Capillary electrochromatography with monolithic stationary phases - III. Evaluation of the electrochromatographic retention of neutral and charged solutes on cationic stearyl-acrylate monoliths and the separation of water-soluble proteins and membrane proteins

This article, which is closely related to part II, is concerned with the evaluation of the retentive properties of cationic stearyl-acrylate monoliths (i.e. cationic C-17 monoliths) over a wide range of elution conditions with various uncharged and charged solutes including proteins. The retention parameters for charged solutes including the retention factor k* observed under capillary electrochromatography conditions and the velocity factor k*(ep), which reflects the electrophoretic process, were measured for weak, moderate and strong basic compounds. These retention parameters allowed the assessment of the respective contributions from electrophoretic and partitioning separation mechanisms. The cationic C-17 monoliths exhibited sufficient hydrophobic interactions with relatively weak basic solutes. Moderate and strong bases showed migration behaviors dominated by their relatively strong electrophoretic mobility with marginal chromatographic partitioning. At low pH, the cationic C-17 monoliths allowed the separation of proteins with minimum electrostatic interactions between proteins and the cationic sites on the surface of the stationary phase. The utility of the cationic C-17 monoliths was demonstrated in the rapid and efficient separation of two crude extracts of membrane proteins, namely galactosyl transferase and cytochrome c reductase. Short capillary columns (8.5 cm effective length) of the cationic C-17 monoliths allowed rapid and efficient separations of neutral and charged pesticides and metabolites, phenylthiohydrantoin amino acids and proteins at the time scale of seconds at relatively high flow velocity. (C) 2003 Elsevier B.V. All rights reserved.