N LINKED OLIGOSACCHARIDE PROFILING

University Of Georgia (Uga), Athens GA

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Abstract

The samples were analyzed for monosaccharide composition by preparing the TMS derivatives of the methylglycosides followed by GC-MS analysis. The samples were acid hydrolyzed then subjected to methanolysis followed by N-acetylation with pyridine and acetic anhydride (for detection of amino sugars). The samples were then treated with Tri-Sil. GC-MS analysis of the TMS methylglycosides was performed on a Hewlett-Packard 5890 GC coupled to a 5970 MSD using a 30-meter DB1 fused silica capillary column. The samples were subjected to glycosyl-linkage analysis using the Hakomori methylation procedure followed by combined GC-MS analysis. The samples were then subjected to acid hydrolysis using 2 M trifluoroacetic acid to effect cleavage of all glycosidic linkages. The glycosyl residues were reduced with NaBD4 and then acetylated using acetic anhydride/pyridine. The resulting partially methylated alditol acetates were analyzed on a 30-meter Supelco 2330 fused silica capil lary column by GC-MS using a Hewlett-Packard 5890 GC coupled to a 5970 MSD.