Microscale Analysis of Glycosphingolipids from Schistosoma mansoni Cercariae

We have developed a microscale approach for structural analysis of glycosphingolipids. The protocol is based on enzymatic digestion by sphingolipid ceramide N-deacylase in a two-phase liquid system. Resulting lyso-glycosphingolipids were recovered from the aqueous phase and analyzed by mass spectrometry, whereas released fatty acids were isolated from organic and aqueous phases and identified as methyl ester derivatives. Application of this methodology to distinct glycolipid fractions obtained from the cercarial life-cycle stage of the human parasitic helminth Schistosoma mansoni revealed clear differences in the ceramide compositions of ceramide monohexoside versus ceramide pentahexoside species carrying a Lewis X carbohydrate epitope.