The trace analyses of biological components have played a key role towards the development of modern biochemistry. One important contributions has been the development of separations techniques capable of performing rapid and quantitative analysis on minute amounts of biogenic materials. High performance liquid chromatography (HPLC) has proved a capable tool in resolving complex mixtures providing robust methods fro the determination of amino acid composition and sequencing. One significant drawback to HPLC is the quantity of protein or peptide required to perform the analysis, since heroic efforts are often required in isolating these components. As a consequence, more sensitive microanalysis separation schemes are being developed to provide increased mass sensitivities and improved resolving capabilities. Among these new techniques, capillary electrophoresis (CE) has proven to be exceptionally capable and cost effective. One of the modes of CE is micellar electrokinetic capillary chromatography (MEKC) which has developed into a powerful technique for the analysis of many species and gives an alternative selectivity from traditional models of electrophoresis. The overall objective of this project is to extend the utility of MEKC towards the routine analysis of amino acids. The proposed studies will utilize """"""""class I"""""""" organic modifiers to induce improvements in resolution for biogenic species over that observed with traditional surfactants alone. More specifically, the aim is to provide a rapid and reproducible separation scheme providing the complete resolution of naturally occurring amino acids with subfemtomole mass sensitivities This objective will be achieved utilizing """"""""class I"""""""" organic modifiers combined with traditional micellular phases to provide resolution of complex sets of amino acids. Amino acids will be derivatized with commercially available fluorophores and detected using laser induced fluorescence detection (LIF). LIF-MEKC schemes directed for the quantitative determination of amino acids will be optimized. This project will provide a better understanding of micelle modifiers and explore their utility in MEKC towards biogenic species and if successful provide a quicker and more sensitive alternative to analysis of amino acids.
