High-performance liquid chromatographic separation of gangliosides, combined with immunological detection and fast atom bombardment mass spectrometric characterization.
A complete strategy for the isolation of individual mono- and disialogangliosides has been elaborated. We have used straight-phase silica gel chromatography or partitioning to obtain a crude ganglioside fraction. This fraction was then peracetylated and run through a second silica gel column. After anion-exchange chromatography the gangliosides were separated by straight-phase high-performance liquid chromatography with chloroform-methanol-water mixtures as eluting solvents. The method is suitable for preparative isolation of gangliosides and subsequent structural characterization by thin-layer chromatography-enzyme-linked immunosorbent assay, fast atom bombardment mass spectrometry and/or gas chromatography-mass spectrometry, which is demonstrated by several examples, including the separation of GalNAc-II3NeuAc-GgOse4Cer from GalNAc-isoII3NeuAc-GgOse4Cer.
Gottfries
J
J
Department of Psychiatry and Neurochemistry, Gothenburg University, St. Jörgens Hospital, Sweden.
Davidsson
P
P
Månsson
J E
JE
Svennerholm
L
L
D000328:Adult; D000818:Animals; D001923:Brain Chemistry; D002528:Cerebellar Neoplasms / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D004912:Erythrocytes / Q000032:analysis; D005732:Gangliosides / Q000097:blood / Q000302:isolation & purification*; D006801:Humans; D007158:Immunologic Techniques; D007223:Infant; D013058:Mass Spectrometry / Q000379:methods; D008527:Medulloblastoma / Q000032:analysis; D051379:Mice; D008819:Mice, Nude
D005732:Gangliosides