One-step fractionation of neutral and acidic glycosphingolipids by high-performance liquid chromatography.
A procedure for a simultaneous separation of ganglioside components and neutral glycolipid components by high-performance liquid chromatography was described. One column packed with DEAE-derivatized controlled-pore glass (DEAE-CPG) was serially connected to two columns of underivatized, controlled-pore glass (CPG). A mixture of gangliosides and neutral glycolipids were loaded on DEAE-CPG and eluted with a mixture of chloroform-methanol-water, with increasing methanol and water (the first-phase gradient elution), followed by elution with increasing concentrations lithium acetate from 0.015 to 0.1 M in a mixture of chloroform-methanol-water (the second-phase gradient elution). Neutral glycolipids, mono- to hexaglycosylceramides , were separated within 80 min of the first-phase gradient elution, and mono- to tetrasialosylgangliosides were separated during the second-phase gradient elution within 60 min. The method has been applied to the determination of glycolipids isolated from rat tissues, and the procedure was found to be highly reproducible.
D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D005732:Gangliosides / Q000302:isolation & purification*; D006028:Glycosphingolipids / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007668:Kidney / Q000201:enzymology; D008297:Male; D051381:Rats; D012997:Solvents
D005732:Gangliosides; D006028:Glycosphingolipids; D012997:Solvents