A microdetection system for glycosphingolipid analysis has been developed using 5-hydroxy-1-tetralone as the fluorescent labeling reagent. The reagents in H2SO4 permit the fluorometric detection of acidic and neutral glycosphingolipids both in test tube and on thin-layer chromatographic plates. Glycosphingolipids can be detected at concentrations as low as 5 pmol on the thin-layer chromatographic plate. The method is a rapid and simple, and feasible for determination of glycosphingolipid from small amounts of biological samples.