Development of a liquid chromatography–tandem mass spectrometry for the determination of enniatins in biological samples from Wistar rats

Cytoprotective effect of resveratrol diastereomers in CHO-K1 cells exposed to beauvericin

Beauvericin (BEA) causes cytotoxicity, lipid peroxidation and reactive oxygen species in CHO-K1 cells. Resveratrol (RSV) is a polyphenol with multiple biological properties, including antioxidant effects. RSV has two forms: trans and cis. The aims of this study were to determine the cytoprotective effect of trans-RSV and diastereomers mixtures (50:50 trans/cis-RSV and 70:30 trans/cis-RSV) incubated alone and in combination with BEA in ovarian (CHO-K1) cells. The results demonstrated that cell viability increases (from 9% to 77%) when they were exposed to low concentration of RSV. Moreover, when the cells were pre-treated with RSV and then exposed to BEA, a cytoprotective effect (from 25% to…

Analysis of thiabendazole and procymidone in fruits and vegetables by capillary electrophoresis–electrospray mass spectrometry

Abstract A capillary electrophoresis–mass spectrometry method for determining procymidone and thiabendazole in apples, grapes, oranges, pears, strawberries and tomatoes is described. Separation is achieved using a buffer of formic acid–ammonium formate at pH 3.5 with 2% of methanol. Fungicide residues present in the sample are preconcentrated by both solid-phase extraction and injection of large sample volumes into the capillary by a stacking technique, to obtain lower detection limits. Ionization is performed at atmospheric pressure in an electrospray type source and detection is carried out using positive ionization and selected ion monitoring modes. The quantitation limits are 0.005 and …

Determination of several aminoglycoside and macrolide antibiotics in animal tissues by liquid chromatography-electrospray ionization-tandem mass spectrometry

Risk assessment associated to the intake of the emerging Fusarium mycotoxins BEA, ENs and FUS present in infant formula of Spanish origin

Abstract Forty-five samples of Spanish follow-up infant formula with different chemical compositions were analyzed determining the emerging Fusarium mycotoxins beauvericin (BEA), enniatins (ENs) (A, A 1 , B, B 1 ), and fusaproliferin (FUS). The samples were extracted three times with ethyl acetate using an Ultra-turrax homogenizer. Mycotoxins were identified and quantified using a liquid chromatography (LC) coupled to a diode array detector (DAD). Results showed that the percentage of the samples contaminated with ENs and FUS were 46.6 and 20.0% respectively, whereas all analyzed samples were free of BEA. The ENs A and B were detected only in one sample with 149.6 and 39.4 mg/kg respectivel…

Oxidative stress, glutathione, and gene expression as key indicators in SH-SY5Y cells exposed to zearalenone metabolites and beauvericin.

The co-presence of mycotoxins from fungi of the genus Fusarium is a common fact in raw food and food products, as trace levels of them or their metabolites can be detected, unless safety practices during manufacturing are carried out. Zearalenone (ZEA), its metabolites α-zearalenol (α-ZEL) and β-zearalenol (β-ZEL) and, beauvericin (BEA) are co/present in cereals, fruits or their products which is a mixture that consumer are exposed and never evaluated in neuronal cells. In this study the role of oxidative stress and intracellular defense systems was assessed by evaluating reactive oxygen species (ROS) generation and glutathione (GSH) ratio activity in a human neuroblastoma cell line, SH-SY5…

Evaluation by HPLC-UV of Polar Pesticides in Rice Fields

Nuts and dried fruits: Natural occurrence of emerging Fusarium mycotoxins

Abstract The occurrence of enniatins (ENs) and beauvericin (BEA) in nuts, dried fruits and dates available in Valencia (Spain) was surveyed in this study. To do this, seventy-four samples of nuts, dried fruits and dates were analyzed for the determination of mycotoxin contamination. Mycotoxins were identified and quantified using an ultrasonic-C18 extraction and LC–MS/MS with a triple quadrupole (QqQ) mass analyzer. The frequencies of contamination of nuts, shell, dried fruits and dates were 50%, 80%, 35.7% and 83.3%, respectively. Enniatin A (ENA) was the most predominant EN found in nuts (45.2%) while ENB was the most common EN found in dates (58.3%). The analytical results of the shell s…

Mycotoxin Analysis of Human Urine by LC-MS/MS: A Comparative Extraction Study

The lower mycotoxin levels detected in urine make the development of sensitive and accurate analytical methods essential. Three extraction methods, namely salting-out liquid–liquid extraction (SALLE), miniQuEChERS (quick, easy, cheap, effective, rugged, and safe), and dispersive liquid–liquid microextraction (DLLME), were evaluated and compared based on analytical parameters for the quantitative LC-MS/MS measurement of 11 mycotoxins (AFB1, AFB2, AFG1, AFG2, OTA, ZEA, BEA, EN A, EN B, EN A1 and EN B1) in human urine. DLLME was selected as the most appropriate methodology, as it produced better validation results for recovery (79–113%), reproducibility (RSDs < 12%), and repeatability (RSDs…

HPLC-UV/Vis-APCI-MS/MS Determination of Major Carotenoids and Their Bioaccessibility from "Delica" (Cucurbita maxina) and "Violina" (Cucurbita moschata) Pumpkins as Food Traceability Markers

Carotenoids are a widespread group of fat-soluble pigments, and their major nutritional importance comes from their pro-vitamin A activity and their antioxidant capacity. In this study, two different pumpkin cultivars (Cucurbita maxima, also named `Delica&rsquo

Congener profile, occurrence and estimated dietary intake of dioxins and dioxin-like PCBs in foods marketed in the Region of Valencia (Spain).

During 2006-2008, a monitoring program was conducted on 29 target compounds, including PCDD/Fs and dl-PCBs, comprising 150 randomly collected individual food samples marketed in the Region of Valencia, Spain, grouped into 8 categories (vegetables, cereals, fats and oils, eggs, milk and dairy products, fish products, meat and meat products and fish oil). For PCDD/Fs, the highest frequency of detection corresponds to 1,2,3,4,6,7,8-HpCDD, OCDD, 2,3,4,7,8-PeCDF; and PCBs 118, 105 and 156 were the more frequent dl-PCBs. The food groups presenting higher contamination, expressed as toxic equivalents (WHO-TEQs), were fish oil (6.38 pg WHO-TEQ g(-1)fat), fish (1.21 pg WHO-TEQ g(-1)w.w.) and milk an…

Toxicity of mycotoxins in vivo on vertebrate organisms: A review.

Mycotoxins are considered to be a major risk factor affecting human and animal health as they are one of the most dangerous contaminants of food and feed. This review aims to compile the research developed up to date on the toxicological effects that mycotoxins can induce on human health, through the examination of a selected number of studies in vivo. AFB1 shows to be currently the most studied mycotoxin in vivo, followed by DON, ZEA and OTA. Scarce data was found for FBs, PAT, CIT, AOH and Fusarium emerging mycotoxins. The majority of them concerned the investigation of immunotoxicity, whereas the rest consisted in the study of genotoxicity, oxidative stress, hepatotoxicity, cytotoxicity,…

Bioaccessibility and decomposition of cylindrospermopsin in vegetables matrices after the application of an in vitro digestion model.

Research on the human exposure to Cylindrospermopsin (CYN) via consumption of contaminated food is of great interest for risk assessment purposes. The aim of this work is to evaluate for the first time the CYN bioaccessibility in contaminated vegetables (uncooked lettuce and spinach, and boiled spinach) after an in vitro digestion model, including the salivar, gastric and duodenal phases and, colonic fermentation under lactic acid bacteria. The results obtained showed that the digestion processes are able to diminish CYN levels, mainly in the colonic phase, especially in combination with the boiling treatment, decreasing CYN levels in a significant way. Moreover, the potential decomposition…

Toxicity evaluation of individual and mixed enniatins using an in vitro method with CHO-K1 cells

Enniatins (ENs) A, A1, B and B1 are produced by Fusarium species. They are known as emerging fusario- toxins, and can cause outbreaks in both humans and animals. ENs elicits a wide range of different biolog- ical properties and toxicological effects, and their co-occurrence may enhance the extent of these hazards. As the potential toxins reach in vitro cells in the same way as they would in vivo, cytotoxicity was studied with CHO-K1, which is considered one of the most sensitive cell lines for preliminary screen- ing of cytotoxicity studies. In this study, individual cytotoxic effects of ENs were evaluated by MTT assay after exposing ENs to CHO-K1 cells for 24, 48, and 72 h. The IC50 values…

Neurotoxicity of zearalenone’s metabolites and beauvericin mycotoxins via apoptosis and cell cycle disruption

Cell cycle progression and programmed cell death are imposed by pathological stimuli of extrinsic or intrinsic including the exposure to neurotoxins, oxidative stress and DNA damage. All can cause abrupt or delayed cell death, inactivate normal cell survival or cell death networks. Nevertheless, the mechanisms of the neuronal cell death are unresolved. One of the cell deaths triggers which have been wildly studied, correspond to mycotoxins produced by Fusarium species, which have been demonstrated cytotoxicity and neurotoxicity through impairing cell proliferation, gene expression and induction of oxidative stress. The aim of present study was to analyze the cell cycle progression and cell …

Determination of quinolone residues in chicken and fish by capillary electrophoresis-mass spectrometry

A specific pressure-assisted CE-MS method is described for the analysis of five quinolone residues. MS using a single quadrupole is compared with multiple-stage MS using a quadrupole IT (QIT-MS(n)). The procedure involves a common sample preparation by SPE on disposable cartridges. The most suitable electrolyte is 60 mM (NH(4))(2)CO(3) at pH 9.2. Single quadrupole does not provide enough fragmentation to confirm identities according to the current legislation. However, QIT-MS(n) achieves selective fragmentation. Using this method, danofloxacin, enrofloxacin, flumequine, ofloxacin, and pipemidic acid are analyzed in fortified samples of chicken and fish. Recoveries at levels of 50 ng/g were …

Surveillance of pesticide residues in fruits from Valencia during twenty months (2004/05)

Abstract The aim of this study was to investigate the pesticide residues in market fruits (oranges, tangerines, nectarines, peaches and khakis) from one Valencian Cooperative (Spain) and to conduct a monitoring of 32 organophosphorous, organonitrogen and organohalogenated pesticides and nine dithiocarbamate fungicides (DTCFs) usually applied on cultures of this area. Extracts were obtained by an official procedure for routine analysis based on ethyl acetate extraction. Residues of pesticides were determined by gas chromatography with nitrogen phosphorous detector (NPD), electron-capture detector (ECD) and mass spectrometry (MS) detectors. Mean recoveries obtained at fortification levels bet…

Solid-phase extraction on C18 in the trace determination of selected polychlorinated biphenyls in milk.

The utility of solid-phase extraction with octadecylsilica for determining fifteen polychlorinated biphenyl (PCB) congeners from milk samples was examined. Recoveries higher than 80% and relative standard deviations better than 10% were obtained for PCBs from different kinds of milk (whole, skimmed, 2%, powdered, breast and evaporated). A comparison with other procedures was made. The described method provides better detection limits than those attainable with the liquid-liquid extractions currently used as standard methods, when capillary gas-liquid chromatography is used for the final determination. A study of the separation was also performed using six different fused-silica capillary co…

Analysis of pyridoquinoline derivatives by liquid chromatography/atmospheric pressure chemical ionization mass spectrometry

A method using liquid chromatography/atmospheric pressure chemical ionization mass spectrometry (LC/APCI-MS) has been developed for the characterization and determination of pyridoquinoline derivatives 4,6-bis(dimethylaminoethylamino)-2,8,10-trimethylpyrido[3,2-g]quinoline, 4,6-bis(dimethylaminoethoxy)-2,8,10-trimethylpyrido[3,2-g]quinoline and 4,6-bis[(dimethylaminoethyl)thio]-2,8,10-trimethylpyrido[3,2-g] quinoline, all with potential antitumor properties. LC separation was performed on a conventional C18 column using a binary mobile phase composed of acetonitrile and 50 mM aqueous ammonium formate at pH 3. The APCI mass spectra obtained showed that proton addition giving [M + H]+ was the…

Occurrence of fumonisins B1, B2 and B3 in maize-products commercialized in Italy and Spain

Mitigation of enniatins in edible fish tissues by thermal processes and identification of degradation products.

Emerging mycotoxins, such as enniatins and beauvericin, are common contaminants in vegetal matrices, but recently, the occurrence of mycotoxins in foodstuffs from animal origin has been also reported as they can be present in edible tissues of animals fed with contaminated feedstuffs. Sea bass, sea bream, Atlantic salmon and rainbow trout from aquaculture analyzed in the present survey showed contamination by emerging Fusarium mycotoxins enniatins (ENs). ENs were extracted from raw and cooked fish with acetonitrile and analyzed by Liquid Chromatography coupled to Mass Spectrometry. In this study, the stability of ENs was evaluated during food processing by the application of different cooki…

Comparative assessment of three extraction procedures for determination of emerging Fusarium mycotoxins in pasta by LC–MS/MS

Abstract A new rapid, sensitive, reproducible and reliable method was developed for the quantitative determination of enniatins A, A1, B and B1, beauvericin and fusaproliferin in dry and fresh pasta by liquid chromatography-triple quadrupole-tandem mass spectrometry. A comparative study of different rapid and economical extraction procedures was performed for the extraction of these mycotoxins in pasta. For this purpose, three different approaches were studied during the extraction step (Ultra-Turrax, ultrasonic bath and microwave). Optimal extraction conditions were reached using Ultra-Turrax with acetonitrile for 3 min without purification step. The chromatographic separation of the six m…

Risk assessment and monitoring programme of nitrates through vegetables in the Region of Valencia (Spain)

This study was carried out to determine current levels of nitrate in vegetables marketed in the Region of Valencia (Spain) and to estimate the toxicological risk associated with their intake. A total of 533 samples of seven vegetable species were studied. Nitrate levels were derived from the Valencia Region monitoring programme carried out from 2009 to 2013 and food consumption levels were taken from the first Valencia Food Consumption Survey, conducted in 2010. The exposure was estimated using a probabilistic approach and two scenarios were assumed for left-censored data: the lower-bound scenario, in which unquantified results (below the limit of quantification) were set to zero and the up…

Analysis of carbamate pesticides and their metabolites in water by solid phase extraction and liquid chromatography: A review

Carbamates are an important, broad class of pesticides that are used extensively as insecticides, fungicides, and herbicides. Sensitive, economical, fast, and environmental friendly procedures are constantly developed to investigate their residues in water samples. The state of the art in methods based on solid phase extraction (SPE) and liquid chromatographic determination are examined here. SPE is presently the most extended method for preconcentration of carbamate pesticide residues and their transformation products from water samples. Advantages and limitations of alkyl bonded-silica, and polymeric sorbents, carbon, and mixed-phases in off-line and on-line procedures are discussed. Beca…

Multiresidue analysis of pesticides in honey by quechers and gas chromatography mass spectrometry in tandem

Multiple-stage mass spectrometric analysis of six pesticides in oranges by liquid chromatography–atmospheric pressure chemical ionization–ion trap mass spectrometry

Abstract Six pesticides were determined by liquid chromatography (LC) with positive ion (PI) atmospheric pressure chemical ionization quadrupole ion-trap tandem mass spectrometry (APCI–MS–MS). Ion fragmentation was studied by MS, MS 2 and MS 3 . Fragmentation of the pesticides produced ions formed by various losses from the side-chains and through heterocyclic ring opening, but without any common fragmentation pathway. Multiple reaction monitoring (MRM) of MS, MS 2 and MS 3 was used to identify and quantify the pesticides. The samples were extracted with ethyl acetate and dried over anhydrous sodium sulfate. Comparison of the three MS modes showed that MS 3 is slightly less sensitive but mu…

The soluble dietary fiber inulin can influence the bioaccessibility of enniatins.

Enniatins (ENs) are bioactive compounds produced by the secondary metabolism of several Fusarium strains and are known to have various biological activities, such as acting as enzyme inhibitors, antifungal antibacterial agents, and immunomodulatory substances. This study investigated the bioaccessibility of the ENs in wheat crispy breads produced with three different inulin concentrations (1, 5 and 10%). The mean bioaccessibility data of the four ENs (A, A(1), B and B(1)) ranged from 68.67% to 84.67 in the experiments carried out without inulin, whereas the data ranged from 51.00 to 74.00% in the experiments carried out with the wheat crispy bread produced with 5 and 10% of the inulin.

Incidence of enterotoxigenic staphylococci and their toxins in foods

Of 504 food samples collected from cafeterias, 19 (3.8%) yielded strains of enterotoxigenic staphylococci, and 10 (52.6%), 4 (21.1%), 3 (15.8%), and 2 (10.5%) of these strains produced enterotoxins C (SEC), D (SED), B (SEB), and A (SEA), respectively. Moreover, SEA, SEB, and SEC were isolated from three hamburger samples. Of 181 food samples collected from four restaurants before the implementation of the hazard analysis and critical control point (HACCP) system, 7 (3.9%) were found to contain enterotoxigenic strains, and SED, SEC, and SEA were produced by 4 (57.1%), 2 (28.6%), and 1 (14.3%) of these strains, respectively. One meatball sample with SEC was detected in a restaurant. After the…

Pressurized liquid extraction followed by liquid chromatography–mass spectrometry for determination of zearalenone in cereal flours

Abstract A method for determination of zearalenone in cereal flour has been developed applying pressurized liquid extraction (PLE) using methanol/acetonitrile (50:50 v/v) as the extraction solvent. The extracted samples were analyzed with liquid chromatography coupled to mass spectrometry (LC–MS) with an electro spray ionisation interface (ESI). The method was validated as a quantitative confirmatory method according to the Eu Commission Decision 2002/657/EC. Recoveries of the extraction step data were satisfactory with values higher then 70%. Quantification limits (LOQ) were 5 μg/kg for ESI (+) and 1 μg/kg for ESI (−). Twenty one flour samples produced in different countries were extracted…

Determination of aminoglycoside and macrolide antibiotics in meat by pressurized liquid extraction and LC-ESI-MS

A simple method for the simultaneous determination of dihydrostreptomycin, spectinomycin, spiramycin, streptomycin, tilmicosin, and tylosin in meat has been developed using pressurized liquid extraction and LC-triple quadrupole MS (LC-ESI-MS/MS). The pressurized liquid extraction operational parameters were optimized and no protein precipitating and fat removing steps were required. A gradient HPLC separation was developed with ion-pair mobile phases consisting of aqueous 1 mM heptafluorobutyric acid water and methanol. Protonated molecules were used as precursor ions for CID. Data acquisition under MS/MS was achieved by applying multiple reaction monitoring of three fragment ion transition…

Micronucleus induction and cell cycle alterations produced by deoxynivalenol and its acetylated derivatives in individual and combined exposure on HepG2 cells.

Mycotoxins are produced by a number of fungal genera spp as e.g. Aspergillus, Penicillium, Alternaria, Fusarium and Claviceps. 3-Acetyl-Deoxynivalenol (3-A-DON) and 15-Acetyl-Deoxynivalenol (15-ADON) which are produced by Fusarium, chemically belong to trichothecenes and occur in significant amounts as modified forms of deoxynivalenol (DON) in various cereal crops and processed grains. This study aims to determine the cytotoxicity, cell cycle and genotoxicity of the mycotoxins DON, 3-A-DON and 15-A-DON on HepG2 cells. Cytotoxic concentration range studied was from 100 to 3.1 μM for DON and 12.5 to 0.04 μM for 3-A-DON and 15-A-DON by the Neutral Red (NR) assay, over 24, 48 and 72 h. Potentia…

Evolution of emerging Fusarium mycotoxins contents throughout the shelf-life period of food

which is cooked by steaming. It has been historically eaten in North African countries but nowadays its consumption is widely extended all around the world. As a cereal-based food, semolina is highly susceptible to contamination by mycotoxin-producing fungi. The presented procedure involves a modified QuEChERSbased extraction of 24 mycotoxins (15-ADON, 3-ADON, AFLAB1, AFLAB2, AFLAG1, AFLAG2, BEA, DON, DAS, ENA, ENA1, ENB, ENB1, FB1, FB2, FB3, FUSX, HT-2, NEO, NIV, OTA, STG, T2, ZEA) produced by Aspergillus, Penicillum and Fusarium fungi. The validation was performed by analyzing recovery samples at three different spiked concentrations with four replicates (n=4) at each concentration. Recov…

Determination of 5-nitrofurylacrylic acid in wines by high-performance liquid chromatography

Capillary zone electrophoresis for the determination of thiabendazole, prochloraz and procymidone in grapes

Capillary zone electrophoresis with UV detection was applied to the simultaneous determination of thiabendazole, prochloraz and procymidone in grapes. Electrolyte conditions such as pH, composition and concentration of the buffer, addition of organic solvent and working voltage were checked to obtain a high-performance separation of the three fungicides (by measurement of separation efficiency and resolution). The most critical parameter was the pH of the running buffer. The best separation was achieved in 4 mM phosphate solution at pH 3.5. The repeatability of the migration times, expressed as RSD, was <0.44%. The three peaks were completely resolved with a separation efficiency up to 100 …

Reactive oxygen species induced by beauvericin, patulin and zearalenone in CHO-K1 cells

The cytotoxic effects of mycotoxins, induction of reactive oxygen species (ROS) and generation of lipid peroxidation products in CHO-K1 cells were determined as function of increasing time of exposure and concentrations of beauvericin (BEA), patulin (PAT) and zearalenone (ZEA). The end points were evaluated after 24h of exposure, by the tetrazolium salt (MTT) and neutral red (NR) assays. The IC(50) values obtained on the MTT and NR assays ranged from 0.69 to 79.40 microM and 4.40 to 108.76 microM, respectively. To determine the intracellular production of ROS, the intensity of fluorescence emitted from the probe H(2)-DCFDA was measured. The relative intensity of fluorescence from cells incu…

Toxicological interactions between the mycotoxins beauvericin, deoxynivalenol and T-2 toxin in CHO-K1 cells in vitro.

Abstract Beauvericin (BEA), deoxynivalenol (DON) and T-2 toxin (T-2) are important food-borne mycotoxins that have been implicated in human health. In this study, the acute toxicity of individual and combined mycotoxins (BEA, DON and T-2) were tested in immortalized hamster ovarian cells (CHO-K1) at 24, 48 and 72 h of exposure, by the tetrazolium salt (MTT) and neutral red (NR) assays. The IC50 values obtained for all mycotoxins by the MTT and NR assays ranged from 0.017 to 12.08 μM and from 0.042 to 17.22 μM, respectively. Both, individual and combined mycotoxins demonstrated a significant cytotoxic effect in CHO-K1 cells in a dose-dependent manner. When mycotoxins were assayed individuall…

Risk assessment of beauvericin, enniatins and fusaproliferin present in follow-up infant formula by in vitro evaluation of the duodenal and colonic bioaccessibility

Abstract In this study, 72 samples of follow-up infant formula of Spanish origin were analyzed for the presence of the mycotoxins beauvericin (BEA), enniatins (ENs) (A, A1, B, B1), and fusaproliferin (FUS). The samples analyzed were extracted three times with ethyl acetate and then the mycotoxins were identified and quantified using a liquid chromatography (LC) coupled to a diode array detector (DAD). The positive samples analyzed in this study were digested through a simulated gastrointestinal digestion model, which permit to simulate the physiological condition of the human gastrointestinal tract (duodenal and colonic compartments) in order to assess the bioaccessibility of the bioactive …

Simultaneous determination of mycotoxin in commercial coffee

Abstract Mycotoxins are secondary metabolites produced by filamentous fungi that usually contaminate food products. Coffee is a natural product susceptible to mycotoxin contamination. The present study evaluates the presence of nivalenol, deoxynivalenol, T-2 and HT-2 Toxin, diacetoxyscirpenol, aflatoxin B 1 , aflatoxin B 2 , aflatoxin G 1 , aflatoxin G 2 , fumonisin B 1 , fumonisin B 2 , ochratoxin A, zearalenone, enniatin A, enniatin A 1 , enniatin B, enniatin B 1 , and beauvericin in coffee samples, using liquid chromatography tandem mass spectrometry (LC-MS/MS). The results show that zearalenone was not present in any sample. In the positive samples the contents of fumonisins ranged from…

Pressurized liquid extraction combined with capillary electrophoresis–mass spectrometry as an improved methodology for the determination of sulfonamide residues in meat

A new analytical method, based on capillary electrophoresis and tandem mass spectrometry (CE-MS2), is proposed and validated for the identification and simultaneous quantification of 12 sulfonamides (SAs) in pork meat. The studied SAs include sulfathiazole, sulfadiazine, sulfamethoxypyridazine, sulfaguanidine, sulfanilamide, sulfadimethoxyne, sulfapyridine, sulfachloropyridazine, sulfisoxazole, sulfasalazine, sulfabenzamide and sulfadimidine. Different parameters (i.e. separation buffer, sheath liquid, electrospray conditions) were optimized to obtain an adequate CE separation and high MS sensitivity. MS2 experiments using an ion trap as analyzer, operating in the selected reaction monitori…

Comparison of microextraction procedures to determine pesticides in oranges by liquid chromatography-mass spectrometry.

A liquid chromatographic-mass spectrometric method has been developed for the determination of bitertanol, carbendazim, fenthion, flusilazole, hexythiazox, imidacloprid, methidathion, methiocarb, pyriproxyfen and trichlorfon. Two procedures, based on stir bar sorptive extraction (SBSE) and matrix solid-phase dispersion (MSPD), have been evaluated for the extraction of these compounds in oranges. Their respective advantages and disadvantages are also discussed. The recoveries obtained by MSPD ranged from 47 to 96% and the relative standard deviations (RSDs) ranged from 1 to 15%, whereas with the SBSE method the recoveries were between 8 and 84% and the RSDs between 4 and 16%. Although, the l…

Determination of polycyclic aromatic hydrocarbons in atmospheric particulate matter of Valencia city

Polycyclic aromatic hydrocarbons (PAHs) were determined in atmospheric particulate matter in 11 sites of the Valencia area and at several times during the year. Sample analysis was carried out by ultrasonic acetonitrile extraction followed by reverse phase HPLC separation and fluorescence detection. The maximum concentration of total PAH developed in winter and spring. Mean values per sampling site varied from 0.193 to 1.668 μg/m3 of filtered air. Environmental noise and temperature were determined at those same 11 sites and correlated with PAH levels.

Solid-phase extraction in multi-residue pesticide analysis of water

The determination of pesticides in water is fundamental to the solution of environmental problems as natural waters are usually contaminated with a large number of pesticides. The selection of an isolation and/or concentration technique depends largely on the class of pesticides to be determined. It is often necessary to determine simultaneously a wide variety of compounds in a water sample. Application of solid-phase extraction techniques offers a solution. The mechanisms of solid-phase extraction, types of sorbents and their application to multi-residue pesticide analysis are reviewed.

Formation of fumonisin B(1)-glucose reaction product, in vitro cytotoxicity, and lipid peroxidation on kidney cells.

Fumonisin B(1) (FB(1)) content in corn products decreases during the heating process in foods containing reducing sugars, mainly because of the formation of N-(carboxymethyl)fumonisin B(1). In this study, a rapid method has been developed for the determination of both compounds in corn products using a high-speed blender, Ultra-Turrax, for solvent extraction and liquid chromatography-tandem mass spectrometry. The kinetics of FB(1) degradation and the formation of the Maillard adduct were studied in a model system constituted by corn bread spiked with FB(1) and heated at 160, 180, and 200 degrees C for 3, 6, 10, 15, and 20 min. FB(1) decreased from 0.96 to 0.3 mg/kg and N-(carboxymethyl)fumo…

Influence of different soluble dietary fibers on the bioaccessibility of the minor Fusarium mycotoxin beauvericin.

Abstract Beauvericin (BEA) is a bioactive compound produced by the secondary metabolism of several Fusarium strains and is known to have various biological activities. This study investigated the bioaccessibility of the BEA tested in concentrations of 5 and 25 mg/L, in a model solution and in wheat crispy breads elaborated with different natural binding compounds as the soluble alimentary dietary fibers β-1,3 glucan, chitosan low molecular weight (L.M.W.), chitosan medium molecular weight (M.M.W.), fructooligosaccharides (FOS), galattomannan, inulin and pectin, added at concentrations of 1% and 5%. The bioaccessibility was determinated by employing a simulated gastrointestinal digestion tha…

Comparison of solid-phase microextraction and stir bar sorptive extraction for determining six organophosphorus insecticides in honey by liquid chromatography-mass spectrometry.

Abstract Two approaches based on sorptive extraction, solid-phase microextraction (SPME) and stir bar sorptive extraction (SBSE), in combination with liquid chromatography (LC)–atmospheric pressure chemical ionization mass spectrometry (MS) have been assayed for analyzing chlorpyriphos methyl, diazinon, fonofos, phenthoate, phosalone, and pirimiphos ethyl in honey. In both, SPME and SBSE, enrichment was performed using a poly(dimethylsiloxane) coating. Significant parameters affecting sorption process such as sample volume, sorption and desorption times, ionic strength, elution solvent, and dilution (water/honey) proportion were optimized and discussed. Performance of both methods has been …

DNA damage and perturbation on cell cycle and mitochondrial membrane potential by alternariol mycotoxin in Caco-2 cells

Fluorescent probes on HepG2 cells treated with emergent Fusarium mycotoxins

Determination of urea-derived pesticides in fruits and vegetables by solid-phase preconcentration and capillary electrophoresis

A multiresidue analytical method based on solid-phase extraction (SPE) enrichment combined with capillary electrophoresis (CE), using micellar electrokinetic capillary chromatography (MEKC), was developed to determine ten substituted urea pesticides in orange and tomato samples. Several factors such as pH, composition and concentration of the buffer, concentration of surfactant, addition of organic solvent, and working voltage were optimized to obtain the best compound separation in the shortest time. Separation can be achieved in 7 min using a micellar aqueous pH 9 buffer composed of 4 mM borate and 35 mM sodium dodecyl sulfate. After an SPE procedure, which provided a 10-fold enrichment, …

Nanoelectrospray with ion-trap mass spectrometry for the determination of beta-casomorphins in derived milk products.

Beta-casomorphins (b-CMs) are bioactive peptides derived from casein with opioid agonist effects similar to morphine. The use of electrospray (ESI) with quadrupole ion-trap mass spectrometry (QIT-MS) for these compounds in two matrices, cheese and milk, was examined. It was compared to a liquid chromatography (LC) coupled to mass spectrometry (LC-MS), and a "soft" ionisation technique, NanoMate, with selected ion monitoring (SIM), which are unreliable for the determination of trace casomorphins in derived milk products. b-CM mass fragmentation pathways were done for the four most common b-CMs: beta-casomorphin (1-5) bovine (b-CM-5), beta-casomorphin (1-7) bovine (b-CM-7), [D-Ala2, D-Pro4,Ty…

Mycotoxins and their consequences in aquaculture: A review

Fish consumption has been increasing worldwide, mainly due to the availability, access and price in relation to other kinds of meat consumption, such as beef, pork, and poultry. Consequently, some concerns begin to emerge, primarily regarding the quality of fish available in the market. Residues could be present in any product of animal origin causing economic losses and putting into a risk human and animal health. Food contamination by mycotoxins is a risk to human and animal health, and it is responsible for significant economic losses. It's very difficult to prove that a disease is a mycotoxicosis, and even when mycotoxins are detected, it is not easy to show that they are the etiologica…

Inhibition of aflatoxin B1, B2, G1 and G2 production by Aspergillus parasiticus in nuts using yellow and oriental mustard flours

Abstract Aflatoxins (AFs) are naturally occurring mycotoxin compounds produced by several species of Aspergillus, as Aspergillus flavus and Aspergillus parasiticus and are mutagenic, teratogenic, and carcinogenic compounds that have been implicated as causative agents in human hepatic and extra hepatic carcinogenesis. In this study, the reduction of the AFs present in dried fruits (peanut, cashew, walnut, almond, hazelnut and pistachio), produced by A. parasiticus CECT 2681, by isothiocyanates (ITCs) generated by the enzymatic hydrolysis of the glucosinolates (GLCs) present in oriental and yellow mustard flours was evaluated. The AFs reduction activity through ITCs application in dried frui…

Antibacterial activity of the enniatins A, A1, B, B1 produced by fusarium tricinctum in liquid culture, and cytotoxicity effects on Caco-2 cells

Validation of a multi-residue UHPLC-HRMS method for antibiotics screening in milk, fresh cheese, and whey

Abstract The aim of this study was to develop a UHPLC–HRMS methodology as a suitable tool for the screening of antibiotics in milk, fresh cheese, and whey. Compounds were analysed using an Orbitrap Exactive™ analyser working at a resolving power of 50,000 FWHM in full scan, both in positive and negative electrospray ionization mode, and a high mass accuracy (

Analysis of trichothecenes in laboratory rat feed by gas chromatography-tandem mass spectrometry

A method for the determination of seven trichothecenes, neosolaniol (NEO), diacetoxyscirpenol (DAS), deoxynivalenol (DON), nivalenol (NIV), fusarenon-X (FUS-X), 3-acetyldeoxynivalenol (3-ADON) and 15-acetyldeoxynivalenol (15-ADON), in laboratory rat feed by GC-MS/MS was developed. Sample extraction and purification was performed by an acidified mixture of acetonitrile/water (80-20% v/v). Limits of quantitation (LOQs) were between 1 and 10 μg kg(-1) for all studied trichothecenes. Eight concentration levels between the LOQ and 100 × LOQ were used for the calibration curves. Matrix-matched calibration was used for quantitation purposes to compensate the detector signal enhancement obtained fo…

Application of capillary electrophoresis-mass spectrometry for determining organic food contaminants and residues.

Food contamination continues to be a serious problem around the world. Surveillance of chemical contaminants in foods is important not only for public health but also because of the negative economic impact of contamination. From the analytical perspective, analysis of contaminants in food is an extremely challenging area. There is a wide variety of questions, ranging from the quantification of extremely low levels of individual components to the detailed assessment and evaluation of the analytical technique possibilities. This review considers the applications of CE coupled to MS detection (CE-MS) for the analysis of organic contaminants in food. Analytical information on sample concentrat…

Current trends in solid-phase-based extraction techniques for the determination of pesticides in food and environment

Solid-phase extraction (SPE) procedures for pesticide residues in food and environment are reviewed and discussed. The use of these procedures, which include several approaches such as: matrix solid-phase dispersion (MSPD), solid-phase micro-extraction (SPME) and stir-bar sorptive extraction (SBSE), represents an opportunity to reduce analysis time, solvent consumption, and overall cost. SPE techniques differ from solvent extraction depending on the interactions between a sorbent and the pesticide. This interaction may be specific for a particular pesticide, as in the interaction with an immunosorbent, or non-specific, as in the way a number of different pesticides are adsorbed on apolar or…

Study of enzymatic activity in human neuroblastoma cells SH-SY5Y exposed to zearalenone's derivates and beauvericin.

Abstract Beauvericin (BEA), α-zearalenol (α-ZEL) and β-zearalenol (β-ZEL), are produced by several Fusarium species that contaminate cereal grains. These mycotoxins can cause cytotoxicity and neurotoxicity in various cell lines and they are also capable of produce oxidative stress at molecular level. However, mammalian cells are equipped with a protective endogenous antioxidant system formed by no-enzymatic antioxidant and enzymatic protective systems such as glutathione peroxidase (GPx), glutathione S-transferase (GST), catalase (CAT) and superoxide dismutase (SOD). The aim of this study was evaluating the effects of α-ZEL, β-ZEL and BEA, on enzymatic GPx, GST, CAT and SOD activity in huma…

SOLID-PHASE EXTRACTION OF PESTICIDES FROM WATER SAMPLES

Transcriptional Changes after Enniatins A, A1, B and B1 Ingestion in Rat Stomach, Liver, Kidney and Lower Intestine

Enniatins (ENs) are depsipeptide mycotoxins produced by Fusarium fungi. They are known for their capacity to modulate cell membrane permeability and disruption of ionic gradients, affecting cell homeostasis and initiating oxidative stress mechanisms. The effect of the acute toxicity of ENs A, A1, B and B1 at two different concentrations after 8 h of exposure was analysed in Wistar rats by a transcriptional approach. The following key mitochondrial and nuclear codified genes related to the electron transport chain were considered for gene expression analysis in stomach, liver, kidney and lower intestine by quantitative Real-Time PCR: mitochondrially encoded NADH dehydrogenase 1 (MT-ND1), mit…

Effects of four carbamate compounds on antioxidant parameters

Abstract The effect of four carbamates, aldicarb and its metabolites (aldicarb sulfone and aldicarb sulfoxide) and propoxur on glutathione content and the activity of the enzymes involved in the sulfur-redox cycle in the mammalian cellular model CHO-K1 cells after 24-h exposure were determined. Carbamate exposure resulted in a depletion of intracellular reduced glutathione (GSH) content, no change was observed in oxidized glutathione (GSSG) and a decrease in GSH/GSSG ratio was detected. After carbamates exposition a GSH/GSSG decreases in ranged from 12.44% to 21.35% of control was observed. Depletion of GSH levels was accompanied by the induction of glutathione reductase (GR) after 24 h exp…

Determination of mycotoxins in multicereal flour by matrix solid phase dispersion and LC–MS/MS

Dietary exposure to trace elements and health risk assessment in the Region of Valencia (Spain). A Total Diet Study

ABSTRACTDietary exposure of the Valencian region population to lead, cadmium, inorganic arsenic (iAs), chromium, copper, tin and methylmercury (meHg) was assessed in a total diet study carried out in the region of Valencia in 2010–11. A total of 8100 food samples were collected and analysed. Occurrence data were combined with consumption data to estimate dietary exposure in adults (> 15 years of age) and young children (6–15 years of age). The estimated intake was calculated by a probabilistic approach. Average intake levels (optimistic scenario) for lead, iAs, chromium and tin were 0.21, 0.08, 1.79 and 1.87 µg kg−1 bw day−1 respectively; for Cd and meHg average intake levels were 0.77 and …

Quantitative determination of octylphenol, nonylphenol, alkylphenol ethoxylates and alcohol ethoxylates by pressurized liquid extraction and liquid chromatography-mass spectrometry in soils treated with sewage sludges.

6 páginas, 1 figura, 2 tablas.

In vitro mechanisms of Beauvericin toxicity: A review.

Beauvericin (BEA) is a mycotoxin produced by many species of fungus Fusarium and by Beauveria bassiana; BEA is a natural contaminant of cereals and cereals based products and possesses a wide variety of biological properties. The mechanism of action seems to be related to its ionophoric activity, that increases ion permeability in biological membranes. As a consequence, BEA causes cytotoxicity in several cell lines and is capable to produce oxidative stress at molecular level. Moreover, BEA is genotoxic (produces DNA fragmentation, chromosomal aberrations and micronucleus) and causes apoptosis with the involvement of mitochondrial pathway. However, several antioxidant mechanisms protect cel…

Multimycotoxin LC-MS/MS Analysis in Tea Beverages after Dispersive Liquid-Liquid Microextraction (DLLME).

The aim of the present study was to develop a multimycotoxin liquid chromatography tandem mass spectrometry (LC-MS/MS) method with a dispersive liquid-liquid microextraction procedure (DLLME) for the analysis of AFs, 3aDON, 15aDON, NIV, HT-2, T-2, ZEA, OTA, ENNs, and BEA in tea beverages and to evaluate their mycotoxin contents. The proposed method was characterized in terms of linearity, limits of detection (LODs), limits of quantification (LOQs), recoveries, repeatability (intraday precision), reproducibility (interday precision), and matrix effects to check suitability. The results show LODs in the range of 0.05-10 μg/L, LOQs in the range of 0.2-33 μg/L, and recoveries in the range of 65…

Toxicological Assessment of Recombinant Xylanase X22 in Wine

Toxicological evaluation of xylanase X(22) from Aspergillus nidulans expressed in a wine yeast strain was carried out. The safety of the X(22) intake was assessed by digestibility, bioinformatic, and mouse short-term repeated dosing studies, although X(22) shows resistance to proteolytic degradation in the gastrointestinal system, is a minority protein component (<0.5 10(-)(6) %) of the produced wine, and shows no significant amino acid sequence homology to any known food allergens. The 4-week oral toxicity study was performed in Swiss mice at a dose level of 0.01, 0.1, or 1 mg/kg/day (these dosages correlate to 8, 80, and 800 times, respectively, the enzyme amount contained in 250 mL of wi…

Solid-phase extraction of quaternary ammonium herbicides

This paper highlights recent advances in the solid-phase extraction (SPE) of quaternary ammonium herbicides in water, soil, plant and biological samples. After a brief introduction summarizing the properties of quaternary ammonium herbicides and the difficulties involved in measuring them, attention is paid primarily to solid supports used for isolation and concentration, pre-treatments required for the different matrices, and eluents applied for quantitative desorption of these analytes. The determination techniques used after SPE and applications of the proposed SPE methodology are also briefly discussed.

Simultaneous determination of imidacloprid, carbendazim, methiocarb and hexythiazox in peaches and nectarines by liquid chromatography–mass spectrometry

Abstract A liquid chromatographic (LC) atmospheric pressure chemical ionization mass spectrometric (MS) method is described for determining imidacloprid, carbendazim, methiocarb and hexythiazox in peaches and nectarines. The samples were extracted with ethyl acetate and anhydrous sodium sulfate. Recoveries yield for spiking samples were ranged from 64±9% (R.S.D) for carbendazim to 108±14% (R.S.D.) for hexythiazox at the concentration of 0.1 mg kg−1 (n=5). The correlation coefficients were greater than 0.998 over the range between 0.02 and 2 mg kg−1. The limits of quantitation (LOQ) were 0.02 mg kg−1 for all the pesticides. The applicability of the method to detect and quantify imidacloprid,…

Effects of soyasaponin I and soyasaponins-rich extract on the Alternariol-induced cytotoxicity on Caco-2 cells

Abstract Alternariol (AOH) is a mycotoxin produced by Alternaria spp. Soyasaponin I (Ss-I) is present naturally in legumes, and it has antioxidant properties. Cytotoxic and genotoxic effects of AOH have been demonstrated previously in vitro. In the present study, the cytotoxicity of AOH, Ss-I, and soyasaponins-rich extract from lentils was investigated; as well as, the cytoprotective effects of Ss-I and lentil extracts against AOH induced-cytotoxicity on Caco-2 cells. Cytotoxicity was carried out using MTT and PC assays (AOH: 3.125–100 µM, Ss-I: 3.125–50 µM, and lentil extracts: 1:0–1:32) during 24 h of exposure. Only AOH showed cytotoxic effect. The reduction in cell proliferation ranged f…

Cytoprotection assessment against mycotoxins on HepG2 cells by extracts from Allium sativum L

Abstract Cytoprotection effects of Allium sativum L garlic extract from a local garlic ecotype from Ferrara (Italy) on hepatocarcinoma cells, HepG2 cells, is presented in this study. This garlic type is known as Voghiera garlic and has been characterized as PDO (Protected designation of Origin) product. Voghiera garlic extract (VGE) was evaluated against beauvericin (BEA) and two zearalenone (ZEA) metabolites (α-zearalenol (α-ZEL) and β-zearalenol (β-ZEL))-induced cytotoxicity on HepG2 cells by the MTT (3–4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay, over 24 h and 48 h. Direct treatment, simultaneous treatment and pre-treatment strategies at the dilution 1:16–1:00 for VG…

In vitro cytotoxicity of patulin, deoxynivalenol, nivalenol and zearalenone on CHO-K1 cells

Transcriptional study after Beauvericin and Enniatin B combined exposure in Jurkat T cells

Simultaneous mycotoxins toxicity is complex and non-predictable based on their individual toxicities. Beauvericin and Enniatins are emerging mycotoxins highly co-occurrent in food and feed, and their cytotoxicity has been reported in several human cell lines. RNA-seq studies of individual exposure in Jurkat cells demonstrated human genome perturbation mainly affecting mitochondrial pathways, however, both mycotoxins showed differences between their toxic responses. This study investigates the transcriptional effects of combined exposure to Beauvericin and Enniatin B (1:1) (0.1, 0.5, 1.5 μM; 24 h) in Jurkat cells by qPCR on 30 selected target genes (10 mitochondrial, 20 nuclear). Gene expres…

Pressurised liquid extraction and capillary electrophoresis–mass spectrometry for the analysis of pesticide residues in fruits from Valencian markets, Spain

Abstract A procedure based in capillary electrophoresis–mass spectrometry (CE–MS) for the analysis of seven pesticides (flutolanil, simazine, haloxyfop, acifluorfen, dinoseb, picloram, and ioxynil) in four Mediterranean summer fruits with high water content (peaches, melon, watermelon, and apricot) is developed. Several conditions were studied for the optimisation of both, the separation and the electrospray (ESI) connection. The best results were obtained using 35 mM ammonium formiate (pH 9.7) as separation buffer, 20 °C as capillary temperature and 23 kV as applied voltage in an uncoated fused-silica capillary with 50 cm total length, 25 cm thermostated length, 25 cm at room temperature l…

Cellular redox status by alternariol in caco-2 cells

A preliminary study in Wistar rats with enniatin : A contaminated feed

A 28-day repeated dose preliminary assay, using enniatin A naturally contaminated feed through microbial fermentation by a Fusarium tricinctum strain, was carried out employing two months-old female Wistar rats as in vivo experimental model. In order to simulate a physiological test of a toxic compound naturally produced by fungi, five treated animals were fed during twenty-eight days with fermented feed. As control group, five rats were fed with standard feed. At the 28th day, blood samples were collected for biochemical analysis and the gastrointestinal tract, liver and kidneys were removed from each rat for enniatin A detection and quantitation. Digesta were collected from stomach, duode…

Natural occurrence of emerging Fusarium mycotoxins in feed and fish from aquaculture.

A new analytical method for the simultaneous determination of enniatins (ENs) and beauvericin (BEA) in fish feed and fish tissues by liquid chromatography coupled to mass spectrometry with linear ion trap (LC-MS/MS-LIT) was developed. Results showed that the developed method is precise and sensitive. The presence of emerging Fusarium mycotoxins, ENs and BEA, was determined in samples of aquaculture fish and feed for farmed fish, showing that all feed samples analyzed were contaminated with mycotoxins, with 100% coexistence. In aquacultured fish samples, the highest incidence was found in edible muscle and liver. As for the exposure assessment calculated, it was found that average consumer i…

Mechanisms of beauvericin toxicity and antioxidant cellular defense

Beauvericin (BEA) is a secondary metabolite produced by many species of fungus Fusarium. This study determines the injury (cell viability, cell proliferation, mitochondrial membrane potential, cell death and DNA damage) and the intracellular defense mechanisms (catalase and superoxide dismutase) in Chinese Hamster ovary (CHO-K1) cells after BEA exposure. The results obtained in this study demonstrated that BEA induces cytotoxicity in a dose- and time-dependent manner in CHO-K1 cells. Moreover, disruption in mitochondrial enzymatic activity and cell proliferation has been observed after BEA exposure, which can lead or be consequence of cell death. BEA inhibits cell proliferation by arresting…

Determination of fungicide residues in fruits and vegetables by liquid chromatography–atmospheric pressure chemical ionization mass spectrometry

Abstract A liquid chromatography (LC) method for the quantitative determination of five fungicide residues (dichloran, flutriafol, o -phenylphenol, prochloraz and tolclofos methyl) in oranges, lemons, bananas, peppers, chards and onions is described. The residues were extracted by matrix solid-phase dispersion (MSPD) using C 8 . Quantitative analysis was performed by isocratic LC coupled to quadrupole mass spectrometer using atmospheric pressure chemical ionization in the negative ionization mode. The limit of quantification was 0.01 mg kg −1 for flutriafol, o -phenylphenol and dichloran, and 0.1 mg kg −1 for prochloraz and tolclofos methyl. The MSPD method is also suitable for LC–UV analys…

Comparative cytotoxicity study of enniatins A, A1, A2, B, B1, B4 and J3 on Caco-2 cells, Hep-G2 and HT-29

Abstract Enniatins (ENs) are ionophoric, phytotoxic, antihelminthic, and antibiotic compounds of hexadepsipeptidic structure produced by several strains of Fusarium spp. The cytotoxicity effect of the ENs A, A 1 , A 2 , B, B 1 , B 4 and J 3 was compared on three tumor cell lines, the human epithelial colorectal adenocarcinoma (Caco-2), the human colon carcinoma (HT-29), and the human liver carcinoma (Hep-G2). The endpoint evaluated was the mitochondrial integrity by using the MTT assays, after 24 and 48 h of incubation. The IC 50 value for EN A 2 on Caco-2 cells, after 24 h exposure, was 18.7 ± 4.5 μM and decrease to 2.6 ± 0.7 μM at 48 h of incubation. However, ENs A, A 1 , B 1 and B 4 exer…

Mycotoxin Incidence in Some Fish Products: QuEChERS Methodology and Liquid Chromatography Linear Ion Trap Tandem Mass Spectrometry Approach

The inclusion of vegetal raw materials in feed for fish farming has increased the risk of mycotoxin occurrence in feed, as well as in edible tissues from fish fed with contaminated feed, due to the carry-over to muscle portions. Therefore, the objective of this study was to evaluate the occurrence of 15 mycotoxins in processed fish products, which are commonly consumed, such as smoked salmon and trout, different types of sushi, and gula substitutes. A QuEChERS method was employed to perform the mycotoxin extraction from fish samples. For mycotoxin identification and quantitation, the selected technique was the liquid chromatography-tandem mass spectrometry linear ion trap (LC-MS/MS-LIT). Sm…

Influence of pro- and prebiotics on gastric, duodenal and colonic bioaccessibility of the mycotoxin beauvericin

Abstract Beauvericin (BEA) is a bioactive compound produced by the secondary metabolism of several Fusarium strains and known to have various biological activities. This study investigates the influence of several dietary fibers (galactomanan, glucomannan, citrus fiber, bamboo fiber, carrot fiber, pie fiber, β-glucan, xilan, and cellulose) and probiotic strains ( Lactobacillus animalis , Lb. casei , Lb. casei , Lb. plantarum , Lb. rhuminis , Lb. casei casei , Bifidobacterium breve , Bf. Adolescents , Bf. bifidum , Corynebacterium vitaeruminis , Streptococcus faecalis , Eubacterium crispatus , and Saccharomyces cerevisiae ) on the minor Fusarium mycotoxin BEA bioaccessibility employing a mod…

On-line preconcentration strategies for analyzing pesticides in fruits and vegetables by micellar electrokinetic chromatography.

Five pesticides (fludioxonil, procymidone, pyriproxyfen, dinoseb and carbendazim) were separated in reversed migration micellar electrokinetic chromatography (RM-MEKC) using 20 mmol l(-1) phosphate buffer at pH 2.3, containing 25 mmol l(-1) sodium dodecylsulfate and 10% methanol. Three on-line concentration strategies, sweeping (SW), normal stacking with reversed migration and a water plug (SRW) and stacking with reverse migration and removal of sample matrix using polarity switching (SRMM), were compared. About 10-, 30- and 50-fold increases in detection sensitivity, compared with standard hydrodynamic injection (5 s at 0.5 psi), were observed with SW, SRW and SRMM, respectively. Limits of…

Oxidative damages in cho-k1 cells treated with beauvericin

Binary and tertiary combinations of 3-ADON, 15-ADON and AOH mycotoxins on HepG2 cells: Evaluation of cytotoxic effects and detection of metabolite products

Detecting residues of urea and carbamate pesticides

Multimycotoxin analysis in water and fish plasma by liquid chromatography-tandem mass spectrometry

High performance liquid chromatography-mass spectrometry was used for the determination of 15 mycotoxins in water and fish plasma samples, including aflatoxins, fumonisins, ochratoxin A, sterigmatocistin, fusarenon-X and emerging Fusarium mycotoxins. In this work, dispersive liquid-liquid microextraction (DLLME) was assessed as a sample treatment for the simultaneous extraction of mycotoxins. Results showed differences in recovery assays when different extraction solvents were employed. Ethyl acetate showed better recoveries for the major part of mycotoxins analyzed, except for aflatoxins B2, G1 and G2, which showed better recoveries when employing chloroform as extractant solvent. Fumonisi…

Clean-up and confirmation procedures for gas chromatographic determination of pesticide residues in contaminated waters. Part I

The effects of sulphuric acid, potassium hydroxide and chromic acid on eleven organochlorine and ten organophosphorus pesticides were investigated. The treatments destroy some pesticides totally or partially, leave others unaltered and have a clean-up effect. These reactions can be used to confirm the presence of an identified pesticide. The clean-up extracts in the environmental samples facilitate the identification of the organochlorine and organophosphorus pesticides and their quantitative analysis. The results obtained for contaminated surface waters show the usefulness of these methods for multi-residue capillary gas chromatography without the need for other additional separative chrom…

Emerging Fusarium mycotoxins in organic and conventional pasta collected in Spain

Abstract One of the main sources of emerging Fusarium mycotoxins in human nutrition is the cereals and cereal products. In this study, an analytical method to determine enniatins A, A1, B and B1 (ENs), beauvericin (BEA) and fusaproliferin (FUS) based on Ultra-Turrax extraction followed by liquid chromatography coupled to triple quadrupole mass spectrometer detector (MS/MS QqQ), was applied for the analysis of pasta. For this purpose, 114 commercial samples of pasta were acquired from supermarkets located in Valencia. The results showed higher frequencies of contamination in organic pasta than in conventional pasta, while the concentration levels were variable for both types of pasta. In pos…

Beauvericin-induced cytotoxicity via ROS production and mitochondrial damage in Caco-2 cells.

The cytotoxicity of beauvericin (BEA) on human colon adenocarcinoma (Caco-2) cells was studied as a function of time. Moreover, the oxidative damage and cell death endpoints were monitored after 24, 48 and 72 h. After BEA exposure, the IC₅₀ values ranged from 1.9 ± 0.7 to 20.6 ± 6.9 μM. A decrease in reduced glutathione (GSH; 31%) levels, as well as an increase in oxidized glutathione (GSSG, 20%) was observed. In the presence of BEA, reactive oxygen species (ROS) level was highly increased at an early stage with the highest production of 2.0-fold higher than the control that was observed at 120 min. BEA induced cell death by mitochondria-dependent apoptotic process with loss of the mitochon…

Determination of macrolide and lincosamide antibiotics by pressurised liquid extraction and liquid chromatography-tandem mass spectrometry in meat and milk

Abstract A total of 155 samples, including meat and milk, were analyzed using a method developed for determining five macrolide antibiotics: tilmicosin (TILM), tylosin (TS), spiramycin (SPM), erythromycin (ERY), and tulathromycin (TULA) and two lincosamide antibiotics: pirlimycin (PIRL) and lincomycin (LM). The method was performed by pressurised liquid extraction (PLE) coupled to LC–MS/MS using electrospray ionization in positive ion mode. 2.5 mL of milk or 2.5 g of meat were dispersed in ethylenediaminetetraacetic acid sodium salt 2-hydrate (EDTA) and sea sand. The PLE conditions as: solvent, temperature, pressure, static time, and cell size were optimized. The following analytical parame…

Study of the potential toxicity of enniatins A, A(1), B, B(1) by evaluation of duodenal and colonic bioavailability applying an in vitro method by Caco-2 cells.

Abstract The bioavailability of the minor Fusarium mycotoxins enniatins (ENs) utilizing an in vitro method which allows the simulation of the small and large intestine tracts has been studied. This method, based on the application of the Caco-2 cells grown alone or in symbiosis with several strains characteristics of the gastrointestinal tract, has permitted to simulate the duodenal and colonic intestinal compartments, respectively. The duodenal bioavailability expressed as absorption value after 4 h of exposure, ranged from 57.7 to 76.8% for EN A, from 68.8 to 70.2% for EN A1, from 65.0 to 67.0% for EN B, and from 62.2 to 65.1% for EN B1. Colonic bioavailability after 48 h of incubation ra…

Oxidative stress of alternariol in Caco-2 cells

Alternariol (AOH) is a mycotoxin produced by fungus Alternaria. It is found in a wide variety of fruits and cereals products. AOH is able to damage human health. The aim of this study was to evaluate the cytotoxicity of AOH in human colon adenocarcinoma (Caco-2) cells. Moreover, some events related to oxidative stress were evaluated: reactive oxygen species (ROS) generated by oxidation of 2',7'-dichlorodihydrofluorescein diacetate; peroxidation of lipid (LPO) by malondialdehyde (MDA) production; and antioxidant enzymatic capability of catalase (CAT) and superoxide dismutase (SOD). Cytotoxicity of AOH (from 3.125 to 100 μM) was determined during 24, 48 and 72 h of exposure by different endpo…

Influence of the making and cooking pasta on enniatins contents

Development of a new method for the simultaneous determination of 21 mycotoxins in coffee beverages by liquid chromatography tandem mass spectrometry

Abstract A new method for the simultaneous detection of 21 mycotoxins (ochratoxin A, aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, sterigmatocystin, nivalenol, deoxynivalenol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, diacetoxyscirpenol, neosolaniol, HT-2 toxin, T-2 toxin, fumonisin B1, fumonisin B2, enniatin A, enniatin A1, enniatin B, enniatin B1, and beauvericin) in coffee beverages was internally validated. The method is based on liquid/liquid extraction with a mixture of ethyl acetate/formic acid (95:5 v/v) and detection using triple quadrupole (QqQ) and ion trap (IT) liquid chromatography tandem mass spectrometry. The limits of detection and quantification were 0.02…

Simultaneous determination of eight underivatised biogenic amines in fish by solid phase extraction and liquid chromatography-tandem mass spectrometry.

Biogenic amines on fish tissue are formed as a result of bacterial contamination and spoilage during storage. A new method based on liquid chromatography (LC) and tandem mass spectrometry (MS/MS) using a triple quadrupole (QqQ) analyser was developed for the analysis of eight biogenic amines (cadaverine, histamine, phenylethylamine, putrescine, spermine, spermidine, tyramine and tryptamine) in fish tissues. Sample preparation was performed by extraction with trichloroacetic acid 5% and solid phase extraction clean up with STRATA X cartridge. The MS/MS method was validated and compared with a method based on the analysis of dansyl derivatives by LC and fluorescence detector (FD). MS/MS achie…

Toxicity assessment of pesticides using the microtox test: application to environmental samples.

In silico methods for metabolomic and toxicity prediction of zearalenone, α-zearalenone and β-zearalenone.

Zearalenone (ZEA), α-zearalenol (α-ZEL) and β-zearalenol (β-ZEL) (ZEA's metabolites) are co/present in cereals, fruits or their products. All three with other compounds, constitute a cocktail-mixture that consumers (and also animals) are exposed and never entirely evaluated, nor in vitro nor in vivo. Effect of ZEA has been correlated to endocrine disruptor alterations as well as its metabolites (α-ZEL and β-ZEL); however, toxic effects associated to metabolites generated once ingested are unknown and difficult to study. The present study defines the metabolomics profile of all three mycotoxins (ZEA, α-ZEL and β-ZEL) and explores the prediction of their toxic effects proposing an in silico w…

Isolation and purification of enniatins A, A1, B, B1, produced by Fusarium tricinctum in solid culture, and cytotoxicity effects on Caco-2 cells

Enniatins (ENs) are antibiotic compounds of hexadepsipeptidic structure produced by several strains of Fusarium spp. The ENs A, A(1), B, B-1 were purified from extracts of Fusarium tricinctum grown on a solid medium of corn, by a low pressure liquid chromatography (LPLC) on reverse phase of Amberlite XAD-7 followed by semipreparative LC. The purity and the structure of the isolated compounds were confirmed by LC-MS/MS. The technique of the purification of the fungal extract enabled complete separation of the ENs A, A(1), B, B-1 with a mean purity of 97% for all the compounds. The cytoxicity of the ENs was tested in the cell lines of human origin (epithelial colorectal adenocarcinoma cells, …

Indirect analysis of urea herbicides from environmental water using solid-phase microextraction.

We described here a solid-phase microextraction procedure used to extract six urea pesticides-- chlorsulfuron, fluometuron, isoproturon, linuron, metobromuron and monuron--from environmental samples. Two polydimethylsiloxanes and a polyacrylate fiber (PA) are compared. The extraction time, pH control, addition of NaCl to the water and the influence of organic matter such as humic acid on extraction efficiency were examined to achieve a sensitive method. Determination was carried out by gas chromatography with nitrogen-phosphorus detection. The proposed method requires the extraction of 2 ml of sample (pH 4, 14.3%, w/v, NaCl) for 60 min with the PA fiber. The limits of detection range from 0…

Antibacterial activity of the enniatin B, produced by Fusarium tricinctum in liquid culture, and cytotoxic effects on Caco-2 cells.

The enniatins (ENs) are bioactive compounds of hexadepsipeptidic structure produced by several strains of Fusarium sp. The EN B was purified from extracts of Fusarium tricinctum growth on liquid culture of potato dextrose broth (PDB), using a semipreparative liquid chromatography (LC) followed by an analytical LC. The purity and the structure of the isolated compound were confirmed by the determination of the extinction coefficient and with electrospray ionization-mass spectrometry (ESI-MS) study. The pure fraction of EN B was utilized to determine the antibiotic effects on several bacterial strains that are considered normally pathogens of the intestinal tract: Escherichia coli, Enterococc…

Comparison of three solid-phase extraction processes in quantification of ciprofloxacin and enrofloxacin in pork meat

Due to strong implications for food safety, control of fluoroquinolones residues in swine meat should be undertaken to verify compliance of the contamination levels with the maximum residue limits recently updated by Commission Regulation (EU) No. 37/2010 of 22 December 2009. Solid-phase extraction is widely used in antibiotic analysis in food of animal origin. In this study, the results of a comparative study using different types of solid-phase extraction columns, HLB, MCX, and MAX, for ciprofloxacin and enrofloxacin analysis, in pork meat, are presented. In addition, diverse sample treatments for defatting, precipitate proteins, eliminate cations, and increase the ionic strength, were us…

Influence of the solvent on the gas chromatographic behaviour of urea herbicides

Degradation products of chlorsulfuron, chlortoluron, diuron, fluometuron, isoproturon, linuron, metabenzthiazuron, metobromuron, and monuron formed in the gas chromatographic injector have been used for identification of the respective herbicides. Mass spectra of the derived compounds were obtained with a quadrupole mass spectrometric detector working in scan mode (20–450 amu). The compounds generated often depended on the solvent used for phenylurea herbicide injection (ethanol, methanol, dichloromethane, and acetonitrile). When methanol and ethanol were used as solvents the major products formed from phenylureas were carbamic acid esters. When acetonitrile or dichloromethane were used the…

Role of quercetin on Caco-2 cells against cytotoxic effects of alternariol and alternariol monomethyl ether.

Molds of the genus Alternaria have been reported as contaminants of a variety of food and feed. Alternaria toxins such as alternariol (AOH) and its naturally occurring monomethyl ether (AME) produce cytotoxicity and oxidative stress in cell cultures. On the other hand, it has been proved that natural polyphenols have antioxidant effect. Quercetin (Quer) is a polyphenol present in berries and other commodities which exhibits these effects. The aims were to evaluate the cytotoxicity of AOH, AME and the binary combination of them, and the cytoprotective effect of Quer exposed simultaneously with AOH, AME and the mycotoxin mixture in human adenocarcinoma (Caco-2) cells. The cytotoxicity and the…

Matrix effects on solid-phase microextraction of organophosphorus pesticides from water

Abstract This study develops a method for solid-phase microextraction (SPME) of eight organophosphorus pesticides, diazinon, fenthion, fenitrothion (sumithion), methyl-parathion, parathion, methyl-trithion, ethion and triazophos, from water. Determination is carried out by gas chromatography with nitrogen-phosphorus detection. To perform the SPME, poly(dimethylsiloxane) and polyacrylate fibers were initially compared on the basis of their absorption capacities for the selected pesticides, and polyacrylate was selected to accomplish the rest of assays. The main factors affecting the SPME process such as memory effect, stirring rate, extraction temperature and absorption-time profile were stu…

Occurrence of mycotoxins in laboratory rat feeds

Evaluation of solid-phase extraction and stir-bar sorptive extraction for the determination of fungicide residues at low-microg kg(-1) levels in grapes by liquid chromatography-mass spectrometry.

Abstract A liquid chromatography–mass spectrometry method has been developed for determining bitertanol, carboxin, flutriafol, pyrimethanil, tebuconazole and triadimefon. The evaluation of both atmospheric pressure interfaces (API), atmospheric pressure chemical ionization (APCI) and electrospray (ESI) using positive and negative ionization modes, clearly shows that the studied pesticides are more sensitive using APCI in positive mode. Two procedures based on solid-phase extraction (SPE) and stir-bar sorptive extraction (SBSE) have been assessed for extracting these compounds in grape. The recoveries obtained by SPE in samples spiked at the limit of quantification (LOQ) level ranged from 60…

Comparative study of two sample treatments for a liquid chromatography–tandem mass spectrometry determination of mycotoxin biomarkers in urine

Monitoring of five postharvest fungicides in fruit and vegetables by matrix solid-phase dispersion and liquid chromatography/mass spectrometry

Abstract A method was developed for monitoring dichloran, flutriafol, o-phenylphenol, prochloraz, and tolclofos-methyl in fruits and vegetables, using matrix solid-phase dispersion and liquid chromatography with mass spectrometry detection. The method was used to determine fungicide content in 200 samples of chards, onions, peppers, bananas, lemons, and oranges. Of the samples examined, 54% contained o-phenylphenol with concentrations ranging from 0.005 to 3.34 mg/kg and 35% showed prochloraz in the range of 0.06–1.95 mg/kg. Dichloran, flutriafol, and tolclofos-methyl were detected only occasionally. Only 4% of the samples exceeded the European Union maximum residue limits. The pesticides i…

Analysis of post-harvest fungicides by micellar electrokinetic chromatography.

A method based on solid-phase extraction (SPE) and micellar electrokinetic chromatography (MEKC) was developed for the simultaneous determination of carbendazim, imazalil, methylthiophanate, O-phenylphenol, prochloraz, procimidone, thiabendazole and triadimefon residues in grape, lettuce, orange and tomato. Selectivity and resolution were studied changing the pH and the concentration of the buffer, the type and concentration of surfactant and the methanol content in the mobile phase. A buffer consisting of 4 mM borate with 75 mM sodium cholate (pH 9.2) gave the best results. The recoveries of the fungicides in spiked fruit and vegetable samples ranged from 30 to 105%, and the limits of dete…

Effects of aldicarb and propoxur on cytotoxicity and lipid peroxidation in CHO-K1 cells

Abstract Cytotoxic effects of aldicarb, its sulfone and sulfoxide, and propoxur, lipid peroxidation and antioxidant parameters in Chinese Hamster Ovary (CHO-K1) cells were determined. d , l -buthionine-( S , R )-sulfoximine (BSO) was assayed to determine the role of GSH in the protection against carbamate cytotoxicity. Pre-treatment with 60 μM BSO, induced a significant decrease in the glutathione reductase (GR; 64–141%), the glutathione peroxidase (GPx; 10–30%) and the glutathione S-transferase (GST; 59–93%) activities, and its GSH levels (79–85%), while the oxidized glutathione (GSSG) levels significantly increased (64–78%) respect to experiment non-BSO-pretreated. Carbamates BSO pre-trea…

Simultaneous determination of bisphenol A, octylphenol, and nonylphenol by pressurised liquid extraction and liquid chromatography–tandem mass spectrometry in powdered milk and infant formulas

Abstract A new analytical method, using pressurised liquid extraction (PLE) and liquid chromatography–tandem mass spectrometry (LC–MS/MS), was developed for the simultaneous determination of bisphenol A (BPA), octylphenol (OP) and nonylphenol (NP) in powdered infant formulas (IF) and powdered skimmed milk (PM). The analytes were extracted by PLE, using this optimised conditions: ethyl acetate as solvent, 70 °C of temperature, reversed-phase silica C18 as dispersing agent and three cycles of extraction. The extracts were then injected in LC–MS/MS using a Gemini C18 column and a mixture of 5% water and 95% methanol/acetonitrile, both with 0.1% ammonia, as a mobile phase. Recoveries at differe…

Simultaneous determination of different classes of antibiotics in fish and livestock by CE-MS

A specific CE-MS method was developed for the simultaneous determination of 12 antibacterial residues (four sulfonamides: sulfamethazine, sulfathiazole, sulfadiazine, and sulfachlorpyridazine; four beta-lactams: amoxicillin, ampicillin, oxacillin, and penicillin V, and four quinolones: danofloxacin, enrofloxacin, ofloxacin, and flumequine) in fish and livestock. Separation conditions, sheath liquid composition and electrospray parameters were optimized to obtain adequate CE separation and a high sensitivity. CE employed a 75 cm long fused-silica capillary (50 cm thermostated plus 25 cm at room temperature) 75 microm id and a 60 mM ammonium acetate separation buffer at pH 8 with 10% of metha…

Capillary electrophoresis for analyzing pesticides in fruits and vegetables using solid-phase extraction and stir-bar sorptive extraction.

Two procedures based on solid-phase extraction (SPE) and stir-bar sorptive extraction (SBSE) in combination with micellar electrokinetic chromatography (MEKC)--diode array detection (DAD) were compared for the simultaneous extraction of acrinathrin, bitertanol, cyproconazole, fludioxonil, flutriafol, myclobutanil, pyriproxyfen, and tebuconazole in lettuce, tomato, grape, and strawberry. Selectivity and resolution of the MEKC procedure were studied changing the pH and the molarity of the buffer, the type and the concentration of surfactant, and the methanol content in the mobile phase. A buffer consisting of 6 mM sodium tetraborate decahydrate with 75 mM of cholic acid sodium solution (pH 9.…

Impact of Pharmacists’ Participation in a Pharmacotherapy Follow-Up Program

Objective. To evaluate the impact of a continuing pharmacy education (CPE) course on Spanish community pharmacists’ participation in a pharmacotherapy follow-up program. Design. Participation in a CPE course offered 4 times over a 4-year period via satellite teleconferencing was monitored and the data analyzed to determine the course’s impact on community pharmacists’ participation in a pharmacotherapy follow-up program. Assessment. Community pharmacists’ participation in the pharmaceutical care CPE course had a slightly positive impact on their participation in the pharmacotherapy follow-up program. In the best profiles, there was a probability of 7.3% that participants would participate i…

Assessment of metal levels in foodstuffs from the Region of Valencia (Spain).

Graphical abstract

Further data on the occurrence of Fusarium emerging mycotoxins enniatins (A, A1, B, B1), fusaproliferin and beauvericin in raw cereals commercialized in Morocco

Abstract In this study, 64 samples of raw cereals (wheat, maize and barley) purchased from local markets in Rabat–Sale area from Morocco were analyzed for the occurrence of six emerging mycotoxins: four enniatins ENs (ENA, ENA1, ENB and ENB1), beauvericin (BEA) and fusaproliferin (FUS). Samples were extracted with a mixture of water/acetonitrile (85/15, v/v) by using an Ultra-turrax homogenizer. Mycotoxins were then identified and quantified with a liquid chromatography (LC) with diode array detector (DAD). Positive samples were confirmed with an LC–MS/MS. Analytical results showed that the frequencies of contamination of total samples with ENs, BEA and FUS were 50, 26.5 and 7.8%, respectiv…

Analysis of Polychlorinated Biphenyls in Aqueous Samples Using C18 Glass Column Extraction

Abstract A method for polychlorinated biphenyl (PCB) trace extraction from contaminated water at the part-pertrillion levels is described. This procedure involves quantitative adsorption of PCBs on Cis glass microcolumns. PCBs are retained on the surface and subsequently eluted with n-hexane (5 mL) before gas chromatography. Recovery of water fortified with PCBs was 83.6-108.5%. Compared to liquid-liquid extraction methods, the Cis glass microcolumns give comparable results, lessen solvent costs, and are less time-consuming.

In vitro effect of organophosphate pesticides, Malathion and chlorpyriphos, on lipid peroxidation and antioxidant enzymes

Bioaccesibility of Cylindrospermopsin from cooked fish muscle after the application of an in vitro digestion model and its bioavailability.

Humans can be exposed to cyanotoxins through the ingestion of contaminated water, food or beverages. In the present work, the bioaccesibility of Cylindrospermopsin (CYN), one of the most relevant cyanotoxins, was evaluated in a pure CYN solution and cooked CYN-contaminated fish muscles (20 μg/mL). An in vitro digestion model including the salivar, gastric, duodenal and colonic phases was performed, being each fraction analyzed by HPLC-MS-MS to evaluate CYN degradation. Moreover, Caco-2/TC7 cells were exposed to the digested duodenal and colonic phases to elucidate the final bioavailability of CYN in an approximation to the real human exposure scenario. The results revealed that CYN bioacces…

Determination of five pesticide residues in oranges by matrix solid-phase dispersion and liquid chromatography to estimate daily intake of consumers.

Abstract Residues of benzoylphenylurea insecticides (diflubenzuron, hexaflumuron, and flufenuxuron), carboxamide acaricides (hexythiazox), and carbamate insecticides (benfuracarb) were determined in 150 orange fruit samples from September 1998 to June 1999, to estimate exposure of the Valencian population to oranges contaminated with these newly developed pesticides. The method for monitoring these residues is based on matrix solid-phase dispersion and liquid chromatography with UV or atmospheric pressure chemical ionization/mass spectrometry (APCI/MS) detection. Orange samples representing 11 varieties were collected from an agricultural cooperative and examined for the 5 pesticides. In 74…

Application of solid-phase microextraction for determining phenylurea herbicides and their homologous anilines from vegetables.

Abstract Residues of metobromuron, monolinuron and linuron herbicides and their aniline homologous were analyzed in carrots, onions and potatoes by solid-phase microextraction (SPME) performed with a polyacrylate fiber. A juice was obtained from food samples that were further diluted, and an aliquot was extracted after sodium chloride (14%) addition and pH control. At pH 4 only the phenylureas were extracted. A new extraction at pH 11 allowed the extraction of phenylureas plus homologous aniline metabolites. Determination was carried out by gas chromatography with nitrogen–phosporus detection (NPD) the identity of the determined compounds was studied by gas chromatography–mass spectrometry.…

Validation of a confirmatory method for the determination of macrolides in liver and kidney animal tissues in accordance with the European Union regulation 2002/657/EC

Abstract This study proposes a simple multiresidue liquid chromatography–diode array detector (LC–DAD) method capable of determining seven macrolide antibiotics in samples of liver and kidney animals at concentrations lower than those allowed by current legislation. Samples were prepared by homogenizing the tissue with EDTA–McIlvaine's buffer and extracted with an Oasis HLB cartridge. The consumption of organic solvent during extraction was minimum. The analytes were detected by LC–DAD and also by liquid chromatography–mass spectrometry with electrospray ionization (LC–(ESI)MS). The method was specific, stable and robust enough for the required purposes. The DAD method was validated in acco…

Determination of organic contaminants in food by capillary electrophoresis

This review addresses recent advances in the analysis of organic contaminants, such as antibiotics, pesticides, biological toxins, and food-borne pathogens, in foods by capillary electrophoresis (CE). Special attention is paid to those aspects that increase sensitivity and/or selectivity, such as sample extraction and concentration, on-line preconcentration techniques (stacking), affinity capillaries or/and specific detectors (laser induced fluorescence (LIF), mass spectrometry (MS)). The various CE modes used to separate the compounds and the quantification strategies are also examined. As a result, this work presents an updated overview on the principal applications of CE, together with a…

Analysis of fumonisins in corn-based food by liquid chromatography with fluorescence and mass spectrometry detectors

Abstract The presented procedure involves an extraction with methanol–water, centrifugation and cleanup with immunoaffinity columns. A comparison study between fluorescence detector, mass spectrometry, and tandem mass spectrometry with a triple quadrupole (QqQ) analyzer using an electrospray ionisation interface for the determination of fumonisin B1 and B2 in corn-based products has been performed. Limits of quantification obtained by the three detectors were lower than the maximum levels established by European Commission. Liquid chromatography coupled to tandem mass spectrometry provides higher sensitivity (12 μg kg−1for fumonisins B1 and B2) when compared to mass spectrometry (40 μg kg−1…

Enniatin B induces expression changes in the electron transport chain pathway related genes in lymphoblastic T-cell line

Abstract Enniatin B is a ionophoric and lipophilic mycotoxin which reaches the bloodstream and has the ability to penetrate into cellular membranes. The purpose of this study was to reveal changes in the gene expression profile caused by enniatin B in human Jurkat lymphoblastic T-cells after 24 h of exposure at 1.5, 3 and 5 μM by next generation sequencing. It was found that up to 27% of human genome expression levels were significantly altered (5750 genes for both down-regulation and up-regulation). In the three enniatin B concentrations studied 245 differentially expressed genes were found to be overlapped, 83 were down and 162 up-regulated. ConsensusPathDB analysis of over-representation…

Control of pesticide residues by liquid chromatography-mass spectrometry to ensure food safety.

Liquid chromatography-mass spectrometry (LC-MS) has become an invaluable technique for the control of pesticide residues to ensure food safety. After an introduction about the regulations that highlights its importance to meet the official requirements on analytical performance, the different mass spectrometers used in this field of research, as well as the LC-MS interfaces and the difficulties associated with quantitative LC-MS determination, are discussed. The ability to use practical data for quantifying pesticides together with the option of obtaining structural information to identify target and non-target parent compounds and metabolites are discussed. Special attention is paid to the…

Further data on the levels of emerging Fusarium mycotoxins enniatins (A, A1, B, B1), beauvericin and fusaproliferin in breakfast and infant cereals from Morocco

Abstract Sixty-eight samples of cereals products, including breakfast cereals ( n  = 48) and infant cereals ( n  = 20), purchased from supermarkets and pharmacies in Rabat-Sale area from Morocco were analysed for the determination of six emerging mycotoxins: four enniatins ENs (ENA, ENA1, ENB and ENB1), beauvericin (BEA) and fusaproliferin (FUS). Samples were extracted with a mixture of acetonitrile:water (85:15, v/v), using an Ultra-Turrax® homogeniser. Mycotoxins were then identified and quantified by liquid chromatography (LC) with diode array detection (DAD). Positive samples were confirmed by LC–MS/MS. Analytical results showed that the percentages of analysed samples contaminated with…

Evaluation of a solid-phase extraction system for determining pesticide residues in milk.

Abstract A simple and rapid procedure based on reversed solid-phase extraction with octadecylsilica was developed for determining chlorinated pesticide residues in milk. The need for an agent that breaks the fat globules was first investigated and the method variables were optimized on whole homogenized milk (3.6% fat content). Recovery experiments performed for 26 organochlorine pesticides at levels of 3–40 μg/l gave > 80% recoveries for all the compounds. The method was validated and evaluated by comparison with two widely used liquid-liquid extraction methods. The performance was checked by analysis of a certified standard with natural low endogenous levels (CMR 187) and different kinds …

Bioaccessibility and bioavailability of bioactive compounds from yellow mustard flour and milk whey fermented with lactic acid bacteria.

Microbial fermentation with lactic acid bacteria (LAB) is a natural food biopreservation method. Yellow mustard and milk whey are optimum substrates for LAB fermentation. The aim of the present study was to evaluate the bioaccessibility and bioavailability of bioactive compounds from yellow mustard flour and milk whey both with and without LAB fermentation. All extracts were subjected to a simulated digestion process. Total polyphenols, DL-3-phenyllactic acid (PLA), lactic acid, and the antioxidant activity were determined in the studied matrices before and after simulated digestion. Yellow mustard flour was significantly richer in total polyphenols, whereas significantly higher concentrati…

Oxidative DNA damage and disturbance of antioxidant capacity by alternariol in Caco-2 cells

Oxidative stress occurs as a consequence of an imbalance between the prooxidant/antioxidant systems, causing an increase of intracellular generation of reactive oxygen species. Alternariol (AOH), a mycotoxin produced by Alternaria sp. can alter the action of glutathione (GSH) and the enzymes involved in the redox system, causing damage to cellular macromolecules such as DNA. The aims of this work were to determine the induction of oxidative stress by the antioxidant defenses imbalance in relation to glutathione (GSH), glutathione reductase (GR), glutathione transferase (GST), glutathione peroxidase (GPx) levels and DNA damage in Caco-2 cells derived from adenocarcinoma human colon. Oxidativ…

Editorial: Mechanism of mycotoxins.

Enniatin B metabolites in liver and tissues of gilthead sea bream (Sparus aurata)

Detoxification of fumonisins by isothiocyanates

Interaction effects of Fusarium enniatins (A, A1, B and B1) combinations on in vitro cytotoxicity of Caco-2 cells

Abstract Foodstuff is usually contaminated by more than one mycotoxin, however toxicological data are lacking as regards the effects in combinations compared to their individual effect. This study investigated the in vitro effects of enniatins (ENs) A, A1, B and B1, alone and in combinations, on Caco-2 cells viability by MTT assay after 24 h of exposure. Cells were treated with concentrations ranging from 0.9 to 15.0 μM, individually and in combination of two, three and four mycotoxins. Dose–response curves were generated for each mycotoxin and the isobologram method was used to determine the interactive effects of tested mixtures. Tested ENs produced significant cytotoxic effects both indi…

Study of the cytotoxic activity of beauvericin and fusaproliferin and bioavailability in vitro on Caco-2 cells.

Abstract Beauvericin (BEA) is a cyclohexadepsipeptide mycotoxin which has insecticidal properties and produces cytotoxic effects in mammalian cells. Fusaproliferin (FUS) is a mycotoxin that has toxic activity against brine shrimp, insect cells, and teratogenic effects on chicken embryos. The aim of this study was to determine the cytotoxicity of BEA and FUS in human epithelial colorectal adenocarcinoma HT-29 and Caco-2 cells, the transepithelial transport and the bioavailability using Caco-2 cells as a simulated in vitro gastrointestinal model of the human intestinal epithelium. The inhibitory concentration (IC 50 ) evidenced by BEA in the Caco-2 cells was 24.6 and 12.7 μM at 24 and 48 h ex…

Bioaccessibility of Enniatins A, A1, B, and B1 in Different Commercial Breakfast Cereals, Cookies, and Breads of Spain

Fusarotoxins enniatins (ENs) can represent a potential risk as natural contaminants of cereal commodities. However, only their bioaccessible fraction can exert a toxicity. The purpose of this study was to determine the ENs A, A₁, B, and B₁ bioaccessibility added in 1.5 and 3.0 μmol/g concentrations in breakfast cereals, cookies, and breads using a simulated in vitro gastrointestinal extraction model. Bioaccessibility values ranged between 40.4 ± 1.9 and 79.9 ± 2.8%. The lower values were 50.1, 40.4, 43.9, and 46.3% in wheat bran with fibers, and the higher values were 79.9, 64.2, 69.8, and 73.6% in white loaf bread for the ENs A, A₁, B, and B₁, respectively. Food composition, compounds stru…

Antioxidant capacity of trans -resveratrol dietary supplements alone or combined with the mycotoxin beauvericin

Trans-resveratrol (trans-RSV) is a polyphenol with multiples biological properties, such as anti-inflammatory, antioxidant, anti-aging, anti-diabetic, and antiplatelet. It occurs naturally in grapes and derivate, peanuts and berries. Beauvericin (BEA) is a mycotoxin present in cereals that produces cytotoxicity, intracellular reactive oxygen species and lipid peroxidation. The general objective of this research was to evaluate whether trans-RSV could be used as a good polyphenol against damages produced by BEA. Because trans-RSV can be ingested through dietary supplements, to reach this goal, the following specific objectives were proposed: to determine a) the trans-RSV content in different…

Survey of mycotoxins in dates and dried fruits from Tunisian and Spanish markets

Abstract In an effort to enforce the knowledge on mycotoxin occurrence and co-occurrence in dates and dried fruits, 228 samples purchased from Tunisian and Spanish markets were subjected to multi-mycotoxin liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis of 16 mycotoxins. At least one mycotoxin was detected in 160 samples (70%). The frequency of contaminated samples was 83%, 80%, 64%, 59% and 26% for dates, dried vine fruits, figs, apricots and plums, respectively but none of the analyzed samples contained detectable levels of fumonisins (FB1, FB2, FB3) or T-2 toxin (T2). The incidence and levels of mycotoxins varied in samples. The most prevalent mycotoxin was enniat…

Determination of abamectin in citrus fruits by liquid chromatography-electrospray ionization mass spectrometry.

Liquid chromatography coupled to electrospray mass spectrometry (LC-ES-MS) with positive ion detection was used to determine abamectin in oranges. MS conditions were optimized to achieve maximum sensitivity. The main ion for abamectin was [M+Na]+ at a fragmentor voltage of 180 V. Abundant structural information can be obtained at different fragmentor voltages. The detection limit for the standard solution was 12 pg injected, and good linearity and reproducibility were observed. Abamectin residues were extracted using matrix solid-phase dispersion. Orange samples were homogenized with C18 bonded silica placed onto a glass column and eluted with dichloromethane. Recoveries of the abamectin fr…

Interactive effects of zearalenone and its metabolites on cytotoxicity and metabolization in ovarian CHO-K1 cells.

Zearalenone (ZEA) is a non-steroidal estrogen mycotoxin with high binding affinity to estrogen receptors. ZEA is rapidly absorbed and metabolized in vivo to α-zearalenol (α-ZOL) and β-zearalenol (β-ZOL). So, mixtures of them may be present in biological systems and suppose a hazard to animals and human health. The aims of this study were to determine the cytotoxic effects of ZEA and its metabolites, alone and in combination in ovarian (CHO-K1) cells during 24, 48 and 72h by the MTT assay; and to investigate the metabolism of the CHO-K1 cells on ZEA, and its conversion into α-ZOL and β-ZOL by CHO-K1 cell after 24 and 48h of exposure. The IC50 value obtained for individual mycotoxins range fr…

Solid-Phase Microextraction Liquid Chromatography/Tandem Mass Spectrometry To Determine Postharvest Fungicides in Fruits

A method to determine five postharvest fungicides (dichloran, flutriafol, o-phenylphenol, prochloraz, tolclofos methyl) in fruits (cherries, lemons, oranges, peaches) has been developed using solid-phase microextraction (SPME) coupled to liquid chromatography (LC) with photodiode array (DAD), mass spectrometry (MS), or tandem mass spectrometry (MS/MS) with ion trap detection. Extraction involved sample homogenization with an acetone/water solution (5:1), filtration, and acetone evaporation prior to fiber extraction. The pesticides were isolated with a fused-silica fiber coated with 50-microm Carbowax/template resin. The effects of pH, ion strength, sample volume, and extraction time were in…

Immunotoxicological in vitro effects of BEA and ENN B on Jurkat cell line

Enterotoxigenic staphylococci and their toxins in restaurant foods

Abstract This review presents an overview of the enterotoxigenic staphylococci and their toxins in restaurant foods, with special reference to the characteristics of these micro-organisms and their enterotoxins. Furthermore, this paper reviews the staphylococcal food poisoning outbreaks, principal sources of contamination and food safety measures that can be applied to eliminate the presence of enterotoxigenic staphylococci in restaurant foods.

Evaluation of immunologic effect of Enniatin A and quantitative determination in feces, urine and serum on treated Wistar rats.

Abstract Study of dietary supplementation with ENN A mycotoxin during 28 days of exposure time on Wistar rats to determinate its levels in serum, urine and feces and, to evaluate the immunologic effect in peripheral blood lymphocytes (PBL) is presented. The first method for ENN A extraction, determination and detection by LC–MS/MS in serum, urine and feces samples is reported. ENN A food dose administrated was detected in serum samples and influenced lymphocyte phenotyping. Levels in serum were founded from the second week of the experiment; reaching values of 4.76 μg/ml on the fourth week, which corresponds to 3.24 μg/ml in blood. PBL as T helper (CD4 + ) were presented in greater percenta…

Monitoring of the pesticide levels in natural waters of the Valencia Community (Spain)

Monitoring Alternaria mycotoxins and pesticide residues in tomato by dispersive liquid–liquid microextraction coupled to gas chromatography–tandem mass spectrometry

Quantitative determination of trichothecenes in breadsticks by gas chromatography-triple quadrupole tandem mass spectrometry.

Breadsticks are pencil-sized sticks of dry bread widely consumed as a pre-meal appetiser. They are basically wheat-based snacks, which makes them a good matrix to evaluate mycotoxin contamination, since wheat is very susceptible to fungal attack. In this sense, the fast, selective and sensitive gas chromatography-triple quadrupole tandem mass spectrometry (GCQqQ- MS/MS) method proposed here allows simultaneous determination of deoxynivalenol (DON), 3-acetyldeoxynivalenol, fusarenon-X, diacetoxyscirpenol, nivalenol, neosolaniol, HT-2 and T-2 toxin in breadsticks after QuEChERS extraction and clean-up. The performance of the method was assessed with respect to European Commission Regulations …

Optimization of a matrix solid-phase dispersion method for the analysis of pesticide residues in vegetables

A multiresidue method based on matrix solid-phase dispersion (MSPD) is studied to determine chlorfenvinfos, chlorpyrifos, fenarimol, iprodione, procimydone, propiconazole, tetradifon, triadimefon and vinclozolin in artichokes, green beans, lettuces and tomatoes. Alumina, silica and Florisil were assessed as extracting phases, and the extracts from Florisil were the cleanest. To facilitate manual extraction, sand was added to the sample together with the dispersing phase. Three eluting systems were then studied, and dichloromethane proved to be the best. Further purification can be performed using solid-phase cleanup after diluting extracts with aqueous solutions. Octyl- and octadecyl-silica…

Determination of Aldicarb, Aldicarb Sulfoxide, and Aldicarb Sulfone in Oranges by Simple Gas–Liquid Chromatography with Nitrogen–Phosphorus Detection

Abstract A reversed-phase liquid chromatographic method, using an acetonitrile–water gradient mobile phase, and a gas-liquid chromatographic method with nitrogen–phosphorus and flame photometric detection were compared for simultaneous and separate determinations of trace quantities of aldicarb and its metabolites aldicarb sulfone and aldicarb sulfoxide. One hundred gram samples of fortified oranges were extracted with water–acetone, water-acetonitrile, or water–methanol, and the extracts were partitioned with methylene chloride. Partitioning with other organic solvents such as chloroform, ethyl acetate, or methyl isobutyl ketone was also investigated. When performed under optimal condition…

BEHAVIOUR OF GRAPHITIZED CARBON BLACK IN THE EXTRACTION OF POLAR NON-IONIC NITROGEN-CONTAINING PESTICIDES. A CHECKING OF HYPOTHESES

Graphitized Carbon Black (GCB) extractive cartridges are evaluated for on-line coupling with a C8 analytical column to determine eleven carbamates and one carboximide pesticide from spiked deionized water at the 1.2 μg/L level. Several experiments were carried out to ascertain whether GCB saturation, pesticide degradation on the surface, existence of by-pass channels, mobility among the bulk cartridge, or strong retention on the surface interfere with the determination of pesticides. Problems in on-line CGB elution are partially solved by modifying the acetonitrile/water gradient to contain a front of 100% acetonitrile for a few seconds. Eluting the same GCB cartridges off-line with dichlor…

Determination of pesticides in soil samples by solid phase extraction disks

A systematic study comparing the methodology and analytical results obtained in an investigation of seven pesticide residues (Molinate, Atrazine, Carbofuran, Pirimicarb, Prometryn, Malathion and Tetrachlorvinphos) in soil samples is reported. Solid-phase extraction (SPE) using glass columns and 47 mm disks of octyl and octadecyl-bonded silica was used in the pesticide analysis. The best extraction efficiency and clearest extracts are obtained with C8 disks. The analyses were carried out by capillary gas chromatography with nitrogen and phosphorus detection. Recovery experiments were performed at ppb levels in spiked soil samples. The average recoveries of the compounds were 53–77%. Detectio…

Alternaria Mycotoxins in Food and Feed: An Overview

Alternariais one of the major mycotoxigenic fungal genera with more than 70 reported metabolites.Alternariamycotoxins showed notably toxicity, such as mutagenicity, carcinogenicity, induction of DNA strand break, sphingolipid metabolism disruption, or inhibition of enzymes activity and photophosphorylation. This review reports on the toxicity, stability, metabolism, current analytical methods, and prevalence ofAlternariamycotoxins in food and feed through the most recent published research. Half of the publications were focused on fruits, vegetables, and derived products—mainly tomato and apples—while cereals and cereal by-products represented 38%. The most studied compounds were alternario…

Analysis of pesticides in fruits by pressurized liquid extraction and liquid chromatography-ion trap-triple stage mass spectrometry.

A multi-residue method using pressurized liquid extraction (PLE) and liquid chromatography-quadrupole ion trap-triple stage mass spectrometry (LC-IT-MS(3)) has been developed for determining trace levels of pesticides in fruits. The selected pesticides can be distinguished in: benzimidazoles and azoles, organophosphorus, carbamates, neonicotinoids, and acaricides. PLE has been optimized to extract these pesticide residues from oranges and peaches by studying the effect of experimental variables on PLE efficiency. Samples were extracted at high temperature and pressure (75 degrees C and 1500psi) using ethyl acetate as extraction solvent and acidic alumina as drying agent. The recoveries obta…

In vitro blood brain barrier exposure to mycotoxins and carotenoids pumpkin extract alters mitochondrial gene expression and oxidative stress.

Abstract Food and feed are daily exposed to mycotoxin contamination which effects may be counteracted by antioxidants like carotenoids. Some mycotoxins as well as carotenoids penetrate the blood brain barrier (BBB) inducing alterations related to redox balance in the mitochondria. Therefore, the in vitro BBB model ECV304 was subcultured for 7 days and exposed to beauvericine, enniatins, ochratoxin A, zearalenone (100 nM each), individually and combined, and pumpkin extract (500 nM). Reactive oxygen species were measured by fluorescence using the dichlorofluorescein diacetate probe at 0 h, 2 h and 4 h. Intracellular ROS generation reported was condition dependent. RNA extraction was performe…

Dietary exposure and risk assessment of polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans and dioxin-like polychlorinated biphenyls of the population in the Region of Valencia (Spain)

Dietary exposure of the Valencia Region population to polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and PCBs was assessed in the Region of Valencia in 2010–2011. A total of 7700 food samples were collected. Occurrence data were combined with consumption data to estimate dietary exposure in adults (&gt;15 years of age) and young people (6–15 years of age). The estimated intake was calculated by a probabilistic approach. Average intake levels (upper-bound scenario) were 1.58 and 2.76 pg toxic equivalent (TEQ) kg−1 body weight (bw) day−1 for adults and young people, respectively. These average intakes are within range of the tolerable daily intake of 1–4 pg W…

Determination of mycotoxins in bee pollen by gas chromatography-tandem mass spectrometry.

Bee pollen, promoted as a natural food supplement, is consumed increasingly by people to maintain a healthy diet. Depending on environmental conditions, pollen can also be an optimum medium for growth of molds such as Fusarium and Penicillium . A quick, easy, cheap, rapid, and safe (QuEChERS) extraction procedure followed by a gas chromatography-tandem mass spectrometry (GC-MS/MS) determination of eight selected Fusarium toxins in bee pollen was developed and optimized. Recovery studies at 20, 80, and 1000 μg/kg showed values between 73 and 95% with relative standard deviations (RSDs) of15% for all studied mycotoxins. Limits of quantitation (LOQ) ranged from 1 to 4 μg/kg. The proposed metho…

Cytotoxic effects of zearalenone and its metabolites and antioxidant cell defense in CHO-K1 cells.

Zearalenone (ZEA) and its metabolites (α-zearalenol; α-ZOL, β-zearalenol; β-ZOL) are secondary metabolites of Fusarium fungi that produce cell injury. The present study explores mycotoxin-induced cell damage and cellular protection mechanisms in CHO-K1 cells. Cytotoxicity has been determined by reactive oxygen species (ROS) production and DNA damage. ROS production was determined using the fluorescein assay and DNA strand breakage by comet assay. Intracellular protection systems were glutathione (GSH), glutathione peroxidase (GPx), catalase (CAT) and superoxide dismutase (SOD). The results demonstrated that all mycotoxins increased the ROS levels up to 5.3-fold the control levels in CHO-K1 …

Alternariol induce toxicity via cell death and mitochondrial damage on Caco-2 cells

Alternariol (AOH), a mycotoxin produced by Alternaria sp, appears as food contaminant in fruit, vegetables and cereal products. Its toxicity has been demonstrated, but the mechanisms involved have not been elucidated yet. In this study, the pathways triggered by AOH and degradation products generated on Caco-2 cells were evaluated. Cells were exposed to AOH sub-cytotoxic concentrations of 15, 30 and 60 μM. Cell cycle disruption, the induction of apoptosis/necrosis and changes in mitochondrial membrane potential (Δψm) after 24 and 48 h was asses by flow cytometry. Also, AOH and its degradation products were evaluated after 24 and 48 h by high-performance liquid chromatography with tandem mas…

Study of the potential toxicity of commercial crispy breads by evaluation of bioaccessibility and bioavailability of minor Fusarium mycotoxins

Abstract Enniatins (ENs) are bioactive compounds produced by the secondary metabolism of several Fusarium strains and known to have several biological activities, such as acting as enzyme inhibitors, antifungal and antibacterial agents, and immunomodulatory substances. This study has investigated the ENs bioaccessibility, spiked in commercial wheat crispy bread at 1.5 and 3.0 μmol/g concentrations, their transepithelial transport and bioavailability using Caco-2 cells as a model of the human intestinal epithelium. The content (%) of the four ENs contained in the gastric fluid has resulted variable from 69% to 91%, considering the two concentrations assayed. The mean bioaccessibility data fo…

Evaluation of the Fate of Aldicarb and Its Metabolites in Oranges

Abstract The accumulation, persistence and fate of systemic pesticide aldicarb was melencholy evaluated in orange crops. The concentration of this pesticide and its two toxic metabolites, aldicarb sulfoxide and aldicarb sulfone was determined in leaves, rind and pulp of three orange varieties (Satsuma, Navelina and Clemetina de Nules) and in the top soil of the orange groves. The groves were located in two different places in the Valencia Community (Spain). The analysis showed that the aldicarb concentration was lower than those of aldicarb sulfoxide and aldicarb sulfone. In all cases, the residues persisted at least 160 days in vegetable samples and between 157 and 227 days in soil samples…

Interaction effects of enniatin B, deoxinivalenol and alternariol in Caco-2 cells.

Enniatin B (ENN B), deoxinivalenol (DON) and alternariol (AOH) are secondary metabolites of filamentous fungi. These mycotoxins are contaminants of vegetables and cereals. They are cytotoxic and their effects are enhanced by their mixtures. The objectives of this study were to compare the cytotoxicity of ENN B, DON and AOH alone or in combination in human adenocarcinoma (Caco-2) cells and to evaluate the type of interactions of mycotoxin mixtures by the isobologram analysis. Cells were treated with concentrations ranging from 1.85 to 90μM (AOH) and from 0.312 to 10μM (for ENN B and DON), individually and in combination of two and three mycotoxins (from 1.85 to 30μM for AOH and from 0.312 to…

Individual and Combined Effect of Zearalenone Derivates and Beauvericin Mycotoxins on SH-SY5Y Cells

Beauvericin (BEA) and zearalenone derivatives, &alpha

Cytotoxicity of deoxynivalenol and cytoprotection of antioxidants in Caco-2 cells

Matrix solid-phase dispersion microextraction and determination by high-performance liquid chromatography with UV detection of pesticide residues in citrus fruit.

A multiresidue method based on matrix solid-phase dispersion (MSPD) microextraction was studied to determine the carbamate, benfuracarb, and urea insecticides, diflubenzuron, flufenoxuron hexaflumuron and hexythiazox, used in control of citrus pests. Optimisation of different parameters, such as the type of solid support for matrix dispersion, elution solvents and the clean-up step were carried out. The method used 0.5 g of orange sample, C8 bonded silica as MSPD sorbent and dichloromethane as eluting solvent. Recoveries, at spiked concentrations below the maximum residue levels established by Spanish Government, were between 74 and 84% with relative standard deviations ranging from 2 to 4%…

Enniatin A1, enniatin B1 and beauvericin on HepG2: Evaluation of toxic effects

Hepatotoxicity of three Fusarium mycotoxins, beauvericin (BEA) and two enniatins (ENNs) ENN A1 and ENN B1, in hepatocarcinoma cells (HepG2) were evaluated and compared. Concentrations used were 1.5 and 3 μM at 24, 48 and 72 h for each mycotoxin. Flow cytometry was used to examine enniatins effects on cell proliferation, to characterize the cell cycle phase where the cells blocked and to study the mitochondria role in ENNs-induced apoptosis. ENN B1 treated cells showed a time dependent G1 blockade at both concentrations used. ENN A1 and BEA decreased the apoptotic-necrotic percentage of cells comparing to control and disrupted the MMP as observed by TMRM and ToPro-3 fluorochromes signal. It …

Solid-phase extraction disks for determining pesticides from soil leachates

Abstract Analytical techniques are essential for identifying and quantifying the chemicals present in environmental samples. A method for extracting pesticides from soil leachates was developed. The proposed method is based on solid-phase extraction and uses 47-mm disks of octyl-bonded silica. Analysis was carried out by gas chromatography with nitrogen–phosphorus and electron-capture detectors. Several factors that can provide better pesticide recoveries with this extraction method are discussed. These factors are: the kind and percentage of organic solvent used in the extraction procedure, the volume and the ionic strength (adding 15% of NaCl) of the aqueous phase which passes through the…

In vivo toxicity studies of fusarium mycotoxins in the last decade: a review.

This review summarizes the information regarding the in vivo studies of Fusarium mycotoxins in the last decade. The most common studies are classified as subacute toxicity, subchronic toxicity, acute toxicity, toxicokinetic studies and teratogenicity in order of importance. The most used animals in in vivo studies are pigs, rats, chickens and mice. Fumonisin B1, deoxynivalenol, zearalenone, nivalenol and T-2 toxin are the most studied fusarotoxins. Studies with combinations of mycotoxins are also frequent, deoxynivalenol generally being one of them. The predominant route of administration is oral, administered mostly in the form of naturally contaminated feed. Other administration routes al…

Solid-phase microextraction-liquid chromatography-mass spectrometry applied to the analysis of insecticides in honey

An approach based on solid-phase microextraction-liquid chromatography-mass spectrometry (SPME-LC-MS) has been developed for determining 12 insecticides (bromophos ethyl, chlorpyrifos methyl, chlorpyrifos ethyl, diazinon, fenoxycarb, fonofos, phenthoate, phosalone, pirimiphos methyl, profenofos, pyrazophos, and temephos) in honey. The influence of several parameters on the efficiency of the SPME was systematically investigated. Under optimal conditions, the procedure provided excellent linearity (>0.990), detection and quantification limits (between 0.001 and 0.1 microg g(-1) and between 0.005 and 0.5 microg g(-1), respectively), and precision (<19% at the quantification limits and from 6 t…

Effect of polyphenols on enniatins-induced cytotoxic effects in mammalian cells

Enniatins (ENs) are fungal secondary metabolites produced by genus Fusarium. The ENs exert antimicrobial and insecticidal effect, and has also been demonstrated cytotoxic effects on several mammalian cell lines. On the other hands, it has been proved that natural polyphenols have antioxidant effect. In this study, cell effects at low levels of exposure of four ENs (A, A(1), B and B(1)) and five polyphenols (quercetin, quercetin-3-β-D-glucoside, rutin, myricetin and t-pterostilbene) present in wine; and the cytoprotective effect of these polyphenols exposed simultaneously with ENs in Chinese Hamster Ovary (CHO-K1) cells, were studied. Cell effects were determined by the MTT test after 24 h o…

Determination of soyasaponins I and βg in raw and cooked legumes by solid phase extraction (SPE) coupled to liquid chromatography (LC)-mass spectrometry (MS) and assessment of their bioaccessibility by an in vitro digestion model.

Legumes contain a rich variety of phytochemicals as soyasaponins, triterpenoidal glycosides that possess multiple health-promoting properties, such as lowering of cholesterol. In this work, the quantification of soyasaponins I and βg in 60 raw and cooked legumes by using a solid phase extraction (SPE) coupled to a liquid chromatography (LC)-mass spectrometry (MS) method was carried out. Results showed that lentils are a good source of soyasaponins, with a content of soyasaponin I that ranged from 636 to 735 mg kg(-1) and of soyasaponin βg from 672 to 1807 mg kg(-1). The cooking process produced a small loss of soyasaponins in water, that is, 4.8-8.7%, and partially converted soyasaponin βg …

Exposure estimates to Fusarium mycotoxins through cereals intake.

Abstract Mycotoxins are harmful substances produced by fungi in several commodities with a widespread presence in foodstuffs. Human exposure to mycotoxins occurs mainly by contaminated food. The quantitation of mycotoxins in cereal-based food, highly consumed by different age population, is of concern. In this survey, 159 cereal-based samples classified as wheat, maize and rice-based, have been evaluated for the occurrence of patulin, deoxynivalenol, 3-acetyl-deoxynivalenol, fusarenon-X, diacetoxyscirpenol, nivalenol, neosolaniol, HT-2, T-2 and zearalenone by gas chromatography–tandem mass spectrometry. Intakes were calculated for average consumers among adults, children and infants and com…

Determination of macrolide antibiotics in meat and fish using pressurized liquid extraction and liquid chromatography–mass spectrometry

We developed a method for determining the quantities of seven macrolide antibiotics in meat and fish by using pressurized liquid extraction (PLE) and liquid chromatography-mass spectrometry with electrospray ionization (LC-(ESI)MS). The PLE was optimized with regard to solvents, temperature, pressure, extraction time and number of cycles. The optimum conditions were: methanol as the extraction solvent; a temperature of 80 degrees C; a pressure of 1500psi; an extraction time of 15min; 2 cycles; a flush volume of 150% and a purge time of 300s. All recoveries for macrolide antibiotics were over 77% at 200mug/kg, except for erythromycin, which was 58%. The repeatability and reproducibility on d…

Beauvericin exposure induces cell cycle arrest, cell death and mitochondrial membrane potential disruption in CHO-K1 Cells

Solid phase techniques in the extraction of pesticides and related compounds from foods and soils

The application of SPE technology to the isolation of pesticides and related compounds from food and soils has grown enormously in the last decade. Much of this growth has been due to the relative ease of sample handling and the wide range of solid supports currently available for a variety of applications. The aim of this review is to present the methods for solid phase extraction (SPE) of pesticide residues from soils and foods. There are three main areas according to the type of approach used to handle the sample: solid phase extraction, solid phase clean-up (SPC), and matrix solid phase dispersion (MSPD). This review covers milk products, fatty foods, fruits, vegetables, and soils. Soli…

Development a mitigation strategy of enniatins in pasta under home-cooking conditions

The effect of pasta cooking on the enniatins (ENs) contents was evaluated in white and whole-grain pasta. The cooking of pasta was performed for 10 min in boiling water. ENs were extracted from raw and cooked pasta by Solid-Liquid extraction with acetonitrile, and from cooking water by Dispersive Liquid–Liquid Micro-Extraction (DLLME) with a mixture of acetonitrile and carbone tetrachloride. Mycotoxin analysis was performed by Liquid Chromatography coupled to Tandem Mass Spectrometry. The effect of pasta cooking has a marked influence on the reduction of ENs levels. Reductions of 98–100% for ENA, 94–95% for ENA1, 14–49% for ENB and 53–65% for ENB1 were achieved after the process. ENs were d…

Production, purification, and mass spectrometry characterization of the cyclohexadepsipeptide enniatin J3and study of the cytoxicity on differentiated and undifferentiated Caco-2 cells

The enniatins (EN) are bioactive compounds of hexadepsipeptidic structure produced by several strains of Fusarium spp. The enniatin J3 (EN J3) was purified from extracts of Fusarium solani growth on solid medium of kamut, using semipreparative liquid chromatography (LC) followed by an analytical LC. The purity and the structure of the isolated compound were confirmed by electrospray ionization-mass spectrometry study-linear ion trap (ESI-MS-LIT). The pure fraction of EN J3 was utilized to study the cytotoxicity on differentiated and undifferentiated Caco-2 cells. The use of both chromatographic techniques allowed us to produce and purify 50 mg of EN J3 completely characterized with the tech…

Solid-Phase Extraction of Organochlorine Pesticides from Water Samples

Abstract A procedure for the accumulation of organochiorine pesticides by reversed-phase adsorption on octadecylsilica glass microcolumns was used for the isolation and concentration from spiked distilled and natural waters. The pesticides were eluted from the octadecylsilica with light petroleum, and analyzed by gas chromatography using an electron capture detector. The average recovery was over 90% in spiked water samples at 0.1 ng/ml. The performance of the octadecylsilica when applied to the analysis of water samples were compared with the Rodier and the APHA solvent extraction methods.

Comparative cytotoxicity effect of zearalenone and its metabolites on the CHO-K1 cells

SPME of 52 pesticides and polychlorinated biphenyls: Extraction efficiencies of the SPME coatings poly(dimethylsiloxane), polyacrylate, poly(dimethylsiloxane)-divinylbenzene, Carboxen-poly(dimethylsiloxane), and Carbowax-divinylbenzene

Fiber/water partition coefficients (Kfw) of 52 pesticides, including triazines, organophosphorus pesticides, organochlorine pesticides, and polychlorinated biphenyls (PCBs) were experimentally calculated for the five polymeric coatings poly(dimethylsiloxane) (PDMS), polyacrylate (PA), poly(dimethylsiloxane)-Divinylbenzene (PDMS-DVB), Carboxen-poly(dimethylsiloxane) (CAR-PDMS), and Carbowax-divinylbenzene (CW-DVB) commercially available for solid-phase microextraction coupled to gas chromatography. The equilibrium-time profiles for the five coatings were previously studied in order to establish the time needed for equilibrium. To calculate Kfw the amount of analyte extracted at equilibrium w…

Reactive oxygen species involvement in apoptosis and mitochondrial damage in Caco-2 cells induced by enniatins A, A1, B and B1

The cytotoxic effects, the generation of reactive oxygen species (ROS) and lipid peroxidation (LPO) as well as the cell cycle disruption, the induction of apoptosis and changes in mitochondrial membrane potential (ΔΨm) as a function of increasing time have been determined in human colorectal adenocarcinoma (Caco-2) cells after exposure to enniatins (ENs) A, A₁, B and B₁. IC₅₀ values obtained by the MTT and Neutral Red assay, after 24, 48 and 72 h of exposure ranged from 0.5±0.1 to >15 μM. A significant increase (p≤0.05) in ROS generation and LPO production, as determined by the fluorescent probe H2-DCFDA and TBARS method respectively, was observed for all mycotoxins tested at 3.0 μM concent…

Comparison of toxic effects on CHO-KL and VERO cells after individual and combined treatment with Fusarium mycotoxins, beauvericin, deoxynivalenol and the T-2 toxin

Determination of organochlorine pesticide residues in honey from the central zone of Portugal and the Valencian community of Spain

In this study nine organochlorine pesticide residues ([alpha]-, [beta]-, and [gamma]-hexachlorocyclohexane (HCH), hexachlorobenzene (HCB), aldrin, p,p'-DDE, p,p'-DDD, o,p'-DDT, and p,p'-DDT) in forty nine samples of honey collected from markets of Portugal and Spain during 2001 and 2002, respectively, were evaluated. For this evaluation, three analytical procedures were studied. The analytical procedure, based on LLE extraction with ethyl acetate followed by gas chromatography-electron-capture detection (GC-ECD) for quantification, and mass spectrometry (GC-MS) for confirmation, has been selected. Recoveries of spiked samples ranged from 68%, for [beta]-HCH, and 126% for p,p'-DDT, for forti…

Mitochondrial transcriptional study of the effect of aflatoxins, enniatins and carotenoids in vitro in a blood brain barrier model

C. maxima (var. Delica), a variety of pumpkin, is well known for its high concentration on carotenoids, possessing dietary benefits and antioxidant properties. Aflatoxins and enniatins are common mycotoxins present in food and feed with an extended toxicity profile in humans and animals. Both types of substances reach a wide range of tissues and organs and have the capability to penetrate the blood brain barrier. Since carotenoids and mycotoxins have been reported to modify diverse mitochondrial processes individually, transcriptional in vitro studies on human epithelial cells ECV 304 were conducted to analyze the relative expression of 13 mitochondria related genes. ECV 304 cells were diff…

Co-occurrence and risk assessment of mycotoxins in food and diet from Mediterranean area.

The contents of 14 mycotoxins were studied in samples of different cereals and cereal products from four countries of the Mediterranean region. Two hundred and sixty-five samples from Spain, Italy, Morocco and Tunisia were analysed. Samples were extracted with matrix solid-phase dispersion (MSPD) and determined by liquid chromatography-tandem mass spectrometry with a triple quadrupole mass analyser. The percentage of total samples contaminated was 53%. The frequency of contaminated samples from Spain, Italy, Tunisia and Morocco was 33%, 52%, 96% and 50%, respectively. Nivalenol and beauvericin were the most predominant mycotoxins. This is the first international report to study the presence…

Disturbance of antioxidant capacity produced by beauvericin in CHO-K1 cells

Glutathione (GSH) levels, glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione-S-transferase (GST) as antioxidant defense system were evaluated in CHO-K1 cells after beauvericin (BEA) exposure. The effect of N-acetyl-cysteine (NAC) pre-treatment was assessed. GSH levels significantly decrease 18% and 29% after 5 μM of BEA in fresh medium and NAC pre-treatment, respectively compared to their controls. The GPx activity increased significantly from 35% to 66% in fresh medium and 20% in NAC pre-treatment. GR activity decreased after 5 μM of BEA up to 43% and 53% in fresh medium and NAC pre-treatment, respectively. The GST activity increased in fresh medium (from 61% to 89%) …

Proteomics evaluation of enniatins acute toxicity in rat liver

Abstract Enniatins (ENs) are emerging mycotoxins produced by Fusarium fungi which are cytotoxic also at low concentrations due to its ionophoric properties. The aim of this study was to evaluate the hepatic toxicity of ENs exposure at different concentrations in Wistar rats through a proteomic approach. Animals were intoxicated by oral gavage with medium (EN A 256, ENA1 353, ENB 540, ENB1 296 μg/mL) and high concentrations (ENA 513, ENA1 706, ENB 1021, ENB1 593 μg/mL) of an ENs mixture and sacrificed after 8 h. Protein extraction was performed using powdered liver. Peptides were analyzed using a liquid chromatography coupled with a quadrupole time-of-flight mass spectrometer. Proteins were …

Multi-mycotoxin analysis in wheat semolina using an acetonitrile-based extraction procedure and gas chromatography–tandem mass spectrometry

A new analytical method for the rapid and simultaneous determination of ten mycotoxins including patulin, zearalenone and eight trichothecenes (nivalenol, fusarenon-X, diacetoxyscirpenol, 3-acetyl-deoxynivalenol, neosolaniol, deoxynivalenol, T-2 and HT-2) in wheat semolina has been developed and optimized. Sample extraction and purification were performed with a modified QuEChERS-based (acronym of Quick, Easy, Cheap, Effective, Rugged and Safe) procedure and determined by gas chromatography (GC) coupled to triple quadrupole instrument (QqQ). This is the first paper on the application of GC-QqQ-MS/MS to analysis of mycotoxins. Careful optimization of the gas chromatography-tandem mass spectr…

Alternariol-induced cytotoxicity in Caco-2 cells. Protective effect of the phenolic fraction from virgin olive oil.

The extra virgin olive oil (EVOO) has been associated to antioxidant effects. The mycotoxin alternariol (AOH) can contaminate olives. The aims of this work were to determine the cytotoxic effects and reactive oxygen species (ROS) produced by AOH, tyrosol and oleuropein (two polyphenols of olive oil) and a real EVOO extract in Caco-2 cells. The MTT assay and the ROS production by the H2-DCFDA probe were used. Results demonstrated that AOH reduces cellular proliferation depending on concentration, whereas tyrosol and oleuropein did not (12.5-100 μM). The combination of AOH + oleuropein (50 μM) increased cell proliferation (24%) whereas, AOH + tyrosol decreased (47%) it. Besides, AOH increased…

Preliminary Estimation of Deoxynivalenol Excretion through a 24 h Pilot Study

A duplicate diet study was designed to explore the occurrence of 15 Fusarium mycotoxins in the 24 h-diet consumed by one volunteer as well as the levels of mycotoxins in his 24 h-collected urine. The employed methodology involved solvent extraction at high ionic strength followed by dispersive solid phase extraction and gas chromatography determination coupled to mass spectrometry in tandem. Satisfactory results in method performance were achieved. The method"s accuracy was in a range of 68%108%, with intra-day relative standard deviation and inter-day relative standard deviation lower than 12% and 15%, respectively. The limits of quantitation ranged from 0.1 to 8 μg/Kg. The matrix effect w…

Determination of dithiocarbamates and metabolites in plants by liquid chromatography–mass spectrometry

Abstract A quantitative matrix solid-phase dispersion and liquid chromatography–atmospheric pressure chemical ionization mass spectrometry (LC–APCI–MS) method is outlined for the simultaneous analysis of dithiocarbamates (DTCs) and their degradation products in plants. Compounds analyzed are dazomet, disulfiram, thiram and the metabolites ethylenthiourea and propylenthiourea. The performance of two different sample preparation protocols, the proposed one and other based on solid-phase extraction, as well as, of both atmospheric pressure ionization sources, APCI and electrospray, were compared. The effect of several parameters on the extraction, separation and detection was studied. Dithioca…

Environmental and food applications of LC-tandem mass spectrometry in pesticide-residue analysis: An overview

An overview is given on pesticide-residue determination in environmental and food samples by liquid chromatography/mass spectrometry/mass spectrometry (LC/MS/MS). Pesticides comprise a large number of substances that belong to many completely different chemical groups, the only common characteristic is that they are effective against pests. They still constitute a challenge in MS because there is no collective pathway for fragmentation. A brief introduction to the theory of tandem MS permits a discussion of which parameters influence the ionization efficiency when the ions are subjected to different actions. Emphasis is placed on the different tandem MS instruments: triple and ion-trap quad…

Mycotoxin contamination in laboratory rat feeds and their implications in animal research

Compound feed is particularly vulnerable to multi-mycotoxin contamination. A method for the determination of 12 mycotoxins; enniatins A, A1, B, B1; aflatoxins B1, B2, G1, G2; OTA; ZEA; T-2 and HT-2 by liquid chromatography-tandem mass spectrometry has been developed and applied for the analysis of laboratory rat commercial feeds. The method trueness was checked by recovery assays at three different spiked levels (n = 9). Recoveries ranged from 73% to 112%, and the intra-day and inter-day precision were lower than 9% and 13%, respectively. Limits of quantitation were lower than 15 μg/kg. Twenty-seven laboratory rats feed samples showed multi-contamination by at least three up to six differen…

Involvement of enniatins-induced cytotoxicity in human HepG2 cells.

Enniatins (ENNs) are mycotoxins found in Fusarium fungi and they appear in nature as mixtures of cyclic depsipeptides. The ability to form ionophores in the cell membrane is related to their cytotoxicity. Changes in ion distribution between inner and outer phases of the mitochondria affect to their metabolism, proton gradient, and chemiosmotic coupling, so a mitochondrial toxicity analysis of enniatins is highly recommended because they host the homeostasis required for cellular survival. Two ENNs, ENN A and ENN B on hepatocarcinoma cells (HepG2) at 1.5 and 3 μM and three exposure times (24, 48 and 72 h) were studied. Flow cytometry was used to examine their effects on cell proliferation, t…

Pesticide residue determination in fruit and vegetables by liquid chromatography–mass spectrometry

An overview is given of pesticide residue determination in fruit and vegetables by liquid chromatography-mass spectrometry (LC-MS). Emphasis is placed on the thermospray, particle beam and atmospheric pressure ionization interfaces including advantages and drawbacks and typical detection limits. The capacity of each interface to provide useful data for identification/confirmation of analytes and the possibility of obtaining structural information for the identification of target and non-target compounds is discussed. Finally, sample preparation techniques are dealt with in relation to their influence on further LC-MS determination.

Risk assessment of mycotoxins in coffee beverages

Effect of soyasaponin I against cytotoxicity and oxidative stress induced by alternariol in Caco-2 cells

Degradation study of enniatins by liquid chromatography–triple quadrupole linear ion trap mass spectrometry

Abstract Enniatins A, A1, B and B1 (ENs) are mycotoxins produced by Fusarium spp. and are normal contaminants of cereals and derivate products. In this study, the stability of ENs was evaluated during food processing by simulation of pasta cooking. Thermal treatments at different incubation times (5, 10 and 15 min) and different pH (4, 7 and 10) were applied in an aqueous system and pasta resembling system (PRS). The concentrations of the targeted mycotoxins were determined using liquid chromatography coupled to tandem mass spectrometry. High percentages of ENs reduction (81–100%) were evidenced in the PRS after the treatments at 5, 10 and 15 min of incubation. In contrast to the PRS, an im…

Effects of technological processes on enniatin levels in pasta

BACKGROUND Potential human health risks posed by enniatins (ENs) require their control primarily from cereal products, creating a demand for harvesting, food processing and storage techniques capable to prevent, reduce and/or eliminate the contamination. In this study, different methodologies to pasta processing simulating traditional and industrial processes were developed in order to know the fate of the mycotoxin ENs. The levels of ENs were studied at different steps of pasta processing. The effect of the temperature during processing was evaluated in two types of pasta (white and whole-grain pasta). Mycotoxin analysis was performed by LC-MS/MS. RESULTS High reductions (up to 50% and 80%…

Chronic cumulative risk assessment of the exposure to organophosphorus, carbamate and pyrethroid and pyrethrin pesticides through fruit and vegetables consumption in the region of Valencia (Spain).

In the present study, the chronic cumulative exposure to organophosphorus (OPs), carbamates (CBs) and pyrethroid and pyrethrin (PPs) pesticides in the region of Valencia through fruit and vegetables consumption is presented. A total of 752 samples and 84 pesticides were studied of which, 52 were OPs, 23 CBs and 9 PPs. Residue data were derived from the Valencia Region monitoring program 2007-2011 and food consumption levels from a questionnaire-based dietary survey conducted in 2010 in the same area. The relative potency factor (RPFs) approach was used to estimate chronic cumulative dietary exposure to OPs, CBs and PPs using acephate, oxamyl and deltamethrin as index compounds, respectively…

Determination of triazines and organophosphorus pesticides in water samples using solid-phase extraction.

Abstract Octadecyl (C 18 )-bonded porous silica was evaluated for the extraction of triazine and organophosphorus pesticides from natural water. The extraction results showed an effective performance when 1 l of water was passed through small glass columns containing 500 mg of 50–100-μm C 18 bonded porous silica. The absorbed compounds were removed with ethyl acetate, evaporated to 200 μl and determined by gas chromatography. The overall average recoveries were greater than 85% except for dimethoate and trichlorfon. Application of this procedure to the analysis of natural water samples gave results that agree well with those obtained by solvent extraction methods.

Optimization of experimental conditions for the identification of pesticide mixtures on six GLC columns

Quantitative analysis of six pesticides in fruits by capillary electrophoresis-electrospray-mass spectrometry.

A method to identify and quantify six pesticide residues - dinoseb, pirimicarb, procymidone, pyrifenox, pyrimethanil, and thiabendazole - in peaches and nectarines using capillary electrophoresis-electrospray ionization-quadrupole ion trap-tandem mass spectrometry (CE-ESI-MS/MS) is described. Separation was carried out using a buffer of 0.3 M ammonium acetate at pH 4 with 10% methanol. Pesticide residues present in peach and nectarine samples were preconcentrated by solid-phase extraction using C(18), eluted with CH(2)Cl(2), concentrated to dryness, and redissolved in buffer to obtain lower detection limits. The recoveries of the analytes ranged from 58 to 99% and the relative standard devi…

Assessment of Pesticide Residues in Honey Samples from Portugal and Spain

Fifty samples of honey collected from local markets of Portugal and Spain during year 2002 were analyzed for 42 organochlorine, carbamate, and organophosphorus pesticide residues. An analytical procedure based on solid-phase extraction with octadecyl sorbent followed by gas chromatography-mass spectrometry (GC-MS), for organochlorines, and by liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (LC-APCI-MS), for organophosphorus and carbamates, has been developed. Recoveries of spiked samples ranged from 73 to 98%, except for dimethoate (40%), with relative standard deviations from 3 to16% in terms of repeatability, and from 6 to 19% in terms of reproducibility. …

Evaluation of Mycotoxin Residues on Ready-to-Eat Food by Chromatographic Methods Coupled to Mass Spectrometry in Tandem

Simultaneous determination of twenty-seven mycotoxins in ready-to-eat food samples using &ldquo

Influence of dissolved humic material and ionic strength on C8 extraction of pesticides from water

A mixture of twelve intermediate polarity pesticides, mostly carbamate-type, have been chromatographied on an octadecyl-silica bonded-phase column using acetonitrile-water gradient mobile phase and UV detection. Pesticides were extracted from aqueous matrix by solid-phase extraction. The extraction system consisted of a glass microcolumn containing octyl-silica solidphase conditioned with methanol and water. The elution was by acetonitrile-dichloromethane (50:50). Studies of extraction factors like volume of sample, addition of 10% sodium chloride and presence of dissolved humic material are investigated.

Evaluation of 10 pesticide residues in oranges and tangerines from Valencia (Spain)

Abstract One hundred and sixteen orange and tangerine samples from an agricultural cooperative of the Valencian Community (Spain) were analyzed for bitertanol, carbendazim, hexythiazox, imazalil, imidacloprid, methidathion, methiocarb, pyriproxyfen, thiabendazole, and trichlorfon by liquid chromatography–mass spectrometry (LC–MS) after a conventional multiresidue extraction procedure. Of 52 samples that contained pesticide residues, carbendazim was detected in 27 (51.9%) in the concentration range of 0.02–0.04 mg kg−1, hexythiazox in 22 (42.3%) in the concentration range of 0.02–0.05 mg kg−1, imazalil in 8 (15.0%) in the concentration range of 0.02–1.2 mg kg−1, imidacloprid in 5 (9.6%) in a…

Effect of different thermal processes in the reduction of enniatins in fish tissues

Analysis of mycotoxins in coffee and risk assessment in Spanish adolescents and adults.

Mycotoxins are toxic compounds produced by fungal secondary metabolism that cause toxicological effects. Coffee is a highly popular beverage that is susceptible to contamination by mycotoxigenic fungi. The aim of the present study was to determine the presence of the following 21 mycotoxins in coffee using liquid chromatography tandem mass spectrometry (LC-MS/MS-IT): aflatoxin B1, B2, G1 and G2; ochratoxin A; nivalenol; deoxynivalenol; 3-acetyldeoxynivalenol; 15-acetyldeoxynivalenol; diacetoxyscirpenol; neosolaniol; T-2 and HT-2 toxin; sterigmatocystin; enniatin A, A1, B, and B1; beauvericin; and fumonisin B1 and B2. We aimed to determine differences by coffee process (coffee maker, electri…

Determination of mycotoxins in fruit berry by-products using QuEChERS extraction method

Abstract Fruit berries by-products are mainly elaborated with berry but most of them mixed with other fruits, grapes, plums or apples. These present favourable properties for the growth of a great range of mycotoxins mould producers. Consequently, alternariol (AOH), alternariol monomethyl ether (AME), tentoxin (TEN), aflatoxins (AFB1, AFB2, AFG1, AFG2) and ochratoxin A (OTA) are the main mycotoxins that should be present in berries by-products. To establish a reliable analytical method of these mycotoxins on two fruit berries by-products (jam and juice), four different QuEChERS extraction method were evaluated. Recoveries obtained were higher than 74% and 76% in berries juice and jam berrie…

Cytotoxic effects of mycotoxin combinations in mammalian kidney cells

The cytotoxicity of three Fusarium mycotoxins (beauvericin, deoxynivalenol and T-2 toxin) has been investigated using the NR assay, after 24, 48 and 72h of incubation. The IC(50) values ranged from 6.77 to 11.08, 3.30 to 10.00 and 0.004 to 0.005 for beauvericin, deoxynivalenol and T-2 toxin, respectively. Once the potential interaction has been detected, a quantitative assessment is necessary to ensure and characterize these interactions, that is, each mycotoxin contributes to the toxic effect in accord with its own potency. Combination of mycotoxins was determined in Vero cells after 24, 48 and 72h of exposure. Isobolograms and median effect method of Chou and Talalay were used to assess t…

Analysis of fumonisins B1, B2 and B3 in corn-based baby food by pressurized liquid extraction and liquid chromatography/tandem mass spectrometry

A sensitive and reliable method using pressurized liquid extraction (PLE) and liquid chromatography (LC)/electrospray ionization (ESI) tandem mass spectrometry with a triple quadrupole (QqQ) analyzer has been developed for the analysis of fumonisin B(1) (FB(1)), fumonisin B(2) (FB(2)) and fumonisin B(3) (FB(3)) in corn-based baby foods. Influence of several extraction parameters that affect PLE efficiency such as temperature, pressure, solvent extraction, number of cycles and dispersant/clean-up agents were studied. The selected PLE operating method was: 3g of sample was packed into 11 ml stainless-steel cell and fumonisins were extracted with methanol at 40 degrees C, 34 atm in one cycle o…

Bioaccessibility and bioavailability of the enniatins A, A1, B, B1 contained in a commercial wheat crispy bread

Dissipation and distribution of atrazine, simazine, chlorpyrifos, and tetradifon residues in citrus orchard soil.

An environmental fate study was conducted in a citrus orchard plot in Valencia (Spain) in the fall of 1993. Dissipation and distribution of atrazine, simazine, chlorpyrifos and tetradifon residues following their controlled addition for agricultural purposes in a mediterranean red soil (Luvic Calcisol, Rhodoxeralf) were evaluated. During a two-month period, the amounts of applied pesticides in different soil layers (0-0.05, 0.05-0.22, 0.22-0.42, and 0.42-0.52 m) were monitored. In addition, information on soils, weather and agricultural practice were collected. Degradation half-lives were calculated, assuming zero-order kinetics: 11 days for atrazine, 12 days for simazine, 10 days for chlor…

Aldicarb residues in citrus soil, leaves and fruits

Aldicarb was applied to the soil of Late Valencia orange field plots. A comparative study between four analytical methods is reported, the NPD-gas chromatography method being the best choice. The work also reports the study of accumulation and persistence of aldicarb and its biologically active metabolites in the soil, leaves and fruits (rind and pulp, separately). Residue content, at the ppb level, was found in the order leaves much greater than rind greater than pulp. Carbamate was found to accumulate mainly in its oxidized forms. Residues had reached a maximum level in about 90 days after application.

Gas chromatographic behaviour of urea herbicides

Gas chromatographic conditions for determining eight phenylurea (chlortoluron, diuron, fluometuron, isoproturon, linuron, metabenzthiazuron, metobromuron and monuron) and one sulfonylurea (chlorsulfuron) herbicides were assessed. Degradation products of the herbicides formed in the injector were used for identification. Most phenylureas formed their respective carbamic acid methyl esters, metabenzthiazuron formed an aminobenzothiazol and chlorsulfuron formed an aminotriazine plus a phenylsulfonamide. On-column injection of standards using a BP10 capillary column was evaluated to identify the chromatographic behaviour. Detection limits ranged from 0.05 ng for chlorsulfuron to 3 ng for monuro…

The role of pumpkin pulp extract carotenoids against mycotoxin damage in the blood brain barrier

Some mycotoxins such as beauvericin (BEA), ochratoxin A (OTA), and zearalenone (ZEA) can cross the blood brain barrier, which is why we tested the anti-inflammatory action of a pumpkin carotenoid extract (from the pulp) against these mycotoxins and their combinations (OTA+ZEA and OTA+ZEA+BEA) on a blood brain barrier model with co-cultured ECV304 and C6 cells using an untargeted metabolomic approach. The cells were added with mycotoxins at a concentration of 100 nmol/L per mycotoxin and pumpkin carotenoid extract at 500 nmol/L. For control we used only vehicle solvent (cell control) or vehicle solvent with pumpkin extract (extract control). After two hours of exposure, samples were analysed…

Determination of Urea Pesticide Residues in Vegetable, Soil, and Water Samples

The main physico-chemical, toxicological, and environmental properties of urea pesticides are summarized. General characteristics of analytical methods for residues of phenylurea herbicides (PUHs), sulfonylurea herbicides (SUHs), and Benzoylurea insecticides (BUIs) in crops, soil, and water samples, employed in the last 5 years are reviewed. Provided it is information about liquid-liquid and solid-phase extraction of the samples and clean-up steps. The applications of gas chromatography (GC), liquid chromatography (LC), and capillary electrophoresis (CE) techniques in the analysis of these compounds are exposed in tabular form and commented on. Sensitivity and instrument conditions of liqui…

Liquid chromatography/atmospheric pressure chemical ionization-mass spectrometric analysis of benzoylurea insecticides in citrus fruits.

A liquid chromatography (LC) method for the quantitative determination of three benzoylurea insecticide residues (diflubenzuron, flufenoxuron and hexaflumuron) in citrus fruits is described. Residues were successfully separated on a C18 column by methanol/water gradient elution. Detection was by negative-ion, selected-ion monitoring atmospheric pressure chemical ionization-mass spectrometry (APCI-MS); the main ions were [M - H]-, and the secondary fragment ions were [M - H - HF]-. Useful confirmatory information can thus be obtained at low extraction voltages from losses of HF. Detection limits for standard solutions were 10 fg injected and good linearity and reproducibility were obtained. …

Applications of flow cytometry to toxicological mycotoxin effects in cultured mammalian cells: a review.

This review gives an overview of flow cytometry applications to toxicological studies of several physiological target sites of mycotoxins on different mammalian cell lines. Mycotoxins are secondary metabolites of fungi that may be present in food, feed, air and water. The increasing presence of mycotoxins in crops, their wide distribution in the food chain, and their potential for toxicity demonstrate the need for further knowledge. Flow cytometry has become a valuable tool in mycotoxin studies in recent years for the rapid analysis of single cells in a mixture. In toxicology, the power of these methods lies in the possibility of determining a wide range of cell parameters, providing valuab…

Cytotoxicity, biaccessibility and transport by Caco-2 cells of enniatins and beauvericin

Further data on the presence of Fusarium emerging mycotoxins enniatins, fusaproliferin and beauvericin in cereals available on the Spanish markets.

In this work, 64 samples of cereals purchased from local markets in the Valencian community (Spain) were investigated for the presence of six emerging mycotoxins: enniatins ENs (ENA, ENA1, ENB and ENB1), beauvericin (BEA) and fusaproliferin (FUS). Samples were extracted with a mixture of water/acetonitrile (85/15, v/v) by using an Ultra-turrax homogenizer. Mycotoxins were then identified and quantified with a liquid chromatography (LC) with diode array detector (DAD). Positive samples were confirmed with an LC-MS/MS. Analytical Results showed that the frequencies of contamination of samples with ENs, BEA and FUS were 73.4%, 32.8% and 7.8%, respectively. ENA1 was the most mycotoxin found and…

Dietary exposure to trace elements and health risk assessment in the region of Valencia, Spain: a total diet study

Dietary exposure of the Valencian region population to lead, cadmium, inorganic arsenic (iAs), chromium, copper, tin and methylmercury (meHg) was assessed in a total diet study carried out in the region of Valencia in 2010–11. A total of 8100 food samples were collected and analysed. Occurrence data were combined with consumption data to estimate dietary exposure in adults (&gt; 15 years of age) and young children (6–15 years of age). The estimated intake was calculated by a probabilistic approach. Average intake levels (optimistic scenario) for lead, iAs, chromium and tin were 0.21, 0.08, 1.79 and 1.87 µg kg−1 bw day−1 respectively; for Cd and meHg average intake levels were 0.77 and 0.54 …