LC-MS/MS analysis of carboxymethylated and carboxyethylated phosphatidylethanolamines in human erythrocytes and blood plasma[S]

• Published in: Journal of lipid research Vol. 51; no. 8; pp. 2445 - 2453
• Main Authors: Shoji, Naoki, Nakagawa, Kiyotaka, Asai, Akira, Fujita, Ikuko, Hashiura, Aya, Nakajima, Yasushi, Oikawa, Shinichi, Miyazawa, Teruo
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.08.2010; American Society for Biochemistry and Molecular Biology; The American Society for Biochemistry and Molecular Biology; Elsevier
• Subjects: advanced glycation end products; Blood Chemical Analysis - methods; Case-Control Studies; Chromatography, Liquid - methods; diabetes; Diabetes Mellitus - blood; Diabetes Mellitus - metabolism; Erythrocytes - chemistry; Female; Glycation End Products, Advanced - blood; Glycation End Products, Advanced - metabolism; Humans; Hyperglycemia - blood; Hyperglycemia - metabolism; lipid glycation; Male; Methods; Middle Aged; Phosphatidylethanolamines - blood; Phosphatidylethanolamines - metabolism; tandem mass spectrometry; Tandem Mass Spectrometry - methods; Young Adult; tandem mass spectrometry; advanced glycation end products; diabetes; lipid glycation
• ISSN: 0022-2275; 1539-7262; 1539-7262
• DOI: 10.1194/jlr.D004564

An amino group of phosphatidylethanolamine (PE) is considered as a target for nonenzymatic glycation, and the potential involvement of lipid glycation in the pathogenesis of diabetic complications has generated interest. However, unlike an early glycation product of PE (Amadori-PE), the occurrence and roles of advanced glycation end products of PE (AGE-PE) in vivo have been unclear. Here, we developed an LC-MS/MS method for the analysis of AGE-PE [carboxymethyl-PE (CM-PE) and carboxyethyl-PE (CE-PE)]. Collision-induced dissociation of CM-PE and CE-PE produced characteristic ions, permitting neutral loss scanning (NLS) and multiple reaction monitoring (MRM) of AGE-PE. By NLS analysis, a series of AGE-PE molecular species was detected in human erythrocytes and blood plasma. In LC-MS/MS analysis, MRM enabled the separation and determination of the predominant AGE-PE species. Between healthy subjects and diabetic patients, no significant differences were observed in AGE-PE concentrations in erythrocytes and plasma, whereas Amadori-PE concentrations were higher in diabetic patients. These results provide direct evidence for the presence of AGE-PE in human blood, and indicated that, compared with Amadori-PE, AGE-PE is less likely to be accumulated in diabetic blood. The presently developed LC-MS/MS method appears to be a powerful tool for understanding in vivo lipid glycation and its pathophysiological consequence.