Direct Thin-Layer Chromatographic Method for Isolation of Gangliosides

A simple procedure is developed for obtaining a ganglioside mixture. The scheme of the method is as follows: the sample was homogenized and extracted twice with cyclohexane (first extraction). The residue was reextracted with a mixture of chloroform /methanol = 1:3 (v/v) (second extraction). The extracts were collected after solvent evaporation. The residue was dissolved and applied onto a thin layer of silica gel according to the requirements of the preparative thin-layer chromatography. After the zone separation the visualized spots were scrapped and transferred to a column and eluted with a mixture of chloroform/methanol with increasing polarity. The eluate was concentrated and controlled for purity and composition by HPTLC. This procedure has been successfully applied to fractionation of ganglioside mixtures from human, calf and rat brains.