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A microdiaiysis junction, based on a microdialysis membrane connecting both a separation capillary and a short, sharply tapered microelectrospray emitter capillary, is demonstrated for on-line combination of capillary isoelectric focusing (CIEF) with electrospray ionization mass spectrometry (ESIMS). The microdialysis junction provides the necessary electrical connection across the dialysis membrane for defining the electric fields needed for the CIEF separation and the electrospray process. Additionally, post-separation acidification of focused protein zones eluted from the CIEF capillary is achieved using the microdialysis junction while maintaining separation efficiency and resolution. A microelectrospray emitter produces a stable electrospray of protein analytes without the need for a makeup liquid flow and eliminates any subsequent sample dilution and reduction in MS sensitivity. The microdialysis junction is advantageous over the coaxial liquid sheath interface as evidenced by the simplicity in operation procedures, the enhancement in

On-line coupling of capillary electrophoresis and liquid chromatography with electrospray ionization mass spectrometry for protein characterization