Định lượng vitamin A.pptx

8/11/2019 nh lng vitamin A.pptx

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Trng i hc Nng Lm TP H Ch Minh Mn: Phng php phn tch thnh phn ha-l

thc phm ----oOo-----

ti:

Nhm thc hin: Nguyn Th Dim My Nguyn Th Kim OanhPhm Th Kim Chi

Nguyn M Xun Nguyn Th Thanh Thy V Th Trm

GVGD: TRN V HUY


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Mc lc I. Gii thiu

1. Cu to, tnh cht v chc nng ca vitaminA2. Mt s phng php phn tch Vitamin Atrong thc phm

II. Tm hiu mt s phng php phn tchvitamin A1.Phng php so mu2.Phng php sc k lng hiu nng cao3.Phng php quang ph 4.So snh cc phng php

III. Cc hng sn xut my phn tch


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Vitamin A cn c tn gi khc l retinol , axerophthol Tn ti trong t nhin gm 2 dng:

Retinol : dng hot ng ca vitamin A, n ng hatrc tip bi c th.Tin vitamin A: l mt tin cht ca vitamin A c bit nhiu di dng beta-carotene c chuyn hathnh vitamin A m c th c th s dng c.

Ngoi ra cn c anpha, gama caroten

I. Gii Thiu 1. Cu to, tnh cht v chc nng ca vitamin A


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a. Cu to


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Vitamin A d b oxi ha.Trong c th di tc dng enzyme, retanol chuyn thnh anehyt retanal.Trong gan tn ti dng este vi acid acetic v

palmitic.B thy phn bi khng kh, kh bn vi acid, kim

v nhit

b. Tnh cht


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Vitamin A c nhiu chc nng quan trng i vi c th con ngi:Th gic: gip cho t bo trong ca mt lun khe mnh.

Quan trng i vi s bin i nh sng thnh cc tnhiu thn kinh trong vng mc.

Cc m: Vitamin A kch thch qu trnh pht trin ca cc biu m nh sng, rut v cc con ng h hp.

c. Chc nng


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nh hng c bit n da, kch thch s lin so v

phng nga cc chng bnh ca da nh trng c.S sinh trng: vitamin A l yu t khng th thiu cho s pht trin phi thai v tr em.C vai tr i vi s pht trin xng, thiu vitamin

A => xng mm v mnh, qu trnh vi ha b ri lon H thng min dch: do hot ng c hiu ca cc t

bo vitamin A tham gia tch cc vo s chng chu bnh tt.


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Chng lo ha: vitamin A ko di qu trnhlo ha do lm ngn chn s pht trin cc gc

t do.Chng ung th: hot ng km hm ca nvi cc gc t do cng dn n ngn chn

c mt s bnh ung th.Sinh sn: vitamin A nh hng ln chc nng v s pht trin ca tinh trng, bung trng vnhau thai=> cn thit cho chc nng sinh sn


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d. Cc ngun thc phm giu vitamin A


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Phng php sc k lng hiu cao nng Phng php sc k khPhng php nh tnhPhng php chun IodPhng php o quang trc tip v sau trit tchvitamin A.

Phng php sc k lng Phng php so mu

2. Mt s phng php phn tch Vitamin Atrong thc phm


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1. Phng php so muTiu chun ny qui nh phng php so mu xc nh vitamin A trong sa bt giy cha t nht 10 IU ( n v quc t) vitamin A trn gam.

II. Tm hiu mt s phng php phn

tch vitamin A


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a. Nguyn tc. Mu th c x phng ha v c chit,

cht khng th x phng ha c th cho phn ng vi axit trifloaxetic o hp th bc song 620 nm.

TCVN 7081 1 : 2010


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b. Thuc th Ch s dng thuc th t cht lng tinh khit phn tch, tr khi c qui nh khc v ch s dng nc ct hoc nc kh hoc nc c tinh khit tng ng. Gm cc loi thuc th sau:


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Etanol (CH3CH2OH) 95% (tnh theo th tch)khng cha aldehyt.Dung dch Natri ascocbat 200 g/lDung dch Kali hydroxit (KOH) 50%Dung dch nc cn Kali hydroxit 30 g/lDu nh Clorofom ( CHCl3)Axit trifluoroaxetic (CF3COOH)Thuc th muDung dch Vitamin A: s dng vitamin A theotiu chun US PharmacopeiaHydroxytoluen butylat ha


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c. Thit b dng c S dng cc thit b phng th nghim thong thng v c bit nh sau:

My so mu quang in hay my quang ph ( c cu quang hc hoc b lc hot ng bc song 620 nm)

Cc c m hoc bnh nnBnh x phng haBnh nh mc Pipet 1 vch Pipet t ng

Bn hi, bp cch thy hoc bp cch kh dung in Ni cch thy: c th duy tr nhit 40 0CPhiu chit: dung tch 50 mlB siu mGiy lc: ng knh 9 cm


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My quang ph My so mu quang in


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bnh nn Cc c m


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Pipet t ng Bnh nh mc

Phiu chit B siu m


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d. Ly mu Ly mu theo TCVN 6400: 1998 (ISO 707)

Tnh t l Am/An i vi tng bc song ni trn. Nu t l khng vt qu cc gi tr 0,593 bc sng 300 nm, 0,537 bc sng 370 nmtng ng th tnh hm lng Vitamin A, W, theon v quc t trn gam.


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Bng cng thc: W=(A m .V.d)/(10.m)

Trong :Am: l gi tr s hc ca hp th cc i thu c bc sng 326 nm.V: l tng th tch Vitamin A dng este phalong c nng t 10 IU/ml n 15 IU/ml.d: l h s chuyn i t hp th c th ca Vitamin A dng este thnh n v quc t trn

gam (d=1900).


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2. Xc nh Vitamin A trong thc phm bng phng php sc k lng hiu nng cao

a .Chun b dng c - H thng my HPLC, Shimadzu- Ct Purospher Star RP-18e( 150x4.6mm,5um)- Cn phn tch c chnh xc 0.1mg- B chng ct c ng sinhhn hi lu - Bnh nh mc 10ml- Pipet cc loi : 0.1ml, 0.5ml,1ml, 2ml, 5ml,

10ml,...- ng Hatch- Dng c thy tinh cc loi: Becher, erlen.

b Ch b h


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b.Chun b ha cht Ha cht phi l loi tinh khit c s dng

phn tch , gm:- Nc ct loi dng cho HPLC- Dietyl ete tinh khit khng c Peroxit

- Etanol- Dung dch Kalihydroxyde (KOH) 2M trong cn

: ha tan 11.2g KOH trong 100ml cn 96o

- Dung dch Kalihydroxyde (KOH) 2% trongnc : Ha tan 2g KOH trong 100ml nc ct - Acetonnitrile, HPLC

- Na 2SO 4 khan.


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c.Cch tin hnh- Xy dng ng chun (dng Vitamin A

palmitat ): Nu dng chun vitamin A th phn tch ngay

trn thit b HPLC. Nu dng vitamin A di dng este, th d

nh vitamin A acetat, vitamin A palmitat,...th phi tin hnh x phng ha gii phngvitamin A ra th t do ri mi tin hnh phntch bng HPLC.


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- Qu trnh x phng ha Cn 18.32 mg vitamin A palmitat tng ng

vi 10mg vitamin A cho vo bnh cu c c nhm, thm 20-40ml dung dch KOH 2Mtrong etanol v 0.5natri ascorbat.

Lp ng sinh hn hi lu . t trn ni cch thy, x phng ha nhit 70 0C trong 30 pht.

Chuyn dung dch cn nng vo mt bnhlng, ra bnh cu nhiu ln bng H2O ct vtp trung nc ra vo bnh lng.


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Ch : Lng nc khng c qu 3 ln lng KOH 2M. Vitamin A t dung dch x phng ha c chit 3 ln bng dietyl ete khng c peroxit 3 ln, mi ln 40ml. Tp trung lp ete chit c vomt bnh lng. Ra lp ete nhiu ln vi nc ct (mi ln 50ml) cho n khi khngcn vt kim (th vi phenolphtalein). Loi nc trong dch chit etebng cch lc qualp Na 2SO 4 khan, ra lp Na 2SO 4 ba ln,mi ln vi 10ml ete.


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Gp tt c lp ete vo bnh qu l.C quayn cn. Cn cn li ha tan trongacetonitrile, chuyn sang bnh nh mc 10ml. nh mc n vch bng acetonitrile. Dung dch ny dng lmdung dch vitamin A chun gc (1000mg/l).

Dung dch chun 100ug/ml: ht chnh xc1ml dung dch vitamin A chun gc 1000ug/ml vo bnh nh mc 10ml. nh mc ti vch bng ACN.


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d.Chun b mu th - Cn khong 10g mu th c nghin nh (

nu cn) cho vo bnh cu thu tinh c c nhmv tin hnh x phng ho nh ghi phn trn.

- Sau khi ra dch chit ete bng 15ml KOH 2% vnhiu ln vi nc ct ( mi ln 50ml) cho n khi khng cn vt kim ( th vi phenoltalein) .Loi nc trong dung dch chit ete bng cch lc qua lp Na 2SO 4 khan , ra lp Na 2SO 4 ba ln,mi ln vi 10ml ete.

- Gp tt c lp ete vo bnh qu l. C quay n cn. Ho tan phn cn li bng chnh xc 1mldung dch pha ng ( vortex 15giy v siu m 3

pht). Lc dch c qua mng lc 13mm, 0.45umv thu dch lc v tin hnh phn tch trn HPLC.


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e.Tin hnh phn tch trn HPLCiu kin phn tch- Pha ng - Tc dng- Detector UV, = 325nmtim cc dung dch mu th nghim

theo th t sau:+ Cc dch chun . Dng ng chun gia nng cc

chun vitamin A v din tch ca cc chun Vitamin A. Tnhton h s tng quan hi quy tuyn tnh.+ Dung dch mu trng + Dung dch mu kim sot. tnh hm lng cc vitamin A

trong dch chit. v trong mu thng qua ng chun xydng bc trn .Tnh thu hi mu kim sot (R).+ Cc dung dch mu th nghim. tnh hm lng vitamin Atrong dch chit thng qua ng chun xy dng bc trn.


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Tnh kt qu Hm lng vitamin A c trong mu c tnh

theo cng thc sau:C = Co/m

Trong :- C l hm lng Vitamin A c trong mu Tnhtheo mg/Kg

- Co l hm lng Vatamin A trong dch chit.(Co) thng qua ng chun, mg/L- m : Khi lng (g)


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3. Phn tch xc nh hm lng vitamin A trongnguyn liu sn xut thuc th y bng phng

php o quang ph ca dung dch.


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-Lng mu: 0.1g nguyn liu c nghin mn.-Ha cht: . ethanol

. KOH khan

. dietyl ether

. Na2SO4

. iso-propanol

. Nc ct - Mu trng: iso-propanol


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Dng c


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Quy trnh:

- Cn 0,1g nguyn liu c nghin mn cho vo bnh nn 250ml; cho 40ml ethanol tuyt i vo;thm 2g KOH khan; gn vo ng sinh hn un cchthy 30 pht; ngui trng ng sinh hn bng

30ml nc ct; sau chuyn vo phu chit; chit 4 ln; mi ln bng 30ml dietyl ether ( ly phn dungdch trn; phn dung dch di em chit tip).


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- Dung dch gi li thm nc ct, lc, chit tip ng thi o pH dung dch t n 7; thu dung dch vo bnh nh mc 100ml; nh mc bng dietylether va 100ml; ly 50ml cho vo bnh nh mc 50ml c cha 2g Na 2SO 4 lc u, lng ht1ml cho vo bnh nh mc 50ml, nh mc bng iso-propanol o 3 bc sng 310, 320, 325 vi mu trng l iso-propanol.


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4 S h h h


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So mu HPLC o quang ph u im Dng c thng

thng ca phngth nghim

nh tnh v nh lng

Kt qu nhanh chnhxc.Dnh lng a s ion vc v hp cht hu c nh phn ng to muphc hp.

C th phn tch cu t vi lng ( nhy cao)

Nhc im Thuc th tinh khit Nc ct ( hoc qua kh trng)

Pha tnh thng s dng silosan hoc alumino c th gyc .Ha cht tinh khit Dung mi ha tantt vi cu t cn

phn tch

Thit b t tin.Ngi s dng phi ctrnh cao.

4. So snh cc phng php


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III. Cc hng sn xut my phn tch

Hin nay c rt nhiu loi my phn tchthc phm nhng chng ta s gii thiu s lc mt s loi my sau y :

1 My phn tch kch thcht


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1.My phn tch kch thc ht a. Tn thit b : my phn tch kch thc ht

b. K m hiu : SALD-2101c. Hng sn xut :Shimadzu- Nht Bn

a. Cc thng s k thut ch yu l :


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g s t utc yuMy chnh phn tch c ht bng laze

Ngun n laze bn dn c tui th trung bnh cao D dng kim tra hiu nng nh chc nng ca phn cng n nh v chnh xc quang hc tt an ton cao: chm tia laze v ngun cp t ng tt

c tnh k thut phn cng Phng php o: Nhiu x laze

Khong o: 0,03 1000 mm n v o: Ngun laze bn dn (vch o 680nm)

Cm bin quang hc: 81 nguyn t Ngun p ng: 100, 110, 120, 200, 220, hoc 240 VAC Kch thc: di rng cao: 46x75x28cm. Nng 51kg

Khong nhit : 10 30 0C m: 20 80%

n v mu T bo t ng

e. Nguyn l hotng: S tng tc ca nng lng ca tia laz


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e. Nguyn l hot ng : S tng tc ca nng lng ca tia lazvt cht c phn tn trong dung dch trong bnh khuy s thay i tnh cht nhiu x ca chm tia . Theo di s thayxc nh kch thc ca vt liu cn o c ht

f. Yu cu c bn i vi mu o : Mu bt, mu rn phi c b sch, khng bay hi, bn. Cc mu phi khng b phn hy h ph v cu trc bi tia laze.

g. Phm vi ng dng v gii hn :* Phm vi ng dng : Cng nghip vt liu, ngnh a cht

nng nghip v thc phm, ngnh Ho hc, nng lng htv in lc, Y hc, Dc v Sinh hc, Phn tch mi trn

* Gii hn : Xc nh c kch thc vt liu rn hoc h ph. Khng xc nh c cc phn t cht lng v cht kh

h. Nm a vo s dng : 2006 k. nh chp ton b ca thit b

h


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2. My quang ph UV-VisTn thit b : thit b o trc quang UV-Visa. K m hiu : UV-vis Spectrophotometer 2800

b. Hng sn xut : Cole Palmer Instruments - Trung Quc.


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c. Cc thng s k thut ch yu:+ Di bc sng 200 1100 nm+ chnh xc bc sng: 1 nm

+ lp li bc sng: 0,1 nm+ phn gii: 0,1 nm+ Ngun n: Tungsten v Deuterium+ Vng gi tr o quang: ABS = 0 5+Ch : o im v qut ph + Cuvet: 1 cm, 2 cm, 5 cm.+o theo s bin i nhit : T phng n 60 0C+ Kt ni my tnh v phn mm o x l

d. Nguyn l hot ng : Thit b hot ng da trn kh nng hp th nh sng ca mu

f. Cc thng s ca mu m thit b c th c c :+ Mu lng

+ Mu rn c phn tn trong dung dch

k


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g. Cc kt qu chnh :+ Xc nh nng cht tan trong dung dch + Xc nh nh hp th cc i ca mu dng dung

dch + Xc nh nng lng vng cm (Eg)

h. Yu cu c bn i vi mu o :+ Mu dng lng : khng cha cn l lng, bc sng cn

o khng chu nh hng bi tp cht.

+ Mu dng rn, kch thc ht nanomt ( hoc khng b lng ng khi phn tn trong pha lng)i. Lnh vc p dng v gii hn ca thit b :

+ Phn tch nh lng cht tan+ trong dung dch + Xc nh Eg ca cc cht rn + Nghin cu hp ph cc cht Chuyn ha enzym

j. Nm s dng : 2009


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Định lượng vitamin A.pptx

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