Romano, Francesca (2025) FROM MOLECULES TO HEALTH: METABOLOMICS APPROACHES TO STUDY NUTRACEUTICALS AND FUNCTIONAL FOODS. [Tesi di dottorato]

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Abstract

Metabolomics is the scientific discipline dedicated to the comprehensive identification and quantification of exogenous and endogenous metabolites within biological systems. Due to their close relationship with phenotype, metabolites provide highly informative readouts of physiological and pathological states, and metabolomics is increasingly applied across biomedical, pharmaceutical, food, and nutritional sciences. The integration of advanced analytical platforms, including Nuclear Magnetic Resonance (NMR) spectroscopy and Mass Spectrometry (MS), together with advanced chemometric and statistical tools, enables the generation of metabolic “fingerprints” capable of elucidating complex biochemical processes. The first part of this thesis is devoted to the theoretical foundations of metabolomics, encompassing the principles of both the analytical techniques and the chemometric approaches. This theoretical framework provides the basis for the practical studies presented in the subsequent sections, which apply these methodologies to address questions in nutraceutical research, food science, and disease models. Building on this foundation, this PhD project employed metabolomics to elucidate the molecular signatures associated with nutraceuticals and food-derived compounds in various experimental settings. The primary aim was to advance the understanding of their functional properties and biological effects, revealing how bioactive molecules and food processing conditions shape metabolic responses and contribute to health-promoting outcomes. The second part of the thesis presents these applications in detail, encompassing five metabolomics studies performed on different sample types. Specifically, the first two papers report complementary preclinical studies investigating the effects of mango (Mangifera indica L.) in inflammatory bowel disease. In Paper II, a reverse translational approach combined ex vivo assays with a T cell transfer model of colitis, complemented by faecal metabolomics, to unravel the immunomodulatory and microbiota-related effects of mango extract. This study highlighted the ability of mango bioactive compounds to attenuate T cell–driven inflammation, modulate gut microbial metabolism, and restore host–microbe homeostasis. Paper V extended this research to a DNBS-induced rat model of colitis, integrating immunological, histological, and behavioural endpoints with untargeted metabolomic profiling of faecal and colonic samples. This integrative design provided novel insights into the biochemical mechanisms underlying the protective effects of mango extract, not only in reducing gut inflammation but also in alleviating visceral pain. The third and fourth papers focus on Iranian Lamiaceae species, namely Salvia and Phlomis. Targeted metabolomics combined with spectrophotometric assays allowed a comprehensive characterization of their phytochemical diversity, revealing species-specific enrichment in polyphenols and other bioactive compounds. Notably, the observed differences between species were much more pronounced than those arising from collection region or altitude, highlighting the dominant influence of genetic factors on metabolite composition. These studies provided insights into the potential therapeutic applications of these plants, ranging from antioxidant to anti-inflammatory activities, and highlighted metabolite signatures that could guide future nutraceutical development. The fifth study (Paper IV) extends the application of metabolomics to animal-derived foods, specifically eggs, to evaluate how different cooking methods affect chemical composition and quality. Both conventional and innovative approaches, including the novel periodic cooking technique, were assessed for their impact on metabolite profiles, texture, and nutritional content. The periodic cooking method appeared to outperform conventional techniques, enhancing not only the sensory and organoleptic properties of the eggs but also their nutritional quality. This work emphasizes the impact of processing methods on the biochemical and nutritional composition of foods, while illustrating the value of metabolomics as a supporting approach in the study and optimization of functional food quality. Taken together, the studies presented in this thesis demonstrate the value of metabolomics as an analytical strategy for nutraceutical and food research. By integrating metabolomic profiling with biological, physiological and statistical data, this work contributes to a more mechanistic understanding of the relationship between food components and health, supporting the rational development of functional foods and nutraceutical applications.

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