Sialic acid thin-layer chromatography

Thin-layer Chromatography (Rf values) of Natural,. V,0-Acylated Sialic Acids and of the C, and C8 Analogs of Neu5Ac and Neu5Gc in System A" and in Gas-Liquid Chromatography (fiT Values, Relative to Neu5Ac = 1.00) in Systems B and C ... 

Thin-layer chromatography of 2-acylamido-2-deoxy-D-mannoses resulting from cleavage of sialic acids with acylneuraininate pyruvate-lyase is described in Ref. 107, together with the RF values of the different 0-acyl-2-acylamido-2-deoxy-D-mannose species. 

Table 4.3.2 Diagnostic workflow of disorders with accumulation of free or oligosaccharide-bound sialic acid. MRI Magnetic resonance imaging, TLC thin-layer chromatography...

Other Analytical Procedures. Thin layer chromatography was carried out with silica gel plates G60 (Merck), using either chloroform-methanol-water (65 5 9) or n-propanol-0.2% CaCl2 in H20 (80 20). The plates were developed with resorcinol spray (15). Sialic acid was determined with thiobarbituric acid after hydrolysis in 0.1 sulfuric acid (16). 

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. 

Acylneuraminic acids can be prepared by preparative thin-layer chromatography. Details of various systems are given in chapter E. Localization of the sialic acids on the thin-layer plate can for instance be achieved using a radioactive Neu5Ac sample and the position of other sialic acids calculated from Revalues and eluted from the support with water after scraping from the plate. Large amounts cannot be prepared in this way, but the method is especially valuable in the preparation of radioactive acylneuraminic acids from metabolic studies. 

E. Colorimetry and Thin-Layer Chromatography of Sialic Acids... 

Quantitative estimation of the O-acyl content of sialic acid preparations can be made using the hydroxamic acid reaction as described by Ludowieg and Dorfman (1960), modified from the method of Hestrin (1949). Calibration with ethyl acetate or other ester standards provides a simple quantitative assay, measuring the chromophore at 520 nm. Volume reductions in the assay procedure allow quantitation of 0.05(i,mole of sialic acid acyl ester (Schauer 1978). Qualitative identification of different acylhydroxamates can be made on thin-layer chromatography (see section III.3.b)). 

A further use of two-dimensional thin-layer chromatography for the identification of sialic acids in oligosaccharide fractions has been demonstrated... 

Schauer, R., and Corfield, A. P., 1982, Colorimetry and thin-layer chromatography of sialic acids, in Sialic Acids—Chemistry, Metabolism and Function (R. Schauer, ed.). Springer, New York, pp. 77-94. 

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