Preparation of pure lipid hydroperoxides

Daigo Ibusuki, Kiyotaka Nakagawa, Akira Asai, Shinichi Oikawa, Yuichi Masuda, Toshihide Suzuki, Teruo Miyazawa

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Increasing evidence of lipid peroxidation in food deterioration and pathophysiology of diseases have revealed the need for a pure lipid hydroperoxide (LOOH) reference as an authentic standard for quantification and as a compound for biological studies in this field. Generally, LOOH is prepared from photo- or enzymatically oxidized lipids; however, separating LOOH from other oxidation products and preparing pure LOOH is difficult. Early studies showed the usability of reaction between hydroperoxide and vinyl ether for preparation of pure LOOH. Because the reactivity of vinyl ether with LOOHs other than fatty acid hydroperoxides has never been reported, here, we employed the reaction for preparation of a wide variety of pure LOOHs. Phospholipid, cholesteryl ester, triacylglycerol, or fatty acid was photo- or enzymatically oxidized; the resultant crude sample containing hydroperoxide was allowed to react with a vinyl ether [2-methoxypropene (MxP)]. Liquid chromatography (LC) and mass spectrometry confirmed that MxP selectively reacts with LOOH, yielding a stable MxP adduct (perketal). The lipophilic perketal was eluted at a position away from that of intact LOOH and identified and isolated by LC. Upon treatment with acid, perketal released the original LOOH, which was finally purified by LC. Using our optimized purification procedures, for instance, we produced 75 mg of pure phosphatidylcholine hydroperoxide (>99%) from 100 mg of phosphatidylcholine. Our developed method expands the concept of the perketal method, which provides pure LOOH references. The LOOHs prepared by the perketal method would be used as "gold standards" in LOOH methodology.

Original languageEnglish
Pages (from-to)2668-2677
Number of pages10
JournalJournal of lipid research
Volume49
Issue number12
DOIs
Publication statusPublished - 2008

Keywords

  • 2-methoxypropene
  • Lipid hydroperoxide standard
  • Lipid peroxidation
  • Oxidative stress

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

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