Carbohydrate analysis of bacterial polysaccharides by high-pH anion-exchange chromatography and online polarimetric determination of absolute configuration.
Carbohydrate analysis of bacterial polysaccharides by high-pH anion-exchange chromatography and online polarimetric determination of absolute configuration.
复制标题
通过高 pH 阴离子交换色谱和在线旋光测定绝对构型对细菌多糖进行碳水化合物分析。
DOI：
10.1006/abio.2002.5582
复制
发表时间：
2002
期刊：
影响因子：
--
通讯作者：
LaCourse,WilliamR
中科院分区：
文献类型：
--
作者：
Stroop,CorneJM;Bush,CAllen;Marple,RonitaL;LaCourse,WilliamR
在复杂碳水化合物，特别是细菌多糖的结构测定中，一个重要问题是确定组成单糖的绝对构型。许多分析方法已经被用于这一目的，但由于复杂多糖中发现的糖的各种化学性质，没有一种方法是通用的。使用手性试剂进行挥发性衍生物的高分辨气相色谱分析是目前应用最广泛的方法。旋光活性虽然直接和简单，但缺乏灵敏度，通常需要大量的纯单糖。我们报道了一种结合高效阴离子交换层析(HPAEC)与电化学脉冲安培检测和在线旋光检测与在线激光偏振检测相结合的方法，用于分析复杂多糖中发现的许多糖。我们表明，应用该方法分析几种革兰氏阳性和革兰氏阴性致病菌的荚膜多糖，同时提供了关于碳水化合物组成和组分糖的对映体构型的有用信息。
A significant problem in structure determination of complex carbohydrates, especially for bacterial polysaccharides, is determination of the absolute configuration of the component monosaccharides. A number of analytical methods have been used for this purpose but, as a result of the wide variety of chemical properties of sugars found in complex polysaccharides, no single method is universally applicable. High-resolution gas chromatography of volatile derivatives with chiral reagents is the most widely used method. Optical activity, although direct and simple, lacks sensitivity generally requiring a large quantity of pure monosaccharide. We report a combination of high-performance anion-exchange chromatography (HPAEC) with combined electrochemical pulsed amperometric detection and in-line detection of optical rotation with an in-line laser polarimeter for analysis of a number of sugars found in complex polysaccharides. We show that application of the method for analysis of capsular polysaccharides of several gram-positive and gram-negative pathogenic bacteria provides useful information simultaneously on carbohydrate composition and the enantiomeric configuration of component sugars.