Analytical and Bioanalytical Chemistry

, Volume 408, Issue 1, pp 265–270 | Cite as

Rapid assessment of singlet oxygen-induced plasma lipid oxidation and its inhibition by antioxidants with diphenyl-1-pyrenylphosphine (DPPP)

  • Mayuko Morita
  • Yuji Naito
  • Toshikazu Yoshikawa
  • Etsuo NikiEmail author
Research Paper


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.


Antioxidant 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.


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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Mayuko Morita
    • 1
    • 2
  • Yuji Naito
    • 1
  • Toshikazu Yoshikawa
    • 2
  • Etsuo Niki
    • 1
    • 3
    Email author
  1. 1.Department of Molecular Gastroenterology and HepatologyKyoto Prefectural University of MedicineKyotoJapan
  2. 2.Department of Gastrointestinal ImmunologyKyoto Prefectural University of MedicineKyotoJapan
  3. 3.Health Research InstituteNational Institute of Advanced Industrial Science and TechnologyTakamatsuJapan

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