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Trng i hc Cng nghip thnh ph H Ch Minh

Khoa Cng ngh ha hc
Chuyn ngnh Ha phn tch
Xc nh Vitamin B1 trong thc phm
bng phng php hunh quang phn t

GVHD : TS. L nh V
SVTH : Phm Bo Qu
MSSV : 11050991

NI DUNG TRNH BY

Tm hiu v Vitamin B1
Quy trnh xc nh Vitamin B1

1.Tm hiu v Vitamin B1

vitamin B1( thiamine)

2-[3-[(4-amino-2-methyl- pyrimidin-5-yl)methyl]- 4-methylthiazol-5-yl] ethanol
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- Cu trc ha hc:
+ Thiamin gm dn cht pyrimidin gn vi
dn cht thiazol qua nhm methylen.
+ Ngoi ra thiamin cn tn ti di dng
photphat este gm thiaminmonophotphat (TMP),
thiamin triphotphat (TTP), thiamin pyrophotphat
(TPP) l dng m v tr OH s c nh vo
hoc 1, 2 hoc 3 gc photphat (R)

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- Tnh cht ha hc:

+Thiamin b oxy ha bi K3[Fe(CN)6]
trong mi trng kim to thnh thiocrom mu
vng pht hunh quang mu xanh da tri, tnh
cht ny c dng xc nh Vitamin B1
bng phng php o hunh quang.
+ Thiamin to ta khi phn ng vi tannin,
thy ngn (II) clorua, axit picric.
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b. Chc nng sinh hc:

- ng ha ng
- Nhn t ngon ming, kch thch cm gic thm n.
- Tham ra iu ha qu trnh dn truyn xung thn
kinh, kch thch cc hot ng tr no.
-Trong c th, vitamin B1 tham gia vo nhiu phn
ng chuyn ha, c bit l chuyn ha glucid.
- Thiu ht vitamin B1 s dn n bnh beriberi
(bnh t ph)
- Nhu cu vitamin B1: thay i ty theo la tui,
ngh nghip, trng thi sinh l ca c th,

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2. Quy trnh xc nh Vitamin B1 trong sa

bng phng php hunh quang
2.1. Nguyn tc xc nh: Chit Vitamin B1 bng dung
dch HCl trn bp cch thy, sau Vitamin B1 b oxy
ha bi Kaliferricyanur mi trng kim to thnh
Thyocrom c hunh quang xanh da tri. Chit
Thyocrom bng isobutanol sau o hunh quang
bc sng kch thch 365nm v bc sng pht hunh
quang 425nm
2.2. Phm vi p dng : Phng php ny p dng cho
xc nh Vitamin B1 trong mu thc phm.

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2.3. Ho cht
- Natri axetat (TK.PT) dung dch 2N: Ho tan 15g natriaxetat
ngm nc vo nc ct v thm nc ct va 1 lt.
- Natri hydroxit (TK.PT) dung dch 15%: Ho tan 15g natri
hydroxit trong nc v thm nc ct va 100 ml.
- Kali ferixyanua (TK.PT) dung dch 1%: Ho tan 1g kali
ferixyanua trong nc ct v thm nc ct va 100 ml. Dung
dch pha ch dng trong ngy.
- Cht oxy ho: Hn hp 4 ml dung dch kali ferixyanua 1% vi
dung dch natri hydroxit 15% n va 100 ml. Dung dch pha
dng trong 4 gi.
- Isobutanol (TK.PT)
- Vitamin B1 (TK.PT) dung dch chun: Cn khong 30-40mg vi
chnh xc 0,0001 g thiamin clohydrat ( lm kh trong bnh
ht m) ho tan trong bnh nh mc 1 lt bng dung dch cn
etylic 20% axit ho bng axit clohydric n pH-3,5-4. Bo
qun nhit 100C trnh nh sng.
- Kali clorua hay natri clorua; TK.HH;
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2.4. Dng c v thit b:

- Cn phn tch, cn k thut;
- T sy;
- Bnh nh mc, dung tch 1 lt, 250 ml;
- Cc c m, dung tch 250 ml;
- ng ong chia , dung tch 5 ml, 100 ml;
- Ni cch thu;
- Phu lc thu tinh, dung tch 250 ml;
- ng nghim nt mi, dung tch 40-50ml;
- Pipet c bu, dung tch 1ml, 5ml;
- My nh nghin hoc ci chy s;
- My lc;
- pH met;
- Hunh quang k.

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2.5. Quy trnh thc hin

2.5.1. Chit vitamin B1 ra khi mu lng thc:
Cn chnh xc mt lng mu nghin nh trn u c
tnh cha khong 20-50mg thiamin cho vo cc c m dung
tch 250ml. Tin hnh song song vi dung dch chun bng
cch ht chnh xc 1ml dung dch chun Vitamin B1 cho vo
cc khc. Thm vo cc cc, mi cc 100ml dung dch HCl
0,1N (pH=1-1,5), y np, un cch thu si 45 pht hoc
un trong ni p sut (t0=110-1200C) trong 35 pht (k t
khi t nhit ). ngui n nhit phng, iu chnh
pH cc cc bng dung dch natri axetat 2N n pH = 4 - 4,5.
Chuyn ton b dch mi cc sau khi iu chnh pH vo
bnh nh mc 250ml v thm nc ct n vch. Lc dung
dch bng giy lc khng hp ph thiamin (hoc giy lc
khng tro) b vi mililit u.

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2.5.2. Oxy ho thiamin thnh thiocrm:

Vi mi dung dch mu nh lng, k c dung dch chun,
c tin hnh song song trn 4 ng nghim nt mi dung tch
40-50ml. Trong mi ng cha sn 3g kali clorua. Ht chnh xc
5ml mi dch lc vo tng ng nghim trong 4 ng, hai ng c
coi l ng trng, hai ng c coi l ng th ca tng mu.
2.5.2.1. i vi ng trng ca cc mu c tin hnh nh sau:
Lc nh ng nghim bng my lc thm ngay 3ml dung dch natri
hydroxit 15%, lc trn cho u, thm nhanh 15ml isobutanol, y
ng, lc mnh trong 15 giy, lng.
2.5.2.2. i vi cc ng th cn li ca cc mu c tin hnh
nh sau:
Lc ng nghim bng my lc, thm ngay 3ml dung dch cht oxy
ho (dung dch kim kaliferixyanua), lc trn u, thm nhanh
15ml isobutanol, y ng, lc mnh trong 15 giy, lng.
Cc ng nghim trn lng cho tch lp trong bng ti trong
vng 20 pht. Sau ht hoc gn ly khong 10ml dung dch
isobutanol c cha hunh quang bn trn.

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2.5.3. o hunh quang:

o mt hunh quang ca dung dch isobutanol bng
my o hunh quang ti bc sng kch thch v pht x l
360 nm v 435 nm (i vi hunh quang k Coleman th dng
2 knh lc s 12.221 v 14.221 tng ng vi 2 bc sng
trn).

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2.6. Tnh ton kt qu

Hm lng Vitamin B1 (X) tnh bng microgam trong 100g
lng thc c tnh theo cng thc sau:

Trong :
Cc: Hm lng Vitamin B1 trong mu chun (mg)
Dct: Mt hunh quang ng chun th
Dctr: Mt hunh quang ng chun trng
Dmt: Mt hunh quang ng mu th
Dmtr: Mt hunh quang ng mu trng
P: Khi lng mu cn (g)

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