| The miniaturization of separation systems has been widely applied to the area of protein analysis. This work focuses on the use of packed capillary columns for liquid chromatography (LC). Both electrokinetic pumping as used in capillary electrochromatography (CEC) and pressure based pumping will be explored. The fabrication of packed capillary with two kinds of stationary phases is addressed. A splitless-flow nano-LC system in combination with laser induced native fluorescence detection was developed, which showed separation efficiency of 9.9x104 for tryptophan and mass detection limit of 5.1 fmol for alpha-lactalbumin. Factors affecting band broadening were investigated: injector, connecting tubing and detector. To contribute to the understanding of the mechanism of protein concentration found in CEC columns packed with nano-porous particles, this thesis explores the behavior of protein in nano-porous stationary phase used for electrokinetic chromatography. Further, protein separation was demonstrated in pressure assisted size-exclusion electrochromatography (pSEEC). |
