FORMULATION OF A NEW NUTRACEUTICAL INGREDIENT USING GREEN TECHNOLOGIES

This PhD thesis is the result of my three-year research in the fields of food chemistry and green extraction, primarily conducted in the Food and Dietary Products Laboratory of the Department of Pharmacy (DIFAR) at the University of Genoa. This work also benefited from collaborations with other research groups at the University of Genoa, the Synthesis Team of AIMPLAS (Valencia, Spain), THEMIS S.p.A. (Milan, Italy), the Green Extraction Team of ANFACO-CECOPESCA (Vigo, Spain), and the Green Extraction Team of Norwegian food research institute NOFIMA (Tromsø, Norway) . Formulating a new and green nutraceutical ingredient involves several steps to ensure it is both effective and environmentally friendly. These steps include identifying the health benefit, selecting sustainable ingredients, implementing green manufacturing practices, developing formulations, and verifying regulatory compliance and possibly clinical testing. Following the preliminary introduction in Chapter 1, this thesis investigates some of these topics to obtain well-known healthy ingredients such as Collagen, Hydrolysed Peptides, Omega-3-rich oils, and Chitin/Chitosan from innovative sustainable sources using green processes. Three case studies are reported, focusing on valorising innovative biomasses from food industry side-streams (fishery, aquaculture, and insect larvae production) through green extraction methods to isolate both bioactive and functional compounds. The study highlights the use of unsorted, pretreated, and unusual by-products from food/feed industrial chains and the application of green extraction techniques (Enzyme-Assisted Extraction, Ultrasound-Assisted Extraction, Green Solvent-Assisted Extraction). The Multivariate Design of Experiments (DoE) was applied, in each case study, to optimize the extraction conditions of the proposed protocols. The extracted compounds were purified, stabilized mainly by spray-drying, chemically characterized, and tested for their potential industrial applications. Chapter 2 describes the extraction of collagen-derived bioactive compounds (Pepsin-Soluble Collagen, Gelatine, and Hydrolysed Gelatine Peptides) from fishery biomasses. Biomasses from undersized or damaged fishes (e.g., Mugil Cephalus) were used after dehydration by lyophilization to stabilize them. This process aimed to show that unwanted catches and/or by-catches can be used to extract bioactive compounds like Pepsin-Soluble Collagen (PSC), demonstrating a sustainable multi-circular economy model in the Liguria region of Italy. The study also involved extracting PSC from dehydrated by-products of the yellowfin tuna canning industry, as part of the Horizon 2020 project, EcoeFISHent (GA 101036428). Chapter 3 describes the implementation of two methods for extracting fish oils from EcoeFISHent fishery biomasses: solid/solvent extraction using green solvents (e.g. thanol and ethyl acetate) and enzyme-assisted extraction (EAE). The study compared these methods in terms of extraction yield, quality of the extracted oils, and their advantages and disadvantages. The novelty lies in using both green technologies and unsorted biomass from industrial filleting processes, which includes various parts like heads, scales, bones, and skin. This biomass was previously dehydrated by an industrial patented process to improve logistics and avoid the costly cold chain, focusing on maintaining food quality by preventing lipid oxidation. Chapter 4 describes the study to isolate chitin/chitosan from both edible larvae and adult insects of Tenebrio Molitor comparing a traditional chemical extraction with an innovative enzymatic-assisted extraction (EAE) method, optimized by design of experiments (DoE). The process involved, deproteinization, and deacetylation to obtain chitosan, which was chemically characterized using Fourier Transform Infrared Spectroscopy (FT-IR), Scanning electron microscopy (SEM) and Thermo-Gravimetric Analysis (TGA). FT-IR results confirmed that the absorption bands of the prepared chitosan matched the standard compound. Finally, the chitosan was preliminarily tested as a potential functional ingredient in potential nutraceutical formulations, with bioactive compounds such as the fish oil and the hydrolysed peptides extracted as described in the previous chapters.