A SPECTROPHOTOMETRIC METHOD FOR THE DETERMINATION OF HEPARAN-SULFATE

A simple and reliable spectrophotometric method for the determination of heparan sulfate is described. The method is based on the 1,9-dimeth ylmethylene blue assay for sulfated glycosaminoglycans. Addition of bo vine serum albumin, together with a specific NaCl concentration and pH , results in a specific decrease of heparan sulfate-based absorbance. The amount of heparan sulfate can be calculated by subtracting the val ues obtained in the presence of albumin from those obtained in its abs ence. The sensitivity is 0.5 mu g heparan sulfate. Two applications ar e given: the quantification of heparan sulfate in urine, including uri ne from patients with mucopolysaccharidosis, and the evaluation of fra ctions from gel filtration and ion exchange column chromatography for isolation of heparan sulfate proteoglycans.