Determination of human urinary hyaluronic acid, chondroitin sulphate and dermatan sulphate as their unsaturated disaccharides by high-performance liquid chromatography

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Abstract

A method for the determination of hyaluronic acid (HA), chondroitin sulphate (CS) and dermatan sulphate (DS) was developed. HA, CS and DS were converted to the corresponding unsaturated disaccharides by digestion with chondroitinase ABC and/or chondroitinase AC-II and determined by high-performance liquid chromatography with fluorimetric detection using 2-cyanoacetamide as a post-column derivatization reagent. The calibration graphs for the unsaturated disaccharides were linear over the range 2 ng − 2 μg for each unsaturated disaccharide. This method was applied to the analysis of normal human urine.

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