TRNG I HC TY

KHOA DC IU DNG

GIO TRNH THC TP

KIM NGHIM DC PHM

(DNG CHO SINH VIN I HC DC)

Cn Th, nm 2015
 2

MC LC

Trang
Bi 1. Kim nghim nguyn liu Calci clorid .................................................................. 5
Bi 2. Kim nghim vin nang Paracetamol .................................................................. 10
Bi 3. Kim nghim thuc tim Vitamin B12................................................................ 13
Bi 4. Kim nghim thuc nh mt Neodex .................................................................. 17
Bi 5. Kim nghim thuc bt Pabemin ........................................................................ 21
Bi 6. Kim nghim vin nn sulfaguanidin 500 mg ..................................................... 27
Bi 7. Kim nghim siro Theralen ................................................................................. 30
Bi 8. Kim nghim vin nn aspirin ............................................................................. 33
Bi 9. Kim nghim vin mt ngh ................................................................................ 35
 3

NI QUI PHNG THC TP KIM NGHIM

1. Sinh vin phi c mt trong phng th nghim sut bui thc tp. Ra ngoi phi xin
php ging vin hng dn.
2. Sinh vin n tr qu 15 pht xem nh vng mt, nu c l do chnh ng phi lm
n xin thc tp b vo bui khc.
3. Sinh vin vng 01 bui thc tp s b cm thi.
4. Nu ngh c l do chnh ng, sinh vin phi lm n xin php ghi tht r rng l do
(vic g? u? thi gian no?) km giy t xc minh l do v phi trc tip n gp
ging vin trong gi hc thc hnh ca t khc gii trnh l do v xin lch hc b.
5. Mi sinh vin ch c ngh/ i 01 bui thc hnh. Nu khng th sp xp i hc
c, sinh vin xin lm n xin tm dng hc v bo lu kt qu hc tp gii quyt
cng vic v khng b cm thi.
6. Nhng trng hp sau y s khng c chp nhn l vng c php, khng c
thc tp b v b cm thi:
- Ngh hc khng vit n xin php
- Gi in thoi/ nhn tin xin php ngh hc/ hc b
- Vit n xin php vi l do bn vic gia nh m khng ghi r vic g? u? thi
gian no? hoc l do em b bnh m khng ghi r bnh g? thi gian pht bnh?
thi gian iu tr?
- Ngh hc ri mi vit n xin php (nu bnh t xut phi vit n nh bn gi h
trc khi kt thc bui hc)
- T i bui hc m khng vit n xin php trc.
- L do khng chnh ng: v qu chi, i du lch, i d tic, qu cha ln kp, gi
nh, trng em, trng chu,
- L do gian di, khng ng s tht, khng c giy t xc nhn.
7. Sinh vin phi mc o blouse c cu vai mu xanh khi vo phng thc tp, cp sch
gn gng, ng ni qui nh.
8. u mi bui hc sinh vin k nhn dng c. Cui bui dng c phi c ra sch
s, sp xp ng qui nh, kim tra v k tr vo s. Tiu nhm thc tp phi chu
trch nhim v s mt mt, h hng, b v dng c ca tiu nhm.
9. Cm ht thuc, a gin, nghe in thoi trong phng th nghim.
10. Khng c ci ni n o, lm vic ring trong khi thc tp. Khng c qua v
 4

tr ca tiu nhm khc gy mt trt t.

11. Khng c n ung trong phng thc tp, nc ung phi bn ngoi.
12. Mi sinh vin phi mang theo 01 quyn gio trnh thc tp. Nu khng em theo
gio trnh s khng c thc tp.
13. Khng c t s dng/ di chuyn nhng dng c, ha cht, my mc ngoi
phm vi bn thc tp ca mnh. Khng c t thc hin nhng th nghim khng
c trong bi thc hnh.
14. Trc khi thc tp, sinh vin phi c k bi thc tp trc nh, v bi l thuyt
lin quan. Chun b nhng dng c c nhn h tr cho bi thc tp hm nh bao
tay, khu trang, khn giy,
15. Cui mi bui thc tp, nhm trng phn cng cc tiu nhm thay nhau trc v
sinh phng (dn dp dng c, ha cht dng chung, lau bn, v sinh cn, qut nh, lau
nh, v sinh cc my mc c s dng, dn lavabo, git khn phi ln ca s, chm y
cc bnh nc ct, rc, ng ca s). Nhm trc b v khng trc s nhn 0 im.
16. Sinh vin np bi bo co vo u mi bui thc tp ca bi k tip vo ng 15
pht u gi. Sau thi im ny s khng nhn bi v sinh vin phi nhn 0 cho bi
bo co .
17. Mi tiu nhm phi mang theo 01 b dng c nhn nh sau:
- 01 khn vi sch, 01 bt khn giy
- Gng tay y t, khu trang y t
- Vit mc, vit ch, thc, bm
- 01 vit lng mu xanh (loi vit ln thy tinh c)
- 01 nhp y t, 01 mung nha sch
- 01 bm tim 1ml, 01 li lam
Cn Th, ngy thng .. nm....
Sinh vin k tn

H tn:..
 5

BI 1

KIM NGHIM NGUYN LIU

CALCICLORID DIHYRAT
(XC NH GII HN TP CHT)
MC TIU
1. Trnh by c cch xc nh gii hn tp cht trong kim nghim nguyn liu.
2. Thc hin c cc phng php xc nh cc gii hn tp cht.
NI DUNG
1. YU CU KIM NGHIM
1.1. Hnh thc: bt kt tinh trng, d ht nc. D tan trong nc, tan trong ethanol
96%.
1.2. nh tnh: phi cho nhng phn ng c trng ca ion calci v clorid.
1.3. trong v mu sc dung dch: c mu khng m hn mu V6 (ph lc 9.2
v 9.3)
1.4. Gii hn acid kim: dung dch ca nguyn liu khng c acid qu hoc
kim qu.
1.5. Cc gii hn tp cht:
1.5.1. Sulfat: khng c qu 0,03% (ph lc 9.4.14)
1.5.2. Nhm
1.5.3. Bari: khng c c hn dung dch gm 10% dung dch chun + 1 ml nc.
1.5.4. Kim loi nng: khng c qu 20ppm (ph lc 9.4.8).
1.5.5. St: khng c qu 10ppm (ph lc 9.4.13).
1.5.6. Magnesi v mui kim: cn khng c qu 0,5%.
1.6. nh lng: hm lng CaCl2.2H2O t 97,0-103,0%.
2. PHNG PHP TH
2.1. Hnh thc: nh ghi trong yu cu kim nghim
2.2. nh tnh
- Dung dch S: Ha tan 10g CaCl2.2H2O trong nc khng c carbon dioxyd (TT) v
thm nc va 100ml (nc ct dun si ngui).
A. Ly khong 2mg ch phm cho vo mt ng nghim, thm 2ml nc ha tan,
acid ha bng dung dch acid nitric 2M (TT), thm 0,4ml dung dch bc nitrat 2%
 6

(TT), lc v yn, s to ta trng ln nhn. Lc ly ta, ra ta 3 ln, mi ln vi

1ml nc, phn tn ta trong 2ml nc v thm 1,5ml dung dch amoniac 10M, ta tan
ra d dng.
B. Ly 20mg ch phm cho vo mt ng nghim, thm 5ml dung dch acid acetic 5M
(TT) lc cho tan, thm 0,5ml dung dch kali ferocyanid (TT), dung dch vn trong, cho
thm khong 50mg amoni clorid (TT), to thnh ta kt tinh trng.
C. Ly 2ml dung dch S cho vo ng nghim, thm vi git dung dch amoni oxalat
4% (TT), to thnh ta trng, ta ny t tan trong dung dch acid acetic 6M (TT),
nhng tan trong acid hydrocloric (TT).
2.3. tan v mu sc ca dung dch: (bi thc tp 2)
2.4. Gii hn acid kim
Ly 10ml dung dch S mi pha, thm 0,1ml dung dch phenolphtalein (TT). Nu dung
dch c mu , thm 0,2ml dung dch acid hydrocloric 0,01M (C), dung dch phi
mt mu. Nu dung dch khng mu, n phi chuyn sang mu khi thm khng qu
0,2ml dung dch natri hydroxyd 0,01M (C).
2.5. Cc gii hn tp cht
2.5.1. Sulfat: khng c qu 0,03% (ph lc 9.4.14). Ly 5ml dung dch S, pha
long vi nc thnh 15ml th.
Chun b ng thi hai ng chun v th theo bng sau:
Bng 1.1. Quy nh chun b ng th v ng chun
ng th ng chun

Bari clorid 25% 1ml 1ml

Dung dch sulfat mu 10 phn triu 1,5ml 1,5ml

Lc v yn 1 pht

Ch phm th 5ml dd S + 10ml nc ct 15ml 0ml

Dung dch sulfat mu 10 phn triu 15ml

Dung dch acid acetic 5M (TT) 0,5ml 0,5ml

Lc v yn 5 pht
c to thnh trong ng th khng c m hn trong ng chun.
2.5.2. Nhm
10ml dd S, thm 2ml dd amoniclorid 10,7% v 1ml dd amoniac 10% (TT) un nng
 7

ti si, dd khng c vn c hay ta. Nu dng ch phm dng pha cc dd thm

tch th n phi t yu cu php th sau y thay cho php th trn: Ti a 1 phn
triu (Ph lc 9.4.9)
Dung dch th: Ha tan 4g ch phm trong 100ml nc, thm 10ml dung dch m
acetat pH 6,0.
Dung dch i chiu: Trn 2ml dung dch nhm mu (2 phn triu), 10ml dung dch
m acetat pH 6,0 v 98ml nc.
Dung dch mu trng: Trn 10ml dung dch m acetat pH 6,0 vi 100ml nc.
2.5.3. Bari
Ly 10ml dung dch S, thm 1ml dung dch calci sulfat (TT). Sau t nht 15 pht, dung
dch khng c c hn dung dch gm 10ml dung dch S v 1ml nc.
2.5.4. Kim loi nng: khng c qu 20 phn triu (ph lc 9.4.8)
Dung dch th: ly 12ml dung dch S th theo phng php 1 ca ph lc 9.4.8 trong
DVN IV. Dng dung dch ch mu 2 phn triu pha ng mu i chiu.
Phng php 1: chun b ng thi 3 ng nghim
Bng 1.2. Quy trnh chun b ng th, ng chun v ng trng
ng th ng ng trng
chun
Ch phm S (ml) 12ml 2ml 2ml

Dung dch ion ch mu 2 phn triu (TT) 10ml

(ml)
Nc ct (ml) 10ml

Dung dch m acetat pH 3,5 (ml) 2ml 2ml 2ml

Dung dch thioacetamid (TT) (ml) 1,2ml 1,2ml 1,2ml

Lc ngay v yn 2 pht

So snh mu to trong ng th v ng chun.

Mu nu trong ng th khng c m hn mu trong ng chun.
ng chun c mu nu nht khi c so snh vi ng trng.
2.5.5. St: khng c qu 10 phn triu (ph lc 9.4.13)
Tin hnh ng thi theo bng sau:
 8

Bng 1.3. Quy trnh chun b ng th v ng chun th gii hn st

ng th (Nessler ng chun
th) (Nessler chun)
Ch phm S (ml) 10 0
Dung dch st mu 1ppm (TT) (ml) 10

Acid citric (ml) 2 ml 2ml

Acid mercapto acetic 10M (ml) 0,1ml 0,1ml

Lc u

Dung dch amoniac 10M (TT) (ml) Kim ha Kim ha

Nc ct (ml) Va 20ml Va 20ml

Mu hng to thnh trong dung dch th khng c m hn mu chun.
2.5.6. Magnesi v mui kim loi kim: khng c qu 0,5%
Ly 20ml dung dch S, thm 80ml nc, 2g amoni clorid (TT) v 2ml dung dch
amoniac 10% (TT). un si. Rt vo dung dch ang si ny mt dung dch ang nng
gm 5g amoni oxalat (TT) ha tan trong 75ml nc. yn trong 4 gi. Pha long
thnh 200ml bng nc ri lc. Ly 100ml dch lc, thm 0,5ml acid sulfuric (TT).
Bc hi cch thy n kh ri nung 600 0C n khi lng khng i. Lng cn
khng qu 5mg.
2.6. nh lng
Cn chnh xc khong 0,735g CaCl2.2H2O (khong 0,05M) cho vo bnh nh mc
100ml. in nc n vch. Lc u. Ht chnh xc 10ml cho vo bnh nn 250ml.
Thm 90ml nc ct, 10ml NaOH 2N, thm 0,1g ch th Muretxit (khong mt ht u
xanh). nh lng bng dung dch EDTA 0,05M.
1ml dung dch EDTA 0,05M tng ng vi 7,35mg CaCl2.2H2O
nh lng Calci clorid theo DVN IV (sinh vin tham kho thm)
Ha tan 0,280g ch phm trong 100ml nc v tin hnh nh lng calci bng
phng php chun complexon (ph lc 10.5).
1ml dung dch Trilon B 0,1M (C) tng ng vi 14,70mg CaCl2.2H2O. Ly mt
lng dung dch ch phm nh ch dn trong chuyn lun cho vo mt bnh nn 500ml
ri pha long vi nc ct thnh 300ml. Thm 6ml dung dch natri hydroxyd 10M
(TT) v khong 15mg hn hp calcon (TT) ri chun bng dung dch trilon B 0,1M
 9

(C) n khi mu ca dung dch chuyn t tm sang xanh hon ton.

1ml dung dch trilon B 0,1M tng ng vi 14,70mg CaCl2.2H2O.
3. BO CO KT QU: vit bo co theo phiu kim nghim.
CU HI
1. Ti sao phi xc nh gii hn acid- base trong kim nghim nguyn liu ?
2. Ti sao phi xc nh gii hn tp cht trong nguyn liu ?
3. nh lng Calci clorid theo nguyn tc no trong php chun th tch.
 10

BI 2

KIM NGHIM THUC VIN NANG

DOPAGAN (PARACETAMOL 500 mg) (TCCS)
MC TIU
Thc hin nhng yu cu k thut c hc trong phn l thuyt ca bi KIM
NGHIM THUC VIN NANG.
NI DUNG
1. YU CU KIM NGHIM
1.1. Hnh thc cm quan: Vin nang cng, mt u , mt u trng. Mt nang kh,
bng, thuc cha trong nang mu trng.
1.2. Thi gian th r: khng qu 15 pht (ph lc 11.6 DVN IV)
1.3. Chnh lch v khi lng: c php 7,5% so vi khi lng trung bnh ca
bt thuc trong mt nang (ph lc 11.3 DVN IV)
1.4. nh tnh: phi c phn ng c trng ca paracetamol v ph IR ca
paracetamol trong nang phi ging vi ph IR ca paracetamol i chiu.
1.5. nh lng: ch phm phi cha 475 525 mg paracetamol trong mt nang tnh
theo khi lng trung bnh ca bt thuc cha trong mt nang.
2. PHNG PHP TH
2.1. Hnh thc cm quan: th bng cm quan ch phm phi t yu cu nu trn
2.2. ng u khi lng: tin hnh vi 20 nang. Cn ring tng nang (c v v
thuc). Sau , dc ht hot cht ra c th dng bng gn lau cho tht sch i vi
nang cng hoc dng ether hay mt dung mi thch hp ra nang cho sch i vi
nang mm. Cn v nang rng, khi lng ca tng nang l hiu s gia hai ln cn.
Xc nh khi lng trung bnh ca nang. Mu th t yu cu khi c khng c qu
2 vin c khi lng lch qu lch phn trm ca khi lng trung bnh v khng
c vin no vt qu 2 ln lch .
Bng 3.1. Bng quy nh ng u khi lng ca vin nang
Khi lng trung bnh % chnh lch so vi khi lng trung bnh
Nh hn 300mg 10
Ln hn hoc bng 300mg 7,5
2.3. nh tnh
- Lc 0,5 gam ch phm trong 10 ml nc, thm vi git dung dch st (III) clorid (TT)
 11

xut hin mu xanh tm hi nu, lu chuyn thnh mu nu.

- Ly 0,5 gam ch phm un trong 2ml dung dch hydroclorid 10% (TT) un si trn
n cn trong 3 pht, thm 10 git nc, ngui, vi 1 git dung dch kali dicromat
5% (TT) xut hin mu tm hi sau 30 pht s chuyn sang tm hon ton, khng
c chuyn sang .
- o ph IR: ly cn trn o ph IR, so snh vi ph IR ca paracetamol chun
2.4. nh lng
2.4.1. Bng phng php o quang
Ly 20 vin tn thnh bt mn. Cn mt lng bt vin tng mg vi 100m
paracetamol, cho vo bnh nh mc 100ml, thm 20ml dung dch NaOH 0,1N (TT),
thm 10ml nc ct, lc 15 pht, thm nc ct ti vch. Lc u, lc qua giy lc
kh, b 20 30 ml dung dch u. ly chnh xc 5 ml dch lc cho vo bnh nh mc
50ml, thm nc ct ti vch, lc u. Ly chnh xc 5ml dunh dch trn cho vo bnh
nh mc 50 ml, thm 5ml dung dch NaOH 0,1N, thm nc ct ti vch, lc u. o
hp thu ca dung dch ny bc sng 257 nm, cc dy 1cm. Mu trng l dung
dch NaOH 0,01N, ly A11 ca paracetamol tinh khit 257 nm l 715.
Kt qu: lng paracetamol c trong mt vin c tnh theo cng thc
Ax 1 m
C x (mg ) 107 tb (mg )
715 100 P

2.4.2. Bng phng php sc k hiu nng cao

iu kin sc k ti u
- My sc k lng hiu nng cao Hitachi L-2000 elite.
- Ct Phenomenex lunar RP C18 (250 x 4,6mm, 5m)
- Pha ng (MP): MeOH: H2O: acid acetic bng = (20:76:4)
- u d UV Vis: pht hin bc sng 257 nm
- Th tch tim mu: 20l
- Tc dng 1ml/pht
Pha dung dch chun i chiu:pha dung dch Paracetamol i chiu trong pha ng.
Cn 50 mg pha ng trong bnh nh mc 50ml vi pha ng, ht chnh xc 2ml pha
long trong bnh nh mc 50ml, lc qua mng lc milipore 0,46m.
Pha dung dch thuc th: ly 20 vin, tn thnh bt mn. cn mt lng bt vin tng
ng vi 100mg paracetamol, cho vo bnh nh mc 100ml. Cho pha ng vo siu
 12

m 10 pht. Lc qua giy lc b 20-30 ml, thu c dung dch B. Ht chnh xc 2ml
dung dch B cho vo bnh nh mc 50 ml. thm pha ng vo ti vch, lc qua mn
lc milipore 0,46m. Dung dch ny s c dng nh lng paracetamol c trong
vin nang bng phng php sc k lng, kt qu: lng paracetamol c trong 1 vin
c tnh theo cng thc
S t mc 100 25 mtb
P ( mg )
S c 50 25 p

St: dt nh paracetamol c trong dung dch mu th

Sc: dt nh paracetamol c trong dung dch mu chun tim vo ct (mg/ml)
m: khi lng trung bnh ca bt thuc trong mt vin nang
p: lng mu th cn pha dung dch th (g)
3. BO CO KT QU: theo phiu kim nghim.
CU HI
1. Nu r ccch kim nghim chnh lch v khi lng ca vin nang?
2. Nu vi phng php nh tnh paracetamol?
3. Nu vi phng php nh lng paracetamol?
4. Thit lp cng thc tnh hm lng paracetamol khi nh lng paracetamol khi
nh lng bng phng php o quang?
 13

Bi 3

KIM NGHIM THUC TIM

VITAMIN B12 1000 (g/ml)
(TC DVN IV)
MC TIU
1. Thc hin c nhng yu cu k thut chung hc trong bi Kim nghim
thuc tim (xem ph lc thuc tim PL27 DVN IV).
2. Thc hin phng php UV-Vis nh tnh v nh lng
NI DUNG
1. YU CU KIM NGHIM
1.1. Tnh cht: ch phm dng lng, trong sut, mu mn, ng trong ng thy
tinh khng mu.
1.2. trong: ch phm phi trong v hu nh khng c tp cht c hc nhn thy
c
1.3. ng u th tch: th tch ca mi ng phi t 100-115% ca th tch ghi
trn nhn.
1.4. pH: 4,0-5,5 (PL 6.2 DVN IV, trang PL144)
1.5. Tp cht lin quan: khng qu 6%
1.6. nh tnh: phi c cc phn ng c trng ca vitamin B12 khi o ph UV-Vis.
1.7. nh lng: hm lng vitamin B12 trong 01 ng phi t t 950-1150 mcg/ml.
2. PHNG PHP TH
Theo chuyn lun thuc tim vitamin B12, DVN IV, trang 203.
2.1. Tnh cht
Quan st cc ng thuc tim di nh sng ban ngy bng mt thng v so snh vi
yu cu.
2.2. trong
Dung dch thuc tim khi soi bng mt thng iu kin thch hp phi trong v hu
nh khng c tp cht c hc (PL 11.8). Thc hin trn 20 ng (V 5 ml), c 02
ng c nhiu nht 01 vt th nhn thy c.
2.3. ng u th tch (PL 11.1)
Ly 6 ng (01 ng trng bm tim, 05 ng o th tch). Kim tra bng cm quan
 14

cc ng th phi c th tch thuc tim gn bng nhau.

Dng bm tim kh sch c kim tim thch hp, c dung tch khng qu 2,5 ln so vi
th tch cn o. Ly thuc vo bm tim sao cho bm tim khng c bt kh v trong
kim tim vn cha y thuc tim. Ln lt ly ht thuc trong tng ng o theo
cch .
Kt qu th tch ca tng ng phi t 100 -115% ca th tch ghi trn nhn.
2.4. pH
Theo PL 6.2
o trn pH k vi in cc Calomel- thy tinh.
Chun ha my vi 2 dung dch m chun 4,01 v 6,86 (250C).
Th tch dung dch o: 10ml
2.5. Tp cht lin quan
Ch phm phi p ng cc yu cu ca chuyn lun thuc tim cyanocobalamin ca
DVN IV trang 204. Tin hnh nh sau:
Ct RP-C18 (150 x 4,6mm, 5m)
Pha ng (MP): MeOH acid phosphoric 0,1% (26: 74)
Bc sng: 361 nm
Th tch tim mu: 20 l
Tc dng: 1 ml/ pht
Dung dch th:
Pha long mu th trong pha ng c 500 g cyanocobalamin/ 1ml.
Dung dch chun a:
Pha long mu th trong pha ng c 30g cyanocobalamin/ 1ml.
Dung dch th b:
Pha long mu th trong pha ng c 10g cyanocobalamin/ 1ml.
Dung dch th c:
Ly 1 th tch mu th c cha 5mcg cyanocobalamin, thm 1ml cloramin T 0,1%
(kl/tt) v 0,1 ml dung dch HCl 0,05M ri thm nc cho va 10ml, lc u yn
5 pht. Pha long 2ml dung dch ny vi pha ng cho va 10ml.
Ch : dung dch chun c phi o ngay sau khi pha, cc dung dch khc khng c
qu 1 gi. Thi gian phn tch mu trn HPLC gp t nht 3 ln thi gian lu ca
cyanocobalamin.
 15

Cch nh gi:
Trong sc k ca dung dch th, tng din tch ca cc pic ph ngoi pic chnh
khng c ln hn din tch pic chnh thu c trong sc k ca dung dch chun
a (3%).
B qua tt c cc pic m din tch ca chng nh hn din tch pic chnh thu c
trong sc k ca dung dch chun b.
Php th ch c gi tr khi trong sc k ca dung dch chun c c 2 pic chnh v h
s phn gii gia 2 pic ny t nht l 1,5.
Sc k ca dung dch chun b c 1 pic chnh v t s din tch ca pic ny so vi
nhiu ca ng nn khng nh hn 5.
2.6. nh tnh
Trong phn nh tnh, ph t ngoi ca dung dch mu th c cc hp th cc i
278 nm, 361 nm v khong 547 n 559 nm. T s hp th cc i 361 nm so
vi hp th cc i khong 547nm n 559 nm t 3,15 n 3,45. T s hp th
cc i 361 nm so vi hp th cc i 278 nm t 1,70 n 1,90.
2.7. nh lng
2.7.1. Phng php UV-Vis
Pha long dung dch ch phm vi nc c nng 0,0020% (20g/ml). Ly chnh
xc 1ml cho vo bnh nh mc 50ml. Thm nc ct n vch pha long. o
hp thu bc sng 361 nm trong cc o dy 1cm, mu trng s dng l nc ct pha
tim (ch trong sut qu trnh nh lng phi trnh nh sng).
Hm lng vitamin B12 tnh theo % hm lng ghi trn nhn c tnh theo cng
thc sau (ly A(1%, 1cm) = 207).

AT .104.50
X
207
Trong :
X: Hm lng ca vitamin B12
AT: hp thu ca mu th
2.7.2. Phng php HPLC
Dung dch th: pha long chnh xc 1 ml ch phm vi nc c nng 0,002%
(20 mcg/ml) trong bnh nhm mc 50 ml.
Dung dch chun: cn chnh xc khong 0,01 g vitamin B12 chun i chiu, pha
 16

trong bnh nh mc 50 ml vi nc, sau pha long 10 ln vi nc c dung

dch chun c nng 0,002% (20 mcg/ml).
iu kin sc k:
Ct RP-C18 (150 x 4,6mm, 5m)
Pha ng (MP): MeOH acid phosphoric 0,1% (26: 74)
Bc sng: 361 nm
Th tch tim mu: 20 l
Tc dng: 1 ml/ pht
3. BO CO KT QU
Vit phiu kim nghim v phiu phn tch. Tr li cu hi.
CU HI
1. V cng thc cu to ca vitamin B12. Tn IUPAC ca cht ny l g? Vitamin B12
cn c tn gi khc l g?
2. iu kin mt cht c th nh lng bng phng php UV-Vis l g? Vitamin
B12 tha c iu kin no?
3. K tn mt s tp cht lin quan trong thuc tim vitamin B12
4. Vitamin B12 khi o quang c bao nhiu nh cc i? Ngi ta hay dng nh no
nh lng? Ti sao?
5. Nu 2 phng php xc nh th tch thuc tim
6. So snh 2 phng php nh lng UV-Vis v HPLC
7. Mc ch ca vic xc nh t s hp thu trong phn nh tnh l g?
8. Khi o pH ca dung dch vitamin B12 phi s dng cp in cc g? Cu to ca
cp in cc trn.
9. Ti sao phi chun my pH vi dung dch m 4,01 v 6,86? Qua ngy hm sau c
cn chun li khng? C ch o pH ca cp in cc trn l g?
10. Ti sao trong qu trnh nh lng vitamin B12 phi trnh nh sng? Vit phng
trnh phn ng xy ra.
11. K tn mt s ch phm c cha vitamin B12.
12. Trnh by cc bc tin hnh qut ph (nh tnh) trn my UV hc
13. Trnh by cc bc tin hnh o im (nh lng) trn my UV hc.
14. V hnh h thng my UV-Vis hc. Ch thch cc b phn.
 17

BI 4

KIM NGHIM THUC NH MT NEODEX

MC TIU
1. Thc hin nhng yu cu k thut c hc trong phn l thuyt ca bi KIM
NGHIM THUC NH MT
2. ng dng c k thut sc k lp mng vo kim nghim ch phm nhiu thnh
phn hot cht.
NI DUNG
1. YU CU KIM NGHIM
1.1. Hnh thc
Dung dch khng mu hoc c mu vng nht, khng mi, v mn, hi ng.
1.2. Chnh lch th tch: + 10% so vi th tch ghi trn nhn.
1.3. trong: phi t theo tiu chun quy nh 52 TCN 315-87.
1.4. pH: 6,0 7,7.
1.5. nh tnh
Phi c phn ng ha hc c trng ca neomycin sulfat v dexamethason natri
phosphat hoc dexamethason.
1.6. nh lng
Ch phm phi cha:
- Neomycin t 306,000 442,000 UI.
- Dexamethason natri phosphat t 90 110% hm lng ghi trn nhn cho 100 ml ch
phm.
1.7. nhim khun
Phi t cc tiu ch quy nh th nghim v khun (PL 13.7 trang PL 266).
2. PHNG PHP TH
2.1. Hnh thc: ch phm phi t cc yu cu nu trn.
2.2. Chnh lch th tch
Ly 5 chai bt k, xc nh th tch thuc nh mt c trong tng chai bng bm tim
chun hoc bng ng ong sch, kh, c chnh xc ph hp. Th tch mi n v
phi nm trong gii hn cho php. Nu c 1 chai khng t th phi kim tra li ln th
2 ging nh ln u. Ch phm ny t yu cu nu trong ln th ny khng c n v
no c th tch nm ngoi gii hn cho php.
 18

2.3. trong
Dng ng nghim c nt y, ra sch v dng nc ct soi m bo khng c bi, tp
cht c hc. nc ra, cho dung dch ch phm vo soi. Ch phm phi trong sut
khng c bi, cn, x bng hay nhng tp cht c hc khc,
2.4. pH: th theo DVN IV, ph lc 6.2 PL 144.
2.5. nh tnh
2.5.1. nh tnh Neomycin sulfat
Ly 2 ml ch phm cho vo ng nghim, thm 0,1 ml Pyridin v vi git dung dch
Ninhydrin 0,1%, un cch thy 65 70 0C trong 5 10 pht s xut hin mu tm
m.
Ly 0,5 ml ch phm cho vo ng nghim, thm 0,5 ml dung dch HCl (TT) v 1 ml
dung dch BaCl2 (TT), lc u, yn trong 10 pht s xut hin ta trng.
2.5.2. nh tnh Dexamethason
2.5.2.1. Bng phn ng ha hc
Ly 2 ml ch phm cho vo ng nghim, thm 1 ml dung dch HCl long (TT) v 2 ml
dung dch INH 0,05% trong MeOH, un cch thy 65 70 0C trong 5 10 pht s
xut hin mu vng.
Ly 2 ml ch phm cho vo ng nghim, thm 3 git dung dch xanh tetrazolium 1%
trong MeOH, 1 ml NaOH/MeOH, 1 ml tetramethylamonium hydroxyd 10%, lc u
v trnh nh sng (hay un cch thy 5 pht) dung dch s c mu hng tm.
2.5.2.2. Bng sc k lp mng
Bn mng 5 x 20 cm Silicagel F254 trng sn.
Dung dch th: ly 2 ml ch phm c cch thy cn khong 0,1 ml, chm khong 20 l
dch ny ln bn mng.
Dung dch i chiu: 10 mg dexamethason natriphosphat ha tan trong 1 ml nc,
chm khong khong 20 l dch ny ln bn mng.
H dung mi khai trin: n-butanol acid acetic nc (4: 1: 5).
Thuc th pht hin: phun H2SO4 10%/cn v thm vi git formol ri sy 110 0C
trong 10 pht (hay h hi iod).
Kt qu: dch chm ca ch phm phi cho 1 vt tng ng v mu sc v Rf vi
dung dch dexamethason natriphosphat chun (c th soi UV).
2.6. nh lng
 19

2.6.1. Phng php UV Vis

Ht chnh xc 1 ml ch phm cho vo bnh nh mc 50 ml, thm nc ct ti vch,
lc u. o mt quang ca dung dch ny bc sng 241 nm trong cc o dy 1
cm, dng nc ct lm mu trng.
% Hm lng dexamethason natri phosphat tnh theo cng thc:
AT 50 5
100
297 5,5
Vi AT: hp thu ca dexamethason natri phosphat trong mu th 241 nm.
297: E1cm ca dexamethason natri phosphat trong mu chun 241 nm.
2.6.2. Phng php HPLC
iu kin sc k ti u
- My sc k lng hiu nng cao Shimadzu
- Ct RP-C18 (150 x 4,6 mm, 5 m).
- Pha ng (MP): CAN: m phosphat (20: 80)
(KH2PO4 0,01 M pH 3,97 trong hn hp ng lng MeOH-H2O)
- u d UV Vis: pht hin bc sng 242 nm
- Th tch tim mu: 20 l.
- Tc dng: 1,4 ml/pht.
Dung dch chun (c pha ch ngay khi dng): dung dch dexamethason natri
phosphat chun trong pha ng c nng chnh xc khong 110 g/ml, tnh theo
dexamethason phosphat.
Dung dch th: ly chnh xc mt lng ch phm tng ng vi khong 1,1 mg
dexamethason phosphat pha long thnh 10 ml bng pha ng v trn u.
Cch tin hnh: kim tra kh nng thch hp ca h thng sc k: tin hnh sc k vi
dung dch chun, lch chun tng i ca cc din tch p ng t 6 ln tim lp
li khng c ln hn 2,0%. Tin hnh sc k ln lt vi dung dch th v dung
dch chun.
% Hm lng dexamethason natri phosphat tnh theo cng thc:
ST 5
CC 10 100
SC 5,5

CC: nng mu chun (g/ml)

ST: din tch nh mu th
 20

SC: din tch nh mu chun

3. BO CO KT QU: theo phiu kim nghim
CU HI
1. Hy k vi dng ch phm nh mt c cha dexamethason trn th trng (ni v
ngoi nhp)
2. Nhm chc no trong phn t dexamethason phn ng vi thuc th Fehling?
3. Nu cc phn ng c trng nh tnh dexamethason.
4. Trnh by cc ch tiu chung ca thuc nh mt cn thc hin theo quy nh (PL
1.14 trang PL-21)
5. So snh kt qu gia hai phng php nh lng.
 21

BI 5

KIM NGHIM THUC BT

PABEMIN TR EM (TCCS)

MC TIU
Thc hin nhng yu cu k thut c hc trong phn l thuyt ca bi KIM
NGHIM THUC BT, CM.
NI DUNG
1. YU CU KIM NGHIM
1.1. Hnh thc cm quan: dng bt i khi ht nh, mu cam, v ng, c mi thm.
Bt ngoi khng kh trong vng 24 gi phi khng c chy nc.
1.2. Chnh lch khi lng ng gi: 5% so vi khi lng ghi trn nhn.
1.3. m: khng qu 2% nu l thuc cm (sy 105oC).
1.4. nh tnh
- Phi c phn ng ca thiamin nitrat, clorpheniramin maleat, paracetamol.
- Phi c ph hng ngoi ca paracetamol trong ch phm ging vi ph paracetamol
chun.
1.5. nh lng: trong 1 gi phi cha:
- Paracetamol: 308,8 341,3 mg
- Thiamin nitrat: 9,0 11,0 mg
- Clorpheniramin maleat: 1,8 2,2 mg
Tnh theo khi lng trung bnh ca 1 gi.
1.6. mn: theo tiu chun c s.
1.7. ng u hm lng: clorpheniramin maleat 1,8 2,2 mg.
2. PHNG PHP TH
2.1. Hnh thc cm quan: thuc phi t yu cu nu.
2.2. Chnh lch khi lng
Cn tng gi trong s 5 gi c ly bt k. Tt c cc gi phi t quy nh 5% so
vi khi lng ghi trn nhn. Nu c 1 gi c khi lng lch ra ngoi quy nh ny
th phi th li vi 5 gi khc. Tt c 5 gi th li ln 2 phi t th mi t.
Ch : sau khi cn phi b ht thuc ra, dng bng lau sch thuc trong v, ri tnh
 22

khi lng tng gi.

2.3. m: sy 1 gam bt 100oC 105oC n khi trng lng khng i, m
khng qu 2%.
2.4. nh tnh
2.4.1. nh tnh clorpheniramin maleat (dng sc k lp mng):
- Chun b mu chun: cn 10 mg Clorpheniramin maleat ha tan vi 10ml cloroform.
- Chun b mu th: ly khong 1/2 gi cm chit vi 10ml cloroform trong mt ng
nghim, lc k, lc vo mt chn s, lm bay hi dch lc cn 1ml.
- Thc hin sc k:
+ Bn mng trng sn lp silica gel F254 dy 0,25mm, kch thc 3cm x 10cm.
+ Chm 10 l mu th v chm 10 l mu chun ln bng mng.
+ Trin khai vi h dung mi: Benzen EtOH NH4OH (85:15:1).
+ Sau khi trin khai, kh bng mng nhit phng, phun thuc th hin mu
Dragendoff, hoc h hi iod. Mu th, mu chun phi cho vt cng Rf (Rf 0,45) v
cng mu (mu vng sm mt nhanh ngoi khng kh).
2.4.2. nh tnh thiamin nitrat
- Ly 1,5 gam bt cm nghin mn cho vo ng nghim, thm vo 10ml nc ct,
lc k, lc vo mt ng nghim khc. Ly 5ml dch lc cho vo ng nghim, thm 2ml
dung dch NaOH (TT) lc k v vi git kaliferrycianid 5% (TT) vi 2ml n-BuOH.
Lc mnh 1-2 pht. yn, lp cn c mu xanh tm khi xem di n t ngoi. Khi
acid ha dung dch, lc mnh, qunh quang s mt i, kim ha dung dch th qunh
quang tr li.
- Th nitrat: ly 3ml dch lc, thm t t 1ml dung dch FeSO4 10% cho cn thn theo
thnh ng nghim 2ml acid sulfuric m c, ngui dung dch, vng tip gip
gia hai cht lng c mt vng mu nu.
2.4.3. nh tnh paracetamol
- Phn ng trong ng nghim: lc 0,5 gam ch phm trong 10ml nc, thm vi git
st (III) clorid (TT) xut hin mu xanh tm hi nu, lu s chuyn sang mu nu.
- Lc k 1,0 gam ch phm vi 10ml aceton, lc ly dch. dch aceton bc hi ti
cn, Ha tan mt t cn trong 2ml dung dch acid hydroclorid 10% (TT), un si trn
n cn trong 3 pht, thm 10 git nc, ngui, thm 1 git dung dch kali
dicromat 5% (TT) xut hin mu tm hi sau 30 pht s chuyn sang tm hon ton,
 23

khng c chuyn sang .

- o ph IR: ly cn trn o ph IR, so snh vi ph IR ca paracetamol chun.
2.5. nh lng
nh lng ng thi ba hp cht bng phng php HPLC
- Chun b mu:
Dung dch clorpheniramin maleat chun gc:
Cn chnh xc khong 20mg clorpheniramin maleat chun, ha tan trong khong 30ml
m phosphat pH 7 cho vo bnh nh mc dung tch 100ml, sao thm m
phosphat pH 7 va , lc u v lc qua mng lc kch thc 0,45m c dung dch
chun gc c nng clorpheniramin maleat khong 200m/ml.
Dung dch thiamin mononitrat chun gc:
Cn chnh xc khong 40mg thiamin mononitrat chun, ha tan trong khong 30ml
m phosphat pH 7 cho vo bnh nh mc dung tch 100ml, sau thm m
phosphat pH 7 va , lc u v lc qua mng lc kch thc 0,45m c dung dch
chun gc c nng clorpheniramin maleat khong 400m/ml.
Dung dch hn hp chun paracetamol, clorpheniramin maleat, thiamin mononitrat:
Cn chnh xc khong 32,5mg paracetamol chun, ha tan trong khong 20ml m
phosphat pH 7 cho vo bnh nh mc dung tch 50ml, sau thm chnh xc 1ml
dung dch clorpheniramin maleat chun gc v chnh xc 2,5ml dung dch thiamin
mononitrat chun gc vo bnh.
Lc u v thm m pH 7 va . Lc u v lc qua mng lc kch thc 0,45m
c dung dch hn hp chun c nng paracetamol khong 650g/ml,
clorpheniramin maleat khong 4g/ml v thiamin mononitrat khong 20g/ml.
Mu th t ch phm Pabemin:
Cho 5 gi Pabemin vo trong ci, nghin trn u. Cn chnh khong 0,125g ch
phm, ha tan trong khong 10ml m phosphat pH 7 cho vo bnh nh mc dung
tch 25ml, sau thm m phosphat pH 7 va , lc u v lc qua mng lc kch
thc 0,45m c dung dch mu th tim sc k.
- iu kin sc k ti u kho st c cho quy trnh nh lng Pabemin:
+ H thng my HPLC Hitachi L-2000, DAD L-2455.
+ Pha tnh: ct Phenomenex Gemini RP-C18 (150mm x 4,6mm, 5m, 110).
+ Th tch tim mu: 20l
 24

+ Nhit ct: nhit phng

+ Chng trnh chy:
Bng 7.2. Chng trnh chy mu ti u kho st c
Thi gian AcN (%) Methanol Nc (%) m pH 7 Tc dng
(pht) (%) (%) (ml/ pht)
0 0,8 7,2 0 92 2
3,5 0,8 7,2 0 92 2
3,6 5 25 0 70 2
20 5 25 0 70 2
+ Bc sng pht hin paracetamol ti 300nm, clorpheniramin maleat ti 225nm,
thiamin mononitrat ti 275nm.
- Tim ln lt cc mu dung dch hn hp chun v dung dch th vo h thng
HPCL/PDA, xc nh din tch nh paracetamol, clorpheniramin maleat v thiamin
mononitrat ca mu hn hp chun v mu th, ri tnh ton hm lng paracetamol,
clorpheniramin maleat trong 2,5g ch phm.
- Hm lng paracetamol (mg) trong 2,5g ch phm c tnh theo cng thc:
2 ST1
Xparacetamol = .m .a .2,5 (1)
mPab SC1 1 1
Hm lng clorpheniramin maleat (mg) trong 2,5g ch phm c tnh theo cng
thc:
1 ST2
Xclorpheniramin = .m .a .2,5 (2)
mPab.50 SC2 2 2
Hm lng thiamin mononitrat (mg) trong 2,5g ch phm c tnh theo cng thc:
2.5 ST3
Xthiamin = .m .a .2,5 (3)
mPab SC3 3 3
Trong :
mPab: l khi lng bt thuc Pabemin (g).
ST1, ST2, ST3: ln lt l din tch nh ca paracetamol, clorpheniramin maleat v
thiamin mononitrat trong mu th.
SC1, SC2, SC3: ln lt l din tch nh ca paracetamol, clorpheniramin maleat v
thiamin mononitrat trong mu chun.
m1, m2, m3: ln lt l khi lng paracetamol chun, clorpheniramin maleat chun v
thiamin mononitrat chun (mg).
 25

a1, a2, a3: ln lt l tinh khit ca cc cht chun paracetamol, clorpheniramin

maleat v thiamin mononitrat.
2.6. ng u hm lng (clopheniramin maleat)
Chun b mu th
Ly ht bt thuc trong 1 gi nghin mn, cn khong 0,125 g ch phm, ha tan trong
khong 10 ml m phosphat pH 7 cho vo bnh nh mc dung tch 25 ml, sau
thm m phosphat pH 7 va , lc u v lc qua mng lc kch thc 0,45 m
c dung dch mu th tim sc k. Lm tng t vi 9 gi na.
nh lng thnh phn clorpheniramin maleat theo phng php nh lng nh trn.
Tnh ng u hm lng clopheniramin theo DDVN IV (phng php 1 ph lc
11.2 trang PL-221):
- Ch phm em kim tra t yu cu php th nu khng qu 1 n v c gi tr hm
lng nm ngoi gii hn 85 115% ca hm lng trung bnh v khng n v no
nm ngoi gii hn 75 125% ca hm lng trung bnh.
- Ch phm khng t yu cu php th nu c qu 3 n v c gi tr hm lng nm
ngoi gii hn 85 115% ca hm lng trung bnh hoc c 1 hay nhiu n v nm
ngoi gii hn 75 125% ca hm lng trung bnh.
- Nu c 2 hoc 3 n v c gi tr hm lng nm ngoi gii hn 85 115% ca hm
lng trung bnh nhng trong gii hn 75 125% ca hm lng trung bnh th th
li trn 20 n v khc, ly ngu nhin. Ch phm kim tra t yu cu php th nu
khng qu 3 n v trong tng s 30 n v em th c gi tr hm lng nm ngoi
gii hn 85 115% ca hm lng trung bnh v khng c n v no nm ngoi gii
hn 75 125% ca hm lng trung bnh.
3. BO CO KT QU: theo phiu kim nghim.
CU HI
1.Vit cng thc khai trin ca paracetamol, thiamin hydroclorid, clorpheniramin
maleat. C th nhn xt rng thiamin hydroclorid v clorpheniramin maleat c cu trc
nh mt base hu c c khng? Ti sao?
2. Gii thch c ch nh lng?
3. Theo DVN IV, tnh sai s khi lng ca dang thuc gi th phi cn bao nhiu
gi v cch tnh ra sao?
4. Vit c ch cc phn ng nh tnh paracetamol, thiamin HCl?
 26

5. Trong quy trnh nh lng, c th thay helianin bng 1 thuc th khc c

khng? C th thay cloroform bng dung mi hu c khc c c khng?
6. Ngoi h m phosphat cn s dng h m no khc khng? Ti sao?
7. Gii thch cng thc tnh hm lng tng hot cht trong ch phm Pabemin?
8. Chng trnh chy trong iu kin sc k trn l gradient, c th chy chng
trnh ng dng c khng? Loi dung mi dng cho chy gradient cng dng cho
chy ng dng c khng?
9. C th phn tch 3 hot cht trn cng mt bc sng? Ti sao?
 27

BI 6

KIM NGHIM THUC VIN NN

SULFAGUANIDIN 500 mg
MC TIU
1. Thc hin nhng yu cu k thut c hc trong phn l thuyt ca bi KIM
NGHIM THUC VIN NN.
2. Thc hin c php chun in th t ng.
3. Hiu c c ch phn ng ca nh lng bng phng php nitrit.
NI DUNG
1. YU CU KIM NGHIM
1.1 Hnh thc cm quan: vin mu trng.
1.2. ng u khi lng: 5% so vi khi lng trung bnh ca thuc trong
vin.
1.3. r: thi gian r khng c qu 15 pht.
1.4. nh tnh: phi c phn ng ca sulfaguanidin.
1.5. nh lng: hm lng sulfaguanidin C7H10N4O2S t 95,0 - 105,0% so vi hm
lng ghi trn nhn.
2. PHNG PHP TH
2.1. Hnh thc: nhn xt bng cm quan, vin phi t yu cu nu.
2.2. r: thc hin theo chuyn lun thuc vin nn, ph lc 1.20 ca DVN IV
(khng thc hin).
2.3. ng u khi lng: thc hin theo chuyn lun Php th ng u
khi lng, ph lc 11.3, phng php 1, ca DVN IV.
Bng 11.3.1. Bng quy nh ng u khi lng cho ch phm n liu
Dng bo ch Khi lng trung bnh (KLTB) % chnh lch so vi KLTB
Vin nn Nh hn hoc bng 80 mg 10
Ln hn 80 mg v nh hn 250 mg 7,5
Bng hoc ln hn 250 mg 5
Cn ring bit 20 n v ly ngu nhin, tnh khi lng trung bnh. Khng c c
qu hai n v c khi lng nm ngoi gii hn chnh lch so vi khi lng trung
 28

bnh quy nh trong bng 11.3.1 (ph lc 11.3 DVN IV) v khng c c n v
no c khi lng vt gp i gii hn .
2.4. nh tnh
A. Ly mt lng bt vin tng ng vi khong 0,2 g sulfaguanidin, thm 5 ml dung
dch natri hydroxyd 10% (TT), un si, s c hi amoniac bay ln.
B. Ly mt lng bt vin tng ng vi khong 50 mg sulfaguanidin, thm 2 ml
dung dch acid hydrocloric 10% (TT), lc k, lc. Lm lnh dch lc trong nc ,
thm 4 ml dung dch natri nitrit 1% (TT), lc u. Ly 1 ml dung dch thu c, thm
5 ml dung dch 2-naphtol trong kim (TT) s xut hin ta thm.
C. Phng php sc k lp mng (Ph lc 5.4)
Bn mng: Silica gel F254
Dung mi khai trin: dicloromethan - methanol - acid formic khan (70 : 20 : 10)
Dung dch i chiu: ha tan 10 mg sulfaguanidin chun trong 5 ml aceton (TT).
Dung dch th: Ly mt lng bt vin tng ng vi 20 mg sulfaguanidin, thm
10 ml aceton (TT), lc k, lc, dng dch lc chm sc k.
Cch tin hnh: chm ring bit ln bn mng 10 l mi dung dch trn. Trin khai sc
k ti khi dung mi i c khong 15 cm, ly bn mng ra kh ngoi khng kh.
Quan st di nh sng t ngoi bc sng 254 nm.
Vt chnh trn sc k ca dung dch th phi ph hp v mu sc, kch thc v gi
tr Rf vi vt chnh trn sc k ca dung dch i chiu .
2.5. nh lng: Tin hnh chun bng nitrit (Ph lc 10.4 DVN IV).
Cn 20 vin, tnh khi lng trung bnh vin v nghin thnh bt mn. Cn chnh xc
mt lng bt vin tng ng vi khong 0,200 g sulfaguanidin, thm 15 ml dung
dch acid hydrocloric 25% (TT) v 50 ml nc. Lc k. Cho thm 2 g kali bromid
(TT), lm lnh trong nc ri chun bng dung dch natri nitrit 0,1 M (C) (phi
u nh dung dch chun ca buret ngp di mt dung dch cn chun ) .
Trong qu trnh chun phi lc bnh lin tc, nh nhng sao cho khng to ra dng
xoy khng kh trong dung dch (tt nht l dng my khuy t). Nh dung dch chun
vi tc lc u chng 2 ml trong mt pht, n trc im tng ng khong
1 ml th nh tng 0,1 ml mt v yn t nht mt pht sau mi ln thm dung dch.
im kt thc xc nh bng chun o in th t ng: thc hin theo SOP nh
lng Sulfaguanidin ca b mn HPTKN
 29

im kt thc xc nh bng ch th mu: thm 4 git ch th Tropeolin 00, 2 git xanh

methylene trc khi tin hnh chun bng dung dch NaNO2 0,1 M, khuy lin tc
trong sut qu trnh chun . Chun kt thc khi dung dch t bnh nh lng
chuyn t mu tm sang xanh. Thc hin song song vi mt mu trng trong cng
iu kin nh trn i chiu s thay i mu ca ch th.
1 ml dung dch natri nitrit 0,1 M (C) tng ng vi 21,42 mg C7H10N4O2S.
3. BO CO KT QU: theo phiu kim nghim
CU HI
1. Thit lp cng thc tnh kt qu nh lng trong 2 trng hp xc nh im tng
ng bng 2 phng php khc nhau. Nhn xt cch tnh v kt qu ca 2 phng
php chun ?
2. Trong 20 vin em tnh ng u v khi lng, c 2 vin nm ngoi khong
cho php - kt lun g i vi l thuc vin nn ny ?
3. Vit c ch phn ng nh lng sulfaguanidin ?
4. Mc ch ca vic v ng o hm bc 1 E/ V theo V (ml) ?
5. Vit cng thc khai trin ca sulfaguanidin?
6. in cc dng trong php chun th l g?
 30

BI 7

KIM NGHIM THUC SIR THRALENE

MC TIU
Thc hin nhng yu cu k thut c hc trong phn l thuyt ca bi KIM
NGHIM THUC DNG SIR.
NI DUNG
1. YU CU KIM NGHIM
1.1. Hnh thc cm quan: cht lng c st, trong, mu nu nht, c nh cam, mi v
v c bit
1.2. Th tch: Chai 90 ml +10% so vi th tch trn nhn (90 -99ml). Chai 125ml +
8% so vi th tch trn nhn (125 135 ml).
1.3. T trng: 1,250 1,300 (200C). (ph lc 6.5 trang PL 148)
1.4. pH: 2,3 -5,5 (ph lc 6.2 DVN IV, trang PL 144.)
1.5. nh lng: sir Thralene phi cha trong 100ml sn phm 46,5 53,5mg
alimemazin baz (C18H22N2S) 0,090 0,110mg acid ascorbic (C6H8O6) .
2. PHNG PHP TH
2.1. Hnh thc cm quan: ch phm phi t yu cu nu trn
2.2. Chnh lch th tch: cho ton b th tch sir cha trong chai vo mt ng lng
c th tch tng ng. Chnh lch th tch cho php c c theo bng sau:
Bng 9.1. Bng ghi gii hn cho php v th tch sir
Loi thuc Th tch ghi trn nhn Gii hn cho php (% chnh lch)
Siro v cao thuc Ti 100 ml +10%
(tr cao ng vt) Trn 100 ml 250 ml +8%
Trn 250 ml +6%

3.3. T trng
o 200C bng picnomet. Cn chnh xc picnomet rng, kh v sch. vo
picnomet mu th chnh nhit thp hn 20 0C, ch ng bt kh. Gi
picnomet 20 0C trong khong 30 pht, dng giy lc thm ht cht lng tha trn
vch mc, lm kh mt ngoi ca picnomet. Cn ri tnh khi lng cht lng cha
trong picnomet. Tip , mu th i, ra sch picnomet, lm kh bng cch trng
EtOH, ri trng aceton, sau thi khng kh nn hoc khng kh nng ui ht hi
 31

aceton ri xc nh khi lng nc cha trong picnomet nhit 20 0C nh trn. T

s gia khi lng mu th v khi lng nc ct thu c l t trng D20 cn xc
nh (Ph lc 6.5 DVN PL 148).
2.4. pH: ly 10ml sir cho vo cc c m 50 ml v thc hin theo ph lc 6.2 DVN,
trang PL 144
2.5. nh tnh
2.5.1. nh tnh alimemazin tartrat
Pha ch dung dch th T: ly 10 ml ch phm cho vo bnh lng, thm vo bnh ny 10
ml dung dch acid hydroclorid 2N, thm 20 ml cloroform, lc u. Chuyn lp
cloroform qua mt bnh lng th 2 cha 5 ml dung dch NaOH 1N, lc mnh xoay
vng trnh to nh, yn. Lc phn cloroform qua gn ri chia lm 2 phn, cho
vo 2 bt s nh. bay hi n cn, lm cc phn ng nh tnh sau:
a. Nh t t vo cn ca chn I 3-4 git acid suluric m c, s xut hin mu
chuyn nhanh sang vng .
b. Nh t t vo cn ca chn II 3-4 git acid sulfuric m c, s xut hin mu
bn vng.
2.5.2. nh tnh acid ascorbic
Ly 5 ml ch phm cho vo mt cc c m 50 ml, thm 4 ml nc, lc u, thm tng
git dung dch dicloro-2,6 phenol indophenol 0,05% trong nc: thuc th mt mu
ngay. Thng th phi iu chnh n pH=4 bng dung dch NaOH 0,1N).
2.6. nh lng
Phng php quang ph UV-Vis
Ly chnh xc 1 th tch siro Thralne tng ng vi khong 1 mg alimemazin base
cho vo bnh lng 125 ml, thm 50 ml nc v 10 ml NaOH 2,5 N, trn ln v chit
vi cyclohexan 3 ln , mi ln 10 ml. Ra ton b dch chit cyclohexan thu c vi
10 ml nc ct, loi b nc ra.
Chit xut dch cyclohexan vi acid hydrocloric 0,1 N 3 ln, mi ln 20 ml. Tp hp
ton b dch chit acid cho vo bnh nh mc 100 ml, thm acid hydrocloric 0,1 N
n vch. Ht chnh xc 25 ml (hoc 50 ml) dch t bnh ny v cho vo bnh nh
mc 50 ml (hoc 100 ml), thm dung dch acid hydrocloric 0,1 N va n vch, lc
u.
Xc nh hp thu bc sng 251 nm, 276 nm ca dung dch cn o cng lc vi
 32

dung dch chun alimemazin tartrat chun trong cng mt mi trng c nng c
bit khong 5 mcg/ml. Dng acid hydrocloric 0,1 N lm mu trng.
Tnh ton hm lng C18H22N2S (mg) trong 100 ml siro theo cng thc:
( At 251 At 276
0,005 50 2 102
Ac 251 Ac 276

At251; At276: hp thu ca mu th

Ac251; Ac276: hp thu ca mu chun
3. BO CO KT QU: theo phiu kim nghim.
CU HI
1.Vit cng thc ca hot cht chnh c trong sir Theralene 2 dng?
2. Trong khi thc hin phn nh tnh, giai on lc CHCl3 vi NaOH hay gp hin
tng to nh, lm cch no ph nh? C bao nhiu cch? gii thch?
3. Nhn xt g v quy trnh nh tnh v quy trnh nh lng. C mu thun nhau
khng? T rt ra kt lun g?
4. Ti sao trong quy trnh nh lng khng dng dung mi chit l cloroform m
dng cyclohexan? Gii thch?
5. Ti sao trong quy trnh nh lng dng 2 bc sng, dng mt bcc sng c c
khng?
6. Gii thch cng thc tnh kt qu?
 33

Bi 8

KIM NGHIM VIN NN ASPIRIN

(TC DVN IV)
MC TIU
1. Thc hin nhng yu cu k thut c hc trong bi Kim nghim vin nn
2. Thc hin phng php chun acid base.
NI DUNG
1. YU CU K THUT
1.1. Hnh thc: Vin mu trng, mt vin nhn, cnh v thnh vin nguyn vn, 1 mt
c ch aspirin, 1 mt c rnh gia
1.2. ng u khi lng: 5% so vi khi lng trung bnh vin.
1.3. nh tnh: phi c phn ng ca acid acetylsalicylic.
1.4. nh lng: hm lng acid acetylsalicylic C9H8O4 phi cha t 90,0% n
110,0% hm lng ghi trn nhn.
2. PHNG PHP TH
2.1. Hnh thc:
Nhn xt bng cm quan, vin phi t yu cu nu.
2.2. ng u khi lng (ph lc 11.3)
Cn 20 vin, tnh khi lng trung bnh vin (KLTB).
Cn ring tng vin, so snh vi KLTB vin.
Khng c c qu 02 n v c khi lng nm ngoi gii hn chnh lch so vi
KLTB qui nh v khng c c n v no c khi lng vt gp i gii hn .
2.3. nh tnh
un si khong 0,3g bt vin trong 2-3 pht vi 10 ml dung dch NaOH 10% (TT).
ngui, thm dung dch H2SO4 10% (TT) cho n khi tha acid, s c ta kt tinh v
mi ca acid acetic. Lc ly ta, ha tan ta trong vi ml nc, thm 2 git dung dch
FeCl3 0,5% (TT) s c mu tm.
2.4. nh lng
Cn 20 vin, loi b lp bao, xc nh khi lng trung bnh vin nhn, nghin thnh
bt mn.
Cn chnh xc mt lng bt vin tng ng vi 0,25g acid acetylsalicylic, thm
chnh xc 15 ml dung dch NaOH 0,5N (C), un si nh trong 10 pht, ri chun
 34

lng NaOH tha bng dung dch HCl 0,5N (C), dng dung dch phenol (TT)
lm ch th.
Song song lm mu trng.
1 ml dung dch NaOH 0,5N (C) tng ng vi 45,04 mg C9H8O4.
Hm lng % acid acetylsalicylic so vi lng ghi trn nhn tnh theo KLTB vin tnh
theo cng thc:

(Vtrang Vthu ).K HCl .45,04 100

X (%) .mTBV .
mthu 81
Vi: K : h s hiu chun ca dung dch HCl 0,5N
Vtrng : th tch dung dch HCl 0,5N dng cho mu trng (ml)
Vth : th tch dung dch HCl 0,5N dng cho mu th (ml)
mth : khi lng mu cn (g)
mTBV : khi lng trung bnh vin (g)
3. BO CO KT QU
Theo phiu kim nghim v phiu phn tch. Tr li cu hi.
CU HI
1. Gii thch c ch nh tnh acid acetylsalicylic. Vit phng trnh phn ng.
2. Gii thch c ch nh lng acid acetylsalicylic. Vit phng trnh phn ng.
3. Ti sao HCl li c h s K? Nu pha t ha cht gc c h s K khng?
4. Nu nhng phng php khc nh lng acid acetylsalicylic.
5. V cng thc cu to ca aspirin. Tn IUPAC ca cht ny l g?
6. K tn mt s thuc c cha hot cht l aspirin.
7. K thut chun trong bi l g? (trc tip/ tha tr/ th). Trnh by nguyn tc
ca k thut .
8. Ti sao khi nh lng aspirin phi cho chnh xc 15ml dung dch NaOH 0,5N?
9. Chng minh cng thc tnh hm lng % ca aspirin. Ti sao khi nh lng phi
loi b lp bao?
 35

Bi 9

KIM NGHIM THUC VIN HON

Vin hon mt ong ngh
MC TIU
1. Trnh by c cc ch tiu khi kim nghim thuc vin hon
2. Thc hin c phng php th cc ch tiu trong kim nghim thuc vin hon
ngh
NI DUNG
1. YU CU K THUT
1.1. Tnh cht: Vin hon phi trn, u, ng nht v hnh dng, mu sc khi bo
qun, c mi c trng ca dc liu.
1.2. ng u khi lng: Quy nh theo bng sau:

Khi lng trung bnh ca mt hon Gii hn sai s cho php

T 0,05g n 1,5g 12%
Trn 1,5g n 5,0g 10%
Trn 5,0g n 9,0g 7%
Trn 9,0g 5%

1.3. r: Khng qu 1 gi
1.4. nh tnh: Ch phm phi cho phn ng c trng ca ngh
2. PHNG PHP TH
2.1. Tnh cht:
Bng cm quan ch phm phi t yu cu nu: hon phi trn u, ng nht v
hnh dng, mu sc, c mi c trng ca dc liu.
2.2. ng u khi lng:
Cn 10 vin, xc nh khi lng trung bnh. Cn ring r tng vin v so snh khi
lng tng vin vi khong chnh lch so vi khi lng trung bnh. Trong khng
c c qu 2 vin vt qu gii hn cho php v khng c vin no gp i gii hn
cho php.
2.4. r: p dng cho vin hon cng
- Tin hnh th trn 6 vin hon trn my th r
- Nhit th: 360C 10C
 36

- Mi trng th: nc, nu khng t th li ln 2 trong mi trng HCl 0,1N

- Thi gian r ca vin: khng qu 1 gi.
2.5. nh tnh: Ngh
Nghin 5 vin hon thnh bt, cho vo cc, thm 20ml ethanol 96%, lc qua giy lc.
Nh 3-4 git dch chit ethanol ln giy lc. kh, trn giy lc cn li vt mu
vng. Tip tc nh tng git dung dch acid boric 5% (TT) ri dung dch acid
hydrochloric long (TT), h nng nh cho kh, vt vng s chuyn thnh mu . Sau
thm 3 git dung dch aminiac (TT), s tip tc chuyn sang mu xanh en.
 37

Ti liu tham kho

1. ng Vn Ha, Vnh nh (2014), Kim Nghim thuc, Nh xut bn Gio dc

Vit Nam.
2. B Y T (2009), Dc in Vit Nam IV, Nh xut bn Y hc, H Ni
3. Gio trnh Thc hnh Kim nghim thuc (2014), B mn HPT-KN, i Hc Y
Dc Cn Th.