MEABOLOMICS BY NMR: APPLICATIONS AND CHALLENGES FROM BIOMEDICINE TO FOOD RESEARCH

Metabolomics is an emerging technology that holds promise of powerful insights into many different fields. Nuclear magnetic resonance spectroscopy represents an optimal tool in modern chemical and biochemical research; for this reason, numerous applications can be found in biology, biomedicine and food-stuff studies. Nuclear magnetic resonance spectroscopy offers many advantages to perform metabolomic analysis. Overall, because of its characteristics, NMR is most suited for metabolomics fingerprinting studies. Metabolomics is defined as the analysis of the complete ensemble of low molecular weight molecules, the metabolites, present in a biological specimen. In this methodological thesis, with the aim of demonstrating the potential of untargeted NMR-based metabolomics approach from biomedicine to food research studies, different topics have been addressed and discussed: i) the use of NMR-metabolomics applied on biological fluids to unravel fingerprints/biomarkers, both for drug response phenotypes, providing an effective means to predict variation in response to treatment; and for the diagnosis/differentiation of pathological states; ii) the characterization of a common metabolic drift in a heterogeneous group of different cellular breast cancer subtypes following the development of drug resistance, providing in vitro models which will eventually lead the way to early prediction of treatment inefficacy on patients with breast cancer (personalized medicine); iii) the application of NMR- based metabolomics to demonstrate the potentiality in determining molecular features (biomarkers descriptors of plant stress and/or organoleptic properties) of olive pre-harvest and post-harvest phases to exploit the possibility for the establishment of a new, large-scale applicable protocol (treatments or additional post-harvest manipulations) which will eventually be utilized by olive mills and olive oil producers to improve quantity and quality of the final products; iv) the use of NMR-metabolomics to assess food products provenance, even at the scale of several different farms in a restricted geographical territory, such as small milk farms in Mugello (Tuscany), or coffee plantations of a restricted area of the Huila district in Colombia, and proving that NMR can be an accurate and automated service for determining food quality based on metabolomic descriptors. Results presented in this thesis, obtained by a combination of biochemistry, analytical chemistry, bioinformatics, and descriptive data, contribute to the demonstration that NMR-based metabolomics can be considered as a whole as a "universal" quantitative analytical technique. Offering an unbiased and untargeted view of samples composition and the ability to simply quantify multiple compounds simultaneously, could be considered a very powerful choice for diagnosis studies as well as for the characterization and quality control of complex biological/natural samples such as biofluids, foods, plants and cellular extracts.