Application of chemiluminescent detection to a capillary electrophoresis system contained on a glass slide and the development of non-flowing electrogenerated chemiluminescence probes.

摘要

This research has involved expanding the applicability of aqueous phase chemiluminescence detection from the mechanically induced flowing streams of flow injection analysis and high performance liquid chromatography to non flowing probe devices and micromachined electrophoresis systems. The chemiluminescent reagent used for most of this research was tris(2,2;An electrogenerated chemiluminescent probe which does not require the pump(s) and tubing associated with a flow system nor the use of a dark box has been constructed. The general concept for this probe was a device which can be dipped into a solution in a fashion similar to the use of a pH electrode. This device incorporates components necessary for all electrogenerated chemiluminescent systems. A self contained three electrode system is required for all electrogenerated chemiluminescence systems. The sample has free access to all electrodes and is able to flow freely throughout the probe to ensure proper electrical contact and reproducible measurements. Ambient light is excluded from the analytical signal. The collection of the light emission is efficient while the overall size of the probe remained relatively small.;While developing electrogenerated chemiluminescence methods, it is often necessary to investigate the emission characteristics of a reaction occurring within the electrogenerated chemiluminescence cell. The typical method for obtaining this information is to interpret the results of several experiments while varying flow rates, cell spacer thickness, and by using stopped flow data. The stopped flow data gives the most direct information about emission characteristics but is a solution phase system whereas electrogenerated chemiluminescence occurs at the surface of an electrode. To more directly obtain information about electrogenerated chemiluminescent processes, a low volume (125 ;A capillary electrophoresis system contained on a micromachined glass slide which integrates chemiluminescence as the detection method has been constructed and demonstrated as a feasible analytical technique. A simple method which does not require highly specialized facilities to manufacture these planar devices has concurrently been developed. This separation and detection system was characterized by separating and detecting mixtures of proline and tripropylamine. To further illustrate the applicability of this device, the anti-anxiety drug buspirone and its major metabolite 1-(2-pyrimidyl)piperazine were added to a human plasma matrix. The drug and metabolite were then separated from this matrix and detected.