Comparative analysis of the longissimus muscle proteome of European wild boar and domestic pig in response to thermal processing.

This study tries to fill the knowledge gap regarding differences in the expression of proteins in the meat of European wild boar (Sus scrofa scrofa) and domestic pig (Sus scrofa domestica), considering the impact of thermally induced degradation. We assessed relative protein changes between cooked longissimus thoracis et lumborum (LTL) muscle proteomes by using mass spectrometry, chemometric, label-free proteomic, and bioinformatic tools. Among 30 differentially abundant proteins identified MyHC-2a, ATPs-α, CK-S, ADP/ATPt1, IDH2, and MyBP-C1 were upregulated (x > 1) whereas NEB, γ-ENO and EPSF were downregulated (x < 1) in wild boar. ShinyGO and KEGG database pathway analyses revealed that these proteins are mainly involved in processes related to muscle contraction and various pathways of glucose metabolism and energy production. Protein expression changes could have been caused by the different muscle activity of wild animals in response to prolonged movement associated with foraging for food in the natural environment.

Changes in physicochemical, textural, and sensorial properties of pork meatballs made with the addition of hemp oil during storage.

This research aimed to evaluate the quality characteristics of cooked and vacuum-packed meatballs reformulated with cold-pressed hempseed oil as a partial pork substitute (0.8%, 2.5%, 4.2%, and 7.5%) during 12 days of storage. The water activity, cooking, and storage losses increased with a higher content of hemp oil (P < 0.05). The total saturated fatty acids were reduced by 37.6%, whereas the polyunsaturated fatty acids content improved by 96.1%. Hemp oil addition decreased protein and lipid oxidation during the storage period (P < 0.05). The inhibition effect on carbonyl content reached 34.9% and on TBARS values reached 17.5%. Sensory analysis revealed no significant changes to the texture, odour, and taste attributes over 12 days of storage in vacuum packaging. The results indicate that cold-pressed hemp oil can be an alternative ingredient for the production of meat products with improved nutritional value, particularly by enriching them with n-3 α-linolenic fatty acid.

Comparing Different Chemometric Approaches to Detect Adulteration of Cold-Pressed Flaxseed Oil with Refined Rapeseed Oil Using Differential Scanning Calorimetry.

Flaxseed oil is one of the best sources of n-3 fatty acids, thus its adulteration with refined oils can lead to a reduction in its nutritional value and overall quality. The purpose of this study was to compare different chemometric models to detect adulteration of flaxseed oil with refined rapeseed oil (RP) using differential scanning calorimetry (DSC). Based on the melting phase transition curve, parameters such as peak temperature (T), peak height (h), and percentage of area (P) were determined for pure and adulterated flaxseed oils with an RP concentration of 5, 10, 20, 30, and 50% (w/w). Significant linear correlations (p ≤ 0.05) between the RP concentration and all DSC parameters were observed, except for parameter h1 for the first peak. In order to assess the usefulness of the DSC technique for detecting adulterations, three chemometric approaches were compared: (1) classification models (linear discriminant analysis-LDA, adaptive regression splines-MARS, support vector machine-SVM, and artificial neural networks-ANNs); (2) regression models (multiple linear regression-MLR, MARS, SVM, ANNs, and PLS); and (3) a combined model of orthogonal partial least squares discriminant analysis (OPLS-DA). With the LDA model, the highest accuracy of 99.5% in classifying the samples, followed by ANN > SVM > MARS, was achieved. Among the regression models, the ANN model showed the highest correlation between observed and predicted values (R = 0.996), while other models showed goodness of fit as following MARS > SVM > MLR. Comparing OPLS-DA and PLS methods, higher values of R2X(cum) = 0.986 and Q2 = 0.973 were observed with the PLS model than OPLS-DA. This study demonstrates the usefulness of the DSC technique and importance of an appropriate chemometric model for predicting the adulteration of cold-pressed flaxseed oil with refined rapeseed oil.

LC-MS Metabolomic Profiling of Five Types of Unrefined, Cold-Pressed Seed Oils to Identify Markers to Determine Oil Authenticity and to Test for Oil Adulteration.

The authenticity of food products marketed as health-promoting foods-especially unrefined, cold-pressed seed oils-should be controlled to ensure their quality and safeguard consumers and patients. Metabolomic profiling using liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (LC-QTOF) was employed to identify authenticity markers for five types of unrefined, cold-pressed seed oils: black seed oil (Nigella sativa L.), pumpkin seed oil (Cucurbita pepo L.), evening primrose oil (Oenothera biennis L.), hemp oil (Cannabis sativa L.) and milk thistle oil (Silybum marianum). Of the 36 oil-specific markers detected, 10 were established for black seed oil, 8 for evening primrose seed oil, 7 for hemp seed oil, 4 for milk thistle seed oil and 7 for pumpkin seed oil. In addition, the influence of matrix variability on the oil-specific metabolic markers was examined by studying binary oil mixtures containing varying volume percentages of each tested oil and each of three potential adulterants: sunflower, rapeseed and sesame oil. The presence of oil-specific markers was confirmed in 7 commercial oil mix products. The identified 36 oil-specific metabolic markers proved useful for confirming the authenticity of the five target seed oils. The ability to detect adulterations of these oils with sunflower, rapeseed and sesame oil was demonstrated.

Heat-stable peptide markers specific to rabbit and chicken liver tissue for meat product authentication testing.

A three-step analysis was used to detect and identify heat-stable peptide markers specific to liver tissue from rabbit and chicken. It involved peptide discovery by liquid chromatography coupled to high resolution mass spectrometer (LC-HRMS), followed by protein identification using Spectrum Mill software and multiple reaction monitoring (MRM) based confirmation of the discovered peptides using a liquid chromatography coupled to triple quadrupole mass spectrometer (LC-TQ). We identified 50 and 91 heat-stable peptide markers unique to chicken and rabbit liver, respectively. The markers were validated in commercial food samples with declared liver tissue contents ranging from 5% to 30%. The best candidate peptides for distinguishing liver tissue from skeletal muscle were selected and then confirmed using MRM-based approach. Limit of detection of liver was found to be in the range of 0.13 to 2.13% (w/w) for chicken liver-specific peptide markers, and from 0.04 to 0.6% (w/w) for rabbit liver-specific peptide markers.

Identification of sunflower, rapeseed, flaxseed and sesame seed oil metabolomic markers as a potential tool for oil authentication and detecting adulterations.

Testing the composition, quality and authenticity of edible oils is crucial to safeguard the consumers' rights and health. The aim of our study was to identify oil-specific markers to enable the differentiation and authentication of sunflower, sesame, flaxseed and rapeseed oils, and to evaluate their antioxidant activity, total phenolic and carotenoid content. A metabolomic approach based on liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry was employed for marker discovery. Spectrophotometric method was used for determination of antioxidant activity, total phenolic and carotenoid content. 76 oil samples from the four different manufacturers were examined. We identified 13 oil-specific markers for sunflower seed oil, 8 for rapeseed oil, 5 for sesame seed oil and 3 for flaxseed oil, their retention times, accurate masses, and characteristic fragment ions are reported. The abundances of the markers for each plant species were found to vary depending on the oil producer and the product batch. Significant differences in antioxidant activity, total phenolic and carotenoid content were also observed both between oils and within oil type. The highest total phenolic content (84.03 ± 4.19 to 103.79 ± 3.67 mg of gallic acid/kg) and antioxidant activity (245.67 ± 7.59 to 297.22 ± 2.32 mg Trolox/kg) were found in sesame seed and flaxseed oils, respectively. Identified metabolic markers can be used as qualitative markers to confirm the authenticity or to detect adulterations of oils. Composition, properties and authenticity testing should be more rigorous for food products marketed as health-promoting.

A systematic review of DNA-based methods in authentication of game and less common meat species.

Despite the numerous studies on food safety and authenticity, especially for meat and meat products, not enough studies have been conducted focusing exclusively on game species and other unusual meat animals. As a result of the European horse scandal, the horse is currently the target of many meat authenticity studies. With this review, we aim to present various DNA-based methods that have been used by researchers to identify, detect, and quantify game, uncommon meat animals, and wildlife species. Polymerase chain reaction (PCR) is considered the standard method for DNA analysis in meat authenticity testing. However, in this paper, we present several other methods that may or may not involve the PCR technique. For this purpose, we systematically reviewed 131 articles selected according to various criteria such as target animal species, method of analysis, year of publication, and so forth. The result of our study shows the most studied game and uncommon meat species, PCR- and non-PCR-based methods for game meat analysis, and DNA-based methods in wildlife conservation. With this study, researchers can find detailed information about frequent game species used as adulterants for regular meat products and the DNA-based techniques to identify them.

Pork liver tissue-specific peptide markers for food authenticity testing and adulteration detections.

A liquid chromatography-mass spectrometry bottom-up proteomic approach was applied for the detection and identification of proteins from liver tissues. We identified 74 unique pork liver peptide markers that are resistant to the thermal processing of food. These peptides are derived from 43 proteins, which perform various functions in the liver. Roasted and sterilised pâté-type products with a pork liver content ranging from 6% to 51% were examined to select the most reliable pork liver peptide markers that survive unmodified in complex processed food matrices. Of the 74 specific heat-stable peptides detected in pure liver tissue, five (GDAPEEEVSLSK, ALTAELEAVGK, TFYLNVLNEEER, AQFGQPEILLGTIPGTGGTQR and VIAPGFNALEQILQSTAGK) were the best candidates to confirm the presence of liver tissue in highly processed meat products. We have identified unique tissue-specific markers that enable rapid and specific identification of pork liver in processed food and may contribute to the development of new methods for testing food authenticity.

Shotgun Lipidomic Analysis for Differentiation of Niche Cold Pressed Oils.

The fast-growing food industry is bringing significant number of new products to the market. To protect consumers' health and rights, it is crucial that food control laboratories are able to ensure reliable quality testing, including product authentication and detection of adulterations. In our study, we applied a fast and eco-friendly method based on shotgun-lipidomic mass spectrometry for the authentication of niche edible oils. Comprehensive lipid profiles of camelina (CA), flax (FL) and hemp (HP) seed oils were obtained. With the aid of principal component analysis (PCA), it was possible to detect and distinguish each of them based on their lipid profiles. Lipidomic markers characteristic ofthe oils were also identified, which can be used as targets and expedite development of new multiplexed testing methods.

The effect of storage method on selected physicochemical and microbiological qualities of wild boar meat.

BACKGROUND: This study investigated the influence of the storage method on the physicochemical characteristics and microbial growth of m. longissimus thoracis et lumborum (LTL), m. biceps femoris (BF) and m. vastus lateralis (VL) of wild boar. Muscles were stored in a vacuum (VAC), in a modified high-oxygen atmosphere (MAP) or meat seasoning cabinet (DRY-AGED) for 21 days. RESULTS: Wild boar meat was characterised by a high protein and low fat content and a good amount of potassium, sodium, calcium, magnesium, zinc and iron. Significantly higher (P < 0.05) pH values were noted for DRY-AGED muscles stored for 21 days (up to 5.89 for VL). On day 21, a significant decrease in pH was noted for all MAP muscles (down to 5.23 for BF). Storage losses due to desiccation and water loss were significantly higher for DRY-AGED samples and ranged from 25.63% to 32.89% on day 21. MAP affected protein and lipid oxidation, which was also reflected in Warner-Bratzler shear force VAC and DRY-AGED had positive results regarding tenderness, whereas on day 21 the MAP-stored meat had toughened significantly (from 35.3 N to 50.7 N in LTL). Lipids were oxidised much faster than proteins during prolonged storage in MAP. Compared to the other methods, DRY-AGED had the best effect on microbial growth. CONCLUSION: These results indicate that the recommended methods for the storage of wild boar meat are either vacuum packing or dry ageing. The high oxygen content of MAP negatively affected the quality of wild boar meat and carried a risk of increased protein carbonylation. © 2022 Society of Chemical Industry.

The Effect of Hemp Cake (Cannabis sativa L.) on the Characteristics of Meatballs Stored in Refrigerated Conditions.

Hemp cake, a by-product of cold pressing oil from hemp seeds, is a nutritious ingredient that could be used for the production of new or reformulated meat products. The aim of this study was to determine the influence of inclusion of 0.9%, 2.6%, 4.2%, and 7.4% (w/w) hemp cake (Cannabis sativa L.) on the physicochemical and textural properties, oxidation, and sensory acceptance of cooked and vacuum-packed meatballs during refrigerated storage. The addition of 7.4% hemp cake enhanced the amount of dry matter and reduced the content of water. Lightness (L*) and redness (a*) values reduced significantly with higher levels of hemp supplementation. Regardless of the amount of hemp additive, pH, color parameters did not differ significantly during the 12 days of storage. Hemp cake significantly decreased protein and lipid oxidation: the inhibitory effect of adding 7.4% hemp cake on protein carbonyl group formation and TBARS values reached 11.16% and 36.5%, respectively, after 10 days of storage. Sensory analysis revealed that meatballs prepared with 0.9% and 2.6% hemp cake gained higher overall scores. The results indicate that hemp cake, a material considered mainly as waste, may be destined for food purposes and be an alternative ingredient for the production of sustainable meat products.

A Practical Approach to Identifying Processed White Meat of Guinea Fowl, Rabbit, and Selected Fish Species Using End-Point PCR.

Among the foodstuff, most often adulterated are white meat and meat products as well as fish and fish products. For this reason, we evaluated in practice the possibilities of identifying selected species of white meat, i.e., guinea fowl and rabbit as well as four fish species, namely, pollock, hake, sole, and panga, in thermally treated samples. The aim was to check whether the previously published in the scientific literature species-specific primers allows for the identification of processed meat using the end-point PCR technique. To identify the six species, the short sequence fragments (from 130 to 255 bp) of 12S rRNA, COX3, mitochondrial ATP synthase Fo subunit 6 (ATP6) gene, pantophysin (Pan I) gene, 5S rRNA gene, and microsatellite markers (locus: Phy01-KUL) were selected. Stability and specificity of the six pair primers were evaluated on cooked and autoclaved meat, and commercially processed food samples such as rabbit and guinea pâtés, ready-made baby food, and breaded, fried, and deep-frozen fish products. The method proved to be useful for the authentication of severely processed food products against fraudulent species substitution and mislabelling and this approach may be an alternative to more advanced and more expensive PCR techniques.

Multispecies Identification of Oilseed- and Meat-Specific Proteins and Heat-Stable Peptide Markers in Food Products.

Consumer demand for both plant products and meat products enriched with plant raw materials is constantly increasing. Therefore, new versatile and reliable methods are needed to find and combat fraudulent practices in processed foods. The objective of this study was to identify oilseed species-specific peptide markers and meat-specific markers that were resistant to processing, for multispecies authentication of different meat and vegan food products using the proteomic LC-MS/MS method. To assess the limit of detection (LOD) for hemp proteins, cooked meatballs consisting of three meat species and hemp cake at a final concentration of up to 7.4% were examined. Hemp addition at a low concentration of below 1% was detected. The LOD for edestin subunits and albumin was 0.9% (w/w), whereas for 7S vicilin-like protein it was 4.2% (w/w). Specific heat-stable peptides unique to hemp seeds, flaxseed, nigella, pumpkin, sesame, and sunflower seeds, as well as guinea fowl, rabbit, pork, and chicken meat, were detected in different meat and vegan foods. Most of the oilseed-specific peptides were identified as processing-resistant markers belonging to 11S globulin subunits, namely conlinin, edestin, helianthinin, pumpkin vicilin-like or late embryogenesis proteins, and sesame legumin-like as well as 2S albumins and oleosin isoforms or selected enzymic proteins.

Peptide markers for distinguishing guinea fowl meat from that of other species using liquid chromatography-mass spectrometry.

The inability to easily identify the animal species in highly processed meat products makes them highly susceptible to adulterations. Reliable methods for detecting the species origin of meat used in processed food are required to ensure adequate labelling and minimize food fraud and allergenic potential. Liquid chromatography high resolution mass spectrometry was employed to identify new heat-stable guinea-fowl-specific peptide markers that can differentiate guinea fowl meat from other commonly consumed animal species, including closely related poultry species, in highly processed food products. We identified 26 unique guinea-fowl-specific markers. The high stability of guinea-fowl-specific peptides was confirmed by analysing food products with guinea fowl meat content ranging from 4% to 100%. The findings indicate that sensitive and reliable LC-MS/MS methods can be developed for the targeted detection and quantification of guinea fowl meat in highly processed meat products.

LIQUID CHROMATOGRAPHY-MASS SPECTROMETRY BOTTOM-UP PROTEOMIC METHODS IN ANIMAL SPECIES ANALYSIS OF PROCESSED MEAT FOR FOOD AUTHENTICATION AND THE DETECTION OF ADULTERATIONS.

This review offers an overview of the current status and the most recent advances in liquid chromatography-mass spectrometry (LC-MS) techniques with both high-resolution and low-resolution tandem mass analyzers applied to the identification and detection of heat-stable species-specific peptide markers of meat in highly processed food products. We present sets of myofibrillar and sarcoplasmic proteins, which turned out to be the source of 105 heat-stable peptides, detectable in processed meat using LC-MS/MS. A list of heat-stable species-specific peptides was compiled for eleven types of white and red meat including chicken, duck, goose, turkey, pork, beef, lamb, rabbit, buffalo, deer, and horse meat, which can be used as markers for meat authentication. Among the 105 peptides, 57 were verified by multiple reaction monitoring, enabling identification of each species with high specificity and selectivity. The most described and monitored species by LC-MS/MS so far are chicken and pork with 26 confirmed heat-stable peptide markers for each meat. In thermally processed samples, myosin, myoglobin, hemoglobin, l-lactase dehydrogenase A and ß-enolase are the main protein sources of heat-stable markers. © 2019 John Wiley & Sons Ltd. Mass Spec Rev.

Proteomic analysis of oilseed cake: a comparative study of species-specific proteins and peptides extracted from ten seed species.

BACKGROUND: In recent years there has been a visible trend among consumers to move away from consuming meat in favor of plant products. Meat producers have therefore been trying to meet the expectations of consumers by introducing new products to the food market with a greater proportion of plant ingredients. Meat products are enriched not only by the addition of vegetable oils but also by ground or whole oilseeds or their preparation. In this study, we present in-solution tryptic digestion and an ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS)-based proteomics approach to investigate specific proteins and peptides of ten oilseed cakes, by-products of cold pressing oil from coconut, evening primrose, hemp, flax, milk thistle, nigella, pumpkin, rapeseed, sesame, and sunflower seeds, for authentication purposes. RESULTS: We identified a total of 229 unique oilseed proteins. The number of specific proteins varied depending on the sample, from 4 to 48 in evening primrose and sesame. Moreover, we identified approximately 440 oilseed unique peptides in the cakes of all the analyzed oilseeds; the largest amounts were found in sesame (107 peptides), sunflower (100), pumpkin, hemp (42), rapeseed (36), and flax cake (35 peptides). CONCLUSIONS: We provide novel information on unique / species-specific peptide markers that will extend the scope of testing the authenticity of a wide range of foods. The results of this peptide discovery experiment may further contribute to the development of targeted methods for the detection and quantification of oilseed proteins in processed foods, and thus to the improvement of food quality. © 2020 Society of Chemical Industry.

Identification of species-specific peptide markers in cold-pressed oils.

In recent years, cold-pressed vegetable oils have become very popular on the global market. Therefore, new versatile methods with high sensitivity and specificity are needed to find and combat fraudulent practices. The objective of this study was to identify oilseed species-specific peptide markers, using proteomic techniques, for authentication of 10 cold-pressed oils. In total, over 380 proteins and 1050 peptides were detected in the samples. Among those peptides, 92 were found to be species-specific and unique to coconut, evening primrose, flax, hemp, milk thistle, nigella, pumpkin, rapeseed, sesame, and sunflower oilseed species. Most of the specific peptides were released from major seed storage proteins (11 globulins, 2S albumins), and oleosins. Additionally, the presence of allergenic proteins in the cold-pressed oils, including pumpkin Cuc ma 5, sunflower Hel a 3, and six sesame allergens (Ses i 1, Ses i 2, Ses i 3, Ses i 4, Ses i 6, and Ses i 7) was confirmed in this study. This study provides novel information on specific peptides that will help to monitor and verify the declared composition of cold-pressed oil as well as the presence of food allergens. This study can be useful in the era of widely used unlawful practices.

LC-QTOF-MS identification of rabbit-specific peptides for authenticating the species composition of meat products.

Rabbit is a healthy meat, with low allergenicity and excellent nutritional properties. The global popularity of rabbit meat makes it a target for food fraud. We present a LC-QTOF-MS/MS approach for detecting and identifying rabbit-specific peptide-markers from thermally processed meat products to differentiate rabbit from other commonly-consumed animal species. We identified 49 heat-stable specific peptides. We selected the most stable markers for testing complex meat matrices by analysing pâtés-type products with a rabbit meat content ranging from 5% to 85%. Of the 49 heat-stable peptides detected in pure cooked rabbit meat, three were consistently detected in all investigated pâté samples i.e., SSVFVADPK, AFFGHYLYEVAR and PHSHPALTPEQK. Monitoring meat species other than rabbit in the examined pâtés using pork-, lamb- and chicken-specific peptides identified the presence of undeclared chicken in two samples. The results confirm that LC-QTOF-MS/MS is a suitable tool for multi-species detection in processed meat products, particularly for authentication purposes.

Nutritional value, protein and peptide composition of edible cricket powders.

There are various indicators, including FAO and EU sources, that edible insects could become one of the solutions to the problem of global food supply. This report was aimed at improving the knowledge on powdered crickets (Acheta domesticus). The analyses of the basic nutritional composition revealed that cricket powders were rich in protein (42.0-45.8% of dry matter) and fat (23.6-29.1% of dry matter). In terms of mineral content, CPs were rich in Ca, Mg and Fe. Most of all, the levels of Cu, Mn and Zn were especially high (2.33-4.51, 4.1-12.5, 12.8-21.8 mg/100 g of dry matter, respectively). Furthermore, the analyses into the proteins indicated that the cricket powders were treated with high temperatures and allowed the determination of four cricket-specific peptides that showed sufficient thermostability to serve as markers for authentication.

Species-specific peptide-based liquid chromatography-mass spectrometry monitoring of three poultry species in processed meat products.

The detection of adulteration and mislabeling of food products, including intensively processed meat, is a challenge which needs urgent solutions to protect consumers' rights. The aim of the study was to demonstrate the feasibility of species-specific peptide-based LC-MS methods for monitoring duck, goose and chicken in processed meat products. Food commodities of various compositions, subjected to various treatments, including homogenization, cooking, roasting, drying, and sterilization during production, were examined to ensure that MS-based methods are resistant to matrix composition changes. A qualitative LC-QQQ multiple reaction monitoring (MRM) method was developed which allows high-confidence monitoring of duck, goose and chicken meat (ten specific peptides), simultaneously with beef and pork (seven peptides), in the presence of turkey meat, in highly processed food. The developed LC-MS methods can be used for food authentication, monitoring of the food composition conformity with label statements and detection of adulteration of poultry-containing food products.