Analysis of complex mixtures of phospholipid classes from cell membranes using two-dimensional thin-layer...

Increasing recognition of the important roles served by membrane phospholipids in cellular metabolic and signal transduction processes has stimulated interest in examining potential phospholipid abnormalities in patients with psychiatric disorders. This report describes a method, based on several novel modifications of existing techniques, for concurrently analyzing nanomolar amounts of nine phospholipid classes in a single aliquot of membrane extract. With this method, diverse phospholipid classes are first separated by two-dimensional thin-layer chromatography, and then determined using two-dimensional scanning laser densitometry. The method is able to quantitate even small amounts of specific phospholipid classes, corresponding to < 10 ng of lipid phosphorus. The sensitivity of this method allows it to be readily applied to clinical studies involving membranes from cell types that are obtainable only in small quantities.