High-performance thin-layer chromatography/mass spectrometry for rapid analysis of neutral glycosphingolipids.
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.
Miyazaki
Masao
M
Tokai University, Institute of Glycoscience, Hiratsuka, Kanagawa 259-1292, Japan.
Yonesige
Azusa
A
Matsuda
Junko
J
Kuroda
Yasuhiro
Y
Kojima
Naoya
N
Suzuki
Akemi
A
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
D000468:Alkalies; D006028:Glycosphingolipids; D007202:Indicators and Reagents; D008055:Lipids