Nuclear Magnetic Resonance (NMR)-Based Drug Metabolite Profiling

The identification of drug metabolites in biofluids such as urine, plasma and bile is an important step in drug discovery and development. Proton nuclear magnetic resonance (¹ H-NMR) spectroscopy can provide detailed information regarding the structural transformation of a compound as a consequence of metabolism. However, successful identification of drug metabolites by ¹ H-NMR spectroscopy is generally compromised by the presence of endogenous metabolites, which can obscure the signals of the drug metabolites in question. Hence, sample clean-up and separation of the metabolites from the biofluid matrix is crucial. This is generally achieved by extraction of the biofluid, solid-phase extraction (SPE), high-performance liquid chromatography (HPLC) or any combination of these. Apart from ¹ H, other NMR-active nuclei, such as ¹⁹ F, can provide a useful handle for metabolite profiling, provided they are not naturally present in the biofluid. Successful studies have shown that the presence of a fluorine-handle on the drug and its metabolites can provide additional qualitative and quantitative data by ¹⁹ F-NMR spectroscopy. This chapter provides guidelines and examples of NMR-based drug metabolite profiling.