Abstract

This dissertation focuses on the development of hand-portable capillary liquid chromatography (LC) instrumentation. In this work, battery-operable nano-flow pumping systems (isocratic and gradient) were developed and integrated with portable UV-absorption detectors for capillary LC. The systems were reduced in size to acceptable weights and power usage for field operation. A major advantage of the pumps is that they do not employ a splitter, since they were specifically designed for capillary column use, thereby greatly reducing solvent consumption and waste generation. UV-absorption detectors were specifically designed and optimized for on-column detection to minimize extra-column band broadening. Initially, an isocratic nano-flow pumping system with a stop-flow injector was integrated with an on-column UV-absorption detector (254 nm). The pumping system gave excellent flow rate accuracy (<99.94%) and low percent injection carry-over (RSD 0.31%) suitable for quantitative analysis. Using sodium anthraquinone-2-sulfonate, the detector gave an LOD (S/N = 3) of 0.13 µM, which was 12 times lower than a commercial UV-absorption detector. Reversed-phase separations of a homologous series of alkyl benzenes was demonstrated. Further miniaturization of UV-absorption detection was accomplished using a 260 nm deep UV LED. The detector was small in size and weighed only 85 g (without electronics). No optical reference was included due to the low drift in the signal. Two ball lenses, one of which was integrated with the LED, were used to increase light throughput through the capillary column. Stray light was minimized by the use of a band-pass filter and an adjustable slit. Signals down to the ppb level (nM) were easily detected with a short-term noise level of 4.4 µAU, confirming a low limit of detection and low noise. The detection limit for adenosine-5'-monophosphate was 230 times lower than any previously reported values. Isocratic separations of phenolic compounds were performed using a poly(ethylene glycol) diacrylate monolithic capillary column. Finally, a novel nano-flow gradient generator integrated with a stop-flow injector was developed. Gradient performance was found to be excellent for gradient step accuracy (RSD < 1.2%, n = 4) and linear gradient reproducibility (RSD < 1.42%, n = 4). Separations of five phenols were demonstrated using the nano-flow gradient system. Efforts to develop a 405 nm laser diode-based UV-absorption detector for hemoglobin analysis were described.

Degree

PhD

College and Department

Physical and Mathematical Sciences; Chemistry and Biochemistry

Rights

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