GLYCOSYL COMPOSITION ANALYSIS OF SACCHARIDE

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Glycosyl-linkage analysis was performed by preparing permethylated alditiol acetates using the Hakomori procedure. The samples were dissolved in DMSO, DMSO anion was added, and the samples were stirred at room temperature for 2 h. An excess of methyliodide was added, and the samples were stirred again from 2 to 4 h at room temperature. Excess methyliodide was removed by evaporation, and the samples were purified using a C18 SepPak cartridge. The resulting samples were hydrolyzed in 2 M TFA, and the monosaccharides were reduced to their alditiols and N-acetylated using acidic anhydride in pyridine. The resulting acetates were then analyzed by GC-MS using a 30 mm SP2330 column from Supelco for the neutral sugars and using a DB1 column from J&W Scientific for the methylated amino sugars.