Determination of amino sugars in synthetic glycopeptides during the conditions of amino acid analysis utilizing precolumn derivatization and high-performance liquid chromatographic analysis.
The growing number of synthetic glycopeptides required an in-house method for the analysis of the final products. A reversed-phase high-performance liquid chromatographic protocol was developed to verify the presence of N-acetylglucosamine and N-acetylgalactosamine in synthetic glycopeptides after hydrolysis and derivatization with 4-dimethylaminoazobenzene-4'-sulphonyl chloride. Different sugar and glycoamino acid standards were used to separate the two carbohydrate moieties, and the location of the derivatized 2-aminoglucose was established by the use of radiolabeled sugar. The utility of the approach is demonstrated by the amino acid analysis of N- and O-glycosylated synthetic peptides and the method could provide an alternative for sugar analysis of glycoproteins.
Wistar Institute of Anatomy and Biology, Philadelphia, PA 19104.
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D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis*; D000606:Amino Sugars / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006020:Glycopeptides / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D004124:p-Dimethylaminoazobenzene / Q000031:analogs & derivatives
D000596:Amino Acids; D000606:Amino Sugars; D006020:Glycopeptides; D007202:Indicators and Reagents; C010572:4-N,N-dimethylaminoazobenzene-4'-sulfonyl chloride; D004124:p-Dimethylaminoazobenzene