DPPH Antioksidan Analizinin Yeniden Değerlendirilmesi

Yıl 2019, Cilt: 9 Sayı: 2, 125 - 135, 25.12.2019

Serap Ayaz Seyhan

Öz

Antioksidanların uygulamaları, oksidatif stresin zararlı etkilerini en aza indirmedeki çeşitli rolleri nedeniyle artmaktadır. DPPH· (1,1-Difenil-2-pikrilhidrazil radikali; C18H12N5O6) antioksidan analizi, farklı bileşiklerin antioksidan aktivitesini değerlendirmek için yaygın olarak kullanılan hızlı, basit ve ucuz bir yöntemdir. Ayrıntılı literatür taraması, çeşitli araştırma gruplarının farklı DPPH· derişimlerinde (22,5–400µM), inkübasyon süresinde (5 dk-120 dk), reaksiyon çözücüsünde (etanol, metanol) ve reaksiyon karışım pH'sında (pH= 3 ve 5.5 tampon ortamı) olan oldukça farklı protokoller kullandıklarını göstermektedir. Farklı laboratuvarların sonuçlarının karşılaştırılması ve sonuçların doğrulanması için standart bir analizin gerekliliği çok önemlidir. Bu çalışmada DPPH· antioksidan analizini etkileyen DPPH· derişimi, standart madde derişimi, reaksiyon/inkübasyon süresi gibi önemli faktörler spektrofotometrik yöntem ile incelenerek standart bir DPPH· antioksidan analiz yöntemi oluşturulmuştur. Bu yöntem kapsamında ölçümler, reaksiyon ortamındaki DPPH· derişimi 25-75 µM aralığında, çalkalayıcılı inkübatörde 30oC’de, 30dk’da ve karanlık ortamda gerçekleştirilmelidir.

Anahtar Kelimeler

DPPH, Antioksidan, Antioksidan aktivite, Yöntem

Destekleyen Kurum

Marmara Üniversitesi Bilimsel Araştırma Projeleri Birimi

Proje Numarası

SAG-A-131113-0423

Teşekkür

Bu çalışma, M. Ü. Bilimsel Araştırma Projeleri Birimi tarafından SAG-A-131113-0423 nolu proje ile desteklenmiştir.

Re-Evaluating of The DPPH• Antioxidant Assay

Öz

Applications of antioxidants are increasing due to
their various roles in minimising harmful effects of oxidative stress. DPPH^·
(1,1-Diphenyl-2-picrylhydrazyl radical) antioxidant assay is a fast, simple,
inexpensive method commonly used to evaluate the antioxidant activity of
different compounds. A detailed literature survey shows that various research
groups have used widely different protocols differed in the concentration of
DPPH^·
(22,5–400µM), incubation time (5 min–120 min), reaction solvent (ethanol,
methanol) and pH of the reaction mixture (pH=3 and 5.5 buffer) . The
requirement of a standard assay is very important in order to compare the
results of different laboratories and validation of the conclusions. In this
study, a standard DPPH• antioxidant analysis procedure was performed by
analyzing the important factors such as DPPH• concentration, standard substance
concentration, reaction / incubation time by spectrophotometric method. Under
this procedure, the measurements should be carried out at DPPH concentration in
the reaction medium in the range of 25-75 µM, 30 ° C in a shaker incubator, 30
minutes and dark medium.

Anahtar Kelimeler

DPPH• radical, Antioxidant, Antioxidant activity, Mthod

Proje Numarası

SAG-A-131113-0423

Kaynakça

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