` Hc vin qun y b mn dc hc qun s ----****----

hng dn thc hnh kim nghim thuc

(Ti liu s dng cho sinh vin i hc Dc).

H ni - 2008.

Bi 1 gii thiu dng c v pha dung dch chun (NaOH 0,1N; HCl 0,1N; AgNO3 0,1N v NaCl 0,1N). I/ Cn: Nhiu loi cn: - Cn phn tch chnh xc 0,1mg. - Cn i hi: + ng: Ngha l khi ta cn 1 vt chun xc th cho kt qu ng vi kt lun ca n. + Nhy: biu din bng s vch chia trn cn. Khi cn thng bng, ch thm 0,1mg lm lch kim ch ca cn. + Tin: Tc l vi mt lng ta cn th cn ln troc v cn ln sau phi cho kt qu lp li. - Cn: + Kim tra cn: M 2 bn lng knh trc 5 - 10 pht lu thng khng kh, kim tra 3 - 5 ln v tr + M hm cn nh nhng. + Mi khi thm, bt mu trn a cn phi hm cn. + Sau khi cn phi hm cn a v v tr s 0 v lm v sinh cn. II/ Dng c o th tch: 1. ng ht: - T th cc dng c o phi thng ng. - Khi ht cht lng bao gi cng ht qu vch 2 3 cm, iu chnh cht lng tha chy ra khi pipet, khi quan st phi vch ca pipet ngang tm mt. - t ng ht vo thnh bnh nh mc ri th vi tc 10ml/pht, git cui cng chm vo thnh bnh nn, tuyt i khng c vy. - Cch c kt qu: i vi dung dch khng mu c phn y ca phn cong, dung dch c mu c phn pha trn ca mt lm. 2. Buret: - T th dng c o phi thng ng. - Bi vaselin vo kho Trnh lp l kho.

- Khi dung dch chun bao gi cng qu vch 2 3 cm, iu chnh cht lng tha chy ra khi buret, khi quan st phi vch ca buret ngang tm mt. - Khi m kho cng iu chnh tc khng qu 10ml/pht, gn ti im tng ng th iu chnh kho cho tc chm li. Th tch dung dch chun tiu tn khng c vt qu dung tch mt buret.aa - Cch c kt qu: i vi dung dch khng mu c phn y ca phn cong, dung dch c mu c phn pha trn ca mt lm. 3. Bnh nh mc: - Cht cn th kh tan: ho tan trong cc c m vi khong 30 40% dung tch bnh, ri chuyn vo bnh nh mc. Trng cc v a thu tinh 3 ln, gp dch trng vo bnh, thm dung mi cho n lc gn ti vch, dng pipet thm dn cho n vch. - Vi dung mi v cc cht rn d tan, ho tan trc tip trong bnh vi 1/2 hay 1/3 dung tch bnh, lc u cho tan hon ton, thm dung mi cho n lc gn ti vch, dng pipet thm dn cho n vch. Ch : Nu gp dung dch to nhit th phi i cho ngui mi chuyn vo bnh nh mc. - Cch c kt qu: i vi dung dch khng mu c phn y ca phn cong, dung dch c mu c phn pha trn ca mt lm. III/ nh Lng bng phng php o th tch: 1. Nguyn tc: - Bt k 1 cht no cn th A th phn ng hon ton vi ng lng gam ca 1 cht B bit, lun c phng trnh sau: NA . VA = NB . VB trong : NA, VA l nng ng lng, th tch cht th. NB, VB l nng ng lng, th tch cht chun. 2. Pha 1 cht chun A: - Pha t ng chun quc gia, K = 1. - Pha t ho cht cha tinh khit, tin hnh nh lng li bng dung dch chun, s dng h s iu chnh.

K=

N1 (Dung dch ta pha)

N chun

IV/ Thc Hnh: - Pha 250 ml dung dch NaOH 0,1N: Cn 1,1g NaOH thm nc 500ml. - Pha 1000 ml dung dch chun HCl 0,1N t ng chun. - nh lng dung dch NaOH va pha, tnh h s hiu chnh K.

BI 2

KIM NGHIM THUC TIM TRUYN NATRI CLORID NG TRNG

NaCl P.t.l. 58,4 L dung dch v khun ca natri clorid trong nc pha thuc tim. Ch phm phi p ng cc yu cu trong chuyn lun "Thuc tim, thuc tim truyn" v cc yu cu sau y: 1. Tnh cht Dung dch trong, khng mu, v mn. pH: 4,5 7,0 2. nh tnh * Xc nh ion natri (kin tp). - Nguyn tc: Na+ + Zn [(UO2 )3(CH3COO-)8] + CH3COO- + 9H2O = NaZn[(UO2)3 (CH3 COO-)9] . 9 H2O
1 git dd Na+ H kh, ngui 1 git TT Streng Tinh th vng, hnh mt nhn (soi kinh hin vi)

- Tin hnh: Trn 0,2 ml dung dch natri clorid 0,9% vi 0,2 ml dung dch km uranylacetat (TT): xut hin ta kt tinh mu vng, soi trn kinh hin vi thy tinh th hnh mt nhn mu vng lc nht * Xc nh ion clo: - Nguyn tc: Cl + AgNO 3 = AgCl + NO 3AgCl + 2 NH3 = [Ag(NH3)2]+ ClPhc tan

- Ly 2 ml ch phm. Acid ho bng vi git acid nitric long (TT). Thm 0,5 ml dung dch bc nitrat (TT): xut hin ta trng ln nhn. Ta khng tan trong cc acid m tan trong dung dch amoniac. 2 ml cp
3 git HNO3 10% 3 git AgNO3 5% trng ln nhn

3. Kim loi nng: < 0,3ppm 4. v khun: t v khun (PL 10.8). 5. Ni c t vi khun: Khng c qu 0,5 EU (PL 10.3). 6. nh lng: - Nguyn tc: phng php kt ta, nh lng trc tip. Cl+ AgNO3 = AgCl (trng) + NO3AgNO3 (tha) + K2CrO4 = Ag2CrO4(hng tht) + 2KNO3 - Tin hnh: Ht chnh xc 5 ml dung dch NaCl 0,9% cho vo bnh nn 100, thm 3 git ch th kali cromat. Chun bng dung dch bc nitrat 0,1N cho n khi xut hin ta hng. (hnh v trang sau).

AgNO3 0,1N

5 ml NaCl 0,9% 3 git CT K2CrO4 mu hng tht

- Tnh ton: 1 ml dung dch AgNO3 0,1N tng ng vi 0,005844 gam NaCl 0,005844 x V(AgNO3) Hm lng = x 100 VNaCl - Yu cu: hm lng NaCl trong ch phm t 0,850% - 0,950%

BI 3

KIM NGHIM THUC TIM TRUYN GLUCOSE 5%

Cng thc:

OH

OH C6H12O6 P.t.l: 180,2 Tn khoa hc: D (+) glucopyranose. Ch phm t yu cu trong chuyn lun "Thuc tim truyn" v cc yu cu sau y: 1. Tnh cht Dung dch trong sut, khng mu. 2. nh tnh. Ly 1ml thuc tim truyn, thm 5 ml thuc th Fehling. un si s xut hin ta ng (I) oxyd c mu gch.

3. Kim loi nng: < 0,5ppm 4. Cht gy st: PL 10.5 5. S nhim tiu phn l: t yu cu ca php th Tiu phn khng nhn thy bng mt thng (PL 8.9). 6. nh lng:
* DVN III: o nng sut quay cc. ** DVN I - Phng php: Phng php oxy ho kh, chun tha tr, ch th h tinh bt. - Nguyn tc: Oxy ho Glucose thnh acid Gluconic trong mi trng kim bng I 2 tha. nh lng I2 tha bng Na2S2O3 0,1N vi ch th h tinh bt trong mi trng acid. Phng trnh phn ng: R-CHO + I2 + H2O = R-COOH + 2HI I2 + H2O HIO + HI I2 + Na2S2O3 = 2 NaI + Na2S4O6 - Tin hnh: 1x 1x C+1 - 2 e -> C+3 I20 + 2 e -> 2 I-

CH2 OH
O

OH
OH

Ly chnh xc 1ml dung dch glucose 5%, chnh xc 10 ml dung dch I 2 0,1N, ly khong 10ml dung dch Na2CO3 5% cho vo bnh nn 250ml. Lc u ri cho vo bng ti khong 10 pht. Ly bnh nn ra, acid ho dung dch nh lng bng 15ml dung dch H2SO4 10%, lc k. Tin hnh nh lng bng dung dch Na2S2O3 0,1N n khi chuyn mu vng rm (vng nht) th thm 3 git CT h tinh bt vo chuyn mu tm en. Tip tc chun n khi dung dch mt mu. Na2S2O3 0,1N

1 ml glucose 5% 10 ml I2 0,1N 10 ml Na2CO3 5%

10 pht

15 ml H2SO4 10%, lc k Khi dd mu vng rm th: 3 git CT h tinh bt Mt mu

Song song lm mu trng: khng c glucose Cng thc tnh: C% = (Vtr - Vth ) 0,009008.K.n.100 Vglc 1 ml I2 0,1N t 0,009008 g glucose Yu cu: Hm lng phi t 95,0 105,0% so vi lng ghi trn nhn.

BI 4 KIM NGHIM THUC TIM CALCI CLORID 10% - Cng thc: CaCl2 P.t.l: 272,7 L dung dch v khun. Ch phm p ng cc yu cu trong chuyn lun "Thuc tim" v cc yu cu sau y: 1. Tnh cht: Dung dch trong, khng mu. 2. nh tnh: Nguyn tc: * Phn ng ca Ca2+: CaCl2 + (NH4)2C2O4 CaC2O4 trng Ta ny khng tan trong CH3COOH, tan trong HCl (TT) * Phn ng ca Cl-: Cl - + AgNO3 = AgCl trng Ta ny khng tan trong HNO3, tan trong NH4OH d (TT) AgCl + NH3 = [Ag(NH3)2] + Clphc tan Tin hnh:
2 ml CaCl2 10%

10 (NH4)2C2O4 4% trng CH3COOH 10% d tan HCl 10% d

2 AgNO3 2% trng NH4OH 10% d

tan

3. pH: 5,0 7,0. 4. nh lng: Dung dch CaCl2 10%. Nguyn tc: Phng php Complexon nh lng trc tip Ca2+ bng dung dch EDTA, phn ng tin hnh trong mi trng kim mnh. Ch th Murexit to phc vi Ca2+ mu , ti im tng ng ch th chuyn sang mu tm hay xanh. Natri ethylendiamintetraacetat (Na2EDTA), cn gi l Complexon III hay Trilon B: NaOOC-CH2 CH2-COOH N-CH2-CH2-N HOOC-CH2 CH2COONa To phc vi Ca2+ O=CO CH2 NaOOC- CH2 N CH2 Ca O-C=O CH2 N CH2 COONa CH2

Tin hnh: Ly chnh xc mt th tch ch phm tng ng vi 0,5g CaCl2, pha long vi nc va 200 ml. Ly chnh xc 25 ml dung dch thu c, chun bng Na2EDTA 0,05M. Khi cn cch im tng ng 2 ml, thm 5ml dung dch NaOH 20% v 3 git Muexit (CT), tip tc chun ti khi dung dch c mu tm hay xanh. 1ml Na2EDTA 0,05M tng ng vi 7,35 mg CaCl2.2H2O Yu cu: Hm lng CaCl2.2H2O t 95 105% so vi hm lng ghi trn nhn.

BI 5 KIM NGHIM THUC TIM PROCAIN HYDROCLORID - Cng thc:

C 2 H5 H2 N COOCH 2 CH 2 N C 2 H5 .HCl

C13H20O2N2.HCl P.t.l: 272,7 - Tn khoa hc: 2-(diethylamino) ethyl p-aminobenzoat monohydroclorid. L dung dch v khun, c ng trng bng cch thm natri clorid. Ch phm p ng cc yu cu trong chuyn lun "Thuc tim" v cc yu cu sau y: 1. Tnh cht: Dung dch trong, khng mu. 2. nh tnh: Nguyn tc: Phn ng to phm mu nit: Ar - NH2 + NaNO2 + H+
HO

(Ar - N N)+ + Na+ + H2O Mui diazoni

HO

(Ar - N+ N)

Ar - N = N

Mui diazoni Phn ng clorid: Cl + AgNO 3

Naphtol = AgCl (trng)

Phm mu nit (mu ) + NO 3-

AgCl + 2 NH3

= [Ag(NH3)2]+ ClPhc tan

Tin hnh: dung dch ch phm 2%.. - 2,5 ml dung dch ch phm thm 1 git HCl 0,5N, 1 git NaNO2 1% (mi pha), sau 1 2 pht, thm 0,5 ml dung dch 10% ca beta - naphtol pha trong NaOH 15% c ta mu cam. - 2 ml dung dch ch phm thm 5 git HNO3 10%, 3 git AgNO3 5% s c ta trng ln nhn. Ta ny tan trong amoniac.
0,1g cp, 5 ml H2O

5 git HNO3 10% 5 git AgNO3 5% trng

1 git HCl 0,5N 1 git NaNO2 0,1M 0,5 ml TT - naphtol/NaOH 10% Mu cam

3. pH: 3,5 5,5. 4. nh lng: Dung dch novocain HCl 2%. * DVN III: Php o nitrit, pp chun o Ampe. * DVN I: - Nguyn tc: Phng php trung ho, xc nh HCl t hp: C13H20O2N2. HCl + NaOH = C13H20O2N2 + NaCl + H2O Novocain base gii phng, do tnh kim, s nh hng ti pH ca mi trng l lp alcol - nc c cha cht cn nh lng, ch th v NaOH nh xung. Cloroform c tc dng ho tan v tch novocain base gii phng t phn ng khi mi trng phn ng, do khng nh hng ti pH ca tng alcol - nc na. Alcol thng acid vi phenolphtalein nn cn trung tnh trc. - Tin hnh: Ht chnh xc 10 ml ch phm cho vo bnh nn 250 ml, thm 10 ml cn 900 trung tnh vi phenolphtalein, thm 10 ml cloroform. nh lng bng NaOH 0,1 N cho ti khi lp nc c mu hng.

NaOH 0,1N

10 ml ethylic 90o 3 git CT phenolphtalein -> trung tnh ho

5 ml CHCl3 10 ml Novocain 2% mu hng bn

- Tnh kt qu: 1 ml NaOH 0,1 N tng ng vi 0,02728 g C13H20O2N2. HCl. - Cng thc tnh: Hm lng novocain hydroclorid trong ch phm: Hm lng 0,02728 x VNaOH x 100 VNov = % Hm lng cloramphenicol so vi hm lng ghi trn nhn: x 100 Hm lng thc t Hm lng ghi trn nhn - Yu cu: Hm lng ca novocain hydroclorid t 95,0 n 105,0% so vi hm lng ghi trn nhn.

BI 8 KIM NGHIM THUC TIM VITAMIN B12

L dung dch v khun ca vitamin B12 (cyanocobalamin) trong nc pha thuc tim. Ch phm p ng cc yu cu trong chuyn lun "Thuc tim, thuc tim truyn" v cc yu cu sau y: 1. Tnh cht: Dung dch trong, mu mn. 2. nh tnh: Ph hp th ca dung dch mu th ghi c phn nh lng phi c 3 cc i hp th 278 1 nm, 361 1 nm, 550 2 nm. T s hp th: A278 / A361 = 0,57 0,02 A550 / A361 = 0,3 0,02 3. pH: 4,0 5,5. 4. nh lng: Dung dch vitamin B12 500 (1000) mcg/ml Nguyn tc: Do c h ni i trong phn t, Vitamin B12 hp th bc x UV VIS. Tin hnh: Pha long dung dch ch phm vi nc c nng 25 mcg/ml. o hp th bc sng 361 nm, mu i chng l nc. Tnh hm lng vitamin B12 theo E1%1cm = 207 max = 361 nm. Ch : trong sut qu trnh o phi trnh nh sng

BI 9 KIM NGHIM THUC M BENZOSALI

COOH

OH COOH

Acid benzoic Cng thc thuc m Benzosali Acid benzoic (bt mn) Acid salicylic (bt mn) Cht nh ho 60g 30g 910g

Acid salicylic

L thuc m dng ngoi da c cha acid benzoic v acid salicylic trong cc cht nh ho thch hp.

Nghin mn dc cht, thm ng lng cht nh ho, nghin tip n ht Ch phm phi p ng cc yu cu trong chuyn lun Thuc m v cc yu cu sau: . 1. Tnh cht Thuc m phi mn, thnh phn phi tht ng nht, khng c chy lng nhit 37oC v phi dnh c vo da hay nim mc. 2. nh tnh Ly 2 g ch phm, thm 20 ml dung dch NaOH 0,1N, va un va khuy nh bng a thu tinh cho chy. ngui, lc, ly 2 ml dch lc, thm 5 git TT FeCl 3, dung dch xut hin mu tm v ta xut hin. 3. nh lng acid salicylic v acid benzoic trong m Benzosali ton phn - Nguyn tc: Acid salicylic v acid benzoic c nhm COOH trong phn t, nhm chc acid, do c th nh lng 2 acid ny bng mt base mnh nh NaOH (phng php acid acid base), ch th phenolphtalein. R COOH + NaOH = RCOONa + H2O - Tin hnh: Ly 2g ch phm cho vo binh non, thm 150 ml nc, un nng nh cho chy, thm 3 git ch th phenolphtalein. Chun bng NaOH 0,1N chun, ti khi dung dch xut hin mu hng.

NaOH 0,1N

2g ch phm 150 ml nc nng 3 git CT phenolphtalein mu hng

- Tnh kt qu: 1ml NaOH O,1N tng ng vi 0,01381 gam acid salicylic 1ml NaOH O,1N tng ng vi 0,01221gam acid benzoic Trong 1 g ch phm c: + 0,06 g acid benzoic + 0,03 g acid salicylic - Cng thc tnh: Theo l thuyt th s ml NaOH 0,1N dng cho 2 gam m : (4,3 + 2,5) x 2 = 13,6 (ml) - Yu cu: s ml NaOH 0,1N dng cho 1 g ch phm l 6,7 7,6 ml 1 g ch phm cha: + 0,057g 0,063g acid benzoic (4,7 5,2 ml NaOH 0,1N) + 0,027g 0,033g acid salicylic (2,0 2,4 ml NaOH 0,1N)

BI 11 KIM NGHIM VIN NN BERBERIN CLORID L vin nnn cha Berberin clorid. Ch phm p ng cc yu cu trong chuyn lun "Thuc vin nn" v cc yu cu sau y: 1. Tnh cht : Vin mu vng. 2. nh tnh : A.
Bt cha 0,1g cp, 10 ml H2O, un nng nh Lc 5ml dch lc + 3 git NaOH Mu cam + 5 git aceton vng aceto berberin Lc 2ml dch lc + vi git HNO3 10% + 3 git AgNO3 5% trng, tan trong NH3

B.

Bt cha 0,05g cp, 2 ml HCl 10%, lc tan. + bt Cloramin T anh o

3. nh lng:

Nguyn tc: Berberin HCl trong mi trng nc c 4 nh hp th UV l

228, 264, 345 v 423 nm. Do vy ta c th s dng phng php o ph hp th t ngoi nh lng Berberin HCl (C th dng nh tnh). Chn = 345nm nh lng Berberin HCl.

Tin hnh:
Dung dch th: Cn 20 vin, tnh khi lng trung bnh v nghin thnh bt mn. Cn chnh xc mt lng bt vin tng ng vi khong 50mg Berberin clorid, thm 10ml nc thm u bt vin, sau thm khong 100ml nc si, khuy k 5 pht, ngui ri chuyn vo bnh nh mc 250ml, thm nc ti vch, lc u. lng t nhin hoc em ly tm. Ly chnh xc 2ml dung dch trong trn em pha long vi nc ct va 50ml.

Dung dch chun c pha tng t dung dch th, dng cht i chiu berberin clorid. o hp th ca dung dch th v dung dch chun trong cng iu kin ti bc sng cc i 345nm (Ph lc 3.1) vi mu trng l nc ct, cc o dy 1cm.

Lu : - tn thi gian ch ly tm hoc ch lng cn th ta tin hnh lc, b

10ml dch lc u tin hnh o dch lc cn li.

- Trnh sai s: Lm 3 mu (Pha t dung dch gc) - Song song lm mu chun: Cn 50mg Ber. HCl chun. 1. Tnh ton kt qu:
Cng thc tnh: Hm lng ca Ber. HCl so vi hm lng ghi trn nhn.
Et x Ec x
mtb mt 50 mc

x 100

Trong : Et: Gi tr o hp th UV ca mu th (Thuc vin). Ec: Gi tr o hp th UV ca mu chun. mth: Khi lng cn thc t ca bt vin (Gm m).
m

: Khi lng trung bnh vin

M m = 20

mchun: Khi lng cn ca mu chun ( 50mg).

Yu cu: Hm lng ca Berberin clorid C20H18NO4Cl t 93,0 n 107,0%

so vi lng ghi trn nhn.