Size fractionation of oligosaccharides by liquid chromatography on a cation-exchange column

Oligosaccharides, labelled with 2-aminopyridine at their reducing ends, were satisfactorily fractionated according to the sugar sizes on Shodex RSpak DC-613, a cation-exchange resin column (Na+ form), with water-acetonitrile as the eluent in the presence of sodium acetate or triethylammonium acetate buffer. For the fractionation of sugar samples, dextran hydrolyzates, chitin oligomers and oligosaccharide moieties of ovomucoid were used. The oligosaccharides were strongly adsorbed to the resin column with solvents containing less water and at lower temperature, and were eluted in order of increasing molecular size above the critical concentration of acetonitrile. Baseline separation of a dextran hydrolyzate up to oligomers having 20 glucose units was observed by gradient elution. The separation efficiency and elution pattern were investigated by changing the buffer concentration, mobile phase pH and temperature.