KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

MT S PHNG PHP TA PROTEIN

I. Phng php ta bng cc tc nhn gy ta l dung mi hu c (Ethanol 80%,

Aceton)
ha tan ca protein trong dung dch ph thuc vo nhiu yu t, mt trong s
l hng s in mi ca dung dch. Nhn chung, nhng phn t dung mi c hng s in
mi ln (nh nc, dimethylsulphoxid) c th n nh tng tc gia chng vi cc phn t
protein v to iu kin thun li cho s ha tan ca protein trong dung dch. Ngc li, cc
dung mi vi hng s in mi nh (aceton, ethanol) ngn cn s phn tn ca cc phn
t protein trong mi trng. Do , ha tan ca nhng phn t protein gim v xy ra kt
ta do s lm gim hng s in mi hin hu ca mi trng. iu ny c c bng cch
thm mt dung mi ha tan trong nc nh aceton vo dung dch cha protein. S ta bng
aceton hoc ethanol 80% c nhiu thun li hn v n tng i r, c sn dng tinh khit
vi t cht tp nhim gy c hay c ch i vi enzym, do nhit bay hi ca dung mi
thp nn d tch b dung mi khi ch phm enzym bng phng php sy nh bng qut
gi.
II. Phng php ta bng mui
Ngi ta c th dng mui (NH4)2SO4, NaCl ta protein v cc mui ny va
lm trung ha in (do cc ion tc ng tng h vi cc nhm tch in tri du), va loi
b lp v hydrat ca phn t protein. Cc protein khc nhau c th b kt ta vi nng
mui khc nhau, v vy c th dng mui tch ring cc protein ra khi hn hp ca
chng.
Phn th nghim
1. Dng c v ha cht
Dng c
Ha cht
Pipet 5ml, 10ml
Nguyn liu thm (da)
Becher 50ml, 100ml, 250ml
Etanol 96%
Bnh tia
Mui (NH4)2SO4
My ly tm
2. Thc hnh
Cn khong 80g n 100g thm, gi nt vt ly nc ri ly tm 6000 vng/pht
trong 15 pht ri ong xem th tch ca nc thm l bao nhiu, ghi nhn th tch V. Sau
chia i th tch V nc thm va thu c thnh hai phn bng nhau ri tin hnh ta
protein bng hai tc nhn ta khc nhau: Ethanol 80% v mui (NH4)2SO4

KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

Quy trnh kt ta enzym Bromelin bng cn 80%

Chi ngn, v qu
Lm sch

Nghin p, xay nhuyn

Lc

Ly tm 6000 v/p, trong

15 pht
cn 80%, lm
lnh 40C

Dch trong

Ta cn 4oC / 2 gi

Ly tm 6000 v/p, trong

20 pht

Thu ta

B
dch
trong

Sy kh

Bromelin
Lu : Phi trn cn vi dch da theo t l 4:1 (1 da : 4 cn)

KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

Quy trnh thu nhn enzym Bromelin bng mui (NH4)2SO4

Chi ngn, v qu
Lm sch
Nghin p, xay nhuyn

Lc

Ly tm 6000 v/p, 15
20pht
Ammonium
sulfate 70%
bo ha

Dch trong

Hn hp dch da,
nhit phng 30 pht
Ly tm 6000 v/p, trong
15 pht
Thu ta

B
dch
trong

Sy kh

Bromelin
Lu : c mui(NH4)2SO4 bo ha 70% ta phi trn vi t l 47,2g mui trong 100ml
nc thm.
3

KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

3. Kt qu: Hy tnh hm lng protein c trong 100g nguyn liu ban u.

III. Phng php ta protein bng im ng in.
1. Nguyn tc
Gi tr pH m ti phn t protein trung ha in gi l im ng in ca protein
(pI), gi tr pH = pI phn t protein trung ha in s khng chuyn dch trong in
trng, phn t protein s km bn nht d kt ta.
Ngi ta li dng tnh cht ny kt ta protein.
2. Dng c v ha cht
Dng c
Ha cht
My khut t
Dung dch NaOH 20%
My ly tm
Dung dch HCl 10%
Cn phn tch
Ete
Bercher 100ml, 250ml
Etanol
Pipet 5ml, 10ml
Giy pH
3. Thc hnh
Tch chit protein c:
Cn 100g c xay nhuyn trong mt bercher ri cho 4 ln nc. (1 c : 4 nc, g/l)
Khuy u ri cho t t NaOH 20% va cho va khuy cho n pH 11 th bng giy pH,
khuy khong 45 pht ri ly tm, gi li dch chit trong mt bercher 250ml, b cn li thm
nc chit tip ln hai ri ly tm ly dch b b. Dch thu c cho vo bercher 250ml gp
chung vi phn dch chit bn trn.
Cho t t HCl 100% vo dch chit va cho va khuy n pH thch hp l 6 s thy
kt ta, ly tm 4000 vng/pht trong 30 pht, ri b dch thu ta protein, ra ta vi ete :
ethanol vi t l 2:1, ta protein sch khi ra ta vi ln vi ete : ethanol. Cn lng ta
protein thu c (mg ta) tnh ra hiu sut protein c trong 100g c.

KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

KHO ST HOT TNH ENZYM BROMELIN

(Xc nh hot tnh enzym Bromelin bng phng php Anson ci tin)
1. Nguyn tc
Phng php ny da trn s thy phn protein casein bng enzym Bromelin c
trong dch nghin cu, tip lm v hot enzym v kt ta protein cha b thy phn bng
dung dch acid trichloracetic. nh lng sn phm c to thnh trong phn ng thy
phn bng phn ng mu vi thuc th Folin. Da vo th chun ca Tyrosin tnh
lng sn phm do enzym xc tc to nn.
2. Dng c v ha cht
Dng c
Ha cht
ng nghim
Dung dch Casein 1%
Pipet 1ml, 2ml, 3ml, 5ml, 10ml
Dung dch TCA 5%
My quang ph
Dung dch NaOH 0,5N
Becher 50ml, 250ml, 500ml
Thuc th Folin
Bnh tia
Dung dch HCl 0,2N
Dung dch Tyrosin chun 1 mM/l trong
dung dch HCl 0,2N
3. Chun b ng chun Tyrosin
Cc bc dng ng chun Tyrosin
Dung dch ha cht
1

ng nghim
3
4

Dung dch Tyrosin chun (ml)

0
0,2
0,4
0,6
Lng Tyrosin tng ng (M)
0
0,2
0,4
0,6
Dung dch HCl 0,2N (ml)
5,0
4,8
4,6
4,4
Dung dch NaOH 0,5N (ml)
10
10
10
10
Thuc th Folin (ml)
3
3
3
3
Lc mnh, sau 10 pht o OD bc sng 660nm

0,8
0,8
4,2
10
3

1,0
1,0
4,0
10
3

ng s 1 l ng th khng (TK), cc ng cn li l ng th nghim (TN). V ng

chun Tyrosin tng quan gia lng Tyrosin (M) v OD (OD = ODTN ODTK).

KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

4. Phng php tin hnh

Ly 6 ng nghim sch, kh, tin hnh lm 3 ng th tht, 3 ng th khng
Cc bc chun b mu enzym o hot tnh
ng nghim
Th tht
Th khng
Dung dch Casein 1% (ml)
5
5
Dung dch TCA 5% (ml)
0
10
Dung dch enzym mu (ml)
1
1
o
Lc u v gi 35,5 C trong 20 pht
Dung dch TCA 5% (ml)
10
0
yn 30 pht, lc ly dch trong
Dung dch ha cht

Ly 2 ng nghim mi khc, cho vo ng th nht 5ml dch lc ca ng th tht v

cho vo ng th hai 5ml dch lc ca ng th khng.
Tip tc thm vo mi ng 10ml NaOH 0,5N v 3ml thuc th Folin, lc mnh, sau
10 pht o OD bc sng 660nm. Tnh OD = OD TT ODTK, sau da vo th chun
suy ra c s M Tyrosin.
5. Tnh kt qu
nh ngha n v Anson: mt n v Anson l lng enzym ti thiu trong iu kin
th nghim (35,50C: pH 7,6 ) thy phn Casein trong 1 pht to thnh sn phm ha tan
trong TCA, phn ng vi thuc th Folin cho ta hp thu OD bc sng 660nm tng
ng vi 1M Tyrosin trong ng chun.
H Protease =

M Tyro sin* V * L
t *m *v

(UI )

Vi: H Protease : hot enzym protease (UI/g)

V : tng th tch hn hp trong ng th tht hoc ng th khng (ml)
v : th tch dch lc em phn tch (ml)
t : thi gian thy phn (pht)
m : khi lng mu enzym em i xc nh hot tnh (g)
L: pha long enzym
M Tyrosin: lng M Tyrosin trong v (ml) suy ra t ng chun

KHO ST HOT TNH INVERTASE

6

KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

I. Nguyn tc:
Invertase l mt enzym thy gii saccharose thnh glucose v fructose.
Trong bi ny ta dng enzym invertase trch trong men bia cho thy gii
saccharose, t tnh c hot tnh ca enzym ny.
II. Dng c v thuc th
Dng c
Ha cht
Becher 100ml 250ml
dung dch saccharose 10%
Erlen 250ml
dung dch NaOH N/10
ng nghim 50ml
Dung dch Iod N/10
Pipet 5ml 10ml
Dung dch H2SO4 N
Buret 25ml
H tinh bt 1%
Phu lc
Dung dch Na2S2O3 N/20
III. Cch tin hnh
iu ch dung dch invertase: cn 0,5g men bia ha tan trong 50ml nc ct. Khuy
u trong 5 pht. lng em lc ly dch lc.
Thc hin ba ng nghim nh sau:
S ng
ml dung dch saccharose 10%
ml nc ct
ml enzym invertase
ml enzym invertase un si

1
10
10

2
10

3
10

10
10

thy gii trong 30 pht nhit phng. Tip , em tt c cc ng un cch

thy trong 2 pht, lm lnh di vi nc.
Ht 5 ml ca hn hp cho vo mt erlen 250ml v nh phn ng glucose nh sau:
cho thm vo bnh 10ml dung dch Iod N/10 v 15 ml dung dch NaOH N/10. Thi gian nh
NaOH phi ko di khong 2 pht (nh tng git). yn trong bng ti 15 -20 pht. Ly ra
thm vo 2ml dung dch H2SO4 N. nh phn lng iod thm cn tha bng dung dch
Na2S2O3 N/20 (dng h tinh bt cho thm vo lc cui phn ng lm cht ch th mu).

IV. Kt qu
Hot tnh ca invertase c biu th bng s mol saccharose thy gii bi lng
enzym invertase c trong 1g men bia trong 1 pht nhit phng th nghim.
7

KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

Gi V1 v V2 l s ml Na2S2O3 N/20 dng nh phn ng s 1 v s 2.

C nhn xt g v kt qu nh phn trong ng s 3?
Hot tnh invertase c tnh theo cng thc sau:
(V1 V2 ) * A * B
mol / g / pht
3
10 20 2 a b * t * m
a: th tch ly ra trong mi ng nh ng glucose (ml)
A: th tch tng cng trong mi ng (ml)
b: th tch dung dch enzym cho vo mi ng (ml)
B: tng th tch dung dch enzym c c t mg men (ml)
t: thi gian thy gii tnh bng pht.

XC NH HOT TNH ENZYM ALPHA AMYLASE

(THEO PHNG PHP SMITH & ROE)
8

KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

I. Nguyn tc:
Hot tnh - amylase biu th kh nng ca enzym amylase xc tc phn ng thy
phn tinh bt n dextrin trong 1 pht 50 0C v c th hin bng s n v ca enzym
trong 1g mu. Khi phn ng thy phn tinh bt xy ra, lng tinh bt cn li cha c thy
phn hy s to phn ng mu vi iod v c so mu bng my so mu quang hc bc
sng 620 nm.
II. Dng c v ha cht
Dng c
Ha cht
Bnh nh mc 50ml, 100ml, 250ml
Cn 960
Bp n nhit
Dung dch tinh bt 1% trong m pH 5,6
Pipet 1ml, 2ml, 3ml, 5ml, 10ml
Dung dch HCl 1N
Becher 50ml, 100ml, 250ml
Dung dch NaCl 3%
My quang ph
Thuc th Lugol
III. Thc hnh
1. Ly trch Amylase
Amylase l 1 tp cht protein khng tan trong ru mnh. Cn 10g la ny mm cho
vo ci vi 30ml nc ct, gi nt la di nc. yn trong 15 pht, em lc. Ly tt c
nc qua lc, thm 4 ln th tch cn 96 0. Khuy u, ta s thy amylase v cc protein khc
ta di dng ni ln.
lng, ly tm ly ta. Ha tan ta trong 20ml nc ct. Ta c dung dch amylase
kh tinh sch. c bit dung dch ny khng c cht ng kh (th 1ml dung dch thuc
th Fehling, un cch thy 3 pht, ta khng c kt ta xut hin)

2. Kho st hot tnh -amylase

Ly su ng nghim chia thnh 2 l, mi l 3 ng nghim gm th tht v th khng
ly trung bnh.
9

KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

Tin hnh th nghim theo bng sau:

ng nghim
ng th tht (t) ng th khng (o)
Nc ct (ml)
0
1
Dch chit - amylase
1
0
Dung dch m (ml)
1
1
Dung dch tinh bt 1% (ml)
1
1
Dung dch NaCl 3% (ml)
0,5
0,5
Dung dch HCl 1N (ml)
0
1
0
em 50 C trong 30 pht
Dung dch HCl 1N (ml)
1
0
Nc ct (ml)
5,5
5,5
Thuc th Lugol (ml)
0,5
0,5
Dung dch ha cht

Lc u hn hp trong ng nghim ri mang i so mu bc sng 620nm

IV. Kt qu:
H A=

(OD0 ODt ) * C * L
(UI )
OD0 * t

Vi OD0: mt quang ca ng th khng

ODt: mt quang ca ng th tht
C: lng tinh bt ban u tham gia phn ng (mg)
T: thi gian phn ng (pht)
L: h s pha long mu enzym.

XC NH M TNG S THEO PHNG PHP KJELDAHL

I. V c ha:
Nguyn tc: S v c ha cht m l s bin i tt c cc cht m d nm di dng
no (hu c, v c, protein) thnh hp cht v c l amonium sulfat (NH 4)2SO4 bng
cch un si vi H2SO4 m c v cht xc tc.
10

KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

H2SO4 m c
Cht m

Xc tc, t0

(NH4)2SO4

Thc hnh: ht chnh xc 1ml nu l nguyn liu lng hay cn chnh xc 1g nu l

nguyn liu kh nghin nhuyn cho ng nht, cho vo mt bnh c cao c dung
tch t 60ml n 100ml (bnh kjeldahl). Thm vo 5ml H 2SO4 m c v khong
0,5g cht xc tc. un si hn hp trong t ht kh c t 2 n 3 gi cho n khi dung
dch tr thnh trong sut v c mu xanh do tri nht. ngui v pha long thnh
100ml vi nc.
Cht xc tc dng y l hn hp xay nhuyn ca: K 2SO4 v CuSO4 c t l theo
trng lng l 9: 1
Thc hin 3 s th tht (c cht m) v 3 s th khng c cht m (thay cht m
bng nc ct)
II. Phng php kjeldahl:
Nguyn tc: Cht m c v c ha nm di dng Amonium sulfat em cho tc
dng vi cht kim mnh nh NaOH s phng thch ra amoniac
(NH4)2SO4 + 2NaOH

2NH4OH + Na2SO4

Sau lng amoniac c hi nc ni cun bng mt dng c my Parnas Wargner

v c dn n mt bnh tam gic c cha mt lng tha H 2SO4. T y cho php chng
ta xc nh c lng amoniac phng thch ra, c ngha l xc nh c lng m c
trong mu nguyn liu.

Dng c v ha cht:
-

Dng c:
My ct m Parnas Wargner
Bnh kjeldahl
Bp un
Bnh xt nc ct

Becher 250 ml, 100ml, 50ml

Erlen 250ml
Bnh nh mc 100ml, 50ml
Buret 25ml
Pipet 1ml, 5ml, 10ml, 20ml.
11

KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

ng ong 25ml

Ha cht:
Dung dch NaOH m c (10N)
Dung dch H2SO4 N/100
Dung dch NaOH N/100 (dung dch
ny sau khi pha xong dng ngy th

ng nng , nhng lu s b
carbonat ha nn nng khng cn
ng nh ban u, v th phi xc
nh li h s chnh l x bng cch
nh phn n vi dung dch acid
chun (acid oxalic, acid phtalic)
Dung dch metil 0,5% pha trong
ru etylic.

Cch thc hnh:

un si bnh nc my Parnas Wargner. t erlen c cha 25ml H 2SO4N/100 v vi
git metil vo phn ui ca my sao cho phn nhn nhng chm vo dung dch. Ht
10ml dung dch v c ha pha long thnh 100ml, em vo my Parnas Wargner;
thm vo 10ml NaOH m c ( ghi nh l khi cho mu cng nh NaOH m c vo
my, phi cho xung t t bng kp Mohr, nu cho chy xung qu nhanh th dung dch s
b my ht ra ngoi). Tip tc un si bnh nc v cho hi nc li cun amoniac sang
erlen trong 3 pht, h erlen xung v tip tc un thm 2 pht na, trng vi ca my bng
mt tia nc ct, tt c u cho chy vo erlen ng H 2SO4 N/100. Ly erlen ra v nh phn
H2SO4 tha bng dung dch NaOH c chun l x N/100 cho u. Khi dung dch i mu
(t mu hng nht sang mu da cam): thc hin 3 s th thut v 3 s th khng ly tr s
trung bnh.
Cch tnh:
Nu nguyn liu l cht lng, thng thng lng m c tnh bng s gam m trong
1 lt, nu nguyn liu l cht rn, th trc ht phi tnh m, sau lng m c tnh
bng phn trm (s gam m c trong 100g nguyn cht liu) theo kh tuyt i)
Gi s nguyn liu l cht lng. Ly 1ml nguyn liu em v c ha v pha long thnh
100ml, sau ht 10ml em chng ct bng my Parnas Wargner.
Gi V0 l th tch dung dch NaOH x N/100 (tr s trung bnh ca 3 ln th khng).
Gi Vt th tch dng dch NaOH x N/100 (tr s trung bnh ca 3 ln th tht).
Vy:
V V0 Vt

l lng NaOH tng ng vi lng amoniac phng thch bi 10ml dung dch v c ha
pha long.
1mol amoniac tng ng vi 1mol NaOH.
Do s mol amoniac phng thch bi 10ml dung dch v c ha pha long l:
12

KHOA CN SINH HC MI TRNG

CM

TH NGHIM HA SINH

Vx(1 / 100)
mol Vx10 5 mol
1000

S gam m c trong 10ml dung dch v c ha pha long l:

14.V .x.10 5 g

S gam m c trong 1000gl dung dch v c ha pha long hay trong 1ml nguyn liu
14.Vx.10 5 .100
14.Vx10 4 g
10

S gam m c trong 1 lt nguyn liu:

14.V .x.10 4 .1000 g / lt 1,4.V .xg / lt

MT S PHNG PHP NH TNH CHT NG

I. Phn ng mu tng qut (phn ng Molish)
Tt c cc cht ng u cho mu tm i vi dung dch Naphthol trong H2SO4
m c. trong ng nghim 1 ml dung dch ng 1% v 2 git dung dch Naphthol
15% trong ru. Lc u, nghing ng v cho H2SO4 m c t t theo thnh ng. C s
to thnh mt lp mu tm hin ra mt ngn cch 2 cht lng.
13

KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

II. Phn bit gia monosaccharid v disaccharid (phn ng Barfoed)

Kh nng kh ca nhng monosaccharid c th ln hn nhiu kh nng kh ca
disaccharid cho nn, thay v s kh mi trng base, s kh mi trng acid yu c th
gip ta phn bit c monosaccharid v disaccharid.
Trong ng nghim ng 2ml thuc th Barfoed, ta thm vo 1ml dung dch ng
mun kho st.
Ta thc hin ng thi ba s th chng vi cc dung dch glucose 1% , lactose 1% v
nc ct. un si cch thy trong 5 pht. Lm lnh, quan st ng c trm hin . un tip
tc 8 pht, quan st ng c trm hin .
Gii thch hin tng.
III. Phn ng vi thuc kh Fehling
Thuc kh ny l dung dch CuSO4 vi dung dch soude m c v tartrat kp
natrium v kalium. Tc dng ca dung dch soude m c ln CuSO 4 cho hydroxyd ng
(CuO, H2O) tan trong dung dch nh s hin din ca tartrat.
Trong mt ng nghim, 1ml thuc th Fehling v 1ml dung dch glucose. un si
cch thy trong 5 pht c trm hin gch Cu 2O nh s kh ng nh thnh ng nht bi
glucose.

KHO ST LIPID
I. Kho st tnh ha tan ca lipid
1. Nguyn tc:
Lipid khng ha tan trong nc do c sc cng b mt rt cao. Nu dng lc c hc
tc ng vo hn hp lipid nc th chng phn tn thnh tng ht nh, nhng ngng tc
dng chng li phn thnh 2 lp: lipid trn v nc di. Nu ta thm vo hn hp cc
14

KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

cht mui kim, x phng, mt.. s c tc dng lm gim sc cng b mt ca cc ht lipid

do vic tr li hai lp kh hn v dung dch lc ny s c mu trng c nh sa (nh
tng), lipid di dng cc ht nh liti cc micell y l hin tng nh ha m. Nu thay
nc bng cc dung mi hu c: acetone, ether, chloroform, benzen th lipid s ha tan vo
cc dung mi ni trn.
2. Ha cht
Du thc vt hay m ng vt
Dung mi hu c: alcohol, acetone, bezen
3. Cch lm
Cho vo ba ng nghim:
ng 1: 2ml alcohol
ng 2: 2ml acetone
ng 3: 2ml benzen
Nh thm vo mi ng 1 git du hoc m, lc nh, quan st s ha tan ca lipid
trong ba dung mi hu c.
II. Kho st mt s ch s c trng ca cht bo
1. Ch s acid:
L s mg KOH dng trung ha cc acid bo t do trong 1g cht bo.
Nguyn tc
Dng KOH 0,1N trong ru trung ha cc cht bo t do trong 1g nguyn liu,
cht ch th mu l phenolphtalein.
Dng c v ha cht
Dng c
Ha cht
Erlen 100ml
KOH 0,1N
Pipet 10ml
Ether ethylic
Becher 100ml
Phenolphtalein 0,5% trong alcohol
Bnh tia
Du kho st
Cch lm
Cn 1g (1ml) cht bo cho vo erlen, thm 10ml ether ethylic, lc u cho tan cht
bo. Cho 1 git phenolphtalein v chun bng KOH 0.1N cho n khi xut hin mu
hng bn trong 30 giy.
Tnh ton kt qu:
Ch s acid = 5.61 * V/m
5,61 l s mg KOH tng ng 1ml KOH 0.1N
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KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

V l s ml KOH
m l trng lng lipid cn (gram)
2. Ch s savon
L s mg KOH cn thit savon ha cc ester cha trong 1g cht bo.
Nguyn tc:
Cho nguyn liu kt hp vi 1 lng tha KOH savon ha cht bo. nh lng
KOH tha bng HCl gip suy ra ch s savon.
Dng c v ha cht:
Dng c
Ha cht
Erlen
KOH 0.5N trong ru
Pipet 10ml
HCl 0.5N
Becher 100ml
Ethanol tuyt i
B chng ct hon lu
Phenolphtalein 0,5% trong alcohol
Cht bo kho st
Cch lm:
Cn chnh xc 500mg cht bo cho vo erlen, thm 10ml KOH 0.5N. un si hon
lu 45, nh phn KOH bng HCl 0.5N vi cht ch th mu l phenolphtalein. Thc hin
mt bnh i chng thay 500mg cht bo bng 500mg nc.
Tnh ton kt qu:
Ch s savon = 28.05 * (V0 V1)/m
28.05 l s mg KOH tng ng 1ml HCl 0.5N
V0 l th tch dung dch HCl 0.5N ca bnh i chng
V1 l th tch dung dch HCl 0.5N ca bnh th nghim
m l trng lng cht bo cn (gram)

NH LNG VITAMIN C
(Phng php nh lng vitamin C bng IOD)
1. Nguyn tc
xc nh nhanh chng hm lng vitamin C trong nguyn liu khi c cht mu
ngi ta thng dng phng php chun iod. Tt c acid ascorbic b oxi ha bi iod.

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KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

Phn iod tha s cho mu xanh vi dung dch tinh bt. iu ni ln l phn ng kt
thc.
2. Dng c v ha cht
Dng c
Ha cht
- Erlen 100ml
- Dung dch HCl 5%
- Pipet 5ml, 10ml
- Dung dch Na2S2O3 0,01N
- Buret
- Dung dch tinh bt 1%
- Bnh nh mc 50ml
- Dung dch iod 0,01N
- Becher
- Ci chy s
- Phu thy tinh
3. Cch thc hnh
a. Chun b mu vt:
Cn N (g) mu vt v nghin nhanh chng trong ci s vi 5ml HCl 5%. Vitamin C
rt d b oxi ha trong khng kh nht l khi c s hin din ca cc ion kim loi (Fe, Cu).
V vy trong khi chun b mu phi ct hoc nghin bng dao khng r v lm nhanh.
Dch chit thu c cho vo bnh nh mc 50ml v thm nc ct (ra ci chy v
b vi ln bng nc ct ri vo bnh), khuy u ri lc.
b. nh phn:
- nh chun li dung dch iod N/100 vi dung dch Na2S2O3 0,01N tnh h s hiu
chnh T ca dung dch iod.
- Chun b hai erlen. Ht vo mi bnh 20ml dch chit c cha vitamin C, 5 -10 git
dung dch h tinh bt 1% v chun ngay bng dung dch iod 0,01N n khi c mu xanh

4. Kt qu
S mg vitamin C trong 1g mu vt c tnh nh sau:
x%

0,00088.a.T .V .100
v.N

a: th tch (ml) dung dch iod 0,01N dng khi chun .

T: h s hiu chnh dung dch iod 0,01N vi dung dch Na2S2O3 0,01N
V: th tch chung ca dch chit (50ml)
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KHOA CN SINH HC MI TRNG

TH NGHIM HA SINH

v: th tch dch chit ly chun (20ml)

N: trng lng mu phn tch
0,00088: s gram vitamin C tng ng vi 1ml dung dch iod 0,01N

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