High-performance liquid chromatographic separation of gangliosides, combined with immunological detection and fast atom bombardment mass spectrometric characterization |
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| Authors: | J Gottfries P Davidsson J E M?nsson L Svennerholm |
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| Affiliation: | Department of Psychiatry and Neurochemistry, Gothenburg University, St. J?rgens Hospital, Sweden. |
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| Abstract: | 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. |
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