A NEW-TYPE OF 2-DIMENSIONAL THIN-LAYER CHROMATOGRAPHY (TLC MAPPING) FOR ANALYSIS OF ACIDIC GLYCOSPHINGOLIPID MOLECULAR-SPECIES

A new approach to the analysis of acidic glycosphingolipid molecular s pecies has been developed by using two-dimensional thin-layer chromato graphy: TLC mapping. TLC mapping was accomplished by use of two types of high-performance thin-layer chromatographic (HPTLC) plates. A NH2-s ilica gel HPTLC plate developed with chloroform:methanol: 1% diethylam ine (50:47:15) was used for the first-dimension chromatography. Acidic glycosphingolipids on the plate were transferred to a normal silica g el HPTLC plate by developing the plate with chloroform:methanol:ammoni a (4:10:6) after the NH2 plate was placed face to face on top of the s ilica gel HPTLC plate. Acidic glycosphingolipids on the HPTLC plate we re then developed with chloroform: methanol:0.2% CaCl2 (60:40:10) for the second-dimension chromatography. The use of the TLC-mapping method allowed us to compare detailed membrane constituents, including minor molecular species with regard to acidic glycosphingolipids. The metho d is rapid and simple and is feasible for determining acidic glycosphi ngolipid from biological samples. (C) 1995 Academic Press, Inc.