IDENTIFICATION OF GLYCOSAMINOGLYCANS USING HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY ON A HYDROXYAPATITE COLUMN

Glycosaminoglycans (heparin, heparan sulfate, dermatan sulfate, chondr oitin sulfate, and hyaluronic acid) were labeled with a fluorescent re agent, 2-aminopyridine. The fluoro-labeled glycosaminoglycans were sub jected to high-performance liquid chromatography on a hydroxyapatite c olumn. The binding property of each glycosaminoglycan to hydroxyapatit e was different. The structural properties of glycosaminoglycans bound to hydroxyapatite were then investigated using chemical desulfated or enzymic depolymerized glycosaminoglycans. This revealed that the sulf ate content and molecular weight of the glycosaminoglycans correlated with their binding properties to hydroxyapatite. Desulfated dermatan s ulfate but not desulfated chondroitin 6-sulfate bound to the hydroxyap atite. These data indicate that iduronic acid residues of glycosaminog lycans are important for the binding property. The method described wh ich uses hydroxyapatite columns facilitates rapid separation and micro analysis of the glycosaminoglycans, especially dermatan sulfate and ch ondroitin sulfate. (C) 1995 Academic Press, Inc.