Phng php DPPH

1.Gii thiu phng php

2.Tin hnh phng php
3.u im , hn ch v tnh

n nh ca phng php

1. Gii Thiu Phng Php

1.1. DPPH l g ?

DPPH
(1, 1-diphenyl-2picrylhydrazyl) l phng php nht
rc gc t do cung cp cc phng
php tip cn u tin nh gi
kh nng chng oxy ha ca mt hp
cht, trch lc hoc cc ngun sinh
hc khc.

1. Gii Thiu Phng

Php
1.2. Ngun gc ca phng php
Phng php ny c pht trin bi Blois (1958)
Vi quan im xc nh cc hot ng chng
oxy ha mt cch nh bng cch s dng mt n
nh gc t do, -diphenyl--picrylhydrazyl
(DPPH; C18H12N5O6, M = 394,33). Xt nghim ny
c da trn vic o lng kh nng nht rc
ca cc cht chng oxy ha i vi n. Cc
electron l ca nguyn t nit trong DPPH c
gim bng cch nhn mt nguyn t hydro t
cht chng oxy ha vi hydrazine tng ng.

2. Tin hnh phng php

2.1. Nguyn tc
Cc cht c kh nng khng oxy ho s trung
ho gc DPPH bng cch cho hydrogen, lm
gim hp th ti bc sng c i v mu
ca dung dnh phn ng s nht dn, chuyn
t tm sang vng nht.
Phng trnh :

2. Tin hnh phng php

2.2. Chun b ha cht

Ha cht :
Dung dnh DPPH
(1, 1-diphenyl-2-picrylhydrazyl ),
C18H12N5O6.
Mu :
c x l qua qu trnh chit lng rn,
h
dung mi l nc , qu trnh chit ny
c thc
hin trn mt lng mu nh v thc
hin phn

2. Tin hnh phng php

Cch tin hnh

Khong
20ln
140l dch chit
trn vi nc ct
t th tch
tng cng 3ml.
Sau thm 1ml
dung dch DPPH
0,2mM, lc u v
yn trong bng
ti 30 pht.
hp thu quang
hc c o
bc sng 517nm

2. Tin hnh phng php

X l kt qu

Kh nng kh gc t do DPPH c xc nh theo

cng thc sau :
DPPH (%) = 100 (ACT - ASP)/ACT
Trong :
ACT: hp thu quang hc ca mu trng khng
cha dch chit.
ASP: hp thu quang hc ca mu c cha dch
chit.
Kt qu bo co bi gi tr IC 50 l nng ca dch
chit kh c 50% gc t do DPPH iu kin
xc nh. Gi tr IC50 cng thp th hot tnh kh
gc t do DPPH cng cao.

3. u im, hn ch ca phng php

u im

- DPPH c th c s dng trong cc dung mi

hu c
v dung dch nc khng phn cc
v c th c s dng kim tra c hai cht
chng oxy ha a nc v a m.
- C chnh xc cao , d dng thc hin v
gi tr kinh t cao.
- DPPH c php phn ng vi ton b mu v
thi gian nht nh trong phng php ny.

3. u im, hn ch ca phng php

Hn ch

- DPPH ch c th c ha tan trong dung mi

hu c v s can thip ca hp th t cc hp
cht mu c th l mt vn i vi cc phn
tch nh lng .
- hp th ca DPPH trong methanol v acetone
gim di nh sng
- Hn ch trong vic phn nh cc phn vng ca
cht chng oxy ha trong cc h thng nh tng
v khng phi l hu ch o lng cc hot
ng chng oxy ha ca huyt tng.