Magnetite nanoparticles-based hydroxyl radical scavenging activity assay of antioxidants using N, N-dimethyl-p-phenylenediamine probe

Authors: ZİYA CAN, BÜŞRA KESKİN, AYŞEM ARDA, EROL ERÇAĞ, MUSTAFA REŞAT APAK

Abstract: Excessive amounts of reactive oxygen species (ROS), unless counterbalanced by antioxidants, can cause cellular damage under oxidative stress conditions; therefore, antioxidative defenses against ROS must be measured. With the development of nanotechnology, nanoparticles have found numerous applications in science, health, and industries. Magnetite nanoparticles (Fe$_{3}$O$_{4}$:MNPs) have attracted attention because of their peroxidase-like activity. In this study, hydroxyl radicals (?OH) generated by MNPs-catalyzed degradation of H$_{2}$O$_{2}$ converted the N,N-dimethyl-p-phenylenediamine (DMPD) probe into its colored DMPD$^{•+}$ radical cation, which gave an absorbance maximum at λ = 553 nm. In the presence of antioxidants, $^{•}$OH was partly scavenged by antioxidants and produced less DMPD$^{•+}$, causing a decrease in the 553 nm-absorbance. Antioxidant concentrations were calculated with the aid of absorbance differences between the reference and sample solutions. The linear working ranges and trolox equivalent antioxidant capacity coefficients of different classes of antioxidants were determined by applying the developed method. In addition, binary and ternary mixtures of antioxidants were tested to observe the additivity of absorbances of mixture constituents. The method was applied to real samples such as orange juice and green tea. Student t-test, F tests, and the Spearman's rank correlation coefficient were used for statistical comparisons.

Keywords: Magnetite (Fe$_{3}$O$_{4}$) nanoparticles, reactive oxygen species (ROS), N,N-Dimethyl-p-phenylenediamine, colorimetric probe, antioxidant activity

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