Chromatography of sialic acids

Wang, W., et al. (1990). High-performance Liquid Chromatography of Sialic Acid-containing Oligosaccharides and Acidic Monosaccharides, . dna/. Biochem. 190 182—187. 

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

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. 

Walkowiak, H., Kedzierska, B., and Starzynski, W., 1%8, Horizontal circular chromatography of sialic acids from serum of certain mammal species. Bull. Acad. Pol. Sci., Ser. Sci. Biol. 16 97. 

Hydrophilic interaction chromatography on Asahipak NH2P or Excel-pak CHA-P44 with pulsed amperometric detection has been used to fractionate malto-oligosaccharides.266 The Asahipak NH2P is a polyvinyl alcohol support with a polyamine bonded phase, and the Excelpak is a sulfonated polystyrene in the Zn+2 form. Amine adsorption of sialic acid-containing oligosaccharides was performed on a Micropak AX-5 column (Varian) using acetonitrile-water-acetic acid-triethylamine.267... 

Naturally occurring oligosaccharides have also been isolated by gel chromatography. Ohman and Hygstedt94 have reported the isolation, from colostrum, of sialic acid oligosaccharides, including di-O-sialoyl-lactose [0-A/-acetylneuraminoyl-(2 — 8)-0-(N-acetylneur-... 

Despite treatment by ion-exchange chromatography, the sialic acid preparation may still contain such impurities as proteins, peptides, or other ionic compounds that may interfere with further analytical or preparative procedures. In these cases, chromatography of the preparation on Sephadex G-10, G-25, or Biogel P2 in water may prove helpful. 

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. 

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. 

Mild acid and neuraminidase treatment and DEAE cellulose chromatography were used to further characterize the Thy-1 active compounds (Table I, previous page). Neuraminidase treatment and mild acid conditions, which result in the removal of sialic acid, destroyed the anti-Thy-1 PFC response to these antigens. Furthermore, both Thy-1.1 and Thy-1.2 glycolipids bound to DEAE cellulose, confirming their acidic nature. 

FIGURE 29.2 High-performance liquid chromatography traces showing the change in concentration of sialic acid (27a), ketodeoxynonulosonic acid (KDN) (27b), and ketodeoxyoctulosonic acid (KDO) (27c) over time for a control incubation. 

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