| The objective of this thesis has been to introduce and develop novel methods for microchip separations for bioanalytical applications.; A novel detection scheme is introduced, involving simultaneous dual amperometric and fluorescence detection on a microchip. Dual detection is shown to increase selectivity and throughput, resolve co-migrating species that may be selectively detected, and provide a convenient means to normalize for the irreproducibility of migration times often encountered in CE applications. Such normalization is expected to facilitate the use of microchip CE to monitor biological samples, which are inclined to exacerbate the irreproducibility of migration times.; The use of electrochemical detection presents a unique and fundamental challenge. An effective method for reproducibly regenerating a clean surface is demonstrated. The method is optimized and utilized to achieve high sensitivity even for highly adsorptive compounds, such as those released from mast cells. The development of an in-situ electrode-cleaning protocol is an essential step toward reliably monitoring cellular release on a microchip CEEC device.; Two novel techniques are presented which are capable of producing disposable microanalytical systems on glass. Electrodes and channels produced with these methods exhibit performance characteristics that are comparable to examples in current literature. These techniques demonstrate the feasibility of manufacturing a disposable glass lab-on-a-chip, which may be used for cellular analysis or as a point-of-use sensor.; Increased interest in analyzing biological samples has led to the development of a wide range of derivatizing agents for biological compounds such as amino acids and peptides. A common tag that is both fluorescent and electroactive is naphthalene-2,3-dicarboxaldehyde (NDA). While there has been much discussion regarding the stability of a similar compound, o-phthalaldehyde, there has been no discussion regarding the stability of NDA derivatives. In this thesis, the degradation of NDA derivatives under a variety of environmental influences is examined by liquid chromatography with mass spectrometric detection. Major degradation products and pathways are suggested and characterized.; This thesis will conclude with a discussion about the future improvements and opportunities envisioned for microchip CEEC as applied to cellular analysis. (Abstract shortened by UMI.)... |
