一种用于测定硫酸乙酰肝素的分光光度法。

A spectrophotometric method for the determination of heparan sulfate.

作者信息

van de Lest C H, Versteeg E M, Veerkamp J H, van Kuppevelt T H

机构信息

Department of Biochemistry, University of Nijmegen, The Netherlands.

出版信息

Biochim Biophys Acta. 1994 Nov 11;1201(2):305-11. doi: 10.1016/0304-4165(94)90055-8.

DOI:10.1016/0304-4165(94)90055-8

PMID:7947946

Abstract

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.

摘要

本文描述了一种简单可靠的分光光度法用于测定硫酸乙酰肝素。该方法基于用于硫酸化糖胺聚糖的1,9-二甲基亚甲蓝测定法。加入牛血清白蛋白，同时控制特定的氯化钠浓度和pH值，会导致基于硫酸乙酰肝素的吸光度出现特定程度的降低。硫酸乙酰肝素的含量可通过用不存在白蛋白时获得的值减去存在白蛋白时获得的值来计算。灵敏度为0.5微克硫酸乙酰肝素。给出了两个应用实例：对尿液中硫酸乙酰肝素进行定量，包括来自黏多糖贮积症患者的尿液，以及评估用于分离硫酸乙酰肝素蛋白聚糖的凝胶过滤和离子交换柱色谱的级分。