Ion-trap tandem mass spectrometric analysis of Amadori-glycated phosphatidylethanolamine in human plasma with or without diabetes

Kiyotaka Nakagawa, Jeong Ho Oak, Ohki Higuchi, Tsuyoshi Tsuzuki, Shinichi Oikawa, Haruhisa Otani, Masatoshi Mune, Hua Cai, Teruo Miyazawa

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

Peroxidized phospholipid-mediated cytotoxicity is involved in the pathophysiology of diseases [i.e., an abnormal increase of phosphatidylcholine hydroperoxide (PCOOH) in plasma of type 2 diabetic patients]. The PCOOH accumulation may relate to Amadori-glycated phosphatidylethanolamine (Amadori-PE; deoxy-D-fructosyl phosphatidylethanolamine), because Amadori-PE causes oxidative stress. However, the occurrence of lipid glycation products, including Amadori-PE, in vivo is still unclear. Consequently, we developed an analysis method of Amadori-PE using a quadrupole/linear ion-trap mass spectrometer, the Applied Biosystems QTRAP. In positive ion mode, collision-induced dissociation of Amadori-PE produced a well-characterized diglyceride ion ([M+H-303]+) permitting neutral loss scanning and multiple reaction monitoring (MRM). When lipid extract from diabetic plasma was infused directly into the QTRAP, Amadori-PE molecular species could be screened out by neutral loss scanning. Interfacing liquid chromatography with QTRAP mass spectrometry enabled the separation and determination of predominant plasma Amadori-PE species with sensitivity of ∼0.1 pmol/injection in MRM. The plasma Amadori-PE level was 0.08 mol% of total PE in healthy subjects and 0.15-0.29 mol% in diabetic patients. Furthermore, plasma Amadori-PE levels were positively correlated with PCOOH (a maker for oxidative stress). These results show the involvement between lipid glycation and lipid peroxidation in diabetes pathogenesis.

Original languageEnglish
Pages (from-to)2514-2524
Number of pages11
JournalJournal of lipid research
Volume46
Issue number11
DOIs
Publication statusPublished - 2005 Nov 1

Keywords

  • Glycation
  • Lipid peroxidation
  • QTRAP mass spectrometer

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

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