Rapid separation of gram quantities of phospholipids from biological membranes by preparative high performance liquid chromatography

Abstract

We present a detailed description of a rapid preparative high performance liquid chromatography (HPLC) procedure for isolation of large scale quantities (a minimum of 200 mg) of rat liver microsomal and mitochondrial phospholipid classes in 40 min on a 25 X 2.5 cm preparative HPLC column of 7-mu silica gel particles using a linear gradient of hexane-isopropanol-water mixtures. A minimum of 1.5 g of phospholipids can be quantitatively separated per day into diphosphatidylglycerol (cardiolipin), phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, sphingomyelin, and a phosphatidylcholine-sphingomyelin fraction. The procedure uses no salts, buffers, acids, or bases, and the isolated phospholipids are suitable for preparing model membranes.