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New studies and developments in capillary electrophoresis : advancements in complex mixture analysis and preparative operation McLaren, David G.

Abstract

New developments in the areas of complex mixture analysis and preparative operation using capillary electrophoresis are presented. Micellar capillary electrophoresis has been applied to the separation of free amino acids in human plasma for the purpose of identifying inborn errors of metabolism. The method developed is capable of separating 32 amino acid conjugates in under 70 minutes and makes use of selected additives to improve the resolution and overall analysis time. A mechanistic investigation into the influence of simple, univalent salts on micelle selectivity and capacity factor is also presented and the use of these additives in the context of complex mixture analysis is explored. New techniques based on the model of flow counterbalanced capillary electrophoresis are described and the optimization of experimental parameters for preparative operation is discussed. Methods for the isolation of individual analytes from multicomponent samples on a micropreparative scale are presented. Under optimum conditions, recoveries as high as 17.5% of the amount present in the original sample can be achieved with exceptional purity. Breakdown of the buffering capacity of the background electrolyte was found to play an important role in determining the maximum recoveries attainable. Methods of mediating electrolytic modification of the buffer during the extended run times necessary for preparative operation are presented and were found to significantly improve the performance of the system. In order to develop new methods for preparative operation of capillary electrophoresis, a new instrument had to be designed. The performance of this specialized instrument is discussed with respect to separation and injection reproducibility, thermal management, sensitivity, limits of detection, and linear dynamic range.

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