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-dimethylmethylene blue assay for sulfated glycosaminoglycans. Addition of bovine 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 values obtained in the presence of albumin from those obtained in its absence. The sensitivity is 0.5 microgram heparan sulfate. Two applications are given: the quantification of heparan sulfate in urine, including urine from patients with mucopolysaccharidosis, and the evaluation of fractions from gel filtration and ion exchange column chromatography for isolation of heparan sulfate proteoglycans.
van de Lest
C H
CH
Department of Biochemistry, University of Nijmegen, The Netherlands.
Versteeg
E M
EM
Veerkamp
J H
JH
van Kuppevelt
T H
TH
D006025:Glycosaminoglycans / Q000032:analysis; D006497:Heparan Sulfate / Q000032:analysis* / Q000652:urine; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008751:Methylene Blue / Q000031:analogs & derivatives; D009083:Mucopolysaccharidoses / Q000652:urine; D011509:Proteoglycans / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine; D012965:Sodium Chloride; D013053:Spectrophotometry / Q000379:methods*
D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D008751:Methylene Blue; D011509:Proteoglycans; D012710:Serum Albumin, Bovine; D012965:Sodium Chloride; C016401:1,9-dimethylmethylene blue