Da. Wiesner et Cc. Sweeley, CHARACTERIZATION OF GANGLIOSIDES BY 2-DIMENSIONAL HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHY, Analytica chimica acta, 311(1), 1995, pp. 57-62
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.