Instrumentation for capillary electrochromatography.

Abstract

One of the reasons for the immense interest in capillary electrochromatography (CEC) is its feature to combine chromatographic selectivity with the high efficiency and the miniaturization potential of capillary electrophoresis (CE). The capability of commercial CE instruments to run CEC has enforced the readiness of users and researchers to work on this separation technique. Nevertheless, to fully exploit the potential of CEC, a routine CE device can certainly not fulfill all requirements. Two different approaches have been made to overcome this problem. The first was to modify commercial CE instruments for various demands. Pressurization of the packed capillary to prevent "air" bubble formation, gradient elution capabilities and thermostating devices allowing a greater flexibility in column designs have been implemented in CE instruments of several manufacturers. A completely different approach is the development of modular laboratory-made instrumentation dedicated to special CEC requirements. In order to increase mobile phase velocity and thus the speed of analysis the availability of voltages higher than 30 kV was accomplished in some of these devices. Gradient elution was achieved by either coupling of gradient LC systems or an electroosmotic generation of the changing eluent composition. When a pressure gradient is applied between both column ends in addition to the voltage gradient, a hybrid between capillary HPLC and CEC results. This chromatographic mode is named pressure-assisted electrochromatography (PEC). Either CE instruments equipped with additional HPLC pumps or modular laboratory-made devices are suitable for PEC. In CEC, sensitivity for UV detection is rather poor due to the short optical path length for on-column detection in capillary separation techniques. A special cell design with enhanced light path is presented and further principles like, e.g., fluorescence detection and coupling to mass spectrometry are discussed.