High-performance liquid chromatographic separation and photometric detection of phospholipids

Abstract

A rapid and efficient method for the separation of (phospho)lipids by high-performance liquid chromatography using n-hexane-2-propanol-water mixtures as the solvent system is described. The lipid separation occurs on silica gel columns and the individual components are monitored directly by UV absorption at 206 nm. Of a total lipid extract from erythrocytes as well as suboesophageal ganglia of the snail Helix pomatia, a complete separation is achieved of cholesterol, phosphatidic acid, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, lysophosphatidylcholine and lysophosphatidylethanolamine, whereas phosphatidylcholine and sphingomyelin are partly separated under these circumstances. In addition to separation of phospholipids in different classes, separation of molecular species can also be achieved in some instances, as is shown for phosphatidylcholines and sphingomyelins.