Separation of reduced disaccharides derived from glycosaminoglycans by high-performance liquid chromatography

A high-performance liquid chromatographic (HPLC) method for the analysis of reduced unsaturated disaccharides derived from enzymatic digestion followed by reduction with sodium borohydride of chondroitin sulfates, dermatan sulfate, heparan sulfate and heparin is described. This method is well suited for the HPLC analysis of glycosaminoglycans (GAGs) because the possibility of obtaining anomeric forms of unsaturated disaccharides is eliminated with provides a major advantage for quantitation. This procedure is more sensitive than existing HPLC methods for the determination of enzymatic degradation products from GAGs. In particular, the resolution of disaccharide products from heparan sulfate is improved after reduction. The applicability of this method for the determination of GAGs in biological samples is demonstrated.