Protein separation by monolithic capillary electrochromatography.
This work presents the separation of model proteins by capillary electrochromatography involving a monolithic stationary phase with C4 functionality. The monolith was fabricated in UV-transparent capillaries by employing a slight modification of a recently published photopolymerization procedure. With the number of theoretical plates per column ranging between 11000 and 33000, the separation efficiency proved to be lower than capillary zone electrophoresis where plate numbers ranged between 18000 and 66000. However, higher resolution was obtained due to the additional chromatographic separation mechanism. Inter- and intra-column reproducibility were evaluated, the latter could be significantly improved when using a rinsing procedure that contained 0.05% sodium dodecylsulfate in the mobile phase. Plate heights became nearly independent of mobile phase velocities higher than 0.5 mm/s indicating that high velocities can be applied without sacrificing efficiency. Furthermore, peak heights showed a dependence on injection times. For proteins, an increase in capacity factors was found when increasing the percentage of organic solvent in the mobile phase.
Bandilla
Dirk
D
Department of Chemistry and Biochemistry, Concordia University, 1455 de Maisonneuve Boulevard Ouest, Montréal, PQ Canada, H3G 1M8.
Skinner
Cameron D
CD
D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods*; D008855:Microscopy, Electron, Scanning; D011506:Proteins / Q000302:isolation & purification*; D015203:Reproducibility of Results
D011506:Proteins