An improved method for the separation of molecular species of cerebrosides

A method employing reversed-phase high performance liquid chromatography (HPLC) has been developed for the analysis of molecular species of underivatized cerebrosides. By using a high carbon (20%) C18-silica bead column, monohexosylceramides isolated from bovine brain and Japanese quail intestine could each be separated into 42 and 56 peaks, respectively, and could subsequently be recovered. Under our analytical conditions, underivatized cerebrosides with 24:1 and 22:0 fatty acids were completely separated by single step HPLC. Each of the molecular species was then analyzed by high performance thin layer chromatography and gas chromatography-mass spectrometry (or fast atom bombardment mass spectrometry). This analytical method permits the complete identification of molecular species of glycosphingolipids.