CUPRAC colorimetric and electroanalytical methods determining antioxidant activity based on prevention of oxidative DNA damage

An unbalanced excess of oxygen/nitrogen species (ROS/RNS) can give oxidative hazard to DNA and other biomacromolecules under oxidative stress conditions. While the ‘comet’ assay for measuring DNA damage is neither specific nor practical, monitoring oxidative changes on individual DNA bases and other...

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Bibliographic Details
Published in:Analytical biochemistry 2017-02, Vol.518, p.69-77
Main Authors: Uzunboy, Seda, Çekiç, Sema Demirci, Eksin, Ece, Erdem, Arzum, Apak, Reşat
Format: Article
Language:English
Subjects:
Antioxidant activity
Antioxidants - analysis
Colorimetry - methods
Copper - chemistry
CUPRAC colorimetry
DNA - chemistry
DNA Damage
DNA sensor
DPV
Electrochemical Techniques - methods
Oxidation-Reduction
Oxidative damage
Thiobarbituric Acid Reactive Substances - analysis
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Summary:An unbalanced excess of oxygen/nitrogen species (ROS/RNS) can give oxidative hazard to DNA and other biomacromolecules under oxidative stress conditions. While the ‘comet’ assay for measuring DNA damage is neither specific nor practical, monitoring oxidative changes on individual DNA bases and other oxidation products needs highly specialized equipment and operators. Thus, we developed a modified CUPRAC (cupric ion reducing antioxidant capacity) colorimetric method to determine the average total damage on DNA produced by Fenton oxidation, taking advantage of the fact that the degradation products of DNA but not the original macromolecule is CUPRAC−responsive. The DNA−protective effects of water-soluble antioxidants were used to devise a novel antioxidant activity assay, considered to be physiologically more realistic than those using artificial probes. Our method, based on the measurement of DNA oxidative products with CUPRAC colorimetry proved to be 2 orders-of-magnitude more sensitive than the widely used TBARS (thiobarbituric acid-reactive substances) colorimetric assay used as reference. Additionally, the DNA damage was electrochemically investigated using pencil graphite electrodes (PGEs) as DNA sensor platform in combination with differential pulse voltammetry (DPV). The interaction of the radical species with DNA in the absence/presence of antioxidants was detected according to the changes in guanine oxidation signal. [Display omitted]
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2016.10.028