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Coppa Piacentina is an Italian protected designation of origin (PDO) dry-cured product obtained from the muscle of pork neck and ripened for at least six months. Metabolomics- and volatilomics-based strategies, combined with a chemical characterization of free amino acids were applied to identify biomarkers of long-ripened Coppa Piacentina PDO. Long ripening induced a significantly increase of total free amino acids, mainly represented by glutamic acid, involved in the umami taste perception. Untargeted metabolomics, performed using UHPLC-HRMS, allowed to identify 32 putative gamma-glutamyl-peptides, known as main contributors to the kokumi taste. Unsupervised and supervised multivariate statistics observed a clear modification of these peptides over the ripening, with gamma-glutamyl-peptides which significantly increased in long-ripened samples. A volatilomics-based strategy, conducted with GCxGC-MS, was then performed, and 93 different compounds were identified, with aldehyde and ketones deriving from the lipid auto-oxidation which increased according to ripening.

Untargeted metabolomics based on ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry was combined with sensory analysis to provide new insights into the impact of the feeding system from mountain regions (grassland deriving from permanent meadows) on the chemical fingerprint of Parmigiano Reggiano PDO hard cheese. In the framework of a representative investigation, two different ripening times (12 and 24 months) were also considered. Multivariate statistics allowed discriminating cheese samples from different feeding regimens according to their metabolomics signatures. Interestingly, mountain grassland-based cheese samples were characterized by a more favourable fatty acid profile, recording also feed-derived compounds (such as terpenoids and linoleic acid derivatives) potentially associated with both beneficial effects on human health and sensory properties. According to the sensory analysis, the impact of herbs and grass enhanced the colour and retro-olfactive complexity of Parmigiano Reggiano PDO cheese, with spicy, umami and intense vegetal aromatic notes representing distinctive features.

The consumption of foods characterized by a wide bioactive-compound profile has been associated with improved health status and reduced risk of developing diseases. Owing to the complexity of food production (e.g. crops and farmed animals), processing (e.g. fermentation), and shelf life (e.g. lipid or protein oxidation), a more comprehensive and holistic analysis is necessary to investigate the intricate metabolic pathways and metabolites involved with or displaying a biological activity. Accordingly, untargeted metabolomics analysis can provide crucial knowledge to support the selection of key metabolites and metabolic pathways to produce foods from animals and crops with a functional value. This short review aims to provide an overview of the applications of metabolomic analysis to screen bioactive compounds in both animal- and plant-based foods.

In the food and feed industry, protein extraction is commonly performed under acid or basic conditions, combined with heat, in order to increase the extraction yield. Under severe processing conditions, proteins may undergo molecular modifications. Here, the effects of heating (30, 60, 90 °C) at different pH values (2, 7, 9, 11, 13) were evaluated on commercial whey proteins, used as a simplified protein model. The main structure and chemical modifications concerning protein aggregation, hydrolysis, insolubilization, amino acid degradation and racemization were investigated in detail. Using in vitro static models, the degree of protein hydrolysis and the released peptides were determined after the digestive process. Accumulation of molecular modifications was mostly observed after basic pH and high temperatures treatments, together with a marked decrease and modification of the digestibility profile. Instead, protein digestibility increased in neutral and acidic conditions in a temperature-dependent manner, even if some modification in the structure occurs.

Plant cell cultures from cloudberry (CL), lingonberry (LI), stone berry (ST), arctic bramble (AB), and strawberry (SB) were studied in terms of their polyphenol and carotenoid composition, antioxidant activity, antihemolytic activity and cytotoxicity effects on cancerous cells. High-resolution mass spectrometry data showed that LI, presented the highest antioxidant activity, contained the highest contents of flavones, phenolic acids, lignans, and total carotenoids, while CL, ST and SB presented the opposite behavior. AB and SB presented the lowest FRAP and CUPRAC values, while AB and CL presented the lowest reducing power. SB presented the lowest antioxidant activity measured by single electron transfer assays and the lowest content of lignans, phenolic acids, and flavones. CL and LI decreased the viability of in vitro mammary gland adenocarcinoma while only LI decreased the viability of in vitro lung carcinoma and showed protective effects of human erythrocytes against mechanical hemolysis.

The interest in the modulation of gut microbiota by polyphenols from olives and derived products is increasing. In this work, phenolic leaf extracts (PLE) were in vitro faecal fermented to evaluate the changes in phenolic profiles and the impact on microbiota, using a commercial extra-virgin olive oil (EVOO) as reference. The in vitro fermentation decreased oleuropein content in PLE, determining an increase of hydroxytyrosol and other phenolic metabolites. An increase (p < 0.05) of hydroxytyrosol (LogFC = 6.02; VIP score = 1.05) was also observed in fermented EVOO. Besides, PLE significantly (p < 0.05) changed amino acids (LogFC = 6.1) and fatty acids (LogFC = 5.9) profile of the faeces. Metagenomic sequencing revealed that Coriobacteriaceae at the family level, and Collinsella at the genus level, were the most affected by PLE fermentation. These findings support the modulation of the gut microbiota exerted by phenolics from PLE and EVOO.

Background: Watermelon (Citrullus lanatus) represents the largest cultivated member of the Cucurbitaceae family, showing great relevance from the economic point of view and largely consumed around the world accounting for about 7% of the world area dedicated to the production of vegetables, with 67% of the total quantity produced in China. It is characterized by several bioactive compounds, showing different chemical structures, such as carotenoids, xanthophylls, phenolic compounds, citrulline, and unsaturated fatty acids. The good amount of total polyphenols, vitamin C, citrulline and the excellent supply of lycopene, about 40% compared to raw tomatoes, give a measure of the importance of this fruit respect to the other crops considering the beneficial effects of these compounds on human health. Scope and approach: In the last decades, this great interest towards natural bioactive compounds led many researchers to study watermelon as natural source of bioactive compounds, mainly when considering some biological properties, including antioxidant, antimicrobial and anti-inflammatory activities, inhibition or induction of enzymes, inhibition of receptor activities, together with induction/inhibition of gene expression. However, the most of scientific literature on this topic was based on in vitro assays, thus limiting the comprehensive understanding of the real health-promoting outcomes. Key findings and conclusions: Therefore, the aim of this review was to present the up-to-date research carried out on watermelon phytochemicals, showing the most important biological activities reported from both in vitro and in vivo trials. Besides, the potential exploitation of watermelon by-products in the green circular economy and food-waste valorization has been discussed.

Milk is a foodstuff widely consumed around the world originating from a variety of different species, animal management and production systems. In recent years, consumers have placed a much greater emphasis on the authenticity and origin of some food products often willing to pay a premium price for such products that is, for example ‘Grass-Fed Dairy’. Therefore, it is important to establish methods to assess both quality and authentication of milk and dairy products for increased food security and consumer protection. Accordingly, NMR-based, GC–MS-based, and LC–MS-based metabolomics have been established as useful tools in the analysis of dairy products, such as raw and processed milk. This short-review provides an updated and critical overview on the most useful metabolomics-based platforms and the most useful multivariate statistical tools available for metabolomic data interpretation.

Moringa (Moringa oleifera Lam.), the so-called ‘miracle tree’, has emerged as an excellent source of nutrients and functional components also in food applications. In this regard, Moringa is worldwide cultivated for its nutritional properties and for a variety of end-uses. The different portions of this plant (leaves, pods, seeds) have been widely studied for their antioxidant, anti-inflammatory, and antihyperglycemic activities. Most of the functional activities reported in the literature are strictly related to its high content of polyphenols, bioactive peptides, carotenoids, and glucosinolates. Besides, a great interest is emerging towards the exploitation of Moringa as a value-added ingredient for both bakery and meat products. On these bases, considering the more recent literature about M. oleifera plant, this short review aims to highlight the different biological and potential food applications of M. oleifera parts and/or extracts as a source of bioactive compounds able to provide health-promoting properties.

Risk-benefit assessment is the comparison of the risk of a situation to its related benefits, i.e. a comparison of scenarios estimating the overall health impact. The risk–benefit analysis paradigm mirrors the classical risk analysis one: risk–benefit assessment goes hand-in-hand with risk–benefit management and risk–benefit communication. The various health effects associated with food consumption, together with the increasing demand for advice on healthy and safe diets, have led to the development of different research disciplines in food safety and nutrition. In this sense, there is a clear need for a holistic approach, including and comparing all of the relevant health risks and benefits. The risk–benefit assessment of foods is a valuable approach to estimate the overall impact of food on health. It aims to assess together the negative and positive health effects associated with food intake by integrating chemical and microbiological risk assessment with risk and benefit assessment in food safety and nutrition. The 2019 Parma Summer School on risk–benefit in food safety and nutrition had the objective was to provide an opportunity to learn from experts in the field of risk–benefit approach in food safety and nutrition, including theory, case studies, and communication of risk–benefit assessments plus identify challenges for the future. It was evident that whereas tools and approaches have been developed, more and more case studies have been performed which can form an inherent validation of the risk–benefit approach. Executed risk–benefit assessment case studies apply the steps and characteristics developed: a problem formulation (with at least 2 scenarios), a tiered approach until a decision can be made, one common currency to describe both beneficial and adverse effects (DALYs in most instances). It was concluded that risk–benefit assessment in food safety and nutrition is gaining more and more momentum, while also many challenges remain for the future. Risk-b

Background: Worldwide, huge amounts of by-products and wastes are generated in agri-food sector annually. This biomass is usually discarded provoking serious environmental problems. However, these residues represent an excellent source of bioactive compounds so that their exploitation can be considered as a promising possibility from an economic and environmental perspective. Conventional extraction technologies using organic solvents have been employed to recover high-value molecules showing several drawbacks such as toxicity, high volatility and non-renewability. Therefore, the search of alternative green solvents for the extraction of bioactives and in order to reduce the environmental impact and to achieve a widespread consumer acceptance is mandatory. Recently, deep eutectic solvents (DESs) and natural deep eutectic solvents (NaDESs) have received great attention for their physicochemical features for bioactive compounds extraction. Scope and approach: This review describes the state of the art of the applications of DESs and NaDESs in the field of the revalorization of agri-food by-products to obtain and isolate bioactive compounds. The synthesis and properties of these avant-garde solvents are described. In particular, the main bioactives extracted using DESs and NaDESs from several residues from citrus, winemaking, olive oil production, onion, seaweeds, animal and fish industries were systematically reviewed and collected. Key findings and conclusions: DESs and NaDESs combined with emerging technologies to recover biomolecules from agri-food by-products show promising results in comparison with traditional methods of extraction using conventional solvents. These emerging solvents present advantages which make them good candidates for being implemented by the industry, considering their low toxicity, recyclability, tunable properties and environmentally friendly. Therefore, this new generation of solvents opens new options in the field of recovering bioactive

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Insects have been proposed as new source of proteins to meet the growing demand connected to the increasing world population. In the EU the inclusion of insect proteins in feed and food is strictly regulated. Hence, analytical methods able to discriminate and identify different insect species in food and feed are a necessity. In this work, a peptidomic approach was applied to determine peptide biomarkers for two edible insect species: lesser mealworm and black soldier fly. Three species specific peptide biomarkers were identified using LC-MS/MS. The two insects were mixed with fish standard feed at different concentrations, to evaluate the feasibility of their use as markers in complex matrices. The detection of marker peptides was confirmed down to 1% insect amount. The data here reported constitutes the first proof of concept for the potential application of the peptide marker approach for the identification and quantification of insect ingredients.

Insects are being explored as novel protein sources in order to overcome the future food demands connected to world growing population. Insects for food/feed uses are currently slowly killed through freezing by most insect rearing companies, and typically, enzymatic browning takes place in the insect proteins fractions. However, very little is known about the influence of these enzymatic reactions on the protein physical, chemical, nutritional and technological properties. In this work a metabolomics and proteomic study was conducted on Black Soldier Fly (Hermetia illucens) prepupae, killed by two different methods: freezing (commonly used), and blanching (with the aim to inhibit the enzymatic activities). Proton nuclear magnetic resonance (1H NMR) metabolomics demonstrated that slow killing method by freezing, compared with blanching, elicits the activation of several enzymatic pathways, among them melanisation with tyrosine consumption, energetic metabolism and lipolysis. These metabolic changes have an impact also on protein nutritional quality, with a loss of cysteine and lysine, likely involved in the process of melanisation and enzymatic browning. A strong effect was also observed on protein extractability: proteins from prepupae killed by blanching were found to be more extractable in milder conditions by chemical methods, and more prone to enzymatic digestion (97% of proteins released in solution upon proteolysis) than proteins from prepupae killed by freezing. All these data indicate that killing by blanching inhibits the browning reaction and other enzymatic changes occurring during slow killing by freezing, increasing the extractability of proteins in aqueous solutions, avoiding essential amino acid loss, and improving enzymatic digestibility.

We formulated and characterised two alginate blends for the encapsulation of stevia extract (SE) via ionic gelation through an extrusion technique. Calcium chloride in SE and calcium chloride solutions were assessed as crosslinkers to overcome phenolic losses by diffusion and increase encapsulation efficiency (EE). Regardless of the blend, all stevia-loaded beads exhibited high EE (62.7–101.0%). The size of the beads decreased as EE increased. Fourier transform infrared analysis showed increased hydrogen bonding between SE and alginates, confirming the successful incorporation of SE within the matrix. Untargeted metabolomics profiling identified 479 free and encapsulated polyphenolic compounds. Flavonoids (catechin and luteolin equivalents) were predominant in SE whereas tyrosols and 5-pentadecylresorcinol equivalents were predominant in all bead formulations. Three-common discriminant compounds were exclusive to each blend and were inversely affected by the crosslinking conditions. Both alginate blends have been shown to be feasible as carrier systems of stevia extracts independent of crosslinking conditions.

Indigenous strains of lactic acid bacteria previously isolated from raw donkey milk samples were used to ferment donkey milk. Each sample was subjected to an in vitro simulated gastrointestinal digestion (SGID) and analysis of the digesta by HPLC-LTQ-Orbitrap XL was performed in order to obtain a comprehensive peptide profile of fermented donkey milk. Functional properties such as ACE-inhibitory, antimicrobial and antioxidant activities of the resulting fermented donkey milks, as well as characterization of the bioactive peptides produced by an in vitro SGID, were assayed. All bioactivities were found to be high in fermented milk and a further significant increase was observed after SGID. The Enterococcus faecium DM33 fermented milk exhibited the strongest antioxidant and the highest antimicrobial activities. The highest ACE-inhibitory activity was observed in milk fermented with Lactobacillus casei DM214. These findings will contribute to the development of a new functional dairy drink with anti-hypertensive, antimicrobial and/or antioxidant activities.

Black soldier fly (BSF, Hermetia illucens) constitutes an economic way to convert residual biomasses into a valuable source of biomolecules, such as proteins, lipids and chitin. The present investigation was undertaken to evaluate the feasibility of applying different extraction protocols, either chemical extractions or enzymatic assisted extraction, to recover pure fat, protein and chitin fractions. First, exact proximate composition, total amino acids, fatty acids profile, and N-acetylglucosamine content of the prepupae samples were determined. BSF prepupae biomass contained, expressed on dry weight, 32% proteins, 37% lipids, 19% minerals, 9% chitin. The lipid fraction was easily recovered by organic solvents, while the most challenging issue was the separation of protein from chitin. The best separation was obtained by alkali extraction of proteins (96% of protein recovered) albeit with loss in their integrity as indicated by the measurement of the degree of hydrolysis with the o-phthaldialdehyde method. To avoid protein damage in alkali media, a stepwise protein extraction adopting milder conditions was also explored based on Osborne fractionation method, allowing the recovery of > 85% of BSF high purity and high quality proteins, and the obtainment of chitin-enriched fraction as well. The possibility of using an enzymatic assisted extraction of proteins was also explored, obtaining a maximum nitrogen solubilisation in the best case (with Bacillus licheniformis protease) of about 60%. In this latter case, the chitin fraction obtained also had a significant residual protein content.