Capillary electrophoresis of carboxylated carbohydrates. Part 2. Selective precolumn derivatization of sialooligosaccharides derived from gangliosides with
7-aminonaphthalene-l,3-disulfonic acid fluorescing tag
The most suitable conditions for selective precolumn derivatization of sialooligosaccharides, derived from gangliosides, with 7-aminonaphthalene-l,3-disulfonic acid (ANDSA) and the subsequent separation of the derivatives by capillary electrophoresis are described. ANDSA-sialooligosaccharide derivatives, which fluoresce at 420 nm when excited at 315 nm, were readily detected in capillary electrophoresis using an on-column lamp-operated fluorescence detector. In addition, the precolumn derivatization described here, which exploited the reactivity of the carboxylic acid group of the sialic acid residue of the oligosaccharides, replaced each weak carboxylic acid group of the parent sugar by two strong sulfonic acid groups. This allowed for electrophoresis over a wide range of pH and improved the resolution of the derivatives when compared to those obtained with underivatized sialooligosaccharides under identical separation conditions. The separation of sialooligosaccharides was best achieved when 75 mM borate, pH 10.0, was used as the running electrolyte. The derivatization and separation conditions described herein are expected to be readily transposed to the capillary electrophoresis of other sialooligosaccharides such as those derived from glycoproteins*.