Chiral separation of acidic drug components by open tubular electrochromatography using avidin immobilized capillaries

Avidin was immobilized covalently onto the inner surface of fused silica capillaries as a stationary phase in an open tubular capillary electrochromatography (OT–CEC) for chiral separations. The modification was attained by a combination of glutaraldehyde with both an amino-silylated fused silica surface and avidin using a Schiff base formation reaction. This method couples the advantage of high efficiency and small consumption of a chiral selector with the possibility of UV detection without limitations of protein absorption. In addition, the prepared capillary was stable for 50 days with over 100 runs. To evaluate the electrochromatographic performance of the prepared capillaries, the chiral separation of abscisic acid and arylpropionic acids were investigated. Effects of the modification condition of protein, pH of running buffer, and an organic modifier on the enantioseparation were also investigated. In addition, the avidin immobilized capillary was employed for the selective chiral analysis with an electrospray ionization mass spectrometry (ESI-MS) detection scheme.