Simultaneous determination of monosaccharides in glycoproteins by capillary electrophoresis.

A rapid, easy, and reproducible capillary electrophoretic method for the simultaneous determination of acidic, neutral, and amino sugars and sugar alcohols was developed. Underivatized mannuronic acid, glucuronic acid, galacturonic acid, N-glycolylneuraminic acid, N-acetylneuraminic acid, glucosamine, galactosamine, mannose, xylose, glucose, galactose, fucose, ribose, mannitol, sorbitol, xylitol, and inositol were simultaneously determined with indirect UV detection using 2,6-pyridinedicarboxylic acid as a background electrolyte. A highly alkaline pH condition was used in order to charge carbohydrates negatively and to promote migration toward the anode. Electroosmotic flow was reversed to the direction of the anode by adding cetyltrimethylammonium bromide to the electrolyte. The separation of the carbohydrates was investigated by optimizing the operating pH value and satisfactory resolution was obtained at pH 12.1. The relative standard deviations of the method for carbohydrates were between 0.02 and 0.33% for migration times and were greater than 2.7% for peak areas (n = 6). The minimum detectable level ranged from 23 to 71 microM with a 6-nl injection at a signal-to-noise ratio of 3. This method was applied to the composition analysis of monosaccharides in glycoprotein. After acid hydrolysis of fetuin under each optimum condition, sialo, neutral, and amino sugars were quantified under the same electrophoretic condition.