Skeletal muscle gangliosides prepared from ten species of animals (human, monkey, bovine, hog, dog, cat, rabbit, guinea pig, mouse, and chicken) were studied by two-dimensional thin-layer chromatography. Densitometric quantification of spots on the chromatograms was carried out with an image analysis system equipped with a computer system. In about thirty-five resorcinol-positive spots, sixteen components could be identified by comparison with authentic standards we had previously obtained. G_M3 ganglioside was found as a major component in all examined animals, but there were remarkable species variations in the minor gangliosides. In bovine and hog muscle gangliosides, more than fifteen minor components were detected owing to the presence of N-glycolyl neuraminic acid species. In contrast to this, muscle gangliosides of human, dog, cat, rabbit, and chicken showed relatively simple patterns because of the absence or the small amounts of N-glycolyl type gangliosides. The presence of lactosamine containing gangliosides was characteristic of muscle gangliosides of the examined species.