Automatic analysis of neutral sugar components in glycoproteins and complex carbohydrates

A simple and sensitive automated method has been developed to separate monosaccharides by reverse-phase chromatography using the amino acid analyzer column in the Li+ form with 90% alcohol as eluent. The column can be regenerated with LiBr and 90% ethanol to achieve highly reproducible resolution of sugar components. Glycoproteins and polysaccharides are hydrolyzed by a combination of methanesulfonic acid/Dowex 50 X 8 resin in water. Glucose and mannose are distinguished by treatment of the hydrolysate with glucose oxidase prior to analysis.