High-Performance Liquid Chromatographic Determination of Xylitole and Hexosaminitols Present in the Reduced Terminal of Glycosaminoglycans

Abstract

A reversed phase isocratic high-performance liquid chromatographic method for the analysis of xylitole and hexosaminitols present in the reduced terminal of glyco-saminoglycans is described. The glycosaminoglycans are covalently bound to protein forming proteoglycans via xylose, galactosamine or glucosamine. Alkaline treatment of the proteoglycan in the presence of NaBH₄ releases the glycosaminoglycan chains which contain in their reduced terminal xylitole, galactosaminitole or glucosaminitole. The obtained glycosaminoglycans are depolymerized with trifluoroacetic acid and the liberated neutral monosaccharides and xylitole are separated from the hexosa-mines and hexosaminitols on a DOWEX 50-X8 microcolumn. The obtained neutral monosaccharides are reduced to alditols by NaBH₄ and separated from xylitole as per-O-benzoylated derivatives on Supelcosil LC-18, using 70 % acetonitrile as eluent, at a flow rate of 1.0 ml/min. Per-O-bcnzoylated derivatives of hexosaminitols are separated from the hexosamines derivatives on the same