Changes in Faecal Microbiota Profile and Plasma Biomarkers following the Administration of an Antioxidant Oleuropein-Rich Leaf Extract in a Rat Model Mimicking Colorectal Cancer.

Oleuropein (OLE), a phenolic compound particularly abundant in the olive leaves, has been reported to have beneficial activities against colorectal cancer (CRC). In vitro studies suggested that these latter could be due to a modulation of the intestinal microbiota. Aiming to evaluate if OLE could affect the intestinal microbiota and the plasma metabolome, an antioxidant oleuropein-rich leaf extract (ORLE) was administered for one week to PIRC rats (F344/NTac-Apcam1137), a genetic model mimicking CRC. ORLE treatment significantly modulated the gut microbiota composition. Plasma metabolomic profiles revealed a significant predictive ability for amino acids, medium-chain fatty acids, and aldehydes. Pathway analysis revealed a significant decrease in phosphatidylcholine accumulation (LogFC = -1.67) in PIRC rats. These results suggest a significant effect of ORLE administration on faecal microbiota profiles and plasma metabolomes, thereby offering new omics-based insights into its protective role in CRC progression.

The Potential of Multi-Screening Methods and Omics Technologies to Detect Both Regulated and Emerging Mycotoxins in Different Matrices.

Mycotoxins are well-known secondary metabolites produced by several fungi that grow and occur in different crops during both pre-harvest and post-harvest conditions. The contamination and occurrence of mycotoxins currently represent some of the major issues in the entire agri-food system. The quantification of mycotoxins in different feeds and foodstuffs is extremely difficult because of the low concentration ranges; therefore, both sample collection and preparation are essential to providing accurate detection and reliable quantification. Currently, several analytical methods are available for the detection of mycotoxins in both feed and food products, and liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) represents the most reliable instrumental approach. In particular, the fast development of high-throughput methods has made it possible to screen and analyze, in the same analytical run and with high accuracy, multiple mycotoxins, such as those regulated, masked, or modified, and emerging ones. Therefore, the aim of this review is to provide an overview of the state of the art of mycotoxins occurrence, health-related concerns, and analyses, discussing the need to perform multi-screening approaches combined with omics technologies to simultaneously analyze several mycotoxins in different feed and food matrices. This approach is expected to provide more comprehensive information about the profile and distribution of emerging mycotoxins, thus enhancing the understanding of their co-occurrence and impact on the entire production chain.

Development of Coated PLA Films Containing a Commercial Olive Leaf Extract for the Food Packaging Sector.

A commercial olive leaf extract (OL), effective against Salmonella enterica, Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus, was added to three different coating formulations (methylcellulose, MC; chitosan, CT; and alginate, ALG) to produce active polylactic acid (PLA) coated films. Evaluation of these coated PLA films revealed significant inhibition of S. aureus growth, particularly with the MC and CT formulations exhibiting the highest inhibition rates (99.7%). The coated films were then tested for food contact compatibility with three food simulants (A: 10% ethanol; B: 3% acetic acid; D2: olive oil), selected to assess their suitability for pre-cut hams and ready-to-eat vegetables in relation to overall migration. However, coated films with active functions exhibited migration values in simulants A and B above legal limits, while promising results were obtained for simulant D2, highlighting the need to deeply investigate these coatings' impact on a real food system. Untargeted metabolomics revealed that the type of coating influenced the selective release of certain phenolic classes based on the food simulant tested. The Oxitest analysis of simulant D2 demonstrated that the MC and ALG-coated PLA films slightly slowed down the oxidation of this food simulant, which is an edible vegetable oil.

Replacement of nitrates and nitrites in meat-derived foods through the utilization of coagulase-negative staphylococci: A review.

Nitrates and nitrites, which are synthetic additives, are traditionally used as curing agents in meat-based products. These synthetic additives are employed in the preparation of fermented meat foods to improve quality characteristics and microbiological safety, develop distinct flavours and red-colour stability, and counteract lipid oxidation. Nitrites also display significant bacteriostatic and bactericidal action against spoilage microorganisms and foodborne pathogens (such as Clostridium botulinum and Listeria monocytogenes). However, meat curing is currently under scrutiny because of its links to cardiovascular diseases and colorectal cancer. Based on the current literature, this review provides recent scientific evidence on the potential utilisation of coagulase-negative staphylococci (CNS) as nitrate and nitrite substitutes in meat-based foods. Indeed, CNS are reported to reproduce the characteristic red pigmentation and maintain the typical high-quality traits of cured-meats, thanks to their arginine degradation pathway, thus providing the nitrite-related desirable attributes in cured meat. The alternative strategy, still based on the NOS pathway, consisting of supplementing meat with arginine to release nitric oxide (NO) and obtain a meat characterised by the desired pinkish-red colour, is also reviewed. Exploiting NOS-positive CNS strains seems particularly challenging because of CNS technological adaptation and the oxygen dependency of the NOS reaction; however, this exploitation could represent a turning point in replacing nitrates and nitrites in meat foods.

Untargeted metabolomics and machine learning unveil quality and authenticity interactions in grated Parmigiano Reggiano PDO cheese.

The chemical composition of Parmigiano Reggiano (PR) hard cheese can be significantly affected by different factors across the dairy supply chain, including ripening, altimetric zone, and rind inclusion levels in grated hard cheeses. The present study proposes an untargeted metabolomics approach combined with machine learning chemometrics to evaluate the combined effect of these three critical parameters. Specifically, ripening was found to exert a pivotal role in defining the signature of PR cheeses, with amino acids and lipid derivatives that exhibited their role as key discriminant compounds. In parallel, a random forest classifier was used to predict the rind inclusion levels (> 18%) in grated cheeses and to authenticate the specific effect of altimetry dairy production, achieving a high prediction ability in both model performances (i.e., ∼60% and > 90%, respectively). Overall, these results open a novel perspective to identifying quality and authenticity markers metabolites in cheese.

Impact of In Vitro Gastrointestinal Digestion on the Phenolic Bioaccessibility and Bioactive Properties of Insect-Containing Beef Burgers.

Mealworm, migratory locust, and house cricket have recently been recognized by the European Commission as novel foods, thus being suitable in different food applications. In this work, we tested their powders as meat extenders at 5% (w/w) inclusion in beef burgers, considering their ability to vehicle phenolic compounds during simulated in vitro static gastrointestinal digestion (INFOGEST). Insect powders were abundant in different phenolic classes, recording the highest values in locust (LP; 314.69 mg/kg), followed by cricket (CP; 113.3 mg/kg) and mealworm (MWP; 51.9 mg/kg). Following a pan-cooking process, LP burgers were confirmed as the best source of phenolics, with a marked abundance of flavonoids and phenolic acids. Interestingly, the insect powders were found to affect the in vitro gastrointestinal bioaccessibility of phenolic compounds when compared with the CTR burger, likely promoted by the interactions between the phenolic compounds and proteins characterizing the tested insect powders. Among the most discriminant phenolic metabolites at the gastrointestinal level, we found several phenolic acids (mainly hydroxycinnamics), recording the highest content for the digested CP-containing burgers. Finally, stilbenes showed significant correlation values at the intestinal level with both antioxidant and enzymatic activities, while total flavonoids were the most correlated with the inhibition of acetylcholinesterase. Taken together, our preliminary findings demonstrated that insect powders added to beef burgers can promote the bioaccessibility and potential bioavailability of phenolics in the distal tracts of the intestine.

The distinctive effect of different insect powders as meat extenders in beef burgers subjected to cooking and in vitro gastrointestinal digestion.

Mealworm (MWP), migratory locust (LP), and house cricket (CP) are novel foods recently authorized by the European Commission. This work tested their powders as meat extenders at 5% inclusion in beef burgers. Insect powders were abundant in phenolics, recording the highest values in LP (1184.9 µg/g). The sensory analysis highlighted a higher visual and olfactory acceptability for MWP-burgers, followed by CP- and LP-burgers, whereas the texture of cooked burgers remained unaffected. Following pan-cooking, MWP-burgers and control exhibited comparable chemical profiles, while a significant down-accumulation of the heterocyclic amine 2-Amino-3,8-dimethylimidazo[4,5-f]quinoxaline was observed in CP-burgers. In vitro gastrointestinal digestion highlighted metabolomic trends like control for MWP- and LP-burgers. In contrast, a reduced accumulation of lipids and increased content of dipeptides like glutaminylarginine (possibly acting as enzyme modulators) was observed for the CP-burgers.

Volatile compounds, gamma-glutamyl-peptides and free amino acids as biomarkers of long-ripened protected designation of origin Coppa Piacentina.

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.

Discrimination of different feed additives and poly-herbal formulations based on their untargeted phytochemical profiles.

INTRODUCTION: Feed additives represents a valid tool in animal nutrition to improve animal performance and livestock productivity under a sustainable perspective; however, there is a paucity of information about their comprehensive metabolomic and bioactive profiles. OBJECTIVE: In this study, we tested the ability of an untargeted metabolomics approach to discriminate nine commercial feed additives and unique blends of botanical extracts used in both ruminant and non-ruminant nutrition, according to their phytochemical profiles and different in vitro bioactive properties. METHODS: An ultra-high-performance liquid chromatography coupled with Orbitrap mass spectrometry and multivariate statistics were combined to search for potential markers, in order to better discriminate the different commercial samples. RESULTS: Several phytochemicals were identified, namely alkaloids, phenolics, organosulfurs, and terpenoids. The polyherbal formulation Zigbir was the best source of phytochemicals, accounting for a cumulative total content of phytochemicals equal to 3.03 mg Eq./g, being particularly abundant in terpenoids, stilbenes, phenolic acids, and small-molecular-weight phenolics. Multivariate statistics allowed to group the different products in 2 bioactive subclusters. The diterpenoid andrographolide recorded the highest abundance in Zigbir and Sangrovit. The most predictive biomarkers were: piperine, isoquercitrin, 6-methylthiohexyldesulfoglucosinolate, 6-methylumbelliferone, benzoic acid, (+)-(1R,2R)-1,2-diphenylethane-1,2-diol, and piperitenone. Flavonoids were highly correlated with both in vitro antioxidant and enzyme inhibition assays. CONCLUSIONS: Our findings provide new insights into the comprehensive phytochemical composition of commercial feed additives and blend of botanical extracts used for both ruminant and non-ruminant nutrition. A great importance of polyphenols in relation to the biological activities was detected.

The effect of common duckweed (Lemna minor L.) extract on the shelf-life of beef burgers stored in modified atmosphere packs: A metabolomics approach.

The impact of duckweed extracts (DEs) on the shelf-life of packaged beef burgers was evaluated through classical assays and untargeted metabolomics. Beef burgers were formulated with an antioxidants-free control (CON), 1 g/kg sodium ascorbate (ASC), and increasing levels of a DEs, namely 1 (DE1), 5 (DE5), and 10 (DE10) g/kg, packaged under modified atmosphere and stored at 4 °C for 19 days. The DEs, abundant in phytochemicals, determined no issues with the hygienic status of the product. DEs modulated the redox status, being ineffective in preserving linolenic acid from peroxidation. However, the oxidation marker 2-nonenoic acid was down-accumulated in the DE10 sample following 19 days of storage, recording a lower glutathione:glutathione disulfide ratio. The accumulation of adipate semialdehyde revealed the inefficiency of DEs in coping with protein oxidation, while DEs prevented the accumulation of biogenic amines. Therefore, this work suggests a potential pro-oxidant role of the formulated DEs.

Untargeted metabolomics provide new insights into the implication of Lactobacillus helveticus strains isolated from natural whey starter in methylglyoxal-mediated browning.

Hard cheeses may occasionally show a brown discolouration during ripening due to multifactorial phenomena that involve bacteria and give rise to pyrazines arising from methylglyoxal. The present work aimed at developing a novel approach to investigate the role of natural starters in browning. To this object, 11 strains of L. helveticus were incubated in a medium containing 10 % rennet casein dissolved in whey, and then growth was monitored by measuring pH and number of genomes/mL. Browning was assessed through CIELab analysis, methylglyoxal production was determined by targeted mass spectrometry, and untargeted metabolomics was used to extrapolate marker compounds associated with browning discoloration. The medium allowed the growth of all the strains tested and differences in colour were observed, especially for strain A7 (ΔE* value 15.92 ± 0.27). Noteworthy, this strain was also the higher producer of methylglyoxal (2.44 µg/mL). Metabolomics highlighted pyrazines and ß-carboline compounds as markers of browning at 42 °C and 16 °C, respectively. Moreover, multivariate statistics pointed out differences in free amino acids and oligopeptides linked to proteolysis, while 1,2-propanediol and S-Lactoylglutathione suggested specific detoxification route in methylglyoxal-producing strains. Our model allowed detecting differences in browning amid strains, paving the way towards the study of individual L. helveticus strains to identify the variables leading to discoloration or to study the interaction between different strains in natural whey starters.

Untargeted metabolomics and conventional quality characterization of rowanberry pomace ingredients in meatballs.

In this study, a rowanberry pomace defatted with supercritical CO2 (2%-AC), its ethanolic extract (1%-E) and extraction residue (2%-R), were tested in meatball preparation. The meatballs with 1%-E demonstrated the highest in vitro radical scavenging capacity. In the case of 1%-E the pH of meatballs was significantly lower compared to the control sample (P = 0.0132) on the 5-day. The lowest cooking loss was achieved when the meatballs contained mainly fibre-rich 2%-R. The UHPLC method detected 184 metabolites, including strong antioxidants, such as chlorogenic acids, 3',4'-methylenedioxy-5,7-dimethylepicatechin, hyperin, isoquercitrin. The 1%-E was particularly effective against the development of unpleasant off-flavours caused by carbonyl compounds. Consistently, the decrease in lipid oxidation, indicated by reduced 7-dodecenal and 2,4-heptadienal contents, has been observed following the addition of rowanberry extract to meatballs. Metabolomics coupled with conventional quality evaluations provided a deeper understanding of the potential utilization and valorisation of different rowanberry pomace extracts as meat ingredients.

Propionibacteria as promising tools for the production of pro-bioactive scotta: a proof-of-concept study.

Dairy propionibacteria are Gram positive Actinomycetota, routinely utilized as starters in Swiss type cheese making and highly appreciated for their probiotic properties and health promoting effects. In this work, within the frame of a circular economy approach, 47 Propionibacterium and Acidipropionibacterium spp. were isolated from goat cheese and milk, and ewe rumen liquor, and characterized in view of their possible utilization for the production of novel pro-bioactive food and feed on scotta, a lactose rich substrate and one of the main by-products of the dairy industry. The evaluation of the Minimum Inhibitory Concentration (MIC) of 13 among the most common antibiotics in clinical practice revealed a general susceptibility to ampicillin, gentamycin, streptomycin, vancomycin, chloramphenicol, and clindamycin while confirming a lower susceptibility to aminoglycosides and ciprofloxacin. Twenty-five isolates, that proved capable of lactose utilization as the sole carbon source, were then characterized for functional and biotechnological properties. Four of them, ascribed to Propionibacterium freudenreichii species, and harboring resistance to bile salts (growth at 0.7-1.56 mM of unconjugated bile salts), acid stress (>80% survival after 1 h at pH 2), osmostress (growth at up to 6.5% NaCl) and lyophilization (survival rate > 80%), were selected and inoculated in scotta. On this substrate the four isolates reached cell densities ranging from 8.11 ± 0.14 to 9.45 ± 0.06 Log CFU mL-1 and proved capable of producing different vitamin B9 vitamers after 72 h incubation at 30°C. In addition, the semi-quantitative analysis following the metabolomics profiling revealed a total production of cobalamin derivatives (vitamin B12) in the range 0.49-1.31 mg L-1, thus suggesting a full activity of the corresponding biosynthetic pathways, likely involving a complex interplay between folate cycle and methylation cycle required in vitamin B12 biosynthesis. These isolates appear interesting candidates for further ad-hoc investigation regarding the production of pro-bioactive scotta.

Bioaccessibility and bioactive potential of different phytochemical classes from nutraceuticals and functional foods.

Nutraceuticals and functional foods are composed of especially complex matrices, with polyphenols, carotenoids, minerals, and vitamins, among others, being the main classes of phytochemicals involved in their bioactivities. Despite their wide use, further investigations are needed to certify the proper release of these phytochemicals into the gastrointestinal medium, where the bioaccessibility assay is one of the most frequently used method. The aim of this review was to gather and describe different methods that can be used to assess the bioaccessibility of nutraceuticals and functional foods, along with the most important factors that can impact this process. The link between simulated digestion testing of phytochemicals and their in vitro bioactivity is also discussed, with a special focus on the potential of developing nutraceuticals and functional foods from simple plant materials. The bioactive potential of certain classes of phytochemicals from nutraceuticals and functional foods is susceptible to different variations during the bioaccessibility assessment, with different factors contributing to this variability, namely the chemical composition and the nature of the matrix. Regardless of the high number of studies, the current methodology fails to assume correlations between bioaccessibility and bioactivity, and the findings of this review indicate a necessity for updated and standardized protocols.

The integrated metabolomics and sensory analyses unravel the peculiarities of mountain grassland-based cheese production: The case of Parmigiano Reggiano PDO.

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.

Untargeted phytochemical profiling and biological activity of small yellow onion (Allium flavum L.) from different regions of Romania.

This study examined the phytochemical profiles (mainly phenolics, carotenoids, and organosulfur compounds) and biological effects of hydroalcoholic extracts of Allium flavum (AF), a species of the Allium genus commonly known as small yellow onion. Unsupervised and supervised statistical approaches revealed clear differences between extracts prepared with samples collected from different areas of Romania. Overall, the AFFF (AF flowers collected from Faget) extract was the best source of polyphenols, also showing the highest antioxidant capacity evaluated through both in vitro DPPH, FRAP, and TEAC anti-radical scavenging assays and cell-based OxHLIA and TBARS assays. All the tested extracts exhibited α-glucosidase inhibition potential, while only the AFFF extract exhibited anti-lipase inhibitory activity. The phenolic subclasses annotated were positively correlated with the assessed antioxidant and enzyme inhibitory activities. Our findings suggested that A. flavum has bioactive properties worth exploring further, being a potential edible flower with health-promoting implications.

Role of Platinum Nanozymes in the Oxidative Stress Response of Salmonella Typhimurium.

Platinum nanoparticles (PtNPs) are being intensively explored as efficient nanozymes due to their biocompatibility coupled with excellent catalytic activities, which make them potential candidates as antimicrobial agents. Their antibacterial efficacy and the precise mechanism of action are, however, still unclear. In this framework, we investigated the oxidative stress response of Salmonella enterica serovar Typhimurium cells when exposed to 5 nm citrate coated PtNPs. Notably, by performing a systematic investigation that combines the use of a knock-out mutant strain 12023 HpxF- with impaired response to ROS (ΔkatE ΔkatG ΔkatN ΔahpCF ΔtsaA) and its respective wild-type strain, growth experiments in both aerobic and anaerobic conditions, and untargeted metabolomic profiling, we were able to disclose the involved antibacterial mechanisms. Interestingly, PtNPs exerted their biocidal effect mainly through their oxidase-like properties, though with limited antibacterial activity on the wild-type strain at high particle concentrations and significantly stronger action on the mutant strain, especially in aerobic conditions. The untargeted metabolomic analyses of oxidative stress markers revealed that 12023 HpxF- was not able to cope with PtNPs-based oxidative stress as efficiently as the parental strain. The observed oxidase-induced effects comprise bacterial membrane damage as well as lipid, glutathione and DNA oxidation. On the other hand, in the presence of exogenous bactericidal agents such as hydrogen peroxide, PtNPs display a protective ROS scavenging action, due to their efficient peroxidase mimicking activity. This mechanistic study can contribute to clarifying the mechanisms of PtNPs and their potential applications as antimicrobial agents.

Phenolic profile, in vitro antimicrobial and in vivo diuretic effects of endemic wild thyme Thymus comosus Heuff ex. Griseb. (Lamiaceae) from Romania.

Thymus comosus Heuff ex. Griseb. (Lamiaceae) is a wild thyme species endemic for Romanian Carpathian areas, frequently collected as substitute for collective herbal product Serpylli herba, cited as antibacterial and diuretic remedy in traditional medicine. The present study aimed to evaluate the in vivo diuretic effect and in vitro antimicrobial properties of three herbal preparations (infusion-TCI, tincture-TCT and an hydroethanolic extract prepared through an optimized ultrasound-assisted method-OpTC) obtained from the aerial parts of T. comosus Heuff ex. Griseb, also evaluating their comprehensive phenolic profile. In vivo diuretic effect was tested using Wistar rats treated orally with each herbal preparation (125 and 250 mg/kg dispersed in 25 ml/kg isotonic saline solution) and quantified based on cumulative urine output (ml), diuretic action and diuretic activity. Additionally, sodium and potassium excretion were monitored using a potentiometric method with selective electrodes. In vitro antibacterial and antifungal activities were assessed using p-iodonitrotetrazolium chloride assay against six bacterial strains and six fungal strains by monitoring minimum inhibitory concentration (MICs), minimum bactericidal concentrations (MBCs) and minimum fungicidal concentrations (MFCs). Finally, phenolic profile of the aforementioned herbal extracts was evaluated using an ultra-high-pressure liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS) method to check the impact of the different preparations on the most abundant and significant compounds. All the extracts exerted a mild diuretic action, TCT and OpTC inducing the most intense diuretic effect. Both herbal preparations produced a statistically significant, dose-dependent and gradual increase of the urine output, the effect being more intense at 24 h (6.63-7.13 ml/24 h). Potentiometric evaluation of urine samples collected from treated rats revealed a clear and mild natriuretic and kaliuretic effect after the administration. In terms of antimicrobial activity, E. coli (MIC-0.38 mg/ml), B. cereus (MIC-0.75 mg/ml)), Penicillium funiculosum and P. verrucosum var. cyclopium (MIC-0.19 mg/ml) showed the greater sensitivity to the tested extracts, respectively. UHPLC-HRMS screening showed that the bioactive potential of T. comosus herbal preparations was likely related to the higher amounts of phenolic acids (including rosmarinic acid), flavonoids (mainly flavones and derivatives) and other phenolics (such as different isomers of salvianolic acids) in their composition. The obtained results support the ethnopharmacological evidence regarding the mild diuretic and antibacterial potentials of the endemic wild thyme T. comosus, this study being the first one that assessed the aforementioned bioactivities for this species.

Evaluation of different blackcurrant seed ingredients in meatballs by using conventional quality assessment and untargeted metabolomics.

Blackcurrants are sources of phenolic compounds, such as anthocyanins, possessing strong antioxidant, antimicrobial and antifungal activity. Therefore, the addition of different blackcurrant pomace ingredients may affect the overall meat quality. The actual chemical profile and bioactivities of blackcurrant pomace ingredients may strongly depend on its preparation; for instance, in our study the highest values of the in vitro antioxidant capacity were determined for blackcurrant seeds after supercritical CO2 extraction. Starting from these background conditions, in this work, we evaluated the ability of three different concentrations (namely 1, 3, and 5% w/w) of blackcurrant (BC) seeds following EtOH/water extraction (BC-AE), before supercritical fluid CO2 extraction (BC-RS), and after supercritical fluid CO2 extraction (BC-ASC) to affect different quality parameters of pork meatballs. These latter were stored considering three different time-points, namely 1, 3 and 6 days at 4 °C packed under modified atmosphere (i.e., 70% N2 and 30% CO2). Untargeted metabolomics allowed to identify several lipid and protein-related oxidation products involved in redox reactions, such as 13-L-hydroperoxylinoleic acid, (12S,13S)-epoxylinolenic acid, 9,10-epoxyoctadecenoic acid, glutathione, glutathione disulfide, L-carnosine, l-ascorbic acid, and tocotrienols. Besides, multivariate statistics applied on the metabolomics dataset confirmed that the chemical profile of meatballs was an exclusive combination of both BC inclusion levels and type of BC-ingredients considered. Our findings showed that the higher the concentration of BC seed ingredients in meatballs, the lower the cooking loss and the higher the fibre content. Also, all the ingredients significantly affected the colour parameters.

Untargeted Metabolomics Combined with Sensory Analysis to Evaluate the Chemical Changes in Coppa Piacentina PDO during Different Ripening Times.

Ripening time is known to drive the chemical and sensory profiles of dry meat products, thus potentially affecting the final quality of the product. Starting from these background conditions, the aim of this work was to shed light, for the first time, on the chemical modifications of a typical Italian PDO meat product-namely, Coppa Piacentina-during ripening, to find correlations between its sensory quality and the biomarker compounds related to the progress of ripening. The ripening time (from 60 to 240 days) was found to deeply modify the chemical composition of this typical meat product, providing potential biomarkers of both oxidative reactions and sensory attributes. The chemical analyses revealed that there is typically a significant decrease in the moisture content during ripening, likely due to increased dehydration. In addition, the fatty acid profile showed that the distribution of polyunsaturated fatty acids significantly (p < 0.05) decreased during ripening, because of their high susceptibility to oxidation and conversion to intermediate and secondary molecules. An untargeted metabolomics approach, coupled with unsupervised and supervised multivariate statistics, highlighted a significant impact (prediction scores > 1) of lipid oxidation during ripening time, with some metabolites (such as γ -glutamyl-peptides, hydroperoxy-fatty acids, and glutathione) being particularly discriminant in predicting the changes observed. The discriminant metabolites were coherent with the progressive increase of peroxide values determined during the entire ripening period. Finally, the sensory analysis outlined that the highest degree of ripening provided greater color intensity of the lean part, slice firmness, and chewing consistency, with glutathione and γ-glutamyl-glutamic acid establishing the highest number of significant correlations with the sensory attributes evaluated. Taken together, this work highlights the importance and validity of untargeted metabolomics coupled with sensory analysis to investigate the comprehensive chemical and sensory changes to dry meat during ripening.