CHARACTERIZATION OF GANGLIOSIDES BY 2-DIMENSIONAL HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHY

A method for the qualitative and quantitative analysis of complex mixt ures of gangliosides by computer-assisted image densitometry is report ed. Gangliosides were applied to silica gel high-performance thin-laye r plates and developed in two dimensions using one of several solvent systems. The plates were developed with a resorcinol reagent and the g angliosides were visualized as bluish-purple bands or spots. The plate s were scanned with a BioImage Visage 110 image analyzer and saved to disk. The densities of the spots were determined and the quantities of gangliosides determined by comparison with an internal standard, GM(4 ). Two-dimensional chromatography using chloroform-methanol-0.2% CaCl2 (50:40:10) in the first dimension and n-propanol-ammonia(aq.) (6:3) i n the second dimension gave linear and reproducible results when analy zed by image densitometry. Gangliosides isolated from human plasma wer e resolved with the above method. We suggest that the use of image den sitometry improves the accuracy of quantitative analyses of complex mi xtures of gangliosides by thin-layer chromatography.