The separation of neutral glycosphingolipids from mammalian erythrocytes by droplet counter-current chromatography (DCC)

Abstract

In a previous paper (Otsuka, H. & Yamakawa, T. (1981) J. Biochem. 90, 247-254), we reported the separation of acidic glycolipids by droplet counter-current chromatography using 500 columns with a commercially available DCC apparatus and described the precise conditions of the separation. In this paper, separation of neutral glycolipids from rabbit and human erythrocytes by DCC is described. For efficient separation of neutral glycolipids, addition of a definite amount of benzene was required. The solvent system of chloroform: benzene: methanol: water = 50: 50: 70: 20 gave clear separation of these glycolipids and further modification of this solvent system to chloroform: benzene: methanol: water = 50: 25: 65: 30 gave satisfactory results for the separation of Globoside I from human erythrocytes due to the difference between the amide linked fatty acids. Also under the same conditions, the clear separation of two peaks with blood group A activity was demonstrated by a hemagglutination inhibition test.