CHNG 2.

KHO ST CARBOHYDRATE (GLUCID)

2.1 KHI QUT V CARBOHYDRATE Cc hp cht carbohydrate gm 3 loi: - Monosaccharide: l n v cu to ca carbohydrate, khng b thy phn. V d: Glucose v fructose. - Oligosaccharide: c t 2 n 10 monose gm: di, tri hay tetrasaccharide V d: Saccharose (ng ma), lactose (ng sa), maltose (ng mch nha). - Polysaccharide: gm 2 loi: + Polysaccharide thun: gm nhng monomer cng loi. V d: tinh bt, cellulose v glycogen. + Polysaccharide tp: Gm nhng monomer khc loi hay dn sut ca monomer hoc c thm cht bo. V d: Heparin, acid hyallycoric. Tnh cht - Monosaccharide: C tnh kh, d b mt nc bi acid v c mnh (H 2SO4, HNO3 m c) cho ra dn sut furfural. Cht ny d kt hp vi cc loi phenol (resorcinol, - naphthol, orcinol...) cho phn ng mu. - Oligosaccharide: B thy phn bng acid hoc enzyme thch hp cho ra cc thnh phn cu to tng ng. Trng hp disaccharide, ty cch kt hp cc monomer s c tnh kh hay khng. Kh nng kh ca oligosaccharide yu hn monosaccharide. Trong c th sinh vt carbohydrate ng vai tr ln, n l thnh phn cu to nn cc c quan v dch gian bo. N cung cp phn ln nng lng cho t bo sng. 2.2 NH TNH MONOSACCHARIDE V TINH BT 2.2.1 Kho st tinh bt Tinh bt l cht d tr ca thc vt, l ngun thc n v ngun cung cp nng lng chnh cho ngi v ng vt. Tinh bt khng tan trong nc lnh. Khi un nng vi nc th trng ln v to thnh h (h tinh bt), cho mu xanh (i khi mu tm ) vi iod, khng c tnh kh. Tinh bt b thu phn bi acid m c (HCl, H2SO4).
CH2OH O CH2OH O O CH2OH O O O CH2OH O

OH

u kh

Cu to mt on amylose ca tinh bt 13

Gio Trnh Thc Tp Sinh Ha

Chun b tinh bt Khoai lang ra sch, mi trn bn mi t trn ry sn trong chu thy tinh. Mi nh tay cho nhuyn, xong vt tht k b khoai lang vi 200 mL nc ct, chuyn nc bt t chu thy tinh vo cc 250 mL, yn cho bt lng. Khi bt lng k, gn b phn nc trn ri tip tc cho nc ct vo khuy u, lng tip tc ri li gn b phn nc trn. Lp li vi ln cho n khi tinh bt trng v sch. Khuy tinh bt vi vi mL nc ct ri cho vo 100 mL nc ct ang si th ta c dung dch h tinh bt. Ly dung dch h tinh bt va nhn c lm cc th nghim sau. Tin hnh a. Cho mu vi iod Tin hnh: Cho vo ng nghim 5 mL dung dch h tinh bt, cho tip 2 git iod. Nhn xt kt qu. Sau xem ng nghim un cch thy khong 5 pht - Nhn xt. ngui - Quan st, nhn xt v gii thch. b. Phn ng thy phn Tin hnh: Dng ng nghim ln cho vo 15 mL dung dch h tinh bt. Cho tip vo 5 mL HCl m c, khuy u. un cch thy, c sau 1 pht ly 1 git dung dch ang thy phn nh ln 1 git iod sn trn a knh ng h. Th nh vy cho n khi khng cho mu vi iod. Nhn xt mu thay i khi th dung dch thy phn vi iod. Kt lun. Lu : Gi dung dch thy phn lm cc phn ng sau. 2.2.2 nh tnh monosaccharide (glucose) v tinh bt a. Phn ng Molish Nguyn tc : Cc loi carbohydrate u cho phc mu tm vi dung dch naphthol trong acid H 2SO4 m c.
CH2OH O

H2SO4 -3H2O

O HOCH2 CHO

-D-glucose

5-hydroxymethyl furfural
OH OH - H2O Napthol O OH

HOCH2

CHO + 2

5-hydroxymethyl furfural

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CH C C CH2OH CH CH

Phc mu tm

Gio Trnh Thc Tp Sinh Ha

Tin hnh Chun b cc ng nghim theo bng sau: ng nghim 1 Glucose 1% 2 mL 3 git 2 H tinh bt 1% 2 mL 3 git 3 H tinh bt thy phn 2 mL 3 git

Ho cht Dung dch Thuc th Molish H2SO4 m c

K thut: cho t t thnh dng chy lin tc theo thnh ng nghim, khng lc, n khi xut hin mu tm th dng li, c th tch dung dch acid dng.

Nhn xt tc phn ng ca mi ng nghim. Gii thch v nu ngha ca phn ng Molish. b. Phn ng vi thuc th Fehling Nguyn tc: Trong mi trng kim un nng, monosaccharide dng enoldiol khng bn, d dng kh cc kim loi nng nh Cu 2+, Ag+, Hg2+. Monosaccharide s kh ng II trong dung dch Fehling thnh ng I c kt ta gch. Cc disaccharide c nhm -OH glycoside t do u c tnh kh:
CHO (HCOH)n CH2OH O CH COONa COOH (HCOH)n O CH COOK CH2OH H O CH COONa H O CH COOK

Cu

Cu2O

o gach

Tin hnh ng nghim Ho cht Glucose 1% Tinh bt thu phn (phi trung ho) Tinh bt khoai lang Fehling (A+B t l 1:1) 1 2 mL 0 0 3 mL 2 0 2 mL 0 3 mL 3 0 0 2mL 3 mL

un cch thy 3 ng nghim trong khong t 3- 5 pht. Quan st hin tng xy ra 3 ng nghim, gii thch v vit phng trnh phn ng. 2.3 NH LNG NG KH Cc phng php ha hc xc nh ng kh u da trn kh nng kh cc hp cht khc nhau ca chng. Trong thc t c rt nhiu phng php xc nh, ty thuc vo hm lng nhiu hay t ngi ta c th s dng mt trong cc phng php thng dng sau y:

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2.3.1 nh lng ng kh theo phng php Bertrand Nguyn tc Trong mi trng kim cc ng kh (glucose, fructose, maltose...) d dng kh ng II thnh ng I theo phn ng Fehling. Kt ta ng I oxyt c mu gch, c kh nng kh vi mui Fe3+ thnh mui Fe2+ trong mi trng acid: Cu2O + Fe2(SO4)3 + H2SO4 2CuSO4 + 2FeSO4 + H2O Fe2+ sinh ra c tnh kh li tc dng vi KMnO 4 l cht oxy ha nn dng KMnO4 chun Fe2+ trong mi trng acid: 10FeSO4 + 2KMnO4 + 8H2SO4 K2SO4 + 2MnSO4 + 5Fe2(SO4)3 + 8 H2O Da vo lng KMnO4 s dng ta c th tnh c lng Cu2O v t tnh c lng ng kh trong dung dch bng cch tra bng t l gia dung dch KMnO4 v ng kh ca Bertrand. 2.5. NH LNG NG SACCHAROSE

ng saccharose cng vi mt s ng kh c nhiu trong cc loi tri cy, rau, c... Saccharose khng c tnh kh nn khng th xc nh trc tip bng cc phng php Bertrand. c th xc nh c saccharose bng phng php ny phi thy phn n thnh cc ng kh glucose v fructose

Nguyn tc: Khi thy phn dung dch saccharose bng acid ta thu c hn hp ng glucose v fructose. nh lng ng kh to thnh cho php tnh c lng sucrose c trong mu phn tch.
C12H22O11 saccharose

H2O

3.3. CC CH S NH GI LIPID 3.3.1. Xc nh ch s x phng nh ngha: Ch s x phng l s miligam KOH cn thit trung ha tt c nhng acid bo t do v kt hp c trong 1 gam cht bo. Nguyn tc: Cho cht bo cn phn tch kt hp vi mt lng KOH tha cht bo chuyn ht thnh x phng. Phn KOH tha c nh phn bng dung dch acid chun vi phenolphtalein lm ch th mu.

Trong :

- Cxp: Ch s x phng - a: S mL HCl 0,5N dng chun bnh th khng - b: S mL HCl 0,5N dng chun bnh th tht (bnh c mu). - m: S gam cht bo ly lm th nghim

3.3.2. Xc nh ch s iod nh ngha: Ch s iod l s gam iod kt hp vi 100 gam cht bo Nguyn tc Trong nhng iu kin th nghim xc nh nhng ni i -CH=CH- cho vi halogen phn ng cng ch khng cho phn ng th. Cho mt lng tha ICl (iodine monochloride) tc dng vi cht bo cn phn tch trong bng ti phn ng cng xy ra hon ton. Lng iod phn ng c xc nh da vo phn ng chun lng iod gii phng ra (sau khi thm KI) bng dung dch Na2S2O3 chun vi tinh bt lm ch th. Nh vy ch s iod xc nh tng qut cc acid bo khng no trong cht bo. Cc phng trnh phn ng c trnh by nh sau:
C C H H

ICl

H C Cl

H C I

ICl + KI KCl + I2 Na2S2O3 + I2 2 NaI + Na2S4O6 Ho cht - Dung dch ICl 0,2M - Dung dch KI 0,1% - Dung dch Na2S2O3 0,1N - Cloroform - Dung dch h tinh bt 1% 26

Gio Trnh Thc Tp Sinh Ha

Tin hnh Cn chnh xc 0,2 gam du cho vo bnh tam gic 250 mL (c nt nhm), thm vo 10 mL cloroform, tip tc cho thm 25 mL dung dch ICl 0,2M. Lc k bnh, yn 1 gi trong bng ti. Tin hnh chun b mu i chng tng t nh mu tht. Trng np v c bnh tam gic vi khong 50 mL nc ct, thm 10 mL dung dch KI 0,1% (w/v) v nh lng iod gii phng ra bng dung dch Na 2S2O3 0,1N n khi c mu vng sm ri thm 1 mL h tinh bt 1%. Nu c mu xanh xut hin th tip tc chun cho n khi dung dch mt mu. Tnh kt qu Ta bit 1mL Na2S2O3 0,1N tng ng 0,01269 g I2
Ci = 0,01269 x 100 x (a - b) m

Trong :

- Ci: Ch s iod - a: S mL Na2S2O3 0,1 N dng chun bnh th khng - b: S mL Na2S2O3 0,1 N dng chun bnh th nghim - m: Lng du dng th nghim (tnh bng gam) - 100: tnh trong 100 gam cht bo

3.3.3. Xc nh ch s acid nh ngha: Ch s acid l s miligam KOH cn thit trung ha ht nhng acid bo t do c trong 1 gam cht bo. Nguyn tc: Ngi ta dng KOH 0,01N trung ha cc acid bo t do c trong cht bo dng phn tch vi phenolphtalein lm ch th mu. Ha cht - KOH 0,01N trong alcol - Alcol tuyt i - Ether ethylic Tin hnh Dng 1 bnh tam gic 100 mL cho vo 5 mL alcol tuyt i v 5 mL ether (theo t l 1:1) cho thm vo bnh ny 2-3 git phenolphtalein v dng KOH 0,01N trong alcol trung ha hn hp n khi xut hin mu hng lt. Sau thm vo hn hp va trung ha 0,5 gam du. em hn hp ny trung ha bng KOH 0,01N trong alcol cho n khi c mu hng bn vng sau 30 giy. c th tch KOH dng trn buret. Tnh kt qu Ta bit 1 mL KOH 0,01N tng ng vi 0,56 mg KOH Ch s acid c tnh theo cng thc sau:

Ca
Trong :

= a x 0,56

- Ca: Ch s acid -a:S mL KOH 0,01N dng trung ha hn hp c du - m: Khi lng du ly lm th nghim (tnh bng gam) 27

Gio Trnh Thc Tp Sinh Ha

3.3.4. Xc nh ch s peroxid nh ngha: Ch s peroxid l s gam iod c gii phng ra bi peroxid c trong 100 gam mu. Nguyn tc: Trong khng kh, cc acid bo c trong cht bo, c bit l cc acdi bo khng no d dng b oxy ha mt phn to thnh peroxid, gy ra hin tng i ha cht bo. Xc nh ch s peroxid da trn phn ng sau:
R1 H C O H C O R2

+ KI + 2 CH3COOH

R1

H C O

H C R

+ I 2 + 2 CH3COOK + H2O

Lng iod gii phng ra c chun bng dung dch Na2S2O3 vi tinh bt lm ch th mu. Na2S2O3 + I2 2 NaI + Na2S4O6 Ho cht - Acid acetic m c - Cloroform - Dung dch KI bo ha - Dung dch Na2S2O3 0,1 N - Dung dch h tinh bt 1% Tin hnh Cn chnh xc khong 2 gam du cho vo bnh tam gic 250 mL, thm vo 10 mL dung dch hn hp acid acetic : cloroform (t l 2: 1) v 1 mL dung dch KI bo ha mi pha. y np, lc k bnh v yn trong bng ti 10 pht. Tin hnh chun b mu i chng tng t nh mu tht, dng 2 mL nc ct thay v du. Cho thm 25 mL nc ct v tin hnh nh lng iod gii phng ra bng dung dch Na2S2O3 0,1N n khi c mu vng sm ri thm 1 mL h tinh bt 1%. Nu c mu xanh xut hin th tip tc chun cho n khi dung dch mt mu. Tnh kt qu Ta bit 1mL Na2S2O3 0,1N tng ng 12,69 mg I2 Ch s peroxid c tnh theo cng thc sau:
Cp = 0,01269 x ( a - b) x 100 m

Trong :

- Cp: Ch s peroxid - a: S mL Na2S2O3 0,1 N dng chun bnh th khng - b: S mL Na2S2O3 0,1 N dng chun bnh th nghim - m: Lng du dng th nghim (tnh bng gam) - 100: tnh trong 100 gam cht bo

3.4. XC NH HM LNG LIPID TH BNG MY SOXHLET Nguyn tc

Da trn kh nng ha tan ca lipid trong dung mi hu c khng phn cc, dng dung mi hu c trch lipid ra khi nguyn liu c nghin nh. Trong qu trnh trch, cc hp cht tan c trong cht bo nh cc sc t, cc vitamin tan trong cht bo... cng b tch ra khi nguyn liu. Do c ln cc tp cht khc, nn thnh phn trch ly c gi l lipid tng s hay lipid th. 28

Gio Trnh Thc Tp Sinh Ha

C 2 phng php xc nh: - Phng php trc tip: Trch lipid ra khi nguyn liu v cn lng lipid c trch

ly. - Phng php gin tip: Xc nh chnh lch khi lng nguyn liu kh trc v sau khi chit xut lipid ra khi nguyn liu, t suy ra khi lng lipid trong mu phn tch.