Simple micro-method for the isolation of gangliosides by reversed-phase chromatography

A simple and convenient technique has been developed for the isolation of gangliosides from small amounts of tissues or cells. A ganglioside fraction obtained by chromatography of the total lipid extract of DEAE-Sephadex was subjected to alkaline hydrolysis and salts and other non-lipid contaminants were removed by reversed-phase chromatography on a C₁₈ Sep-Pak cartridge. The purified gangliosides were then obtained by chromatography on a small Iatrobeads or Unisil column. This procedure yields a quantitative recovery of gangliosides that are free of contaminants which interfere with thin-layer chromatographic analysis. The procedure was used for the quantitative isolation of gangliosides from human brain white matter and human erythrocytes.