Monosaccharide Analysis by HPAEC
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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).
KeywordsReductive Amination Monosaccharide Composition Pump Head Monosaccharide Analysis Gradient HPLC System