DETERMINATION OF HYALURONAN AND GALACTOSAMINOGLYCAN DISACCHARIDES BY HIGH-PERFORMANCE CAPILLARY ELECTROPHORESIS AT THE ATTOMOLE LEVEL - APPLICATIONS TO ANALYSES OF TISSUE AND CELL-CULTURE PROTEOGLYCANS

A rapid, highly sensitive and reproducible HPCE method is described fo r the determination of all non- and variously sulphated disaccharides present in hyaluronan and vertebrate chondroitin sulphates and dermata n sulphates. Following chondroitinase digestion of glycosaminoglycans or proteoglycans, the non-, di- and tri-sulphated Delta-disaccharides are completely separated and readily determined within 14 min on a fus ed-silica capillary in 15 mM sodium dihydrogen orthophosphate, pH 3.00 , using reversed polarity at 20 kV and detection at 232 nm. The determ ination of the various Delta-disaccharides derived from either glucuro nic or iduronic acid and the presence of glucuronic and iduronic clust ered structures in dermatan sulphate can also easily be made, using di gests with chondroitinase AC or B. A linear detector response was obta ined for the entire interval tested (up to 10 mg/l of Delta-disacchari des). Concentrations as small as 32, 65, 100 and 250 pmol/l (22, 38, 5 0 and 98 ng/l) of tri-, di- and nonsulphated Delta-disaccharides, resp ectively, can be reliably detected.