Chemoselective tags for nuclear magnetic resonance spectroscopy-based metabolic profiling
Abstract
Metabolic profiling has received increasing recognition as an indispensable complement to genomics and proteomics for probing biological systems and for clinical applications. It provides important information to aid the full understanding of composition and function of biochemical networks and may lead to solutions of many important questions such as human disease diagnosis, prognosis and therapeutic development. Nuclear magnetic resonance (NMR) and mass spectrometry (MS) are the major techniques used in this field to detect up to hundreds of small molecules in parallel. MS is highly sensitive but not well reproducible and quantitative. 1H NMR is highly reproducible and quantitative but lacks sensitivity. For metabolic profiling, spectral overlap has seriously impeded the identification and quantitation of metabolites in complex mixtures such as human urine and blood serum/plasma. In this thesis, the use of chemoselective isotopic tags allows the analysis of metabolites with certain functional groups using 1H-15N and 1H-13C 2D NMR techniques at much improved resolution. While signals of less than 40 metabolites are usually recognized from the conventional 1H 1D NMR spectra of human biofluids, well over 100 clearly resolved 2D signals are typically observed using the new approach with high reproducibility.
Degree
Ph.D.
Advisors
Raftery, Purdue University.
Subject Area
Analytical chemistry|Biochemistry
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