A high-efficiency silica gel, (type HLF) thin-layer chromatography plate (HETLC), linear high-performance thin-layer chromatography plate (HPTLC) and densitometry method has been devised in order to resolve the major lipid classes obtained from rat brain tissues. This methodology, which has largely overcome prior problems, enhances the opportunity for assessing the glycerophospholipid and glycolipid compositions of tissues. DEAE-Sephadex column chromatography was used to separate the crude lipids extract into neutral and acidic lipid fractions. The lipid fractions were then spotted on separate HPTLC and HETLC plates and chromatographed in one dimension using one solvent system. Quantitation was by in situ densitometry with the absolute quantity of the lipid classes determined from co-chromatographed standards. Sensitivity was increased by using cupric sulfate reagent, which was found to be more sensitive than the conventional cupric acetate reagent. This method is applicable to a broader separation of lipid classes and has improved sensitivity.