title: High-performance thin-layer chromatography/mass spectrometry for rapid analysis of neutral glycosphingolipids.
abstract: Direct coupling of high-performance thin-layer chromatography (HPTLC) to matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry (MS) was shown to be a reliable and reproducible method to obtain structural information and fundamental properties of glycosphingolipids (GSLs). We report a protocol for the preparation of neutral GSL extracts from mouse tissues and demonstrate the applicability of HPTLC/MS to these preparations. The protocol consists of lipid extraction and ion exchange chromatography followed by a mild alkaline treatment and a reversed-phase cartridge extraction. Possible structures for each GSL are proposed based on HPTLC/MS analyses. This fast and simple method can be used to screen neutral GSL extracts obtained from tissues and cells without isolation and purification into individual GSLs.
authors.affiliation: Tokai University, Institute of Glycoscience, Hiratsuka, Kanagawa 259-1292, Japan.
authors.forename: Masao
authors.initials: M
authors.lastname: Miyazaki
authors.forename: Azusa
authors.initials: A
authors.lastname: Yonesige
authors.forename: Junko
authors.initials: J
authors.lastname: Matsuda
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Kuroda
authors.forename: Naoya
authors.initials: N
authors.lastname: Kojima
authors.forename: Akemi
authors.initials: A
authors.lastname: Suzuki
mesh_terms: D000468:Alkalies; D000818:Animals; D002852:Chromatography, Ion Exchange; D002855:Chromatography, Thin Layer; D003720:Densitometry; D006028:Glycosphingolipids / Q000032:analysis* / Q000378:metabolism; D007202:Indicators and Reagents; D008055:Lipids / Q000737:chemistry / Q000302:isolation & purification; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D014018:Tissue Distribution
chemical_list: D000468:Alkalies; D006028:Glycosphingolipids; D007202:Indicators and Reagents; D008055:Lipids