Simultaneous Analysis of Eight Phospholipid Classes by Liquid Chromatography/Mass Spectrometry: Application to Human HDL

Abstract

We developed a method to separate and simultaneously quantitate eight phospholipid classes, phosphatidylglycerol (PG), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidic acid (PA), phosphatidylcholine (PC), sphingomyelin (SM), and lysophosphatidylcholine (LPC). A chloroform/methanol/1‐propanol/ammonium hydroxide/water solvent system was used as the HPLC mobile phase. This condition was set to fulfill a fit in a calibration curve and adequate separation of phospholipids. We evaluated our analytical method for precision, accuracy, and recovery of the phospholipids for human high‐density lipoprotein (HDL). The precision, accuracy, and recovery ranged from 1.4 to 22.6%, 26.9 to +30.4%, and 56.0 to 99.3%, respectively.

Keywords:

• Phospholipid
• Simultaneous analysis
• Human HDL

Acknowledgment

We thank Ms. K. Nakano, Mr. K. Yamada, Dr. S. Yamamoto, Dr. T. Ono, and Dr. K. Hanasaki for the preparation of the human HDL samples.