Extraction, purification and evaluation of structures and physico-chemical properties of glycosaminoglycans

Heparin was extracted and purified from beef intestina! muco¬sa. The two components, fast moving heparin and slow moving heparin were purified by selective precipitation as barium salts. Heparan sulfate was extracted and purified from beef spleen. Dermatan sulfate was purified from beef intestina! mucosa and chondroitin sulfate from bovine trachea.The purity of the purified glycosaminoglycans wase by agarose-gel and cellulose polyacctatc electrophoresis and by spe-cific optical rotation. The relative molecular masses of glycosa¬minoglycans were estimated by high performance-size exclusion chromatography and thè sull'ale to carboxyl ratio by titrimetric analysis. The disaccharide pattcrn of heparin, fast moving and slow moving heparins and heparan sulfate were determined by specific enzymatic clcavage using hcparinase I, li and III; thè disaccharide composition of dcrmatan sulfate and chondroitin sulfate was evaluated by cleavage by chondroitinasc ABC. The disaccharides obtained by enzymatic cleavage were qualitative!} and quantitatively analysed by strong anion exchange-high performance liquid chromatography. The sulfate to carboxyl ra-tios of glycosaminoglycans were also determined by this tccnique and compared with thè values obtained by titrimetric analysis.