Preparation and evaluation of an imidazole-coated capillary column for the electrophoretic separation of aromatic acids

Abstract

An imidazole-coated capillary column for electrophoresis has been prepared by means of organosilanization. With mesityl oxide as neutral marker, the results indicated that the electroosmotic flow of the bonded phase displays a dramatic difference in pH dependence in comparison with that of the bare fused-silica column. The presence of positive charges on the coating surface and the anionic exchange property, due to the cationic property of the imidazole group at pH values below 6, allows the separation of geometric isomers that are very similar in ionic mobility. Separation parameters including buffer composition and concentration, pH, applied voltage, and the influence of other additives were investigated. By using acetate buffer (100 m M, pH 5.2) and an applied voltage of −15 kV with UV detection at 212 nm, the separation of 11 aromatic acids including mono-, di-, tri- and tetra-carboxylic acids could be achieved in less than 14 min. The average plate number was 3×10 5/m. With acetate buffer (25 m M, pH 5.5) and an applied voltage of −25 kV, the addition of silver nitrate or β-cyclodextrin significantly improved the resolution of some more highly charged carboxylic acids.