Separation and quantitation of phospholipids and lysophospholipids by high-performance liquid chromatography

We describe a comprehensive approach to the separation, quantitation, and c haracterization of phospholipids and lysophospholipids present in complex b iological samples. The central feature is a normal-phase HPLC separation of individual phospholipid and lysophospholipid classes. In this single chrom atographic step, phospholipids and lysophospholipids are separated and reco vered for quantitation by organic phosphate assay and characterization by a cyl-group composition. Recovery of phospholipids and lysophospholipids from HPLC averages 80-90%. Isolated phospholipid and lysophospholipid fractions are available for separation of individual molecular species by second-dim ension reverse-phase HPLC and characterization of individual molecular spec ies by mass spectrometry. (C) 2000 Academic Press.