Separation of the dimethyl ester derivative of lysophosphatidic acid into component molecular species was performed. This approach permits assessment of the metabolism of species of a given lysophospholipid class in studies that employ in vivo labeling where fatty acid analysis by gas chromatography is not useful. The relative utility of argentation thin-layer chromatography compared to reversed-phase high-performance liquid chromatography in biologically relevant model systems is discussed. Separation of 1,acyl- from 2,acyl- isomers of methylated lysophosphatidic acid was accomplished as was the analysis of monoglycerides by enzymatic radioactive phosphorylation.