cyclizine, cyclobenzaprine, dantrolene, desipramine, diazepam, diclofenac, diflunisal,
 diltiazem, diphenhydramine, diphenidol, diphenoxylate, dipyridamole, disopyramide, do-
butamine, doxapram, doxepin, droperidol, encainide, ethidium bromide, ethopropazine,
fenoprofen, fentanyl, flavoxate, fluoxetine, fluphenazine, flurazepam, flurbiprofen, fluvox-
amine, furosemide, glutethimide, glyburide, guaifenesin, haloperidol, homatropine, hy-
dralazine, hydrochlorothiazide, hydrocodone, hydromorphone, hydroxychloroquine, hydro-
xyzine, ibuprofen, imipramine, indomethacin, ketoconazole, ketoprofen, ketorolac,
labetalol, levorphanol, lidocaine, loratadine, lorazepam, lovastatin, loxapine, mazindol,
mefenamic acid, meperidine, mephenytoin, mepivacaine, mesoridazine, metaproterenol,
metformin, methadone, methdilazine, methocarbamol, methotrexate, methotrimeprazine,
methoxamine, methyldopa, methylphenidate, metoclopramide, metolazone, metoprolol,
metronidazole, midazolam, moclobemide, morphine, nadolol, nalbuphine, naloxone, na-
phazoline, naproxen, nifedipine, nizatidine, norepinephrine, nortriptyline, oxazepam, ox-
ycodone, oxymetazoline, paroxetine, pemoline, pentazocine, pentobarbital, pentoxifylline,
perphenazine, pheniramine, phenobarbital, phenol, phenolphthalein, phentolamine,
phenylbutazone, phenyltoloxamine, phenytoin, pimozide, pindolol, piroxicam, pramoxine,
prazepam, prazosin, probenecid, procainamide, procaine, prochlorperazine, procyclidine,
promazine, promethazine, propafenone, propantheline, propiomazine, propofol, propran-
olol, protriptyline, quazepam, quinidine, quinine, racemethorphan, ranitidine, remo-
xipride, risperidone, salicylic acid, scopolamine, secobarbital, sertraline, sotalol, spirono-
lactone, sulfinpyrazone, sulindac, temazepam, terbutaline, terfenadine, tetracaine,
theophylline, thiethylperazine, thiopental, thioridazine, thiothixene, timolol, tocainide,
tolbutamide, tolmetin, trazodone, triazolam, trifluoperazine, triflupromazine, trimepra-
zine, trimethoprim, trimipramine, verapamil, warfarin, xylometazoline, yohimbine,
zopiclone

KEYWORDS
some details of plasma extraction

REFERENCE
Koves, E.M. Use of high-performance liquid chromatography-diode array detection in forensic toxicology.
J.Chromatogr.A, 1995, 692, 103-119

SAMPLE
Matrix: solutions
Sample preparation: Inject a 10 |xL aliquot of an aqueous solution.

HPLCVARIABLES
Column: 250 X 4 5 |xm Lichrosphere CN
Mobile phase: 2% acetic acid
Flow rate: 1
Injection volume: 10
Detector: UV 320

CHROMATOGRAM
Retention time: 12.8

OTHER SUBSTANCES
Simultaneous: caffeic acid

REFERENCE
Uang, Y-S.; Kang, R-L.; Hsu, K.-Y. Determination of caffeic acid in rabbit plasma by high-performance
liquid chromatography. J.Chromatogr.B, 1995, 673, 43-49

SAMPLE
Matrix: solutions
HPLCVARIABLES
Guard column: 30 X 3.2 7 |xm SI 100 ODS (not commercially available)
Column: 150 X 3.2 7 (xm SI 100 ODS (not commercially available)
Mobile phase: MeCN: buffer 31.2:68.8 (Buffer was 6.66 g KH2PO4 and 4.8 g 85% phos-
phoric acid in 1 L water, pH 2.3.)
Flow rate: 0.5-1
Detector: UV 211; UV 245

CHROMATOGRAM
Retention time: 1.6
Internal standard: 5-(4-methylphenyl)-5-phenylhydantoin (7.3)

OTHER SUBSTANCES
Also analyzed: aspirin, caffeine, carbamazepine, chlordiazepoxide, chlorprothixene, clona-
zepam, diazepam, doxylamine, ethosuximide, furosemide, haloperidol, hydrochlorothia-
zide, methocarbamol, methotrimeprazine, nicotine, oxazepam, procaine, promazine, pro-
pafenone, propranolol, salicylamide, temazepam, tetracaine, thiopental, verapamil,
zolpidem, zopiclone

REFERENCE
Below, E.; Burrmann, M. Application of HPLC equipment with rapid scan detection to the identification
of drugs in toxicological analysis. J.Liq.Chromatogr., 1994, 17, 4131-4144

SAMPLE
Matrix: urine
Sample preparation: Inject an aliquot of urine directly onto column A with mobile phase
A, after 1 min backflush the contents of column A onto column B with mobile phase B.

HPLCVARIABLES
Column: A 20 X 2.1 Hypersil ODS-C18 30 \xm; B 250 X 4 Hypersil ODS-C18 5 |xm
Mobile phase: A Water; B Gradient. MeCN: buffer 15:85 for 1.5 min then to 80:20 over 8
min. Keep at 80:20 for 2.5 min then re-equilibrate with 15:85. (Buffer was 50 mM
NaH2PO4 + 1 . 4 mL propylamine hydrochloride per liter adjusted to pH 3 with concen-
trated phosphoric acid.)
Flow rate: 1
Injection volume: 50
Detector: UV 230

CHROMATOGRAM
Retention time: 6.9
Limit of detection: 7 ng/mL

OTHER SUBSTANCES
Extracted: acetazolamide, amiloride, bendroflumethiazide, bumetanide, chlorthalidone, cy-
clothiazide, ethacrynic acid, furosemide, hydrochlorothiazide, probenecid, spironolactone

KEYWORDS
column-switching

REFERENCE
Campins-Falco, P.; Herraez-Hernandez, R.; Sevillano-Cabeza, A. Column-switching techniques for
screening of diuretics and probenecid in urine samples. Anal.Chem., 1994, 66, 244-248

SAMPLE
Matrix: urine
Sample preparation: Inject 5 jxL urine onto column A in mobile phase A, after 1 min elute
the contents of column A onto column B with mobile phase B

HPLCVARIABLES
Column: A 20 X 2.1 30 ^m Hypersil ODS-C18; B 125 X 4 5 jxm HP-LiChrospher 100 RP
18
Mobile phase: A water; B Gradient. A was MeCN. B was 3.45 g NaH2PO4-H2O + 700 ^L
propylamine hydrochloride in 500 mL water, pH adjusted to 3 with concentrated phos-
phoric acid. A:B from 0:100 to 50:50 after 4 min then hold at 50:50.
Flow rate: 1
Injection volume: 5
Detector: F ex 230 em 430

CHROMATOGRAM
Retention time: 5.27
Limit of detection: 5 ng/mL

KEYWORDS
column-switching

REFERENCE
Campins-Falco, P.; Herraez-Hernandez, R.; Sevillano-Cabeza, A. Determination of triamterene in urine
by HPLC using fluorescence detection and column-switching. Chromatographia, 1994, 38, 29-34

SAMPLE
Matrix: urine
Sample preparation: Condition a 1 mL 100 mg Bond-Elut C8 SPE cartridge with 500 |xL
MeOH and 500 [xL water. 2 mL urine + 300 \xL MeOH5 add to the SPE cartridge, wash
with 500 JULL water, elute with 500 |xL MeOH, filter (0.45 |xm) the eluate, inject a 5 |xL
aliquot.

HPLCVARIABLES
Column: 125 X 4 5 |xm HP-LiChrospher 100 RP 18
Mobile phase: Gradient. MeCN: buffer from 0:100 to 30:70 over 5 min, maintain at 30:
70. (Buffer was 3.45 g NaH2PO4-H2O + 700 |JLL propylamine hydrochloride in 500 mL
water, adjust pH to 3 with concentrated phosphoric acid.)
Flow rate: 1
Injection volume: 5
Detector: UV 230

CHROMATOGRAM
Retention time: 3.8
Internal standard: triamterene

OTHER SUBSTANCES
Extracted: chlorthalidone
Simultaneous: atenolol, oprenolol, reserpine, spironolactone

KEYWORDS
SPE; triamterene is IS

REFERENCE
Campins-Falco, P.; Herraez-Hernandez, R.; Sevillano-Cabeza, A. Simple and sensitive reversed-phase
liquid chromatographic assay for analysis of chlorthalidone in urine. J.Liq.Chromatogn, 1993, 16,
2571-2581
SAMPLE
Matrix: urine
Sample preparation: Buffer urine to 4.9 by mixing with an equal volume of pH 4.9 200
mM sodium phosphate buffer. Inject a 40 (xL aliquot onto column A with mobile phase A,
after 3 min backflush the contents of column A onto column B with mobile phase B and
start the gradient. At the end of the run re-equilibrate for 10 min.

HPLC VARIABLES
Column: A 20 X 4 5 jxm Hypersil octadecylsilica ODS; B 200 X 4.6 5 |xm Shiseido SG-120
polymer-based C18
Mobile phase: A water; B Gradient. MeCN: buffer from 7:93 to 15:85 over 3.5 min, to
50:50 over 8.5 min, maintain at 50:50 for 11 min (Buffer was 6.9 g NaH2PO4-H2O in 1
L water, pH adjusted to 3.1 with phosphoric acid.)
Flow rate: 1
Injection volume: 40
Detector: UV 360

CHROMATOGRAM
Retention time: 11.6
Limit of detection: 500 ng/mL

OTHER SUBSTANCES
Extracted: acetazolamide, amiloride, bendroflumethiazide, benzthiazide, bumetanide, caf-
feine, carbamazepine, chlorothiazide, chlorthalidone, clopamide, dichlorfenamide, ethac-
rynic acid, furosemide, hydrochlorothiazide, metyrapone, probenecid, spironolactone,
trichlormethiazide

KEYWORDS
column-switching; optimum detection wavelengths vary for each drug

REFERENCE
Saarinen, M.; Siren, H.; Riekkola, M.-L. A column switching technique for the screening of diuretics in
urine by high performance liquid chromatography. J.Liq.Chromatogr., 1993, 16, 4063-4078

SAMPLE
Matrix: urine
Sample preparation: 5 mL Urine + 50 JJLL 100 jjig/mL 7-propyltheophylline in MeOH +
200 |JLL ammonium chloride buffer + 2 g NaCl, extract with 6 mL ethyl acetate by rocking
at 40 movements/min for 20 min and centrifuging at 800 g for 5 min, repeat extraction,
combine organic layers, evaporate to dryness at 40 under a stream of nitrogen. Recon-
stitute in 200 |xL MeCN: water 15:85 and inject 20 |JLL aliquots. (Ammonium chloride
buffer was 28 g ammonium chloride in 100 mL water with the pH adjusted to 9.5 with
concentrated ammonia solution.)

HPLCVARIABLES
Column: 75 X 4.6 3 |mm Ultrasphere ODS
Mobile phase: Gradient. MeCN: buffer from 10:90 to 15:85 over 2 min, to 55:45 over 3
min, to 60:40 over 3 min, maintain at 60:40 for 1 min, to 10:90 over 1 min, equilibrate
at 10:90 for 2 min. (Buffer was 100 mM ammonium acetate adjusted to pH 3 with con-
centrated phosphoric acid.)
Flow rate: 1
Injection volume: 20
Detector: UV 270
CHROMATOGRAM
Retention time: 3.7
Internal standard: 7-propyltheophylline (4.5)

OTHER SUBSTANCES
Simultaneous: acetazolamide, amiloride, bendroflumethiazide, benzthiazide, bumetanide,
buthiazide, caffeine, canrenone, chlorthalidone, clopamide, cyclothiazide, dielofenamide,
ethacrynic acid, furosemide, hydrochlorothiazide, mesocarb, morazone, piretanide, poly-
thiazide, probenecid, spironolactone, torsemide, xipamide

REFERENCE
Ventura, R.; Nadal, T.; Alcalde, P.; Pascual, J.A.; Segura, J. Fast screening method for diuretics, pro-
benecid and other compounds of doping interest. J.Chromatogr.A, 1993, 655, 233-242

SAMPLE
Matrix: urine
Sample preparation: Make 5 mL urine alkaline (pH 9-10), add 2 g NaCl, extract twice
with 6 mL ethyl acetate. Combine the organic layers and evaporate them to dryness under
a stream of nitrogen, reconstitute the residue in 200 |xL MeCN/water, inject a 10-20 |xL
aliquot.

HPLCVARIABLES
Column: 100 X 4 5 jxm SGE 100 GL-4 C18P (Scientific Glass Engineering)
Mobile phase: MeCN: MeOH: water: trifluoroacetic acid 4.5:10.5:85:0,5
Flow rate: 0.8 or 1
Injection volume: 10-20
Detector: MS, ZAB2-SEQ (VG), PSP source coupled to LC, source 250, probe 240-260,
scan m/z 200-550; UV 270

CHROMATOGRAM
Retention time: 4.0
Limit of detection: 50 ng (by MS)

OTHER SUBSTANCES
Extracted: amiloride, bendroflumethiazide, benzthiazide, chlorthalidone, furosemide

REFERENCE
Ventura, R.; Fraisse, D.; Becchi, M.; Paisse, O.; Segura, J. Approach to the analysis of diuretics and
masking agents by high-performance liquid chromatography-mass spectrometry in doping control.
J.Chromatogr., 1991, 562, 723-736

SAMPLE
Matrix: urine
Sample preparation: 2 mL Urine + 0.5 g solid buffer I (pH 5-5.5), vortex 15 s, add 4 mL
ethyl acetate, agitate for 10 min, centrifuge at 600 g for 5 min. Remove organic layer and
vortex it with 2 mL 5% aqueous lead acetate for 10 s, centrifuge at 600 g for 5 min,
remove and keep organic phase. 2 mL Urine + 0.5 g solid buffer II (pH 9-9.5), vortex 15
s, add 4 mL ethyl acetate, agitate for 10 min, centrifuge at 600 g for 5 min. Remove
organic layer and combine it with previous organic layer. Evaporate to dryness at 50
under a stream of nitrogen, reconstitute in 300 u,L 50 |xg/mL p-hydroxyethyltheophylline
in MeOH, inject 5 |xL aliquot. (Solid buffer I was KH2PO4: Na2HPO4 99:1, solid buffer II
was NaHCO3IK2CO3 3:2.)

HPLCVARIABLES
Column: 250 X 4.6 5 ^m HP Hypersil ODS (A) or HP LiChrosorb RP-18 (B)
Mobile phase: Gradient. MeCN: buffer from 15:85 at 2 min to 80:20 at 20 min (Buffer was
50 mM NaH2PO4 containing 16 mM propylamine hydrochloride, adjusted to pH 3 with
concentrated phosphoric acid.)
Flow rate: 1
Injection volume: 5
Detector: UV 230;UV 275

CHROMATOGRAM
Retention time: 6.8 (A)5 7.6 (B)
Internal standard: P-hydroxyethyltheophylline (3.7 (A), 4.4 (B))
Limit of detection: 500 ng/mL

OTHER SUBSTANCES
Extracted: acetazolamide, amiloride, bendroflumethiazide, benzthiazide, bumetanide, can-
renone, chlorothiazide, chlorthalidone, cyclothiazide, dichlorphenamide, ethacrynic acid,
furosemide, hydrochlorothiazide, hydroflumethiazide, methyclothiazide, metolazone, po-
lythiazide, probenecid, quinethazone, spironolactone, trichloromethiazide
Noninterfering: acetaminophen, aspirin, caffeine, diflunisal, fenoprofen, ibuprofen, indo-
methacin, methocarbamol, naproxen, phenylbutazone, sulindac, tetracycline, theobro-
mine, theophylline, tolmetin, trimethoprim, verapamil
Interfering: flumethiazide

REFERENCE
Cooper, S.F.; Masse, R.; Dugal, R. Comprehensive screening procedure for diuretics in urine by high-
performance liquid chromatography. J.Chromatogr., 1989, 489, 65-88

SAMPLE
Matrix: urine
Sample preparation: 20-100 |xL Urine + 1 mL 500 fjig/mL hydroflumethiazide in MeCN,
vortex, centrifuge. Remove the supernatant, inject a 2 JJLL aliquot.

HPLCVARIABLES
Column: 300 X 4 10 jxm Micro Pak MCH reverse phase
Mobile phase: MeCN: 0.02% phosphoric acid 13:87 adjusted to pH 5.3 with NaOH
Flow rate: 2
Injection volume: 2
Detector: F ex 365 em 440

CHROMATOGRAM
Retention time: 8.6
Internal standard: hydroflumethiazide (7.0)
Limit of quantitation: 40 ng/mL

OTHER SUBSTANCES
Extracted: metabolites

KEYWORDS
pharmacokinetics

REFERENCE
Sorgel, R; Lin, E.T.; Hasegawa, J.; Benet, L.Z. Liquid chromatographic analysis of triamterene and its
major metabolite, hydroxytriamterene sulfate, in blood, plasma, and urine. J.Pharm.Sci., 1984, 73,
831-833

ANNOTATED BIBLIOGRAPHY
Oertel, R.; Richter, K.; Dobrev, D.; Berndt, A.; Gramatte, T. Eine empfindliche HPLC-Methode zur Bes-
timmung von Triamteren im Serum [A sensitive HPLC-method for determination of triamterene in
serum]. Pharmazie, 1994, 49, 700-702 [LOQ 2 ng/mL]
Bonet-Domingo, E.; Medina-Hernandez, M.J.; Garcia-Alvarez-Coque, M.C. A micellar liquid chromato-
graphic procedure for the determination of amiloride, bendroflumethiazide, chlorthalidone, spirono-
lactone and triamterene in Pharmaceuticals. J.Pharm.Biomed.Anal., 1993, 11, 711 716
Herraez-Hernandez, R.; Campins-Falco, P.; Sevillano-Cabeza, A. Improved screening procedure for di-
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