ENHANCED THIN-LAYER CHROMATOGRAPHIC-SEPARATION OF G(M1B)-TYPE GANGLIOSIDES BY AUTOMATED MULTIPLE DEVELOPMENT

Enhancement in separation of gangliosides on silica gel precoated high -performance TLC plates has been obtained by automated multiple develo pment chromatography. A less polar mixture of the standard solvent chl oroform-methanol-20 mM aqueous CaCl2 (120:85:20, v/v) was used. Loweri ng the water content achieved separation of two complex monosialogangl ioside fractions, isolated from murine YAC-1 T lymphoma and MDAY-D2 ly mphoreticular cells. Three-fold chromatography in the solvent chlorofo rm-methanol-20 mM aqueous CaCl2 (120:85:14, v/v) resulted in TLC separ ation of G(MIb)-type gangliosides, substituted with C-24 and C-16 fatt y acids and with Neu5Ac and Neu5Gc as well, which could not be achieve d by unidirectional standard chromatography. Compared to conventional single chromatography, the technique described allows high-resolution separation of extremely heterogenous ganglioside mixtures and offers a convenient tool for both analytical and preparative TLC.