A Rapid Method For Phospholipid Class Separation by HPLC using an Evaporative Light-Scattering Detector

Abstract

Abstract A high-performance liquid chromatographic method for the separation and quantification of major natural phospholipid classes using an evaporative light-scattering detector is described. Separation and detection of cholesterol, palmitic acid, phosphatidylethanolamine, phosphatidylserine, phosphatidylcholine and sphingomyelin was achieved in less than 15 minutes using a Spherisorb 10 cm long silica column. A simplified ternary mobile phase gradient of hexane, isopropanol and water was used to separate these six standards reproducibly and easily. A crude sample of egg yolk was also analyzed using this method and found to contain mostly cholesterol with small amounts of phosphatidylethanolamine, phosphatidylcholine and sphingomyelin. The evaporative detector gave a stable flat baseline with detection limits for most compounds in the low nanogram range.