Improved procedure for the separation of phospholipids by high-performance liquid chromatography.

We describe a rapid and efficient high-performance liquid chromatography procedure for the separation of phospholipids. The separation is accomplished on a microparticulate silica gel column using isocratic elution and UV detection at 203 nm. The solvent mixture is acetonitrile--methanol--85% phosphoric acid(130:5:1.5, v/v). Complete separation is achieved within 30 min of phosphatidylinositol, phosphatidylserine, phosphatidylethanolamine, phosphatidyldimethylethanolamine, lysophosphatidylethanolamine, phosphatidylcholine, lysophosphatidylcholine and sphingomyelin. The method is suitable for the analysis of phospholipids in tissue extracts.