Journal of Lipid Research, Vol 28, 1016-1018, Copyright © 1987 by Lipid Research, Inc.
Rapid separation of gram quantities of phospholipids from biological membranes by preparative high performance liquid chromatography
JS Ellingson and RL Zimmerman
Department of Pathology, Jefferson Medical College of Thomas Jefferson University, Philadelphia, PA 19107.
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.