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Scope: Epidemiological evidence suggests that a reduced risk of colorectal cancer (CRC) is correlated with high consumption of fruits and vegetables, which are major sources of fiber and phytochemicals, such as flavan-3-ols. However, it remains unknown how these phytochemicals and their specific gut-related metabolites may alter cancer cell behavior. Methods and results: A focused screening using native (poly)phenols and gut microbial metabolites (GMMs) on 3D HCT116 spheroids is carried out using a high-throughput imaging approach. Dose–responses, IC50, and long-term exposure are calculated for the most promising native (poly)phenols and GMMs. As a result, this research shows that (poly)phenol catabolites may play a key role in preventing cancer propagation. Indeed, µM concentration levels of (4R)-5-(3ʹ,4ʹ-dihydroxyphenyl)-γ-valerolactone significantly decrease spheroid size at early stages of spheroid aggregation and gene expression of matrix metalloproteinases. Conclusion: A chronic exposure to (4R)-5-(3ʹ,4ʹ-dihydroxyphenyl)-γ-valerolactone may lead to a reduced CRC risk. Daily intake of monomeric, oligomeric, and polymeric flavan-3-ols may increase the colonic concentrations of this metabolite, and, in turn, this compound may act locally interacting with intestinal epithelial cells, precancerous and cancer cells.

Simple Summary Glioblastoma is the most lethal primary adult brain tumor. The great number of mutations involved and the aggressiveness of glioblastoma render this type of cancer especially difficult to investigate. To address this problem, cerebral organoids have emerged as promising tools to investigate brain biology and to recapitulates the major steps involved in glioblastoma tumorigenesis. This review focuses on methods of cerebral organoid development, describes the protocols used for inducing glioblastoma, the approach used to derive glioblastoma organoids directly from patients’ biopsies and discusses their limitations and potential future direction. Abstract Glioblastoma (GB) is the most frequent and aggressive type of glioma. The lack of reliable GB models, together with its considerable clinical heterogeneity, has impaired a comprehensive investigation of the mechanisms that lead to tumorigenesis, cancer progression, and response to treatments. Recently, 3D cultures have opened the possibility to overcome these challenges and cerebral organoids are emerging as a leading-edge tool in GB research. The opportunity to easily engineer brain organoids via gene editing and to perform co-cultures with patient-derived tumor spheroids has enabled the analysis of cancer development in a context that better mimics brain tissue architecture. Moreover, the establishment of biobanks from GB patient-derived organoids represents a crucial starting point to improve precision medicine therapies. This review exemplifies relevant aspects of 3D models of glioblastoma, with a specific focus on organoids and their involvement in basic and translational research.

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-benefit is on the verge of really enrolling into the risk assessment and risk analysis paradigm. The interaction between risk–benefit assessors and risk–benefit managers is pivotal in this, as is the interaction with risk–benefit communicators. Highlights: - Risk-benefit assessment goes hand-in-hand with RB-management and RB-communication; - Holistic approach in food safety and nutrition, comparing health risks and benefits; - Risk-benefit assessment estimates the overall impact of food on health; - Risk-benefit characteristics: problem formulation, tiered approach, common currency. info:eu-repo/semantics/publishedVersion

Scope Several studies suggest that regular coffee consumption may help preventing chronic diseases, but the impact of daily intake and the contribution of coffee metabolites in disease prevention are still unclear. The present study aimed at evaluating whether and how different patterns of coffee intake (one cup of espresso coffee/day, three cups of espresso coffee/day, one cup of espresso coffee/day and two cocoa-based products containing coffee two times per day) might impact endogenous molecular pathways. Methods and results A three-arm, randomized, cross-over trial was performed in 21 healthy volunteers who consumed each treatment for one month. Urine samples were collected to perform untargeted metabolomics based on UHPLC-IMS-HRMS. A total of 153 discriminant metabolites were identified. Several molecular features were associated with coffee consumption, while others were linked with different metabolic pathways, such as phenylalanine, tyrosine, energy metabolism, steroid hormone biosynthesis and arginine biosynthesis and metabolism. Conclusion This information has provided new insights into the metabolic routes by which coffee and coffee-related metabolites may exert effects on human health. This article is protected by copyright. All rights reserved.

The life sciences are currently being transformed by an unprecedented wave of developments in molecular analysis, which include important advances in instrumental analysis as well as biocomputing. In light of the central role played by metabolism in nutrition, metabolomics is rapidly being established as a key analytical tool in human nutritional studies. Consequently, an increasing number of nutritionists integrate metabolomics into their study designs. Within this dynamic landscape, the potential of nutritional metabolomics (nutrimetabolomics) to be translated into a science, which can impact on health policies, still needs to be realized. A key element to reach this goal is the ability of the research community to join, to collectively make the best use of the potential offered by nutritional metabolomics. This article, therefore, provides a methodological description of nutritional metabolomics that reflects on the state‐of‐the‐art techniques used in the laboratories of the Food Biomarker Alliance (funded by the European Joint Programming Initiative "A Healthy Diet for a Healthy Life" (JPI HDHL)) as well as points of reflections to harmonize this field. It is not intended to be exhaustive but rather to present a pragmatic guidance on metabolomic methodologies, providing readers with useful "tips and tricks" along the analytical workflow.

In the present work the possibility of using an untargeted metabolomic strategy to discriminate between common and durum wheat lipidome for an authenticity purpose was explored. A first study was conducted by analyzing 52 samples from two durum and common wheat varieties. Afterwards, an extended and independent sample set (173 samples and five varieties) was used as a confirmatory study to verify the stability and consistency of the models obtained. Putatively identified markers were evaluated applying ROC curves resulting in individual marker AUC >90% both in preliminary and confirmatory study. In addition, digalactosyl diglyceride (DGDG) 36:4 was shown to be an effective marker differentiating between authentic durum wheat and its adulterated admixture down to 3% adulteration level, which is the maximum contamination level allowed by Italian legislation. The results demonstrated that untargeted lipidomics, in conjunction with chemometric tools has a significant potential for screening and detection of wheat fraud.

Structures and formation pathways of compounds responsible for blue-green discoloration of processed garlic were studied in model systems. A procedure was developed for isolation of the color compounds and their tentative identification by high-performance liquid chromatography coupled to a diode array detector and tandem mass spectrometry. It was found that the pigment is a mixture of numerous pyrrole-based purple/blue and yellow species. Experiments with isotope-labeled precursors revealed that two molecules of an amino acid are involved in the formation of each color compound. In the purple/blue species (λmax = 565-600 nm), both amino acid molecules are incorporated into two 3,4-dimethylpyrrole-derived rings linked together by a propenylidine bridge. On the other hand, the yellow compounds (λmax = 420-450 nm) contain only one N-substituted 3,4-dimethylpyrrole ring, to which the second amino acid is bound via a propenylidine side chain.

Tiger nut (Cyperus esculentus L.) is a crop traditionally grown in Valencia Region (Spain) and other temperate and tropical regions in the world, where its tubers are commonly consumed as tiger nut milk (horchata). Because of their nutritive potential and original taste, these products are beginning to spread internationally and, as consequence, analytical procedures to assess nutritional profiles, quality control issues are acquiring increasing relevance. The main objective of this study was to use an advance analytical method and chemometrics tools to determine if the ultra-high temperature (UHT) treatment necessary to extend the shelf life of tiger nut milk would affect the profile of nutrients when compared to fresh product. A cold solvent extraction followed by liquid chromatography coupled with high resolution mass spectrometry (UHPLC-HRMS) was used. Datasets obtained from UHT and fresh tiger nut milk data were analyzed through an untargeted metabolomics approach to compare chemical patterns, highlighting differences in citric acid esters of mono- diglycerides (CITREM) and monoacylglycerol (MAG) used as emulsifiers of UHT products, and a remarkably higher abundance of biotin, phosphatidic acid (PA) and L-arginine in fresh products. These results showed that untargeted metabolomics through high resolution tandem mass spectrometry allowed fine differences between food products to be found, therefore, the nutrient lost caused by UHT treatment was clearly discerned. (C) 2017 Elsevier B.V. All rights reserved.

In order to explore the early detection of mycotoxins in wheat three standardized approaches (Fusarium disease severity, PCR assays for Fusarium spp. identification and mycotoxin quantification) and a novel untargeted metabolomics strategy were jointly assessed. In the first phase of this research, standardized approaches were able to quantify mycotoxins and identify Fusarium spp. Then, an UHPLC-QTOF metabolic fingerprinting method was developed to investigate plant-pathogen cross-talk. At the same time, chemometrics analysis demonstrated to be a powerful tool in order to distinguish low and strong infection levels. Combining these results, the cross-talk plant pathogen related to the early detection of mycotoxins was discovered. As a rapid response to fungal infection an overexpression of phosphatidic acids was discovered. By contrast, when the infection became stronger an increase of oxylipins and diacylglycerols was revealed.

Dietary patterns, microbiome dysbiosis, and gut microbial metabolites (GMMs) have a pivotal role in the homeostasis of intestinal epithelial cells and in disease progression, such as that of colorectal cancer (CRC). Although GMMs and microorganisms have crucial roles in many biological activities, models for deciphering diet–microbiome–host relationships are largely limited to animal models. Thus, intestinal organoids (IOs) have provided unprecedented opportunities for the generation of in vitro platforms with the sufficient level of complexity to model physiological and pathological diet–microbiome–host conditions. Overall, IO responses to GMM metabolites and microorganisms can provide new insights into the mechanisms by which those agents may prevent or trigger diseases, significantly extending our knowledge of diet–microbiome–host interactions.

This study was focused on distinguishing of genetically modified organism (Monsanto 89788 variety) and conventional soybeans by employing high-resolution mass spectrometry (HRMS) techniques for non-target screening of sample extracts. Two hyphenated instrumental platforms represented by (i) ultrahigh-performance liquid chromatography (U-HPLC) coupled to quadrupole/time of flight and (ii) ambient mass spectrometry with direct analysis in real time (DART) ion source-coupled OrbitrapMS were used. The statistical processing of generated data (metabolomic fingerprints) was performed by multivariate data analysis; principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) showed that both employed techniques enabled correct classification of genetically modified and conventional soybeans. In addition, some phosphatidylcholines and sugars were identified as the most significant markers.

In this case study, a food supplement with declared content of sea buckthorn oil was investigated since it was suspected to be adulterated by sunflower oil. Polar and non-polar fractions extracted from oil supplement were analyzed and compared with authentic sea buckthorn and sunflower oils examined in the same way. Three different analytical platforms were used in order to characterize these samples: (i) ambient mass spectrometry consisting of direct analysis in real time (DART) ion source coupled to quadrupole time-of-flight mass spectrometry (Q-TOF MS); (ii) ultra performance liquid chromatography (UHPLC) coupled to quadrupole time-of-flight mass spectrometry, and (iii) high-performance liquid chromatography coupled to diode array detector (HPLC-DAD). The fingerprints of polar and non-polar extracts, regardless they were obtained by DART-HRMS and/or UHPLC-HRMS, were almost identical with those of sunflower “reference” oil. The last of employed techniques, HPLC-DAD, provided complementary information on the occurrence of various visible light-absorbing compounds. While a rich carotenoid profile was shown in sea buckthorn oil, dominated by β-carotene, lutein, and lycopene, the color of the suspected sample was caused only by high concentration of β-carotene. In principle, these techniques proved to be suitable and complementary tools for oil authentication and allowed an oil supplement to be rapidly verified.

The transient receptor potential (TRP) channels superfamily are a large group of proteins that play crucial roles in cellular processes. For example, these cation channels act as sensors in the detection and transduction of stimuli of temperature, small molecules, voltage, pH, and mechanical constrains. Over the past decades, different members of the TRP channels have been identified in the human gastrointestinal (GI) tract playing multiple modulatory roles. Noteworthy, TRPs support critical functions related to the taste perception, mechanosensation, and pain. They also participate in the modulation of motility and secretions of the human gut. Last but not least, altered expression or activity and mutations in the TRP genes are often related to a wide range of disorders of the gut epithelium, including inflammatory bowel disease, fibrosis, visceral hyperalgesia, irritable bowel syndrome, and colorectal cancer. TRP channels could therefore be promising drug targets for the treatment of GI malignancies. This review aims at providing a comprehensive picture of the most recent advances highlighting the expression and function of TRP channels in the GI tract, and secondly, the description of the potential roles of TRPs in relevant disorders is discussed reporting our standpoint on GI tract–TRP channels interactions.

Due to unique phytochemicals contained, Vaccinum berries are known to have a number of positive health effects. In this context, lingonberries (Vaccinium vitis-idaea) are considered to be the most effective, thus finding many uses. Recently, fraud suspicion on lingonberries-based products has been reported, partial or even total replacement by less valued cranberries (Vaccinium macrocarpon) was found. In this study, metabolomic fingerprinting employing instrumental platform consisting of U-HPLC-HRMS/MS was investigated for discrimination between the two Vaccinum berries species. Methanolic extracts of 33 authentic samples from two harvest years were analyzed and chemometric evaluation was performed to identify significant marker compounds, their stability during drying process was assessed, too. The characteristic markers most contributing to berries classification were representatives of polyphenols and phospholipids. Peonidin 3-O-arabinoside and myricetin 3-O-glucoside, not occurring in lingonberries, enabled to discover the presence of cranberries in prepared admixtures down to 1% (w/w).

Our daily food intake provides the nutrients to maintain health. However, in addition to the nutritional values, food can promote health and be beneficial in preventing diseases. Human milk is a unique food source that contains essential nutrients in the right balance and other bioactive factors that make it the ideal food for all healthy term infants. Human milk oligosaccharides (HMOs) play an important role in health, at several levels: acting as prebiotics promoting the growth of beneficial bacterial strains, preventing the growth of pathogenic bacteria in the intestine, and modulating the immune response against bacterial infections. However, despite their biological relevance and the advances made in the analytical field, very few studies have been carried out to better understand HMOs bioactivity mechanisms or to examine human metabolic response to dietary supplementation. This review describes the state-of-the-art of glycomics strategies, recent analytical methods, and future trends for the identification and discovery of bioactive sugars, the known mechanisms of action, and discusses findings of some recent human intervention trials.

Currently, the interest in microalgae as a source of biologically active components exploitable as supplementary ingredients to food/feed or in cosmetics continues to increase. Existing research mainly aims to focus on revealing and recovering the rare, cost competitive components of the algae metabolom. Because these components could be of very different physicochemical character, a universal approach for their isolation and characterization should be developed. This study demonstrates the systematic development of the extraction strategy that represents one of the key challenges in effective algae bioprospecting, which predefines their further industrial application. By using of Trachydiscus minutus as a model microalgae biomass, following procedures were tested and critically evaluated in order to develop the generic procedure for microalgae bioprospecting: (i) various ways of mechanical disintegration of algae cells enabling maximum extraction efficiency, (ii) the use of a wide range of extraction solvents/solvent mixtures suitable for optimal extraction yields of polar, medium-polar, and non-polar compounds, (iii) the use of consecutive extractions as a fractionation approach. Within the study, targeted screening of selected compounds representing broad range of polarities was realized by ultra-high performance liquid chromatography coupled with high resolution tandem mass spectrometric detection (UHPLC–HRMS/MS), to assess the effectiveness of undertaken isolation steps. As a result, simple and high-throughput extraction-fractionation strategy based on consecutive extraction with water–aqueous methanol–hexane/isopropanol was developed. Moreover, to demonstrate the potential of the UHPLC–HRMS/MS for the retrospective non-target screening and compounds identification, the collected mass spectra have been evaluated to characterize the pattern of extracted metabolites. Attention was focused on medium-/non-polar extracts and characterization of lipid species present in the T. minutus algae. Such detailed information on the composition of native (non-hydrolyzed) lipids of this microalga has not been published yet.

Tiger nuts and tiger nut milk are well-known Valencian products, and step-by-step these tubers and the tuber-based beverage are becoming more and more relevant products in international markets. However, the increasing demand and success of Valencian tiger nuts did not allow protected designation of origin (PDO) tuber to supply the domestic and international markets. Therefore, the verification of the geographical origin is highly required. In this research, the main objective was to combine an advance analytical method and chemometrics tools in order to decipher the geographical origin of 45 tiger nut samples from (i) 'Xufa de València' PDO and (ii) African samples. The analytical method, based on solid-liquid extraction followed by ultra-high performance liquid chromatography coupled to high resolution tandem mass spectrometry (UHPLC-HRMS) metabolomics approach, highlighted sensitivity and wide linear dynamic range in order to simultaneously analyse polar and non-polar metabolites. After data processing, a pronounced sample clustering according to the geographical origin was clearly observed using unsupervised models, and supervised models revealed that tiger nuts lipidome was associated with the geographical origin. As a result, African samples highlighted an overexpression of phospholipids, such as phosphatidylethanolamine 34:1, and triacylgricerols crosslinked to environmental stress and alteration of membrane lipid compositions.

Food authenticity becomes a necessity for global food policies, since food placed in the market without fail has to be authentic. It has always been a challenge, since in the past minor components, called also markers, have been mainly monitored by chromatographic methods in order to authenticate the food. Nevertheless, nowadays, advanced analytical methods have allowed food fingerprints to be achieved. At the same time they have been also combined with chemometrics, which uses statistical methods in order to verify food and to provide maximum information by analysing chemical data. These sophisticated methods based on different separation techniques or stand alone have been recently coupled to high-resolution mass spectrometry (HRMS) in order to verify the authenticity of food. The new generation of HRMS detectors have experienced significant advances in resolving power, sensitivity, robustness, extended dynamic range, easier mass calibration and tandem mass capabilities, making HRMS more attractive and useful to the food metabolomics community, therefore becoming a reliable tool for food authenticity. The purpose of this review is to summarise and describe the most recent metabolomics approaches in the area of food metabolomics, and to discuss the strengths and drawbacks of the HRMS analytical platforms combined with chemometrics.

This study evaluated the presence of bacteria and anisakid parasites in 45 samples of raw anchovies in vinegar, a dish widely eaten in Spain, and in 227 samples of cooked fish and cephalopods served in Spanish food service establishments. Our analysis showed that, according to European and Spanish regulation, 14 to 30% of the prepared fish and cephalopod dishes exceeded the maximum allowable level for mesophilic aerobic counts, and 10 to 40% of these samples exceeded the allowable levels for Enterobacteriaceae. None of the studied samples showed evidence of anisakid parasites, Escherichia coli, Staphylococcus aureus, Salmonella, or Listeria monocyto genes. These results indicate that application of hazard analysis and critical control points, food safety training courses, and routine inspections in compliance with current European and Spanish legislation help protect consumer health.

Our daily food intake provides the nutrients to maintain health. However, in addition to the nutritional values, food can promote health and be beneficial in preventing diseases. Human milk is a unique food source that contains essential nutrients in the right balance and other bioactive factors that make it the ideal food for all healthy term infants. Human milk oligosaccharides (HMOs) play an important role in health, at several levels: acting as prebiotics promoting the growth of beneficial bacterial strains, preventing the growth of pathogenic bacteria in the intestine, and modulating the immune response against bacterial infections. However, despite their biological relevance and the advances made in the analytical field, very few studies have been carried out to better understand HMOs bioactivity mechanisms or to examine human metabolic response to dietary supplementation. This review describes the state-of-the-art of glycomics strategies, recent analytical methods, and future trends for the identification and discovery of bioactive sugars, the known mechanisms of action, and discusses findings of some recent human intervention trials.

Humans can be exposed to mycotoxins through the food chain. Mycotoxins are mainly found as contaminants in food and could be subsequently excreted via biological fluids such as urine or human breast milk in native or metabolised form. Since breast milk is usually supposed as the only food for new-borns, the occurrence of mycotoxins in thirty-five human milk samples was evaluated by a newly developed method based on QuEChERS extraction and UHPLC-HRMS detection. The method described here allows the detection of target mycotoxins in order to determine the quality of this initial feeding. The method has been fully validated, with recoveries ranging from 64% to 93% and relative standard deviations (RSD, %) being lower than 20%. Using the method described, non-metabolised mycotoxins such as ZEA, NEO, NIV, ENA, ENA1, ENB, ENB1 and metabolites, such as ZEA metabolites, HT-2, DOM and T-2 triol were detected in human milk samples. Results obtained help to estimate the exposure of mothers and infants to mycotoxins. Moreover, to the best of our knowledge, this is the first work describing the simultaneous detection, quantification and screening of mycotoxins and their metabolites in human mature milk.

A survey in African snacks was carried out in order to evaluate the intake of 23 mycotoxins. The African snack samples were purchased from street vendors within Lagos metropolis (Nigeria) and evaluated for the presence of 23 mycotoxins using a modified QuEChERS procedure coupled with liquid chromatography-triple quadrupole linear ion trap mass spectrometer. The snacks included akara, baked coconut, coconut candy, donkwa, groundnut cake (kulikuli), lafun, milk curd (wara), fresh and dried tiger-nuts, and yam flour. Only three mycotoxins were detected in 23.8% of the studied snacks, and at concentrations ranging from 6 to 54 μg kg−1. The concentrations of aflatoxin B1 (AFB1) and AFB2 reached 23 μg kg−1 and 3 μg kg−1, respectively. Moreover a sample of baked coconut contained α-zearalenol (α-ZOL), which was up to 54 μg kg−1 in coconut candy. As considers prevalence, aflatoxins and α-ZOL were not detected in lafun and groundnut-based snacks (donkwa and kulikuli), whereas each of the three mycotoxins contaminated 12.5% (1/8) of the coconut-based samples. This is the first report of α-ZOL in cassava and coconut, and their products. AFB1 and total aflatoxins (TAFs) concentrations exceeded the maximum allowable limit recommended by National Agency for Food and Drug Administration and Control Nigeria (NAFDAC) in one sample of baked coconut (AFB1 = 23 μg kg−1 and TAFs = 26 μg kg−1) and donkwa (AFB1 = 19 μg kg−1 and TAFs = 21 μg kg−1).

The quantification of short-chain and medium-chain fatty acids is becoming more and more relevant in fecal and plasma samples due to their biological impact, which has been associated with colon rectal cancer and fiber consumption. For these reasons, a fast, cost-effective, and reproducible analytical method is highly required. In this research, a gas chromatography–mass spectrometry method based on full scan and multiple reaction monitoring (MRM) acquisition modes were optimized and validated for the analysis of short-chain and medium-chain fatty acids in three biological samples: human fecal water, fecal fermentation supernatants, and human plasma. Several extraction solvents (acidified water, diethyl ether, dichloromethane, ethyl acetate, and methyl tert-butyl ether (MTBE) were further evaluated, demonstrating that the latter was clearly the most suitable solvent with recoveries from 75.4 to 124.4% and coefficient of variations lower than 20%. The applicability of the GC–MS method was tested, for instance, acetic acid was quantified by using samples of plasma and feces from healthy donors at mean values of 66.9 μM and 24.5 mM, respectively. The optimized protocol could successfully find applications within multi-compartment human studies. In parallel, a second pilot experiment on fecal fermentation supernatants indicated that the proposed protocol is suitable to follow the formation of SCFAs during in vitro fermentation by the human gut microbiota. In summary, the present work provided an improved GC–MS method for precise and accurate quantification of SCFAs and MCFAs in human feces and plasma.

A fast and effective method for quantitative determination of fumonisins B1 and B2 (FB1 and FB2) in different cereal-based products was developed. Fumonisins were extracted combining a solid liquid extraction (SLE) pre-treatment and immunoaffinity columns (IACs), which were applied for an effective clean-up. Consecutively, high pressure liquid chromatography (HPLC) coupled to hybrid triple quadrupole-linear ion trap mass spectrometer (QTrap®) was used for the separation, detection and quantification of targeted mycotoxins. The analytical method showed acceptable recoveries ranged from 89.7% to 99.1%, as well as a relative standard deviation (%, RSD) lower than 12%. A total of 1250 samples from France, Germany and Spain were analysed. The incidence of fumonisin B1 and B2 in organic samples were 11.4% and 11.3%, respectively. By contrast, the occurrence of conventional samples was 3.6% of FB1 and 3.5% of FB2. The highest values were obtained in Spanish samples of flour, as an example toasted corn, called gofio that was contaminated with 1201.7 and 1010.5 μg kg(-1) of FB1 and FB2, respectively.

The present study aims to focus on the bioprospecting of marine macroalgae of Turbinaria species, plenteous biomass of the world ocean. Three types of solvents, i.e. water, methanol:water (80:20, v/v) and hexane:isopropanol (50:50, v/v), were used for extraction. Both the biological activity, as well as the pattern of present chemicals were characterized. For the cell proliferation assay, the human embryonic kidney 293 cells, cervix / breast / pancreatic adenocarcinoma, and osteosarcoma cells were used. For the antioxidant activity determination, both intracellular assay with human embryonic kidney and cervix adenocarcinoma cells, as well as the biochemical DPPH test, were employed. To complete the information about macroalgae composition, organic compounds were characterized by the liquid chromatography coupled with high resolution tandem mass spectrometry. Attention was concentrated mainly on the lipidomic profile characterization. In spite the fact that any significant antiproliferative effect was not observed for cancer cells, both the Turbinaria species were shown to be good protectors against the oxidative stress of the non-cancer cells. Most of the antioxidants were determined in the hexane-isopropanolic extract. As regards the lipids identified, most of them belonged to the triacylglycerols followed by sphingomyelins, diacylglycerols, and polar (lyso)phospholipids. Additionally to fatty acids with 14, 16 and 18 carbons, also those with odd carbons number were frequently present. This article is protected by copyright. All rights reserved.

A previous developed matrix solid-phase dispersion (MSPD) extraction method was applied for the routine analysis of aflatoxins (AFs), ochratoxin A (OTA), fumonisins (FB 1 and FB 2 ), beauvericin (BEA), nivalenol (NIV), deoxynivalenol (DON), the toxin T-2 (T-2), toxin HT-2 (HT-2), diacetoxyscirpenol (DAS) and zearalenone (ZEN) in tiger-nuts by liquid chromatography–triple-quadrupole linear ion trap (HPLC–QTRAP ® ). The extraction solid support used was C 18 , while the elution solvent was acetonitrile/methanol (50/50, v/v) 1 mM ammonium formate. Using matrix-matched calibration, recoveries and repeatabilities were in the range 67–89% and 2–11% relative standard deviation (RSD), respectively. The method was applied to determine the occurrence of the fourteen selected mycotoxins in a total of 83 tiger-nut samples purchased from different local markets of Valencian Community (Spain) during (March–June 2010 and March–May 2011). DON, OTA, AFs and BEA were detected in 26 samples of the total number of samples.

This article describes the validation of an analytical method for the detection of 21 mycotoxins in baby food. The analytical method is based on the simultaneous extraction of selected mycotoxins by matrix solid-phase dispersion (MSPD) followed by liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS) using a hybrid triple quadrupole-linear ion trap mass spectrometer (QTRAP®). Information Dependent Acquisition (IDA), an extra confirmation tool for samples that contain the selected mycotoxins, was used. The matrix effects were evaluated, and the corrections for the matrix effects were performed using two calibration approaches: external matrix-matched calibration and internal standard calibration. Matrix-matched calibration was ultimately used for accurate quantification, and the recoveries obtained were generally higher than 70%. The analytical method was applied to the analysis of 35 samples of commercial baby foods. No sample exceeded the maximum limit (ML) fixed by the European Union for these mycotoxins in baby food. However, this survey highlighted the occurrence of mycotoxins in cereal-based infant foods.

This study was carried out to evaluate the microbiological quality of orange juice obtained from squeezing machines in foodservice establishments. The samples included fresh squeezed orange juice and juice which is maintained in metal jugs until consumption. According to the European Commission Regulation (No. 2073/2005 and No. 1441/2007) and Spanish microbiological criteria (No. 3484/2000), 12% and 43% of the total examined lots exceed the adopted limits of mesophilic aerobic counts and Enterobacteriaceae, respectively. Possibly, this contamination is caused by incorrect handling of oranges and juices and also by inadequate cleaning and sanitization of squeezing machine and metal jugs. Furthermore, 0.5 and 1% of all the examined lots were positive for the presence of Salmonella spp. and Staphylococcus aureus , respectively. However all lots were negative for Escherichia coli and Listeria monocytogenes . These results emphasize the need for applying and maintaining good hygienic practices in the restaurants.

A simple and rapid method effective for quantitative determination of deoxynivalenol (DON), T-2 toxin (T-2), HT-2 toxin (HT-2), zearalenone (ZEN), ochratoxin A (OTA), aflatoxins (AFs) B(1), B(2), G(1) and G(2) and fumonisins FB(1) and FB(2) in urine was developed. The urine was diluted with phosphate buffer solution (PBS) and thoroughly mixed. For clean-up and extraction, the mixture was loaded on a MYCO 6in1 IAC. Hybrid triple quadrupole-linear ion trap mass spectrometer (QTrap) was used for the detection. Extra tools for confirmation of selected mycotoxins in positive samples, Information Dependent Acquisition (IDA) experiments, were also developed. The use of immunoaffinity columns followed by the LC-MS/MS analysis showed acceptable average recoveries between 83% and 116% and reached acceptable precision values (relative standard deviation (RSD) ≤ 14%). In a pilot study with 27 volunteers, OTA, DON and AFG(2) were detected. However, this study needs to be extended in order to understand the relation between the mycotoxins intake and mycotoxin levels in human urine.

A method based on Matrix Solid-Phase Dispersion (MSPD) has been developed for the determination of 5 mycotoxins (ochratoxin A and aflatoxins B and G) in different cereals. Several dispersants, eluents and ratios were tested during the optimization of the process in order to obtain the best results. Finally, samples were blended with C 18 and the mycotoxins were extracted with acetonitrile. Regarding to matrix effects, the results clearly demonstrated the necessity to use a matrix-matched calibration to validate the method. Analyses were performed by liquid chromatography–triple quadrupole-tandem mass spectrometry (LC–QqQ-MS/MS). The recoveries of the extraction process ranged from 64% to 91% with relative standard deviation lower than 19% in all cases, when samples were fortified at two different concentrations levels. Limits of detection ranged from 0.3 ng g −1 for aflatoxins to 0.8 ng g −1 for OTA and the limits of quantification ranged from 1 ng g −1 for aflatoxins to 2 ng g −1 for OTA, which were below the limits of mycotoxins set by European Union in the matrices evaluated. Application of the method to the analysis of several samples purchased in local supermarkets revealed aflatoxins and OTA levels.

Hulled, or ancient, wheats were the earliest domesticated wheats by mankind and the ancestors of current wheats. Their cultivation drastically decreased during the 1960s; however, the increasing demand for a healthy and equilibrated diet led to rediscovering these grains. Our aim was to use a non-targeted metabolomic approach to discriminate and characterize similarities and differences between ancient Triticum varieties. For this purpose, 77 hulled wheat samples from three different varieties were collected: Garfagnana T. turgidum var. dicoccum L. (emmer), ID331 T. monococcum L. (einkorn) and Rouquin T. spelta L. (spelt). The ultra high performance liquid chromatography coupled to high resolution tandem mass spectrometry (UHPLC-QTOF) metabolomics approach highlighted a pronounced sample clustering according to the wheat variety, with an excellent predictability (Q2), for all the models built. Fifteen metabolites were tentatively identified based on accurate masses, isotopic pattern, and product ion spectra. Among these, alkylresorcinols (ARs) were found to be significantly higher in spelt and emmer, showing different homologue composition. Furthermore, phosphatidylcholines (PC) and lysophosphatidylcholines (lysoPC) levels were higher in einkorn variety. The results obtained in this study confirmed the importance of ARs as markers to distinguish between Triticum species and revealed their values as cultivar markers, being not affected by the environmental influences.

Saffron is one of the oldest and most expensive spices, which is often target of fraudulent activities. In this research, a new strategy of saffron authentication based on metabolic fingerprinting was developed. In the first phase, a solid liquid extraction procedure was optimized, the main aim was to isolate as maximal representation of small molecules contained in saffron as possible. In the second step, a detection method based on liquid chromatography coupled with high-resolution mass spectrometry was developed. Initially, principal component analysis (PCA) revealed clear differences between saffron cultivated and packaged in Spain, protected designation of origin (PDO), and saffron packaged in Spain of unknown origin, labeled Spanish saffron. Afterwards, orthogonal partial least square discriminant analysis (OPLS-DA) was favorably used to discriminate between Spanish saffron. The tentative identification of markers showed glycerophospholipids and their oxidized lipids were significant markers according to their origin.

Ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (MS) and an alternative technology represented by direct analysis in real time coupled with quadrupole time-of-flight MS were investigated for metabolic fingerprinting of 343 red and white wine samples. Direct injection of pure wine and an extraction procedure optimized for isolation of polyphenols were used to compare different analytical and data handling strategies. After data processing and data pretreatment, principal component analysis was initially used to explore the data structure. Initially, the unsupervised models revealed a notable clustering according to the grape varieties, and therefore supervised orthogonal partial least squares discriminant analysis models were created and validated for separation of red and white wines according to the grape variety. The validated orthogonal partial least squares discriminant analysis models based on data (ions) recorded in positive ionization mode were able to classify correctly 95 % of samples. In parallel, authentication parameters, such as origin and vintage, were evaluated, and they are discussed. A tentative identification of markers was performed using accurate mass measurement of MS and MS/MS spectra, different software packages and different online libraries. In this way, different flavonol glucosides and polyphenols were identified as wine markers according to the grape varieties.

This article describes the application of an analytical method for the detection of 25 mycotoxins in wheat grain based on simultaneous extraction using matrix solid-phase dispersion (MSPD) followed by liquid chromatography coupled to tandem mass spectrometry, a hybrid triple quadrupole-linear ion trap mass spectrometer (QTrapÒ). Information Dependent Acquisition (IDA), an extra confirmation tool for samples that contain the target mycotoxins, was used. The analysis of 40 Syrian and 46 Italian wheat grain sam- ples interestingly showed that Syrian samples were mainly contaminated with ochratoxin A and aflatox- ins, whereas Italian samples with deoxynivalenol and 15-acetyldeoxynivalenol. Emerging Fusarium mycotoxins were predominant in Italian samples compared to the Syrian. Among the analysed samples, only one was found containing zeralenone with level above the maximum European recommended con- centration (100 ppb). These results confirm that climatic differences between Syria and Italy, both in Mediterranean basin, play a key role in the diversity of fungal genera and mycotoxins in wheat grains.

In this work, an existent solid-phase extraction (SPE) procedure was used to study the occurrence of mycotoxins in different European beers. HPLC-QqQ-MS/MS and ultra high resolution mass analyser have been optimized for the analysis of 18 mycotoxins: the methods were validated according to the EU Commission Decision 2002/657/EC guidelines. In this sense, matrix-matched calibration was performed for each type of beer, obtaining an effective quantification. The recoveries ranged from 63 to 91% and repeatability and reproducibility expressed as relative standard deviation (RSD%) were lower than 17%. On one hand, HPLC-LTQ-Orbitrap ® was used for unambiguous identification of target mycotoxins, as well as for screening non-target mycotoxins, such as enniatins, fusaproliferin and deoxynivalenol-3-glucoside. On the other hand, the quantification was carried out using HPLC-QqQ-MS/MS instrument. At the end, Ochratoxin A, fumonisins, HT-2 and T-2 toxins were detected and quantified in European beers. Moreover, pale lager beer showed higher mycotoxin incidence than the other studied beers.

This study was carried out to evaluate the microbiological quality of plant-based foods obtained from foodservice establishments. The samples included cereals, legumes, fruits and vegetables. According to the European Commission Regulation (No. 2073/2005 and No. 1441/2007) and Spanish microbiological criteria (No. 3484/2000), vegetables were the plant-based dishes where more samples exceed the adopted limits of mesophilic aerobic counts and Enterobacteriaceae. Furthermore, Staphylococcus aureus and Escherichia coli were also found in several vegetable dishes. E. coli and Salmonella spp. were detected in 6.6% and 0.7% of lettuce samples, respectively. However, all the samples were negative for Listeria monocytogenes. Salads ingredients are eaten as fresh vegetables without cooking processes and lettuce was one of the most contaminated samples. As ready-to-eat salads which have been previously cleaned, this means that contamination might be caused by incorrect cleaning practices or inadequate hygienic handling. These results emphasize the necessity to apply and to maintain good hygienic practices in the restaurants.

The history of proteomics dates back to the discovery of two-dimensional gels in the 1970s, which provided the first feasible way of displaying hundreds or thousands of proteins on a single gel. Despite mass spectrometry being restricted for a long time to small and thermostable compounds, the development in the late 1980s of two techniques for the routine and general formation of molecular ions of intact biomolecules changed this situation and mass spectrometry has become an indispensable tool for proteomics research. The aim of this chapter is to review the major types of MS instruments used in proteomics analysis and to discuss strategies for the analysis of whole proteins and peptides obtained after degradation. Finally, major applications of mass spectrometry-based proteomics in food safety are summarized.

The effectiveness of four extraction methods (modified QuEChERS, matrix solid-phase dispersion (MSPD), solid-liquid extraction (SLE) and solid-phase extraction (SPE) clean-up) were evaluated for simultaneous determination of 32 mycotoxins produced by the genus Fusarium, Claviceps, Aspergillus, Penicillium and Alternaria in barley by ultra high pressure liquid chromatography coupled to ultra-high resolution mass spectrometry (UHPLC-Orbitrap(®) MS). The efficiency and efficacy of extraction methods were evaluated and compared in number of extracted mycotoxins and obtained recoveries. From the one point of view, QuEChERS procedure was fast and easy, as well as it was able to successfully extract all selected mycotoxins. On the other hand, SLE method, MSPD and SPE clean-up method did not extract adequately all selected mycotoxins and recoveries were not suitable enough. Thereby, method employing QuEChERS extraction connected with UHPLC-Orbitrap(®) MS was developed to quantify 32 mycotoxins in barley within this study. Analytical method was validated and recoveries ranged from 72% to 101% for selected mycotoxins with only one exception nivalenol (NIV) and deoxynivalenol-3-glucoside (D3G), which were lower than 67%. Relative standard deviations (RSD) were lower than 17.4% for all target mycotoxins. The lowest calibration levels (LCLs) ranged from 1 to 100 μg/kg. Validated method was finally used for monitoring mycotoxins in a total of 15 Czech barley samples, when only Fusarium toxins representatives were detected in 53% of samples and the mycotoxins with the highest incidence were enniatins.