title: Carbohydrate analysis of bacterial polysaccharides by high-pH anion-exchange chromatography and online polarimetric determination of absolute configuration.
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.
authors.affiliation: Department of Chemistry & Biochemistry, University of Maryland Baltimore County, 1000 Hilltop Circle, Baltimore, Maryland, 21250, USA.
authors.forename: Corné J M
authors.initials: CJ
authors.lastname: Stroop
authors.forename: C Allen
authors.initials: CA
authors.lastname: Bush
authors.forename: Ronita L
authors.initials: RL
authors.lastname: Marple
authors.forename: William R
authors.initials: WR
authors.lastname: LaCourse
mesh_terms: D001419:Bacteria / Q000737:chemistry*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D008968:Molecular Conformation; D011134:Polysaccharides / Q000032:analysis*
chemical_list: D011134:Polysaccharides