AIMTo develop a rapid and sensitive method for monosaccharide composition analysis.

METHODSGlycoprotein was first hydrolyzed to monosaccharides, which were subsequently reacetylated (amino monosaccharides), tagged with 2-aminopyridine and then separated and monitored in selected ion mode by CapGCC-LC/MS. The use of tetradeuterium labeled-aminopyridyl-monosaccharides prepared by tagging monosacahrides with hexadeuterium labeled 2-aminopyridine as internal standards improved the linearity and reproducibility in quantification.

RESULTSThis method was successfully applied to monosaccharide composition analysis of model glycoproteins, fetuin and erythropoietin down to 1 pmol monosaccharides.

CONCLUSIONThis method has been shown to be highly sensitive and is applicable to monosaccharide composition analysis of glycoproteins.

Animals ; Chromatography, Liquid ; methods ; Erythropoietin ; chemistry ; Glycoproteins ; chemistry ; Hydrolysis ; Isotope Labeling ; Monosaccharides ; analysis ; Sensitivity and Specificity ; Spectrometry, Mass, Electrospray Ionization ; alpha-Fetoproteins ; chemistry