HPLC determination of carbohydrate content

An effort was made to better quantitate this suggested preponderance of N-acetylglucosamine and/or N-acetylgalactos-amine in the carbohydrate portion of microbubble glycopeptide surfactant by performing direct HPLC analysis of the monosaccharides contained in the surfactant. This analysis began with the (carbohydrate) hydrolysis of a partially purified preparation containing approximately 30 pg of the proteinaceous microbubble 

Nonetheless, direct verification of the presence of covalently bound carbohydrate in the proteinaceous microbubble surfactant was provided earlier by the enzymatic degradation experiments (see Section 4.2.3), and this finding has the added usefulness of providing a more accurate interpretation of the sodium dodecyl sulfate/polyacrylamide-gel electrophoretic data summarized earlier. 

This form of electrophoresis is not directly applicable to molecular weight determinations of glycoproteins. Glycoproteins and glycopeptides containing more than 10% carbohydrate behave 

5 Gel-filtration column chromatography Determination of average molecular weight and the NH7-terminus 

NH2-terminal amino acid identification of the molecular subclasses Composing (dansylated) microbubble glycopeptide surfactant. (Taken from ref. 322.) 

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