The separation of four proteins including RNase A, cytochrome C, lysozyme and myoglobin was investigated by reversed-phase gradient pressurized capillary electrochromatography (p-CEC) with 1.5 microm non-porous silica C18 stationary phase. This mode was compared with micro-high performance liquid chromatography (mu-HPLC) and the effects of applied voltage, stationary phase and concentration of ion-pairing agent (trifluoroacetic acid, TFA) on the gradient p-CEC were also studied. This separation was performed rapidly on a new CEC instrument Trisep 2010 GV. The results showed that the retention mechanism of proteins in p-CEC mode is based on both chromatographic partitioning and electrophoretic migration. The results also demonstrated that p-CEC may have great potential for fast and efficient separation of proteins.
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