Gangliosides were isolated from bovine adrenal medulla by DEAE-Sephadex A-25 and Iatrobeads column chromatography. The disialo fraction contained five major gangliosides, A to E; together they accounted for 4.6% of the total lipid-bound sialic acid. On the basis of sugar analysis, permethylation, mild acid hydrolysis and neuraminidase degradation studies, the structures of these five gangliosides are characterized as follows: (formula: see text). The major fatty acids of the disialogangliosides were C16:0, C18:0, C22:0, C23:0, C24:0, and C24:1, which were characteristic of extraneural gangliosides.