AMINO ACID ANALYSIS USING MICELLAR ELECTROKINETIC CAPILLARY CHROMATOGRAPHY

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The trace analysis of biological components has been a critical key towards the development of modern biochemistry and medicine. One particularly powerful contribution has been the development of separations techniques capable of performing rapid and quantitative analysis on small amounts of biogenic materials. High performance liquid chromatography (HPLC) has proven capable of resolving all naturally occurring amino acids in a relatively short time frame providing a robust method for the determination of amino acid composition of proteins and peptides. One significant drawback to HPLC, as well as many other currently available analysis methods, is the quantity of biological sample required to perform the analysis since heroic efforts are often required to isolate the components(s) of interest. As a conseuence, 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 proved to be an exceptionally capable and cost effective tool. The overall objective of this project is to extend the utility of micellar electrokinetic capillary chromatography (MECC), a mode of CE, towards the microanalysis of biological materials. The proposed studies will evaluate a novel series of organic additives used to promote changes in the electrochromatographic selectivity towards biogenic molecules in micellar systems at low concentrations. More specifically, the aim is to provide a rapid and reproducible separation scheme that produces complete resolution of all naturally occurring amino-acids with femto- mole mass sensitivities. This objective will be approached by evaluating a novel series of organic additives combined with traditional micellar phases to determine their effects on electrochromatographic retention and selectivity through a series of systematic studies altering each chromatographic parameter independently. Upon a firm understanding of the behavior of these additives in micellar systems, optimization of a MECC method for the quantitative determination of derivatized amino acids will be addressed. This project will provide a better understanding of the utility of modified micellar phases in MECC towards biogenic species and if successful provide a cost effective alternative to microanalysis of amino acids.