Novel modes of capillary electrophoresis for fast separations and reaction-based chemical analysis
Abstract
Modern analytical chemists strive to analyze increasingly smaller sample volumes with increased speed. Capillary electrophoresis has proven to be an effective mode by which sub-nanoliter samples may be separated at high speed. To date, capillary electrophoresis has not become completely integrated into the biological laboratories due to its lack of ability to perform on-column assays that equal those of polyacrylamide gel electrophoresis coupled to the blotting techniques. In this thesis, novel modes for capillary electrophoresis are presented for fast separations and reaction-based chemical analysis. Chapter 1 describes moving boundary capillary electrophoresis (MBCE) as a capillary electrophoretic mode for fast separations and is introduced by the description of the theory of electrophoresis and joule heating effects. MBCE is theoretically described in terms of the concentration profile and concentration gradient profile of the moving boundary as well as the electric field effects. MBCE electropherograms of proteins, amino acids and nucleotides are presented to show that MBCE is an effective way of performing fast efficient separations. The remainder of the thesis describes a capillary electrophoresis experimental setup for reaction-based chemical analysis. The electrophoretically mediated microanalysis (EMMA) procedure is described in chapter 2 in terms of the reagent-filled capillary approach and the plug-plug approach. Constant potential electropherograms of many different enzyme systems are shown for both the analysis of the enzyme and the substrate. The zero potential mode is qualitatively and quantitatively described and is shown to be effective for the determination of small amounts of enzymes. Chapter 3 mathematically describes the EMMA process using the ADH-ethanol system as an experimental model. The EMMA process is used to analyze for the calcium ion in chapter 4 through a complexation reaction with o-cresolphthalein complexone. A dynamic computer simulation of EMMA is presented in chapter 5 and shown to agree with experimentally obtained results.
Degree
Ph.D.
Advisors
Regnier, Purdue University.
Subject Area
Analytical chemistry
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