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Carbohydrate Analysis of Bacterial Polysaccharides by High-pH Anion-Exchange Chromatography and Online Polarimetric Determination of Absolute Configuration

Authors
  • Stroop, Corné J.M.
  • Bush, C.Allen
  • Marple, Ronita L.
  • LaCourse, William R.
Type
Published Article
Journal
Analytical Biochemistry
Publication Date
Jan 01, 2002
Volume
303
Issue
2
Pages
176–185
Identifiers
DOI: 10.1006/abio.2002.5582
Source
Elsevier
Keywords
License
Unknown

Abstract

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.

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