| Capillary electrophoresis (CE) offers excellent separation efficiency for the analysis of complex mixtures. Also, the small dimensions of capillary tubes offer opportunities for the analysis of extremely small amount of samples, specially those of biological interest. In this thesis, capillary zone electrophoresis (CZE) and micellar electrokinetic capillary chromatography (MECC) are used to separate fluorescently labeled amino acids (chapter 2), peptides (chapter 3), saccharides (chapter 4 and 5), and chemiluminescently labeled amino acids (chapter 6).;Post-column laser-induced fluorescence (LIF) detection in a sheath flow cuvette is the most sensitive technique for capillary electrophoresis. In this thesis, a low-cost and low-power helium-neon laser, operating in the green at 543.5 nm, is used to xcite fluorescently labeled biomolecules. The relatively long excitation wavelength, the low light scatter of the sheath flow cuvette, and the low excitation power results in a very low background signal, which, in combination with high efficiency collection optics, produces detection limits as low as 100 molecules. Two pre-column fluorescent dyes, tetramethylrhodamine isothiocyanate (TRITC) and 5-carboxy-tetramethylrhodamine succinimidyl ester (TRSE), are used to label 20 common amino acids, six monosaccharide standards for oligosaccharide sequencing, and synthetic oligosaccharide substrates for glycosyltransferase assays.;Incomplete pre-column fluorescent labeling of ;A post-column chemiluminescence detector using a sheath flow cuvette is designed for capillary electrophoresis separation of isoluminol thiocarbamyl derivatives of 20 amino acids. This system is optimized and linear calibration curves are constructed. The dynamic range of the instrument is at least three orders of magnitude. Detection limits for isoluminol and isoluminol thiocarbamyl derivative of valine are 40 and 500 attomole, respectively. |
