Development of HPLC-NMR methods for the characterization of low concentration metabolites in bio-fluids
Abstract
Nuclear Magnetic Resonance (NMR) spectroscopy is a powerful analytical method for the identification and quantification of a large number of small molecules in biological mixtures. The complexity that characterizes samples used for bioanalytical investigations has resulted in the need to utilize the two complimentary analytical techniques; liquid chromatography (LC) and Nuclear Magnetic Resonance (NMR). This is especially the case in bio-fluid analysis in which a typical 1H NMR spectrum is characterized by resonances arising from the myriad of molecules present. Thus the use of a separation technique prior to NMR analysis is potentially beneficial as it simplifies the resulting NMR spectrum consequently making data analysis less cumbersome. Additionally the use of sample preconcentration methods prior to NMR detection has extended the limits of detection of several molecules in biological matrices. A comprehensive discussion of some of the advancements that have been made in order to address the major limitations of NMR methods for biological investigations is presented highlighting applications to single molecule detection as well as multiple metabolites' identification after stable isotope labeling in bio-fluids. The use of selective total correlation spectroscopy (TOCSY) for improved metabolomic studies is also reexamined with focus on improved quantitation.
Degree
Ph.D.
Advisors
Raftery, Purdue University.
Subject Area
Analytical chemistry|Biochemistry
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