Gangliosides thin-layer chromatography

J. Muthing, High resolution thin-layer chromatography of gangliosides, J. Chromatogr. A., 720(1-2) (1996) 3-25. 

Glycolipid analysis. Gangliosides were extracted with chloroform-methanol (2 1), purified on Sephadex G-25 columns and separated into individual species by thin-layer chromatography as described previously (12). 

This problem was approached by incorporating radioactive percur-sors into the glycolipids of both brain and lymphoma cells of the Thy-1.2 and 1.1 types. We have used C-palmitate as a per-cursor of ceramide, and - C-N-acetylmannosamine as a percursor of sialic acid (7). Glycolipids were isolated and the radioactive gangliosides were resolved by two-dimensional thin layer chromatography in two different solvent systems followed by autoradiography. 

V. B. Ivleva, Y. N. Elkin, B. A. Budnik, S. C. Moyer, P. B. O Connor, and C. E. Costello, Coupling thin-layer chromatography with vibrational cooling matrix-assisted laser desorption/ ionization Fourier transform mass spectrometry for the analysis of ganglioside mixtures, Anal Chem., 76 (2004) 6484-6491. 

The homogeneity of glycolipids separated by the various procedures already described should be determined by thin-layer chromatography or HPTLC in several solvent systems. Mixtures of chloroform/methanol/water in different ratios, such as 60 35 8 v/v or 55 45 10 v/v as well as those with added calcium (chloroform/methanol/0.02% CaCl2.2H20, 60 40 9 v/v) or ammonia (chloroform/methanol/15M ammonia/water 60 35 1 7 v/v ) are particularly useful for the separation of gangliosides.Neutral and acetylated glycolipids are separated by chloroform/methanol/water (60 25 4 v/v) and 1,2-dichloromethane (DCE)/methanol/water (88 12 0.1 v/v) respectively. 

Some specific isolations are as follows. Hemato-side (Gms) can be obtained from rat liver by Folch partition, dialysis, Sephadex G-50 filtration and preparative thin-layer chromatography on silica gel G. Chloroform-methanol-2.5MNH4OH (60 35 8, by vol.) is used as solvent and the zones detected with iodine vapour or, better, with a non-destructive spray such as primuline (Skipski, 1975). Hematoside is the least polar of the gangliosides and has a Rf of about 0.5. It can be eluted from the silica gel with chloroform-methanol-water (60 35 8, by vol.). 

Ivleva, V.B., Sapp, L.M., O Connor, P.B., and Costello, C.E. (2005) Ganglioside analysis by thin-layer chromatography matrix-assisted laser desorption/ ionization orthogonal time-of-flight mass spectrometry. J. Am. Soc. Mass Spectrom., 16,1552-1550. 

A., Meisen, I., Vukelic, Z., Peter-Katalinic,)., Hfllenkamp, F., and Berkenkamp, S. (2005) Analysis of gangliosides directly from thin-layer chromatography plates by infrared matrix-assisted laser desorption/ ionization orthqional time-of-flight mass spectrometry with a glycerol matrix. Anal. Chem., 77, 4098-4107. 

Wiesner, D. A., and Sweeley, C. C. (1995). Characterization of gangliosides by two-dimensional high-performance thin-layer chromatography. Anal. Chim. Acta 311 57-62. 

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