Metabolite Analysis of Biological Fluids and Tissues by Proton Nuclear Magnetic Resonance Spectroscopy

NMR-based biochemical profiling of natural products has become popular due to the development of high-resolution instruments (>400 MHz) and cryogenically cooled probes/preamplifiers, by increasing the sensitivity of NMR instruments several fold and reducing instrument noise levels. NMR provides a rapid, nondestructive, high-throughput method that requires minimal sample preparation, therefore maintaining the biological integrity of the sample. One-dimensional (1D) solution-state ¹ H NMR is used in untargeted sample screening (metabolomics/metabonomics) to gain insight into spectral pattern changes associated with samples of different origins. Metabolomics and metabonomics contextually explains the systematic and quantitative measurement of metabolites that are produced from the biochemical reactions of living systems. This chapter describes some commonly used ¹ H NMR experiments for identification and quantification of small molecular weight, water soluble metabolites in biological samples, some considerations for choosing the correct NMR experiment, and sample preparation protocols for isolating metabolites from a number of biological sample types.