Abstract
Two methods for the measurement of antioxidant capacity are described: one based on a chronometric variation of a hemoglobin ascorbate peroxidase assay and the other based on electron paramagnetic resonance (EPR) spectra collected upon alkaline treatment of ethanolic samples. The involved chemical mechanisms are discussed, alongside the most important benefits and shortcomings; the assays offer new qualitative and quantitative information on samples of biological as well as synthetic origin.
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References
Niki E (2010) Assessment of antioxidant capacity in vitro and in vivo. Free Radical Bio Med 49:503–515
Jones DP (2006) Redefining oxidative stress. Antioxid Redox Signal 8:1865–1879
Huang D, Ou B, Prior RL (2005) The chemistry behind antioxidant capacity assays. J Agric Food Chem 53:1841–1856
Prior RL, Wu X, Schaich K (2005) Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements. J Agric Food Chem 53:4290–4302
Frankel EN, Finley JW (2008) How to standardize the multiplicity of methods to evaluate natural antioxidants. J Agric Food Chem 56:4901–4908
López-Alarcón C, Denicola A (2013) Evaluating the antioxidant capacity of natural products: a review on chemical and cellular-based assays. Anal Chim Acta 763:1–10
Carocho M, Ferreira ICFR (2013) A review on antioxidants, prooxidants and related controversy: natural and synthetic compounds, screening and analysis methodologies and future perspectives. Food Chem Toxicol 51:15–25
Procházková D, Boušová I, Wilhelmová N (2011) Antioxidant and prooxidant properties of flavonoids. Fitoterapia 82:513–523
Mot AC, Coman C, Miron C et al (2014) An assay for pro-oxidant reactivity based on phenoxyl radicals generated by laccase. Food Chem 143:214–222
Dunne J, Svistunenko DA, Alayash AI et al (1999) Reactions of cross-linked methaemoglobins with hydrogen peroxide. Adv Exp Med Biol 471:9–15
Cooper CE, Silaghi-Dumitrescu R, Rukengwa M et al (2008) Peroxidase activity of hemoglobin towards ascorbate and urate: a synergistic protective strategy against toxicity of hemoglobin-based oxygen carriers (HBOC). Biochim Biophys Acta 1784:1415–1420
Reeder BJ, Svistunenko DA, Cooper CE, Wilson MT (2004) The radical and redox chemistry of myoglobin and hemoglobin: from in vitro studies to human phatology. Antiox Redox Signal 6:954–966
Reeder BJ, Grey M, Silaghi-Dumitrescu RL et al (2008) Tyrosine residues as redox cofactors in human hemoglobin: implications for engineering nontoxic blood substitutes. J Biol Chem 283:30780–30787
Silaghi-Dumitrescu R, Reeder BJ, Nicholls P, Cooper CE, Wilson MT (2007) Ferryl haem protonation gates peroxidasic reactivity in globins. Biochem J 403:391–395
Vollard NB, Reeder BJ, Shearman JP et al (2005) A new sensitive assay reveals that hemoglobin is oxidatively modified in vivo. Free Radical Bio Med 39:1216–1228
Vollaard NB, Sherman JP, Cooper CE (2005) Exercise-induced oxidative stress: myths, realities and physiological relevance. Sports Med 35:1045–1062
Mot AC, Damian G, Sarbu C, Silaghi-Dumitrescu R (2009) Redox reactivity in propolis: direct detection of free radicals in basic medium and interaction with hemoglobin. Redox Rep 6:267–274
Venkataraman B, Fraenkel GK (1955) Proton hyperfine interactions in paramagnetic resonance of semiquinones. J Am Chem Soc 77:2707–2713
Ashworth P, Dixon WT (1972) Secondary radicals in the autoxidation of hydroquinones and quinones. J Chem Soc Perkin Trans 2:1130–1133
Pedersen JA (1973) Electron spin resonance studies of oxidative processes of quinones and hydroquinones in alkaline solution; formation of primary and secondary semiquinone radicals. J Chem Soc Perkin Trans 2:424–431
Roginsky V, Barsukova T (2000) Kinetics of oxidation of hydroquinones by molecular oxygen. Effect of superoxide dismutase. J Chem Soc Perkin Trans 2:1575–1582
Eslami AC, Pasanphan W, Wagner BA, Buettner GR (2010) Free radicals produced by the oxidation of gallic acid: an electron paramagnetic resonance study. Chem Central J 4:15–19
Antonini E, Brunori M (1971) Hemoglobin and myoglobin in their reaction with ligands. North-Holland, Amsterdam
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Mot, A.C., Bischin, C., Damian, G., Silaghi-Dumitrescu, R. (2015). Antioxidant Activity Evaluation Involving Hemoglobin-Related Free Radical Reactivity. In: Armstrong, D. (eds) Advanced Protocols in Oxidative Stress III. Methods in Molecular Biology, vol 1208. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1441-8_18
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DOI: https://doi.org/10.1007/978-1-4939-1441-8_18
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Publisher Name: Humana Press, New York, NY
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