Pseudo-electrokinetic packing of high efficiency columns for capillary electrochromatography

Abstract

An improved and easy electrokinetic packing procedure is presented for the production of stable capillary columns suitable for capillary electrochromatography (CEC). In pseudo-electrokinetic packing a high electric field is used in conjunction with a hydrodynamic flow. The packing of silica-based reversed-phase columns can be achieved with basic, commercially available capillary electrophoresis (CE) equipment in approximately 15 min. The procedure is robust and a high success rate is achieved. No steps which might damage the stationary phase are involved and only a minimum amount of packing material is required. Columns packed according to the developed procedure are operated at high electric field strengths during the CEC separation, without the application of a stabilising pressure. Columns are stable for at least hundred runs and were tested using mixtures of polycyclic aromatic hydrocarbons and positively charged drugs. Separations were performed in a relatively high conducting ammonium acetate buffer, with efficiencies of up to 283 000 plates/m.