High-performance liquid chromatography of long-chain neutral glycosphingolipids and gangliosides

We describe a method for analyzing the perbenzoyl derivatives of both neutral glycosphingolipids and gangliosides with a single high-performance liquid chromatography system. Use of this system, combined with endo- and/or exoglycosidase treatment of glycosphingolipids, provides a sensitive method for obtaining structural information on these compounds. This system has two advantages over previously published chromatography procedures: (i) it uses a commercially available column, and (ii) this single column can be used to analyze gangliosides and their neutral glycosphingolipid products generated by neuraminidase treatment. With this method, we have studied 24 different glycosphingolipids, containing one to ten sugars and one or two sialic acid residues, and have demonstrated its usefulness in evaluating the gangliosides present in human leukocytes.