Carbohydrate Analysis of Bacterial Polysaccharides by High-pH Anion-Exchange Chromatography and Online Polarimetric Determination of Absolute Configuration
✍ Scribed by Corné J.M. Stroop; C.Allen Bush; Ronita L. Marple; William R. LaCourse
- Publisher
- Elsevier Science
- Year
- 2002
- Tongue
- English
- Weight
- 108 KB
- Volume
- 303
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
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.