Graduation Semester and Year

2018

Language

English

Document Type

Dissertation

Degree Name

Doctor of Philosophy in Chemistry

Department

Chemistry and Biochemistry

First Advisor

Kevin A Schug

Abstract

In recent years, the innovation in biomarkers and biotherapeutics has increased the demand for state-of-the-art analytical platforms to achieve reliable and accurate discoveries and quantitative analysis methods. The challenges in their determinations are not limited to the large number of small and large biomolecules but also involve the great heterogeneity in chemical properties and abundance of those analytes. Liquid chromatography and mass spectrometry have advanced significantly to be the most common platform for analyzing biological samples. However, there are still many unmet challenges that that require application of novel bioanalytical methods to fill many gaps in online sample preparation, chromatographic separation, and mass spectrometric detection of different analyte sets. Frontal chromatography was used to investigate the trapping efficiency and capacity of online restricted access media trap columns to streamline sample preparation for online enrichment of small molecule analytes in complex biological samples. To enhance the sensitivity and selectivity for determination of 14 biomarkers of exposure to BTEX in urine, we developed a novel one pot charge reversal derivatization kit, optimized its performance through multivariate analysis using central composite design, and then validated its performance by using isotope dilution mass spectrometry. We further developed a novel high pH reversed phase chromatographic method for orthogonal separation of intact proteins using mass spectrometry compatible buffers. Finally, we adopted this method as the first dimension in a comprehensive two-dimensional reversed phase x reversed phase-mass spectrometry platform for intact protein separations.

Keywords

Bulk derivatization, Biological matrices, On-line sample preparation, Quantitative analysis, Derivatization reactions, Frontal chromatography, Solid phase extraction, Biomarkers, BTEX, Central composite design, Occupational exposure, Metabolites, High pH, Reversed phase liquid chromatography, Intact proteins, Top-down proteomics, Altered selectivity, Comprehensive chromatography, LC x LC method development, High pH separations, Triple quadrupole mass spectrometry

Disciplines

Chemistry | Physical Sciences and Mathematics

Comments

Degree granted by The University of Texas at Arlington

Included in

Chemistry Commons

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