Phng php xc nh polyphenol tng s trong ch Xc nh polyphenol tng s trong ch theo phng php ISO 145021 :2005 1.

Chit xut polyphenol Cn chnh xc 0.2 +- 0.001 g ch kh ( m khong 5%, kch thc 0.5 1mm, phi trn u ly i din) cho vo ng nghim hoc ng chit (10 ml). Cho bnh tam gic c cha dung mi methanol 70% vo b n nhit 70C trong 30 pht. t ng nghim cha mu vo b n nhit 70C, khong 1 pht ri cho 5ml methanol 70% 70C vo ng chit. Sau , ly ra v khuy trn trn Vortex. Tip tc nng nhit ng chit trong b n nhit (70C) v khuy trn li trn Vortex cc thi im 5 v 10 pht trch ly. Sau khi chit ( 10 pht), dch chit c h xung nhit phng bng phng php t nhin v c ly tm tc 3500 vng/pht trong 10 pht. Gn cn thn phn dch trong pha trn ca ng chit v mt ng nghim khc 10ml ( hoc bnh nh mc 10ml). Phn b tip tc c chit li mt ln na vi th tc nh trn. Sau dch chit sau ly tm ca ln chit 2 c dn vi ln chit 1 ( trong bnh nh mc 10 ml) v dng methanol 70% nhit thng ln th tch ti vch.

2. Pha long Ly 1 ml dch chit trn + 99ml nc ct 2 ln, lc u ( chun b trong bnh nh mc 100mml). Dch pha long ny c s dng xc nh hm lng polyphenol tng s. 3. Xy dng ng chun acid gallic ng chun acid gallic c xy dng trong khong nng t 10 n 50g/ml ( dung dch ch s dng trong ngy) 4. Xc nh hm lng polyphenol Dng pipette chuyn 1 ml nc ct vo ng nghim 10 ml ( nhc li 2 ln) mu trng. Dng pipette chuyn 1 ml dch chit mu pha long vo cc ng nghim 10 ml ( nhc li 2 ln) mu phn tch. Dng pipette cho 5 ml dung dch Folin-ciocalteu 10% vo mi ng nghim v lc u. Sau khi cho Folin-ciocalteu 5 pht, tip tc cho 4 ml Na2CO3 7.5% vo mi ng nghim, nt ming ng , lc u. cc ng phn ng nhit phng trong 60 pht sau o mt quang 765 nm (cuvet 10 mm, ca i chng l nc ct).

5. Tnh kt qu Hm lng pomyphenol tng s ( WT, %CK) c tnh theo cng thc:

Phn tch polyphenol theo phng php xanh ph ( Prussian Blue) 1. Nguyn tc Polyphenol s phn ng vi thuc th v to nn dung dch c mu xanh ph. T cng mu ca mi th nghim v th ng chun, ta s suy ra c nng polyphenol c trong mu cn phn tch. Ha cht: Dung dch FeCl3 0.02M trong HCl 0.1M; dd K3Fe(CN)6 0.016M Cht n nh : nc ct : H3PO4 85% : gum arabic 1% ( t l th tch 3:1:1) Cht chun: acid garlic trong nc ct ( 25 50 100 200 300 400 500 ppm) 2. Cch tin hnh Ly 0.1 ml dch ( cht chun hoc mu cn o c pha long n nng thch hp vo ng nghim, cho tip 3ml nc ct, lc u, cho nhanh 1ml K3Fe(CN)6 v 1ml FeCl3 , lc u dung dch. yn dung dch trong 15 pht. B sung cht n nh cho 10 ml, o hp thu A ti bc sng 700nm. Dng ng chun vi acid garlic cc nng nu trn.

3. Tnh kt qu Da vo ng chun tnh v so snh hm lng polyphenol trong cc dch ct