van den Hoogen B M, van Weeren P R, Lopes-Cardozo M, van Golde L M, Barneveld A, van de Lest C H
Department of Veterinary Basic Sciences, Graduate School of Animal Health, Utrecht University, The Netherlands.
Anal Biochem. 1998 Mar 15;257(2):107-11. doi: 10.1006/abio.1997.2538.
The amount of uronic acid residues in samples containing glycosaminoglycans or pectin is an important parameter in the quantitative and structural analysis of these complex carbohydrates. This paper describes a method to determine the content of uronic acids in biological samples, using conventional polystyrene microtiter plates and microtiter plate-reading equipment with standard interference filters (i.e., 540 or 492 nm). This assay is a modification of a commonly used procedure, viz. hydrolysis of uronic acid containing carbohydrate polymers in 80% sulfuric acid containing tetraborate ions at 80 degrees C followed by a coloring step with an m-hydroxydiphenyl reagent at room temperature. The use of microtiter plates has several practical advantages: (i) less risk of handling hot, concentrated sulfuric acid is present; (ii) an accurate estimate of background absorbance by multiple reading of the plates is possible; and (iii) many samples can be assayed in one series without errors due to fading of the final color. The validity of the assay was checked for the quantification of hyaluronic acid in equine synovial fluid samples. We consider this the method of choice when a large number of samples must be analyzed for their content of uronic acid residues.
在含有糖胺聚糖或果胶的样品中，糖醛酸残基的含量是对这些复杂碳水化合物进行定量和结构分析的一个重要参数。本文描述了一种使用传统聚苯乙烯微量滴定板和配备标准干涉滤光片（即540或492 nm）的微量滴定板读数设备来测定生物样品中糖醛酸含量的方法。该测定法是对一种常用程序的改进，即：在80℃下，在含有四硼酸根离子的80%硫酸中水解含糖醛酸的碳水化合物聚合物，然后在室温下用间羟基联苯试剂进行显色步骤。使用微量滴定板有几个实际优点：（i）处理热的浓硫酸时风险较小；（ii）通过多次读取滴定板可以准确估计背景吸光度；（iii）可以在一系列中检测许多样品，而不会因最终颜色褪色而产生误差。对该测定法在马滑液样品中透明质酸定量的有效性进行了检查。当必须分析大量样品中糖醛酸残基的含量时，我们认为这是首选方法。
