Release of O-sulfate groups under mild acid hydrolysis conditions used for estimation of N-sulfate content.
The treatment of chondroitin sulfate isolated from cultured B16 mouse melanoma cells with 0.04 M HCl at 100 degrees C for 90 min released up to 45% of O-sulfate residues as free inorganic sulfate. In addition to the release of inorganic sulfate, extensive degradation of this polysaccharide as well as of cartilage chondroitin sulfate, pig rib cartilage proteoglycan, heparin and hyaluronic acid was also evident under these conditions. The above hydrolysis conditions are used for characterizing 35S-labeled heparan sulfates synthesized by cultured cells and to calculate ratio of N- and O-sulfates in these molecules. Our results suggest that caution is necessary in interpreting the results of mild acid hydrolysis of glycosaminoglycans.
D000818:Animals; D002460:Cell Line; D002807:Chondroitin / Q000031:analogs & derivatives; D002809:Chondroitin Sulfates / Q000302:isolation & purification; D006493:Heparin / Q000302:isolation & purification; D006820:Hyaluronic Acid / Q000302:isolation & purification; D008545:Melanoma / Q000032:analysis; D051379:Mice; D009374:Neoplasms, Experimental / Q000032:analysis; D013464:Sulfuric Acids / Q000032:analysis*
D013464:Sulfuric Acids; D006820:Hyaluronic Acid; D006493:Heparin; D002807:Chondroitin; D002809:Chondroitin Sulfates