Glycosphingolipid analysis by liquid chromatography-mass spectrometry

Abstract

Glycosphingolipids (GSLs) are an essential component of the microdomains that provide suitable microenvironments for membrane molecules and support their functions. The characterization of GSLs is the first step in the functional analysis of GSLs. The structures of the glycan chains and lipid parts of GSLs can be characterized by a sensitive method using liquid chromatography–mass spectrometry (LC-MS) and mixtures of GSLs. GSLs are minor components of the total lipids of tissues and cells. Therefore, sample preparation methods suitable for LC-MS analysis are critical. This chapter describes a protocol for the ganglioside analysis of mouse cerebellum, which consists of lipid extraction, solvent partition, and LC-MS analysis with C30 reversed-phase chromatography.