| Small molecule analysis has been critical in developing an understanding of biological processes. Small molecule metabolites are the products enzyme activity within an organism and study of these molecules, collectively called the metabolome, paints a clearer picture of the phenotype of an organism than genomics, transcriptomics or proteomics. In addition, study of drug metabolism is critical in diagnosing efficacy and safety of potential drug candidates. Recent advances in detection technology have facilitated highly sensitive analysis of the metabolome from samples as small as single cells. This work has revealed a complexity far beyond the simple metabolism charts depicted in biochemistry textbooks. Because of the complexity of the metabolome, a separation technique is often utilized to simplify the sample before analysis. Throughput is of particular interest to metabolomics research, as biological variation between the metabolome of individual organisms is high, requiring studies with large sample sizes and significant instrument run-time.;In this work, I present capillary electrophoresis as a complementary separation technique to high-performance liquid chromatography and gas chromatography for the separation of metabolite samples. In particular, I focus on demonstrating the usefulness of capillary electrophoresis for performing rapid separations lasting less than ten minutes. Capillary electrophoresis is especially well suited for the analysis of small, polar metabolites because of its high resolution and aqueous running conditions. In addition, I demonstrate and investigate new technologies for coupling capillary electrophoresis to mass spectrometry and ionizing molecules for mass spectrometric analysis and detection. As a final demonstration of the capability of capillary electrophoresis to rapidly analyze the metabolome, I present a study of the metabolism of early Xenopus laevis embryos. |
