Separation and on-line preconcentration by sweeping of charged analytes in electrokinetic chromatography with nonionic micelles

Abstract

The separation and on-line preconcentration by sweeping of charged analytes in electrokinetic chromatography using a neutral pseudostationary phase is described. Under neutral or basic conditions, the electrophoretic migration of anionic analytes towards the anode is overcome by a high, cathode-directed electroosmotic flow; hence, they experience net migration towards the cathode, and the system is run at positive polarity mode. The separation and the retention factor, k, are dependent on both the analyte’s electrophoretic mobility and its interaction with the pseudostationary phase. The versatility of the sweeping mechanism is then shown in this system. The charged analyte, prepared in a matrix free of the pseudostationary phase, penetrates the pseudostationary phase zone upon application of voltage. Analyte molecules are consequently accumulated and concentrated. As a demonstration, the separation and preconcentration of phenol derivatives using nonionic surfactants of the alkyl polyoxyethylene ether type (Brij 35 and Brij 58) yielded peak height enhancements up to 100-fold. The efficiency of sample stacking was also found to be improved with the use of a high viscosity background solution.