Rapid assessment of singlet oxygen-induced plasma lipid oxidation and its inhibition by antioxidants with diphenyl-1-pyrenylphosphine (DPPP)
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Recent studies suggesting the involvement of singlet oxygen in the pathogenesis of multiple diseases have attracted renewed attention to lipid oxidation mediated by singlet oxygen. Although the rate constants for singlet oxygen quenching by antioxidants have been measured extensively, the inhibition of lipid oxidation mediated by singlet oxygen has received relatively less attention, partly because a convenient method for measuring the rate of lipid oxidation is not available. The objective of this study was to develop a convenient method to measure plasma lipid oxidation mediated by singlet oxygen which may be applied to a rapid assessment of the antioxidant capacity to inhibit this oxidation using a conventional microplate reader. Singlet oxygen was produced from naphthalene endoperoxide, and lipid hydroperoxide production was followed by using diphenyl-1-pyrenylphosphine (DPPP). Non-fluorescent DPPP reacts stoichiometrically with lipid hydroperoxides to give highly fluorescent DPPP oxide. It was found that plasma oxidation by singlet oxygen increased the fluorescence intensity of DPPP oxide, which was suppressed by antioxidants. Fucoxanthin suppressed the oxidation more efficiently than β-carotene and α-tocopherol, while ascorbic acid and Trolox were not effective. The present method may be useful for monitoring lipid oxidation and also for rapid screening of the capacity of dietary antioxidants and natural products to inhibit lipid oxidation in a biologically relevant system.
KeywordsAntioxidant Carotenoid Diphenyl-1-pyrenylphosphine Plasma lipid oxidation Singlet oxygen
The kind gift of α-tocopherol from Tama Biochemical Co. Ltd. is highly acknowledged.
Compliance with ethical standards
The animal experiments and care were approved by the Institutional Animal Care and Use Committee of Kyoto Prefectural University of Medicine, approved on March 31, 2015, as No. M25-163.
Conflict of interest
The authors declare that they have no competing interests.
- 2.Foote CS (1976) Photosensitized oxidation and singlet oxygen: consequences in biological systems. In: Pryor WA (ed) Free radicals in biology, vol. 2. Academic, New York, pp 85–133Google Scholar
- 10.Umeno A, Shichiri M, Ishida N, Hashimoto Y, Abe K, Kataoka M, Yoshino K, Hagihara Y, Aki N, Funaki M, Asada Y, Yoshida Y (2013) Singlet oxygen induced products of linoleates, 10- and 12-(Z, E)-hydroxyoctadecadienoic acids (HODE), can be potential biomarkers for early detection of type 2 diabetes. PLoS One 8, e63542CrossRefGoogle Scholar
- 16.Di Mascio P, Murphy ME, Sies H (1991) Antioxidant defense systems: the role of carotenoids, tocopherols, and thiols. Am J Clin Nutr 53:194S–200SGoogle Scholar
- 17.Ouchi A, Aizawa K, Iwasaki Y, Inakuma T, Terao J, Nagaoka S, Mukai K (2010) Kinetic study of the quenching reaction of singlet oxygen by carotenoids and food extracts in solution. Development of a singlet oxygen absorption capacity (SOAC) assay method. J Agric Food Chem 58:9967–9978CrossRefGoogle Scholar
- 21.Wagner JR, Motchnik PA, Stocker R, Sies H, Ames BN (1993) The oxidation of blood plasma and low density lipoprotein components by chemically generated singlet oxygen. J Biol Chem 268:18502–18506Google Scholar
- 35.Sugawara T, Baskaran V, Tsuzuki W, Nagao A (2002) Brown algae fucoxanthin is hydrolyzed to fucoxanthinol during absorption by Caco-2 human intestinal cells and mice. J Nutr 132:946–951Google Scholar