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
License
This work is licensed under a Creative Commons Attribution-NonCommercial-Share Alike 4.0 International License.
Recommended Citation
Baghdady, Yehia Zakaria, "ONLINE BIOANALYTICAL MASS SPECTROMETRY METHODS FOR ANALYSIS OF SMALL AND LARGE BIOMOLECULES USING ONE- AND TWO-DIMENSIONAL LIQUID CHROMATOGRAPHY" (2018). Chemistry & Biochemistry Dissertations. 179.
https://mavmatrix.uta.edu/chemistry_dissertations/179
Comments
Degree granted by The University of Texas at Arlington