SUSTAINABLE RECOVERY OF BIOACTIVE AND FUNCTIONAL COMPOUNDS FROM NATURAL SOURCES

THE GROWING DEMAND FOR SUSTAINABLE FUNCTIONAL COMPOUNDS, BIOACTIVE MOLECULES OBTAINED FROM RENEWABLE PLANT SOURCES, HAS DRIVEN THE DEVELOPMENT OF ECO-FRIENDLY EXTRACTION TECHNOLOGIES. IN THIS FRAMEWORK, THE EXPLORATION OF WILD PLANTS AND UNCONVENTIONAL PARTS OF COMMON CROPS AS VALUABLE SOURCES OF FUNCTIONAL COMPOUNDS IS STRONGLY ENCOURAGED BY EU DIRECTIVES, THE UNITED NATIONS SUSTAINABLE DEVELOPMENT GOALS (SDGS) AND THE GLOBAL TRANSITION TOWARD A CIRCULAR ECONOMY. DURING THE FIRST YEAR OF THE PHD PROJECT, A SOFTWARE-ASSISTED UNTARGETED UHPLC-HRMS/MS WORKFLOW WAS DEVELOPED AND COMBINED WITH IN VITRO SCREENING ASSAYS TO ASSESS THE ANTIOXIDANT AND ENZYME INHIBITORY POTENTIAL OF VARIOUS PLANT MATRICES. FOLLOWING A FUNNEL-SHAPED STRATEGY, DATA WERE INTEGRATED WITH LITERATURE FINDINGS, AGRI-FOOD VALUE CHAIN RELEVANCE, AND SUSTAINABILITY CRITERIA, ENVIRONMENTAL PRESERVATION AND RESOURCE OPTIMIZATION, TO IDENTIFY THE MOST PROMISING NATURAL SOURCES FOR SUSTAINABLE EXTRACTION. THIS ANALYSIS LED TO THE SELECTION OF CERATONIA SILIQUA L. (CSL) AND GLYCYRRHIZA GLABRA L. (GGL) LEAVES, UNDERUTILIZED BIOMASSES OF TRADITIONAL ITALIAN CROPS, AS VALUABLE RESOURCES FOR THE RECOVERY OF HEALTH-PROMOTING COMPOUNDS. FOR THEIR RICH METABOLIC PROFILES AND STRONG IN VITRO ACTIVITIES, BOTH MATRICES PROVED TO BE PROMISING CANDIDATES FOR THE DEVELOPMENT OF NOVEL FUNCTIONAL FOODS AND NUTRACEUTICAL FORMULATIONS. THE SECOND YEAR FOCUSED ON GREEN EXTRACTION TECHNIQUES, SELECTED, AND CAREFULLY OPTIMIZED FOR THE SELECTIVE AND EFFICIENT RECOVERY OF CSL AND GGL COMPOUNDS. PRESSURIZED HOT WATER EXTRACTION (PHWE) AND ULTRASOUND PROBE-ASSISTED EXTRACTION (PROBE-UAE) WERE CHOSEN FOR THE EXTRACTION OF CSL POLAR AND MEDIUM-POLAR COMPOUNDS, AND SUPERCRITICAL CO2 EXTRACTION (SFE) FOR GGL APOLAR CONSTITUENTS. PRELIMINARY UNIVARIATE STUDIES AND SUBSEQUENT RESPONSE SURFACE METHODOLOGY (RSM) OPTIMIZATION ENABLED EFFICIENT, SOLVENT-SAVING AND ENVIRONMENTALLY FRIENDLY EXTRACTION. THESE GREEN METHODS OUTPERFORMED CONVENTIONAL TECHNIQUES IN TERMS OF YIELD, SELECTIVITY, QUALITY AND SUSTAINABILITY AND PROVIDE A SOLID BASIS FOR FUTURE PILOT-SCALE TRIALS AND POTENTIAL INDUSTRIAL EXPLOITATION OF THESE HEALTH-PROMOTING NATURAL SOURCES. BUILDING ON THESE RESULTS, THE THIRD-YEAR RESEARCH INTEGRATED PROCESS OPTIMIZATION WITH BIOLOGICAL VALIDATION. CSL EXTRACTS WERE TESTED IN CELL-BASED MODELS (HEPG2 CELLS AND ERYTHROCYTES) TO CONFIRM ANTIOXIDANT EFFICACY UNDER OXIDATIVE STRESS, PROVIDING REALISTIC EVIDENCE OF THEIR BIOACTIVITY AND CYTOCOMPATIBILITY. IN PARALLEL, A PHWE-BASED SPRAY-DRIED FORMULATION WAS DEVELOPED TO ENHANCE COMPOUND STABILITY AND BIOACCESSIBILITY, WHICH WERE FURTHER ASSESSED THROUGH IN VITRO GASTROINTESTINAL DIGESTION. FOR GGL, ANTIOXIDANT AND ANTI-AGING PROPERTIES OF THE EXTRACTS WERE INVESTIGATED USING A SACCHAROMYCES CEREVISIAE MODEL OVEREXPRESSING HUMAN Α-SYNUCLEIN, ENABLING THE EVALUATION OF BOTH OXIDATIVE STRESS RESISTANCE AND LIFESPAN EXTENSION. OVERALL, THE FINDINGS FROM BOTH PLANT BIOMASSES UNDERLINE THE EFFECTIVENESS OF A SUSTAINABLE, MULTIDISCIPLINARY APPROACH THAT INTEGRATES GREEN EXTRACTION, METABOLITE CHARACTERIZATION, FORMULATION DESIGN AND BIOLOGICAL VALIDATION. BOTH C. SILIQUA AND G. GLABRA LEAVES REPRESENT PROMISING, UNDEREXPLOITED SOURCES THAT CAN SERVE AS HEALTH-BENEFICIAL CONSTITUENTS FOR THE DEVELOPMENT OF NOVEL FOODS, FUNCTIONAL INGREDIENTS, AND NUTRACEUTICAL FORMULATIONS.