Preparation of Lysohematoside (Neuraminyl-galactosyl-glucosylsphingosine) from Hematoside of Equine Erythrocyte and Its Chemical and Hemolytic Properties

Abstract

Deacylated hematoside prepared from hematoside of equine erythrocyte by a partial alkaline hydrolysis was purified by Silica Gel H-Hyflo Super Cel column chromatog-raphy and its homogeneity was detected by a thin layer chromatogram showing one single spot. The chemical structure of the deacylated hematoside was found to be neuraminyl (2→3) galactosyl (1→4) glucosylsphingosine by the following procedures: chemical analysis of each component of the sample, gas-liquid chromatography of methyl glycosides obtained after methanolysis of the permethylated one and formation of lactosyl sphingosine from the sample by mild acid hydrolysis releasing a neuraminic acid. Hemolytic activity of the deacylated hematoside was about twice stronger than that of psychosine, sphingosylphosphorylcholine, lysolecithin et al. as a calculated L₅₀ value per red cell. Thus the deacylated hematoside may be reasonably called “lysohematoside” belonging to the so called “lysosphingolipids.” It was found that the lysosphingolipids including the lysohematoside did not produce any O-methyl-sphingosine as artifact by anhydrous methanolysis contrary to sphingolipids. A hypothetical mechanism of the formation of O-methyl-sphingosine from sphingolipids by the anhydrous methanolysis was suggested.