Pre-packed reverse phase columns for isolation of complex lipids synthesized from radioactive precursors.

Abstract

Pre-packed reverse phase columns (Bond Elut) were used for the separation of complex lipids, such as phosphatidylcholine, cerebrosides, sulfatides, and gangliosides, from their respective water-soluble radioactive precursors after their in vitro biosynthesis. After an incubation in vitro, the entire reaction mixture is passed through the column, where complex lipids are retained and the hydrophilic radioactive precursors are washed away from the column. The retained lipids are then eluted with a more nonpolar organic solvent. The procedure is shown to be simpler and more efficient than the normally used Folch partitioning method or other procedures.