Panin G, Naia S, Dall'Amico R, Chiandetti L, Zachello F, Catassi C, Felici L, Coppa G V
Clin Chem. 1986 Nov;32(11):2073-6.
We describe a simple, rapid, precise, and sensitive spectrophotometric method for measuring urinary glycosaminoglycan (GAG) sulfate excretion. The GAG sulfates are precipitated with cetylpyridinium chloride, resuspended in water, and mixed with the basic dye 1,9-dimethylmethylene blue to produce a complex with the polyanionic molecule of sulfated GAGs. Absorbance is read at 535 nm. The standard curve for reaction was linear up to 12 micrograms of the different GAGs: dermatan sulfate, heparan sulfate, keratan sulfate, chondroitin 4-sulfate, and chondroitin 6-sulfate. Within- and between-run precision (CV), measured at three different GAG concentrations (normal and pathological), varied from 1.6% to 2.5% and from 1.8% to 4.5%, respectively. Analytical recovery ranged from 71% to 107%. Urinary GAG excretion, measured by this procedure, correlates (r = 0.837; p less than 0.001) with the values obtained with the borate-carbazole reaction (Anal Biochem 1962;4:330-4).
我们描述了一种用于测量尿中硫酸糖胺聚糖（GAG）排泄量的简单、快速、精确且灵敏的分光光度法。硫酸GAG用十六烷基吡啶氯化物沉淀，再悬浮于水中，并与碱性染料1,9 - 二甲基亚甲蓝混合，以与硫酸化GAG的聚阴离子分子形成复合物。在535nm处读取吸光度。不同GAG（硫酸皮肤素、硫酸乙酰肝素、硫酸角质素、硫酸软骨素4 - 硫酸盐和硫酸软骨素6 - 硫酸盐）反应的标准曲线在高达12微克时呈线性。在三种不同GAG浓度（正常和病理浓度）下测定的批内和批间精密度（CV）分别在1.6%至2.5%和1.8%至4.5%之间变化。分析回收率在71%至107%之间。用该方法测定的尿GAG排泄量与用硼酸盐 - 咔唑反应获得的值相关（r = 0.837；p小于0.001）（《分析生物化学》1962年；4：330 - 4）。
