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A more recent version of this article appeared on November 1, 2005

Papers In Press, published online ahead of print September 8, 2005
J. Lipid Res., doi:10.1194/jlr.D500025-JLR200
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Submitted on July 15, 2005
Revised on August 9, 2005
Accepted on August 17, 2005

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, and Teruo Miyazawa

Gratuate School of Agricultural Science, Tohoku University, Sendai 981-8555

Corresponding Author: miyazawa{at}biochem.tohoku.ac.jp

Peroxidized phospolipids-mediated cytotoxity involves 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 (deoxy-D-fructosyl PE, namely Amadori-PE), since Amadori-PE cause oxidative stress. However, an occurrence of lipid glycation products including Amadori-PE in vivo is still unclear. Consequently, we developed an analysis method of Amadori-PE using quadrupole linear ion trap mass spectrometer, Applied Biosystems QTRAP. Under positive ion mode, collision-induced dissociation of Amadori-PE produced a well-characterized diglyceride ion ([M+H-303]+) permitting neutral loss scan and multiple reaction monitoring. When lipid extract from diabetic plasma was directly infused to QTRAP, Amadori-PE molecular species could be screened out by a neutral loss scan. Interfacing liquid chromatography with QTRAP enabled the separation and determination of predominant plasma Amadori-PE species with sensitivity around 0.1 picomole/injection in multiple reaction monitoring. 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 showed the involvement between lipid glycation and lipid peroxidation in diabetes pathogenesis.


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