NMR-based metabolic profiling procedures for biofluids and cell and tissue extracts

Abstract

Metabolomic studies offer a wealth of information on cells, tissues, and biofluids. The phenotype representation through the metabolic profiling is a valuable tool for direct diagnosis, therapeutic strategies, and system’s biology studies. Nuclear magnetic resonance (NMR) spectroscopy provides a nondestructive and extremely reproducible method allowing simultaneous detection of a large number of known and unknown chemical substances. Sample collection and preparation and experimental conditions are critical for the reliability of the subsequent analysis. The pre-analytical phase is decisive as it could generate biased spectral data misleading the following analysis. The formulation of standard operating procedures is thus of crucial importance in order to access meaningful samples and results. In this protocol, we provide standardized operations and routine procedures from sample preparation to determine the measurement details for the acquisition of NMR spectra highlighting major methodological issues.