I HC QUC GIA H NI TRNG I HC KHOA HC T NHIN KHOA SINH HC

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BO CO THC TP : Ha sinh cng nghip

Sinh vin thc hin: Hong Ngc Anh Phm Nht Khi Giang Huy Dim Nguyn Thanh Tm Lp : K54B Cng ngh sinh hc

Nm 2012

BI 3. NH LNG NG KH THEO PHNG PHP BERTRAND

1/ Nguyn tc: Tt c cc ng c nhm andehit hay nhm xeton t do trong nhng iu kin nht nh c kh nng tham gia vo phn ng oxi hoa kh vi ion kim loi c gi l ng kh. Phng php Bertrand da vo kh nng kh dung dch Cu2+ trong mi trng kim nh lng cc loi andoz, xetoz hay cc disacarit c tnh kh nh mantose, lactose). Cu2+ b kh thnh Cu1+ theo phn ng Fehling. Kt ta Cu1+ c mu gch, c kh nng kh vi mui Fe3+ thnh mui Fe2+ trong mi trng axit: Cu2O + Fe2(SO4)3 + H2SO4 2CuSO4 + 2FeSO4 + H2O Fe2+ sinh ra c tnh kh li tc dng vi KMnO4 ( l cht oxi ha), dng KMnO4 chun Fe2+ trong mi trng axit : 10FeSO4 + 2KMnO4 + 8H2SO4 K2SO4 + 2MnSO4 + 5Fe2(SO4)3 + 8H2O Da vo lng KMnO4 s dng ta c th tnh c lng Cu2O v t tnh c lng ng kh trong dung dch qua t l gia dung dch KMnO4 v ng kh ca Bertrand 2.Nguyn liu v ha cht - Bt go, bt ng hoc khoai lang ti, - Dung dch HCl 20-25%, - Dung dch NaOH 10%, - Dung dich Fe2(SO4)3 - Dung dch KMnO4 0,1N - Thuc th Fehling. 3.Thit b, dng c Ci, chy s, bnh nn, giy lc, bnh nh mc 250ml, ng ong 100ml, cc cc thy tinh, pipet thy tinh, a thy tinh Bnh nn (V = 100ml), buret, phu Buchner

4. Cch lm: - Thu dch lc tinh bt : cn 1g bt go (hoc 2g khoai lang ti nghin nh) cho vo cc, thm 100ml nc ct, khuy lin tc 15-30 pht. Lc qua giy lc, trng cc cn thn chuyn ton b nguyn liu vo phu, ra nguyn liu trong phu nhiu ln (4-5 ln) bng nc ct, mi ln 20-30ml loi hon ton ng kh (kim tra nc ra bng phn ng vi thuc th Fehling). t phu cng vi giy lc c cha nguyn liu trn bnh nn hoc bnh cu. Dng a thy tinh nh chc thng giy, ra ton b nguyn liu trn giy lc vo bnh bng nc ct (khong 80ml). Thm va mt lng HCl 20% vo bnh nng axit cui cng trong dung dch l 2%. Sau khi thy phn xong, trung ha dung dch thy phn bng NaOH 10% (th bng giy qu t mi trng axit yu). Chuyn ton b dung dch sang bnh nh mc, dn nc ct n vch nh mc ca bnh 250ml. Khuy u, lc c dung dch lc trong sut. Dch lc ny dng xc nh ng kh theo phng php Bertrand. Tin hnh th nghim Cho vo bnh nn 10ml dung dch ng, thm 10ml h hp thuc th Fehling. y bnh v un trn bp. un si khong 3 pht k t khi bt u xut hin bt u tin. Kt ta gch ca Cu2O xut hin trong bnh. Ly bnh ra ngui. Ra kt ta Cu2O vi ln bng nc m un si (mc ch loi O 2) cho n khi dung dch ra khng cn phn ng kim trn giy pH. Qu trnh ra c tin hnh trn phu lc Buchner vi giy lc dy. ha tan kt ta Cu2O, t phu Buchner ln bnh nn c cha kt ta. vo phu 15ml dung dich Fe2(SO4)3 chy t t xung bnh, lc. Quan st xem kt ta trong phu v trong bnh ha tan ht cha. Nu cha tan ht th cn phi thm dung dch Fe2(SO4)3. Sau khi ha tan ht kt ta, ra phu bng nc nng cho n khi nc ra khng c phn ng axit trn giy o pH. Chun dung dch trong bnh (Fe2+) bng dung dch KMnO4 0,1N cho n khi xut hin mu hng bn trong 30 giy.

5.Tnh kt qu T lng KMnO4 dng tnh ra lng ng. T lng ng tra bng tnh ra lng ng tng ng. T kt qu trn tnh phn trm ng trong dung dch X (mg%) = a x 100/10 a: s mg ng tra c t bng Lng KMnO4 0,1N s dng chun l : 13,3ml S mol Cu2+ s dng cho phn ng l

nCu2+ = nFe2+ = 5nKMnO4 = 5.CM.V = 5.0,1.13,3 = 6,65 (mmol) Khi lng Cu s dng cho phn ng l: mCu2+ = n.M = 6,65.64 = 0,4256 mg Lng ng tng ng l: a = Phn trm ng trong dung dch l: X (mg%) = a x 100/10

BI 4. XC NH HM LNG PROTEIN THEO PHNG PHP LOWRY

1. Nguyn tc Protein tc dng vi thuc th Folin to thnh sn phm mu xanh. y l sn phm kh ca phophomolipden-photphovolfeamat bi cht ng protein v c hp th cc i bc sng 750nm. Phng php c nhy cao c s dng cho vic xc nh nhiu di protein dng tng i thun khit. Tuy nhin c nhc im l mu ca phc cht c th b nh hng nu trong dung dch c mt s cht nh EDTA, m Tris. ng, 2. Nguyn liu v ha cht: Dung dch protein cn nghin cu. Dung dch A: Na2CO3 ha trong NaOH 0,1N Dung dch B: CuSO4.5H2O 0,5% pha trong dung dch natri xitrate 1% (hoc kali tactate 1%). Dung dch C: ch pha trc khi dng: trn 1 th tch dung dc B vi 49 th tch dung dch A. Dung dch Folin pha long 2 ln. 3. Cch lm Ht chnh xc vo ng nghim sch 0,1 ml dung dch nghin cu, thm vo 0,5 ml dung dch C, lc u, gi nhit phng 10 pht. Sau thm chnh xc 0,05 ml dung dch Folin pha long 2 ln, lc u 30 pht, em so mu trn my so mu bc sng 750 nm. Song song vi ng th nghim, lm ng i chng trong thay dung dch protein bng nc ct v tip tc lm nh cc bc trn. Lp th chun protein: chun b cc dung dch chun ( albumin 1mg/ml ) c cc nng pha long khc nhau cha mt lng protein tng ng l 20, 50, 100, 200 v 400 g/ml protein t mt dung dch gc ban u.

Nng cui (g/ml) 20 50 100 200 400

Th tch albumine (ul) 2 5 10 20 40

Th tch H2O (ul) 98 95 90 80 60

Tin hnh lm phn ng mu vi cc dch chun theo th t, thnh phn v t l cc ha cht nh vi dch nghin cu. Dng th biu din s tng quan gia mt quang hc v nng protein. T s c ca mu th nghim, i chiu vi th chun tnh ra hm lng protein trong 1 ml dch nghin cu, t tnh ra hm lng % protein trong nguyn liu. 4. Kt qu STT 1 2 3 4 5 6 7 8 Nng protein 20 g/ml 50 g/ml 100 g/ml 200 g/ml 400 g/ml Mu i chng Protein ban u (pha long 2 ln) Mt quang hc 97 79 74 69 62 132 56

Protein ban u 74