Fluorescent Components in Human Urine. Characterization by High Performance Liquid Chromatography With Fluorescence Detection (Hplc)
Baer, Charles Sherman
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https://hdl.handle.net/2142/70193
Description
Title
Fluorescent Components in Human Urine. Characterization by High Performance Liquid Chromatography With Fluorescence Detection (Hplc)
Author(s)
Baer, Charles Sherman
Issue Date
1982
Department of Study
Chemistry
Discipline
Chemistry
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Chemistry, Analytical
Abstract
This study has characterized the fluorescent components of the human urine matrix using a unique HPLC-fluorescence detection system. The detection system provided characterization information based on the results returned by a reverse file-search of trapped chromatographic peak excitation and emission spectra searched against a fluorescence library containing 1230 spectra. The best candidates returned by the search offered structural suggestions for use in characterization of the unknown compound. Secondary instrumentation was used to verify the characterizations made by the fluorescence file-search system. However, the methods employed were not sensitive enough to provide any new information.
To perform this study extraction and chromatographic methods had to be developed. Various procedures were tried before optimal conditions were chosen for the study.
Hydrolysis of conjugate urinary metabolites was explored using both acidic and enzymatic conditions. The enzymatic method was found to be preferable due to its less severe reaction conditions. The resulting free fluorescent urine components were characterized using the results from the reverse file-search system.
The system and methodology developed was applied to a study of a set of fluorescent pharmaceuticals spiked into urine samples. Detection limits of 100 ng/mL and below were achieved with this system. Various extraction and chromatographic procedures were explored to demonstrate the applicability of this study to future fluorescence pharmaceutical urinalysis studies.
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