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Monosaccharide Analysis by HPAEC

  • Elizabeth F. Hounsell
  • Michael J. Davies
  • Kevin D. Smith
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Once the presence of monosaccharides has been established by chemical methods (see  Chapter 108), the next stage of any glycoconjugate or polysaccharide analysis is to find out the amount of sugar and monosaccharide composition. The latter can give an idea as to the type of oligosaccharides present and, hence, indicate further strategies for analysis (1). Analysis by high-pH anion-exchange chromatography (HPAEC) with pulsed electrochemical detection, that is described here, is the most sensitive and easiest technique (2). If the laboratory does not have a biocompatible HPLC available that will withstand high salt concentrations, a sensitive labeling technique can be used with gel electrophoresis (3), e.g., that marketed by Glyko Inc. (Navato, CA) or Oxford GlycoSciences (Abingdon, UK), to include release of oligosaccharides/monosaccharides and labeling via reductive amination with a fluorescence label (4).

Keywords

Reductive Amination Monosaccharide Composition Pump Head Monosaccharide Analysis Gradient HPLC System 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Hounsell, E. F. (1993) A general strategy for glycoprotein oligosaccharide analysis, in Methods in Molecular Biology, vol. 14: Glycoprotein Analysis in Biomedicine (Hounsell, E. F., ed.), Humana, Totowa, NJ, pp. 1–15.CrossRefGoogle Scholar
  2. 2.
    Townsend, R. R. (1995) Analysis of glycoconjugates using high-pH anion-exchange chromatography. J. Chromatog. Library 58, 181–209.CrossRefGoogle Scholar
  3. 3.
    Linhardt, R. J., Gu, D., Loganathan, D., and Carter, S. R. (1989) Analysis of glycosaminoglycan-derived oligosaccharides using reversed-phase ion-pairing and ion-exchange chromatography with suppressed conductivity detection. Anal. Biochem. 181, 288–296.PubMedCrossRefGoogle Scholar
  4. 4.
    Davies, M. J. and Hounsell, E. F. (1996) Comparison of separation modes for high performance liquid chromatography for the analysis of glycoprotein-and proteoglycan-derived oligosaccharides. J. Chromatogr. 720, 227–234.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Elizabeth F. Hounsell
    • 1
  • Michael J. Davies
  • Kevin D. Smith
  1. 1.School of Biological and Chemical SciencesBirkbeck University of LondonUK

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