Determination of Superoxide Radical Scavenging Activity and Total Antioxidant Capacity by Modified DMPD Method

Year 2024, Volume: 10 Issue: 3, 506 - 519, 30.09.2024

Ferda Dondurmacıoğlu

https://doi.org/10.28979/jarnas.1382201

Abstract

Among the reactive oxygen species, Superoxide radicals can produce dangerous species that cause lipid peroxidation. Therefore, the determination and scavenging of superoxide radicals is critical. Our study is based on the interaction of the superoxide radical produced from the β-Nicotinamide adenine dinucleotide reduced disodium salt hydrate and phenazine methosulfate (NADH-PMS) system with N, N-dimethyl-p-phenylenediamine dihydrochloride (DMPD) to form the pink colored DMPD-quinone (DMPDQ) radical. In the presence of scavengers with superoxide radical scavenging activity (antioxidants, herbal teas) the color intensity decreases due to reduced DMPDQ radical production. The absorbance of the colored reference solution and the sample solution containing the radical scavenger was measured at 552 nm. The difference in absorbance (ΔA) between the reference solution and the sample solution was found. ΔA is proportional to the scavenger concentration. In the study, the superoxide radical scavenging effect of trolox (TR) and different AOXs was investigated. The superoxide radical scavenging effect of three different herbal tea infusion solutions was measured with this method. From the graph drawn between herbal tea concentrations and percentage inhibition values, 50% inhibition (EC50) values of herbal teas were found. EC50 method values were compared with the EC50 values of the nitroblue tetrazolium (NBT) and the 2,2'-azino-bis(3-etilbenzotiyazolin-6-sülfonik asit (ABTS) method. In addition, ABTS, cupric reducing antioxidant capacity (CUPRAC), and this study total antioxidant capacity (TAC) values of herbal tea infusions were calculated and compared.

Keywords

Antioxidant capacity, NADH-PMS, N N-dimethyl-p-phenylenediamine dihydrochloride, superoxide radical, percentage inhibition

Supporting Institution

TÜBİTAK, Grant number: 114Z089 and Office of Scientific Research Projects Coordination at İstanbul University-Cerrahpaşa, Grant number: FDP-2017-24832

Modifiye DMPD Yöntemi ile Süperoksit Radikali Süpürme Aktivitesi ve Toplam Antioksidan Kapasite Tayini

Abstract

Reaktif oksijen türleri içinde yer alan süperoksit radikalleri lipid peroksidasyonuna neden olan tehlikeli türler üretebilir. Bu nedenle süperoksit radikali tayini, süpürülmesi çok önemlidir. Çalışmamız β-Nicotinamid adenin dinükleotid indirgenmiş disodyum tuz hidratı ve fenazin metosulfat (NADH-PMS) sisteminden üretilen süperoksit radikalinin N, N-dimetil-p-fenilendiamin dihidroklorür (DMPD) ile etkileşimi sonucu pembe renkli DMPD-yarıkinon (DMPDQ) radikali oluşturmasına dayanmaktadır. Süperoksit radikali giderme aktivitesine sahip süpürücüler (antioksidan, bitki çayları) varlığında daha az DMPDQ radikali üretimi nedeniyle renk yoğunluğu azalır. Renkli referans çözeltisinin ve radikal süpürücü içeren örnek çözeltisinin absorbansı 552 nm’de ölçülmüştür. Referans çözelti ile örnek çözeltisinin absorbans farkı (ΔA) bulunmuştur. ΔA ile süpürücü derişimi orantılıdır. Çalışmada, trolox (TR) ve farklı antioksidanların (AOX) süperoksit radikal süpürücü etkisi araştırılmıştır. Yöntemle üç farklı bitki çayı infüzyon çözeltisinin süperoksit radikal süpürücü etkisi ölçülmüştür. Bitki çayı derişimleri ile yüzde inhibisyon değerleri arasında çizilen grafikten bitki çaylarının %50 inhibisyon (EC50) değerleri bulunmuştur. Bulunan EC50 değerleri süperoksit radikal tayininde yaygın olarak kullanılan nitroblue tetrazolium (NBT) ve 2,2'-azino-bis(3-etilbenzotiyazolin-6-sülfonik asit (ABTS) yöntemi EC50 değerleriyle karşılaştırılmıştır. Ayrıca bitki çayı infüzyonlarının ABTS, bakır(II) iyonu indirgeme antioksidan kapasite (CUPRAC) yöntemi ve çalışılan yöntemde toplam antioksidan kapasite değerleri (TAC) hesaplanıp, karşılaştırılmıştır.

Keywords

Antioksidan kapasite, NADH-PMS, N N-dimetil-p-fenilendiamin dihidroklorür, süperoksit radikali, yüzde inhibisyon

Supporting Institution

TÜBİTAK, Proje No: 114Z089 ve İstanbul Üniversitesi-Cerrahpaşa Bilimsel Araştırma Projeleri Koordinasyon Birimi, Proje No: FDP-2017-24832

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