Physico-chemical characterisation of whole meal flours from three wild chickpea varieties and their technological performance in Gluten Free Bread.

The present study investigated the physico-chemical characteristics of whole-meal flours from three wild chickpea varieties (white chickpea - WC, red rough chickpea - RRC, red smooth chickpea - RSC) compared to a modern chickpea variety (MC) and their bread-making performances in 30% (w/w flour) substituted GF breads. Wild chickpea flours showed the highest ash, total dietary fiber (TDF), and total antioxidant capacity (6.3%, 13.4%, and 9.5% increase for WC, RRC, and RSC flour compared to MC flour) values compared to MC sample, and red varieties (RRC- and RSC-samples) showed the highest total phenolic content (15.5% and 17.0% increase compared to MC flour). Significant differences were also found in protein content and techno-functional properties. Bread specific volume and crumb hardness were significantly affected by chickpea variety, with red varieties (RRC- and RSC-samples) revealing the lowest impact. 1H NMR proton molecular mobility significantly changed as a function of chickpea variety, and these differences might be associated to the different macroscopic bread quality. Overall, the tested wild chickpea flours revealed valuable chemical composition, and differed in the techno-functional and bread-making performances, with red varieties showing the most promising results to improve GF breads.

Key molecular compounds for simultaneous origin discrimination and sensory prediction of cocoa: An UHPLC-HRMS sensomics approach.

Cocoa-based and chocolate mono-origin products are increasingly gaining market share because they are perceived by consumers as more valuable and high quality. A comprehensive characterization of the sensory profile of a specific geographical area is complex and different analytical and sensorial strategies have been adopted. This study focused on identifying molecular markers capable of discriminating between different origins and, at the same time, predicting their sensory attributes adopting a sensomics approach. The aim is to provide a useful tool for chocolate producers to effectively screen the origins of cocoa, controlling and optimizing the gustative properties and processing flow. An untargeted method was adopted, based on the coupling of UHPLC-HRMS, followed by the application of chemometric tools for the selection of 71 discriminating molecular markers for six origins. These markers, via OPLS-Regressions, also demonstrated a strong global correlation with the sensory descriptors, evaluated by trained assessors, allowing their prediction.

Unravelling the Influence of Extraction Techniques on Protein Yield and Nutritional Value in Lesser Mealworm Larvae.

In the present work, chemical and enzymatic assisted techniques were compared for protein extraction from lesser mealworm larvae (LM, Alphitobius diaperinus), recently approved as a novel food in the European Union. All extracts showed appreciable nutritional quality, with quantities of essential amino acids above the reference standard. Conventional alkali extraction allowed the isolation of only 73% of the protein, preserving the amino acid composition but potentially causing denaturation or racemisation. The "stepwise" method, following the Osborne fractionation, improved protein recovery to 91% by isolating four fractions with different solubility properties. Additionally, enzymatic hydrolysis using Bacillus licheniformis proteases was also tested, and it provided hydrolysates with an average degree of hydrolysis of 14%, making them a potential hypoallergenic solution. Overall, these findings indicate the ability to tailor the composition of LM protein to meet specific needs, offering promising prospects for the use of insect protein ingredients in various applications.

Integration of LC-HRMS and 1H NMR metabolomics data fusion approaches for classification of Amarone wine based on withering time and yeast strain.

Two untargeted metabolomics approaches (LC-HRMS and 1H NMR) were combined to classify Amarone wines based on grape withering time and yeast strain. The study employed a multi-omics data integration approach, combining unsupervised data exploration (MCIA) and supervised statistical analysis (sPLS-DA). The results revealed that the multi-omics pseudo-eigenvalue space highlighted a limited correlation between the datasets (RV-score = 16.4%), suggesting the complementarity of the assays. Furthermore, the sPLS-DA models correctly classified wine samples according to both withering time and yeast strains, providing a much broader characterization of wine metabolome with respect to what was obtained from the individual techniques. Significant variations were notably observed in the accumulation of amino acids, monosaccharides, and polyphenolic compounds throughout the withering process, with a lower error rate in sample classification (7.52%). In conclusion, this strategy demonstrated a high capability to integrate large omics datasets and identify key metabolites able to discriminate wine samples based on their characteristics.

Development of a protocol for fractionating and characterising fibres from lignocellulosic food waste.

This study aims to explore an advanced protocol for characterising dietary fibre (DF) fractions to meet the growing demand for accurate and reliable data. Although current enzymatic-gravimetric approaches, e.g., AOAC and Van Soest analysis, provide information about soluble and insoluble DF quantification, they present limitations related to the lack of fractions characterisation. To overcome these limitations, the proposed protocol integrates the official AOAC 991.43 method with the sequential fibre fractionation by exploiting the different resistance of the fibre fractions to acid hydrolysis treatments (TFA and H2SO4), utilising hazelnut shells as a case-study. Each hydrolysed fraction was quantified and characterised through GC-MS analysis of monosaccharides. The data obtained for hemicellulose, cellulose, and lignin fractions were then discussed and compared with the Van Soest method. This approach yields a comprehensive procedure applicable to different food and nutraceutical products, emphasising the importance of DF characterisation for a deeper understanding of their bio-functional properties.

Oil and protein extraction from fruit seed and kernel by-products using a one pot enzymatic-assisted mild extraction.

This research evaluated the application of a one-pot enzymatic extraction by using a protease for the concomitant and sustainable extraction of oils and proteins from fruit seeds/kernels of different species of stone, citrus and exotic fruits. The proteolysis improved the oil solvent-extractability of seeds/kernels of some fruit species compared to the use of acid and/or organic solvents and led to directly recover fat (10-33%) from mango, lemon and pumpkin seeds. Good protein extraction yields were obtained compared to conventional solvent extractions and with a good hydrolysis degree (almost 10%) in the case of lemon and pumpkin seed protein hydrolysates. The nutritional quality of all the protein hydrolysates was quite low, because of their limiting amino acids (histidine, methionine and lysine). On the contrary, the fruit seed/kernel oils resulted with high nutritional value, as they were mostly rich in unsaturated fatty acids, primarily oleic acid (>25%) and linoleic acid (till 40%).

Hazelnut (Corylus avellana L.) shells as a potential source of dietary fibre: impact of hydrothermal treatment temperature on fibre structure and degradation compounds.

BACKGROUND: Hemicellulose extraction from lignocellulosic biomasses has gained interest over the years, and hydrothermal treatment is one of the most common methods employed for this purpose. This work aimed to deeply study hazelnut (Corylus avellana L.) shells as a new source of dietary fibre, evaluating the effect of hydrothermal treatment temperatures on the type and structure of fibre extracted, but also on the formation of side-products derived from lignocellulose degradation. RESULTS: Different process temperatures led to diverse polysaccharides in the hydrothermal extract. Pectin was identified for the first time in hazelnut shells when experimenting with extraction at 125 °C, whereas at 150 °C a heterogeneous mixture of pectin, xylan, and xylo-oligosaccharides was present. The highest yield in terms of total fibre was gained at 150 and 175 °C, and then decreased again at 200 °C. Finally, more than 500 compounds from different chemical classes were putatively identified and they appeared to be present in the extracted fibre with a different distribution and relative amount, depending on the heat treatment severity. A generally high content of phenols, phenyls, oligosaccharides, dehydro-sugars, and furans was observed. CONCLUSIONS: Modulation of the hydrothermal treatment temperature allows fibre extracts with very different compositions, and therefore different potential end uses, to be obtained from hazelnut shells. A sequential temperature-based fractionation approach, as a function of the severity of the extraction parameters, can also be considered. Nevertheless, the study of the side-compounds formed from lignocellulosic matrix degradation, as a function of the applied temperature, needs to be fully addressed for a safe introduction of the fibre extract within the food chain. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Inhibition of DPP-IV Activity and Stimulation of GLP-1 Release by Gastrointestinally Digested Black Soldier Fly Prepupae.

The beneficial effects of an insect-based diet on human health and, in particular, the regulatory ability of digested insects' proteins on the glycaemic response in humans are topics that need to be investigated deeper. In this work, we performed an in vitro study on the modulatory activity of gastrointestinal digested black soldier fly (BSF) prepupae on the enterohormone GLP-1 and its natural inhibitor, DPP-IV. We verified whether actions intended to valorise the starting insect biomass, i.e., insect-optimised growth substrates and prior fermentation, can positively impact human health. Our results highlight that the digested BSF proteins from all the prepupae samples had a high stimulatory and inhibitory ability on the GLP-1 secretion and the DPP-IV enzyme in the human GLUTag cell line. Gastrointestinal digestion significantly improved the DPP-IV inhibitory capacity of the whole insect protein. Moreover, it was seen that optimised diets or fermentation processes preceding the digestion, in any case, did not positively affect the efficacy of the answer. BSF was already considered one of the edible insects more suitable for human consumption for its optimal nutritional profile. The BSF bioactivity here shown, after simulated digestion, on glycaemic control systems makes this species even more promising.

A multiplatform metabolomics/reactomics approach as a powerful strategy to identify reaction compounds generated during hemicellulose hydrothermal extraction from agro-food biomasses.

Hydrothermal treatment is commonly used for hemicelluloses extraction from lignocellulosic materials. In this study, we thoroughly investigated with a novel approach the metabolomics of degradation compounds formed when hazelnut shells are subjected to this type of treatment. Three different complementary techniques were combined, namely GC-MS, 1H NMR, and UHPLC-IM-Q-TOF-MS. Organic acids, modified sugars and aromatic compounds, likely to be the most abundant chemical classes, were detected and quantified by NMR, whereas GC- and LC-MS-based techniques allowed to detect many molecules with low and higher Mw, respectively. Furans, polyols, N-heterocyclic compounds, aldehydes, ketones, and esters appeared, among others. Ion mobility-based LC-MS method was innovatively used for this purpose and could allow soon to create potentially useful datasets for building specific databases relating to the formation of these compounds in different process conditions and employing different matrices. This could be a very intelligent approach especially in a risk assessment perspective.

Feeding Lactic Acid Bacteria with Different Sugars: Effect on Exopolysaccharides (EPS) Production and Their Molecular Characteristics.

Exopolysaccharides (EPS) are complex molecules produced by some microorganisms and used in foods as texturizers and stabilizers, their properties depending on their chemical structure. In this work, three different lactic acid bacteria (LAB), were tested for their ability to produce EPS, by using five different mono- and disaccharides as their sole carbon source. The growth and acidifying ability were analysed, the EPSs were quantified by the official method AOAC 991.43, and their chemical structure was investigated. The amount of EPS varied from 0.71 g/L to 2.38 g/L, and maltose was the best sugar for EPS production by Lacticaseibacillus paracasei 2333. Lacticaseibacillus rhamnosus 1019 produced the highest amount when fed with lactose, whereas the EPS amount of Lactobacillus bulgaricus 1932 was not significantly different depending on the sugar type. The EPS chains consisted of fructose, galactose, glucose, mannose, ribose, glucosamine, galactosamine, and in some cases rhamnose in different proportions, depending on the strain and carbon source. The molecular weight of EPS ranged from <10 KDa to >500 KDa and was again highly dependent on the strain and the sugar used, suggesting the possibility of growing different strains under different conditions to obtain EPS with different potential applications in the food system.

1H NMR Metabolomics on Pigs' Liver Exposed to Antibiotics Administration: An Explorative Study.

An untargeted Nuclear Magnetic Resonance (NMR) spectroscopy-based metabolomics approach was applied as a first attempt to explore the metabolome of pigs treated with antibiotics. The final goal was to investigate the possibility of discriminating between antibiotic-treated (TX group) and untreated pigs (CTRL group), with the further perspective of identifying the authentication tools for antibiotic-free pork supply chains. In particular, 41 samples of pig liver were subjected to a biphasic extraction to recover both the polar and the non-polar metabolites, and the 1H NMR spectroscopy analysis was performed on the two separate extracts. Unsupervised (principal component analysis) and supervised (orthogonal partial least squares discriminant analysis) multivariate statistical analysis of 1H NMR spectra data in the range 0-9 ppm provided metabolomic fingerprinting useful for the discrimination of pig livers based on the antibiotic treatment to which they were exposed. Moreover, within the signature patterns, significant discriminating metabolites were identified among carbohydrates, choline and derivatives, amino acids and some lipid-class molecules. The encouraging findings of this exploratory study showed the feasibility of the untargeted metabolomic approach as a novel strategy in the authentication framework of pork supply chains and open a new horizon for a more in-depth investigation.

Production of xylo-oligosaccharides (XOS) of tailored degree of polymerization from acetylated xylans through modelling of enzymatic hydrolysis.

Xylo-oligosaccharides (XOS) are emerging prebiotics that have recently been gained a great interest in the market of functional foods. Since their beneficial activity strictly depends on their chemical structure and on their degree of polymerization (DP), in this work an enzymatic method was developed to produce XOS with variable and modellable DPs, involving a combination of a commercial endo-ß-1,4-xylanase M3 from Trichoderma longibrachiatum and a deacetylase, using a commercial acetylated standard xylan as substrate. A Design of Experiment (DoE) was developed and through the variation of some hydrolysis conditions, some experiments allowed to obtain significant amounts of XOS with DP 7-10, up to 11%, despite XOS with DP 2-4 were always the most abundant (60-96% of total XOS). The most impacting parameter on the XOS distribution was the order of addition of the xylanase and deacetylating enzyme, while pH showed to have a great influence on the total yield. The method was also tested on an acetylated xylan extracted from grape stalks, structurally similar to the commercial standard xylan. The model was found to work in a very similar way also on the non-purified xylan sample, allowing the manipulation of enzymatic hydrolysis on a low-cost by-product, with the potential to obtain on a large scale XOS with high added value and with a specific DP, depending on the final application.

Assessing food authenticity through protein and metabolic markers.

This chapter aims to address an issue of ancient origins, but more and more topical in a globalized world in which consumers and stakeholders are increasingly aware: the authenticity of food. Foods are systems that can also be very complex, and verifying the correspondence between what is declared and the actual characteristics of the product is often a challenging issue. The complexity of the question we want to answer (is the food authentic?) means that the answer is equally articulated and makes use of many different analytical techniques. This chapter will consider the chemical analyses of foods aimed at guaranteeing their authenticity and will focus on frontier methods that have been developed in recent years to address the need to respond to ever-increasing guarantees of authenticity. Targeted and non-targeted approaches will be considered for verifying the authenticity of foods, through the study of different classes of constituents (proteins, metabolites, lipids, flavors). The numerous approaches available (proteomics, metabolomics, lipidomics) and the related analytical techniques (LC-MS, GC-MS, NMR) are first described from a more general point of view, after which their specific application for the purposes of authentication of food is addressed.

Molecular composition of lipid and protein fraction of almond, beef and lesser mealworm after in vitro simulated gastrointestinal digestion and correlation with the hormone-stimulating properties of the digesta.

The current production of meat presents many disadvantages for the environment and much research focuses on alternative protein sources. Insects are novel protein sources highly valued for their nutritional and sustainable potential. However, many aspects concerning biological and nutritional properties of the insects after digestion, in comparison with other protein sources, are still overlooked. In this work, a comparative study on three different protein sources, namely almond, lean beef and insect Alphitobius diaperinus (lesser mealworm), was performed after in vitro simulated gastrointestinal digestion. An in-depth characterization of the chemical composition of the solubilized protein and lipid fractions of the digesta was performed by applying different analytical techniques, including chromatographic methods coupled to mass spectrometry and 1H NMR spectroscopy. Beef and insect were proven to be very similar in amino acid composition and protein solubilization after digestion, when considering the proper corrections for the chitin content. Lipid fraction from insects was solubilized during digestion as the one of almonds, but with a fastest kinetics. Thus, lesser mealworms are a good source of both lipids and highly nutritional proteins. Then, the amino acid composition of raw and digested protein fraction from the three sources was related to the PYY, ghrelin, GLP-1 and CCK release and rats' food intake. The composition of amino acids in insect digesta was found to be related to specific effects on enterohormone release, and the modulation of food intake in rats.

Dynamic changes in molecular composition of black soldier fly prepupae and derived biomasses with microbial fermentation.

Black soldier fly (BSF) is being increasingly used for agro-food by-products valorisation. Adult flies, puparia, and excess of prepupae are the by-products of this process, which could be further valorised. Lactic fermentation of BSF biomasses with two different strains (L. rhamnosus and L. plantarum) has been used for this purpose. Deep changes in the molecular composition were observed, without significant differences related to the different strains used. The lipid and protein fractions were the most impacted. Fermentation enriched the biomass in monounsaturated and polyunsaturated fatty acids and essential amino acids, significantly improving the nutritional properties of the substrates. Although not particularly marked, a proteolytic activity of lactobacilli was observed on the BSF muscular and cuticular proteins, especially in the samples of adult flies and puparia, where fermentation resulted more effective. Conversely, there was no evidence of chitinolytic activity.

Protease-Assisted Mild Extraction of Soluble Fibre and Protein from Fruit By-Products: A Biorefinery Perspective.

By-products from the fruit supply chain, especially seeds/kernels, have shown great potential to be valorised, due to their high content of macronutrients, such as lipids, protein, and fibre. A mild enzymatic assisted extraction (EAE) involving the use of a protease was tested to evaluate the feasibility of a cascade approach to fractionate the main fruit by-products components. Protease from Bacillus licheniformis (the enzyme used in the AOAC 991.43 official method for dietary fibre quantification) was used, and besides protein, the conditions of hydrolysis (60 °C, neutral pH, overnight) allowed us to dissolve a portion of soluble fibres, which was then separated from the solubilized peptide fraction through ethanol precipitation. Good protein extraction yields, in the range 35-93%, were obtained. The soluble fibre extraction yield ranged from 1.6% to 71% depending on the by-product, suggesting its applicability only for certain substrates, and it was found to be negatively correlated with the molecular weight of the fibre. The monosaccharide composition of the soluble fibres extracted was also diverse. Galacturonic acid was present in a low amount, indicating that pectin was not efficiently extracted. However, a predominance of arabinose and galactose monomers was detected in many fractions, indicating the isolation of a fruit soluble fibre portion with potential similarity with arabinogalactans and gum arabic, opening up perspectives for technological applications. The residual solid pellet obtained after protease assisted extraction was found to be an excellent fibre-rich substrate, suitable for being subjected to more "hard" processing (e.g., sequential pectin and hemicellulose extraction) with the objective to derive other fractions with potential great added economic value.

Study on the Effect of Ensiling Process and Ruminal Digestion on the Synthesis and Release of Cyclopropane Fatty Acids in Cow Feeding.

Cyclopropane fatty acids (CPFA) were found in milk fat from cows fed maize silage and suggested to be synthesized by lactic acid bacteria during ensiling. This study aimed to elucidate some gaps of knowledge about the microbial synthesis of CPFA, to strengthen the current authentication method based on their detection in cheese fat and performed for Parmigiano Reggiano (UNI11650), whose Specifications forbid the use of silage. CPFA were screened in different ensiled cows' feeding by gas chromatography-mass spectrometry, and the effect of feed ingredients and ruminal digestion on CPFA microbial production were further examined by in vitro tests. Results showed that solely the environmental conditions developed in silos for specific plant materials (e.g., maize) are essential for the bacterial synthesis of CPFA, whereas rumen activity did not affect CPFA levels in feeds. This supports the suitability of using CPFA as biomarkers of a crop silage-based diet forbidden by certain PDO feedstock regulations.

Effect of the Rearing Substrate on Total Protein and Amino Acid Composition in Black Soldier Fly.

Insects are becoming increasingly relevant as protein sources in food and feed. The Black Soldier Fly (BSF) is one of the most utilized, thanks to its ability to live on many leftovers. Vegetable processing industries produce huge amounts of by-products, and it is important to efficiently rear BSF on different substrates to assure an economical advantage in bioconversion and to overcome the seasonality of some leftovers. This work evaluated how different substrates affect the protein and amino acid content of BSF. BSF prepupae reared on different substrates showed total protein content varying between 35% and 49% on dry matter. Significant lower protein contents were detected in BSF grown on fruit by-products, while higher contents were observed when autumnal leftovers were employed. BSF protein content was mainly correlated to fibre and protein content in the diet. Among amino acids, lysine, valine and leucine were most affected by the diet. Essential amino acids satisfied the Food and Agricultural Organization (FAO) requirements for human nutrition, except for lysine in few cases. BSF could be a flexible tool to bio-convert a wide range of vegetable by-products of different seasonality in a high-quality protein-rich biomass, even if significant differences in the protein fraction were observed according to the rearing substrate.

Cyclopropane Fatty Acids as Quality Biomarkers of Cheeses from Ewes Fed Hay- and Silage-Based Diets.

This study aimed to investigate cyclopropane fatty acids (CPFAs) as quality biomarkers of forage feedings in cheese fat obtained from ewe's milk, based on two different dietary treatments (hay and silage). The gas chromatography-mass spectrometry (GC-MS) analysis detected CPFAs in most cheese samples, both from hay and silage-based diets. CPFA levels in cheese fat from hay feeding were positively correlated to the total trans-monounsaturated fatty acids (MUFAs) and n-6 polyunsaturated fatty acids (PUFAs), whereas they were negatively correlated to cis-MUFAs, odd- and branched-chain fatty acids (i.e., C13:0 anteiso, C16:0 iso, and C17:1), and C22:5n-3, which are mainly associated with a low starch intake and grass pasture. Overall, the presence of CPFAs in ovine cheese fat suggests the use of silage, but it can also be an indicator of poor-quality hay forages. This approach confirmed the reliability of CPFAs as biomarkers of forage quality, especially in relation to the use of conserved forages and good livestock practices.

Potential Valorization of Hazelnut Shells through Extraction, Purification and Structural Characterization of Prebiotic Compounds: A Critical Review.

Hazelnuts are one of the most widely consumed nuts, but their production creates large quantities of by-products, especially shells, that could be upcycled into much more valuable products. Recent studies have shown that hazelnut shell hemicellulose is particularly rich in compounds that are potential precursors of xylooligosaccharides and arabino-xylooligosaccharides ((A)XOS), previously defined as emerging prebiotics very beneficial for human health. The production of these compounds on an industrial scale-up could have big consequences on the functional foods market. However, to produce (A)XOS from a lignocellulosic biomass, such as hazelnut shell, is not easy. Many methods for the extraction and the purification of these prebiotics have been developed, but they all have different efficiencies and consequences, including on the chemical structure of the obtained (A)XOS. The latter, in turn, is strongly correlated to the nutritional effects they have on health, which is why the optimization of the structural characterization process is also necessary. Therefore, this review aims to summarize the progress made by research in this field, so as to contribute to the exploitation of hazelnut waste streams through a circular economy approach, increasing the value of this biomass through the production of new functional ingredients.