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

J Lipid Res. 2010 Aug;51(8):2445-53. doi: 10.1194/jlr.D004564. Epub 2010 Apr 12.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood Chemical Analysis / methods*
  • Case-Control Studies
  • Chromatography, Liquid / methods*
  • Diabetes Mellitus / blood
  • Diabetes Mellitus / metabolism
  • Erythrocytes / chemistry*
  • Female
  • Glycation End Products, Advanced / blood
  • Glycation End Products, Advanced / metabolism
  • Humans
  • Hyperglycemia / blood
  • Hyperglycemia / metabolism
  • Male
  • Middle Aged
  • Phosphatidylethanolamines / blood*
  • Phosphatidylethanolamines / metabolism*
  • Tandem Mass Spectrometry / methods*
  • Young Adult


  • Glycation End Products, Advanced
  • Phosphatidylethanolamines
  • phosphatidylethanolamine