Abstract
An automated anion-exchange chromatographic procedure for the separation of uronic acids and uronic acid containing oligosaccharides is described. The method has been applied, in particular, to the separation and quantitation of l-iduronic and d-glucuronic acid in hydrolyzates of dermatan sulfate, and the hydrolysis conditions required for optimum yield of uronic acids have been determined. Whereas uronic acids and aldobiuronic acids were successfully separated by elution with formic acid, hyaluronic acid oligosaccharides and, especially, the highly charged chondroitin sulfate oligosaccharides were more conveniently resolved in a chloride system by elution with a nonlinear LiCl gradient. This system was also employed for the separation of unsaturated and saturated disaccharides obtained by chondroitinase digestion of chondroitin and chondroitin sulfate. The effluent from the columns was analyzed by the automated orcinolsulfuric acid method of the Technicon sugar chromatography system.
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