Determination of unsaturated glycosaminoglycan disaccharides by spectrophotometry on thin-layer chromatographic plates.

Chondroitin sulfate isomers and hyaluronate, digested with chondroitinase AC II and separated as unsaturated disaccharides by thin-layer chromatography on cellulose, were quantitated with scanning spectrophotometry using reflectance measurement at 232 nm. The sensitivity of the assay was increased by one order of magnitude as compared to previous modifications of the disaccharide analysis on cellulose. The method allowed quantitation of 0.2-20 micrograms of glycosaminoglycan uronic acid. Since the elution of the disaccharides from the plate could be omitted, the speed and reproducibility of the assay was enhanced.