Developing Methods in Capillary Electrophoresis for the Detection of Carbon Monoxide Poisoning

摘要

This project demonstrates the use of capillary electrophoresis as an improvement to existing methods of analysis for carbon monoxide (CO) in human hemoglobin (Hb). Current methods for CO analysis are CO-oximetry and tonometry/ gas chromatography, which involve lengthy sample manipulation followed by spectroscopic or gas displacement measurements. These methods are time-consuming and technically difficult because of the many manipulations they require. Capillary electrophoresis (CE), a modern analytical technique, is presented as a faster and easier method to quantify CO in Hb. Several steps towards the development of a CE procedure to detect CO in blood have been completed. The first step of this analysis involved isolating heme, the CO binding site in Hb, from the Hb molecule. Initially, mixtures of reduced heme and heme-CO were successfully isolated from Hb standards and separated using CE. Finally, reduced heme and heme-CO were isolated from three blood samples of CO-related accident victims and analyzed using CE. Differences in the heme-CO signals from blood samples known to contain fatal and non-fatal levels of CO were observed. These differences were quantified by measuring the areas under the peaks representative of reduced heme and heme-CO. Results indicate that an improvement was made to the current methods of CO analysis.