Application of matrix solid phase dispersion to the determination of imidacloprid, carbaryl, aldicarb, and their main metabolites in honeybees by liquid chromatography–mass spectrometry detection

Abstract A method based on matrix solid phase dispersion (MSPD) using C18 as dispersant and dichloromethane–methanol as eluent and liquid chromatography–atmospheric pressure chemical ionization–mass spectrometry (LC–APCI–MS) has been developed for the simultaneous determination of imidacloprid, 6-chloronicotinic acid, carbaryl, aldicarb, aldicarb sulfoxide, and aldicarb sulfone in honeybees. The proposed method was compared with liquid–liquid extraction (LLE) combined with LC–APCI–MS analysis. Spiked blank samples were used as standards to counteract the matrix effect observed in the chromatographic determination. Recovery studies were performed at different fortification levels. Average re…

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 …

FoodSimplex as a Mean to Improve Portuguese Restaurants’ Goods Manufacturing Practices - Audit and Microbial Assessment

Background: The consumption of meals at resturants has increased mass catering establishments. Such food services may present several risk factors for foodborne diseases, which have a substantial negative impact on public health. The smallest business with no technical or financial support struggles to comply with food safety European Regulation. The aim of this four years study was to assess Good Manufacture Practices (GMP) in Portuguese restaurants applying a designed food safety methodology, FoodSimplex, through audit data to determine good manufacture practices, and microbiological sampling of the meals before and after its implementation, by food groups according to their risk. Method…

Inhibitory effect of sweet whey fermented by Lactobacillus plantarum strains against fungal growth: A potential application as an antifungal agent

Abstract: The presence of mycotoxigenic fungi such as Aspergillus, Penicillium, and Fusarium genera represents a problem in food preservation and consequently, its spoilage. During the fermentation process with lactic acid bacteria, a range of secondary metabolites associated with beneficial health effects were released. In the present study, goat whey fermented by Lactobacillus plantarum (CECT 220, 221, 223, and 748) species has shown a satisfactory inhibitory effect against 28 fungi, showing for certain species of Fusarium genus and also, for Aspergillus steynii, a value of minimum inhibitory concentration until 1.95 g/L. In addition, phenyllactic acid was identified in each sample of fer…

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

Optimization of LC–MS/MS using triple quadrupole mass analyzer for the simultaneous analysis of carbosulfan and its main metabolites in oranges

This paper describes an analytical method involving a simple solvent extraction for the simultaneous liquid chromatography coupled to quadrupole tandem mass spectrometry (LC-MS/MS) determination of carbosulfan, its most toxic metabolite--carbofuran--, and its other main metabolites--3-hydroxycarbofuran, 3-ketocarbofuran, 3-hydroxy-7-phenolcarbofuran, 3-keto-7-phenolcarbofuran, 7-phenolcarbofuran and dibutylamine--in oranges. Chromatography was performed on a Zorbax Bonus-RP (150 mm x 2.1 mm, 5 microm). The mobile phase was a ternary gradient water-methanol-acetonitrile with 1.0 mM ammonium acetate at flow rate of 0.2 ml min(-1). The LC separation and MS/MS optimization were studied to selec…

Occurrence of fumonisins B1 and B2 in broa, typical Portuguese maize bread

Fumonisin B1 (FB1) and fumonisin B2 (FB2) are mycotoxins mainly produced by Fusarium verticillioides, and Fusarium proliferatum, fungi species most commonly isolated from maize. The natural occurrence of FB1 and FB2 in broa, typical Portuguese maize bread, was evaluated in 30 samples. Twenty five were found positive with levels ranging from 142 to 550 [mu]g kg- 1. The limit established by the European regulations was exceeded by 27% of the samples. The tolerable daily intake for fumonisin B1, and B2, alone or in combination, for all of the analysed samples, was lower than 2 [mu]g kg- 1 body weight per day established by the European Commission. http://www.sciencedirect.com/science/article/B…

Analysis of Chlorpyrifos in Water, Fruit Juice, and Honeybee Extract by Chemiluminescent Elisa

Abstract The suitability of competitive enzyme-linked immunosorbent assays (ELISAs) with chemiluminescent detection-based immobilized antigen (indirect assay) for rapid and accurate determination of chlorpyrifos in various food matrices was tested. The limit of detection (LOD) values were 1–1.75 ng mL−1, the standard curve midpoint (IC50) was 3.5 ng mL−1, and the assay duration was 1.5 h. Assay application to the analysis of honeybee extract resulted in chlorpyrifos recoveries varying between 62 and 83% in 5–15 ng mL−1 herbicide concentration range.

Occurrence of ochratoxin A in bread consumed in Morocco

Abstract One hundred samples of commercial bread purchased from January to October (2006) from retail bakeshops in five different cities (Rabat, Temara, Sale, Casablanca and Meknes) in Morocco were surveyed for the presence of ochratoxin A (OTA) using liquid chromatography coupled to fluorescence detection. The identification of OTA in positive bread samples was confirmed by methyl ester derivatization. Analytical results showed that forty eight (48%) samples were positive with OTA greater than the limit of quantification (LOQ = 0.051 ng/g). Levels of OTA in positive samples ranged between 0.14 and 149 ng/g. The average contamination of bread samples with OTA was 13 ± 1.5 ng/g. The highest …

Antifungal activity of natamycin and development of an edible film based on hydroxyethylcellulose to avoid Penicillium spp. growth on low-moisture mozzarella cheese

Abstract This study aimed to investigate natamycin's in vitro antifungal activity against Penicillium strains and determine its antifungal effect on Mozzarella cheese slices contaminated with P. commune. Besides, the residual content of natamycin in the samples was assessed during the storage. Natamycin was either sprayed on the cheese surface or incorporated into an edible hydroxyethylcellulose film at concentrations of 0, 0.25, 0.5, and 1 mg/dm2. Natamycin showed an antifungal effect against all Penicillium strains with MIC values ranging from 1.6 to 3.1 μg/mL and MFC values ranging from 12.5 to 200.0 μg/mL. Moreover, both applied techniques significantly (p ≤ 0.05) reduced the population…

Antibacterial effect of the bioactive compound beauvericin produced by Fusarium proliferatum on solid medium of wheat.

To obtain the bioactive compound beauvericin (BEA), Fusarium proliferatum CECT 20569 was grown on a solid medium of wheat, utilizing the technique of the solid state fermentation (SSF), being this mycotoxin purified by high performance liquid chromatography (HPLC) with a reverse phase semi-preparative column using as the mobile phase acetonitrile/water in gradient condition. The purity of the BEA was verified by analytical HPLC and liquid chromatography tandem mass spectrometry (LC/MS-MS). The pure fractions of BEA were utilized to determinate the antibiotic effects on several bacterial strains that are considered normally pathogens of the intestinal tract as: Escherichia coli, Enterococcus…

Capillary electrophoresis for the determination of pesticide residues

Capillary electrophoresis (CE) is becoming an advantageous tool for determining pesticide residues in environmental matrices because of its simplicity and high separation efficiency. However, inadequate limits of detection (LODs) and a lack of selective detectors limit the technique. The present review gives an overview of current developments in off-column and on-column approaches to trace enrichment in analysing real samples, and summarises the determination of pesticides residues by conventional CE as well as by the emerging techniques, such as CE-mass spectrometry (MS). The usefulness of these approaches in environmental applications is documented.

Influence of the heat treatment on the degradation of the minor Fusarium 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 investigated the degradation of the minor Fusarium mycotoxin BEA present in the concentration of 5 mg/kg in a model solution and in different crispy breads produced with different flours typologies (corn, hole, wheat, durum wheat, soy and rice) during the heat treatment carried out in an oven at three different temperatures of 160, 180 and 200 °C and at 3, 6, 10, 15 and 20 min incubation. The concentration of the bioactive compound studied, analyzed with the technique of the liquid chromatography tandem mass spectrome…

Aflatoxins levels in dried fruits and nuts from Rabat-Salé area, Morocco

Abstract The occurrence of aflatoxins (AFs) in dried fruits and nuts available in Rabat-Sale area (Morocco) was surveyed in this study. One hundred samples of dried fruits and nuts, purchased from retail shops and local markets from January to October 2006, were analyzed for AFs content by immunoaffinity (IAC) clean-up with liquid chromatography and fluorescence detection. Results showed that the incidences of total aflatoxins (AFT) and aflatoxin B1 (AFB1) in peanut, dried raisins, dried figs, walnut, and pistachio were 5%, 20%, 30%, 30% and 45% and 5%, 20%, 5%, 30% and 45%, respectively. The highest contamination levels of AFB1 were found in one walnut sample (2500 μg/kg) and one pistachio…

Occurrence of Aflatoxins in Tigernuts and Their Beverages Commercialized in Spain

A method based on matrix solid phase dispersion extraction was applied to determine aflatoxins B(1), B(2), G(1), and G(2) from tigernuts and tigernut beverages. Recoveries of each aflatoxin from tigernut (spiked at 10 microg/kg level) and from tigernut beverages (spiked at 10 microg/L level) ranged from 72.3 to 82.1% and from 74.0 to 86.3%, respectively. The limits of quantification ranged from 0.21 to 1.49 microg/kg (for tigernuts) and from 0.13 to 0.57 microg/L (for tigernut beverages) studied using liquid chromatography with fluorescence detection. The proposed extraction method followed by liquid chromatography-fluorescence detection determination was applied to 37 and 25 samples of tig…

Off-line solid-phase microextraction and capillary electrophoresis mass spectrometry to determine acidic pesticides in fruits.

A method based on solid-phase microextraction (SPME) and capillary electrophoresis/mass spectrometry (CE/ MS) is described for determining simultaneously five acidic pesticides (o-phenylphenol, ioxynil, haloxyfop, acifluorfen, picloram) in fruits. The CE device is coupled to an electrospray interface by a commercial sheath-flow adapter. Emphasis is placed on fulfillment of the speed and sensitivity requirements. The best separation is achieved using 32 mM ammonium formate/acid formic buffer at pH 3.1, with a working voltage of 25 kV. The MS detection of the five pesticides was performed in negative ionization mode. Full-scan spectra with base peaks corresponding to [M-H]- were obtained exce…

Multi-Mycotoxin Analysis in Durum Wheat Pasta by  Liquid Chromatography Coupled to Quadrupole  Orbitrap Mass Spectrometry.

A simple and rapid multi-mycotoxin method for the determination of 17 mycotoxins  simultaneously  is  described  in  the  present  survey  on  durum  and  soft  wheat  pasta  samples.  Mycotoxins included in the study were those mainly reported in cereal samples: ochratoxin-A  (OTA), aflatoxin B1 (AFB1), zearalenone (ZON), deoxynivalenol (DON), 3-and 15-acetyl-deoxynivalenol  (3-AcDON and 15-AcDON), nivalenol (NIV), neosolaniol (NEO), fusarenon-X, (FUS-X), T-2 toxin  (T-2) and HT-2 toxin (HT-2), fumonisin B1 and B2 (FB1 and FB2), and four emerging mycotoxins:  three  enniatins  (ENA,  ENA1,  and  ENB),  and  beauvericin  (BEA).  Twenty-nine  samples  were  analyzed to provide an overview on…

Food-Based Dietary Guidelines around the World: A Comparative Analysis to Update AESAN Scientific Committee Dietary Recommendations

Food-Based Dietary Guidelines (FBDG) include dietary recommendations based on food groups according to the general and accepted nutrition principles and current scientific evidence. Adoption of FBDG contributes to the prevention of malnutrition in all its forms, promotes human health, and reduces environmental impact. The present review aims to perform an international comparative analysis of the FBDG adopted in different countries from three different continents (America, Asia, and Europe), with particular reference to the Spanish Food Safety and Nutrition Agency (AESAN, Agencia Española de Seguridad Alimentaria y Nutrición) Scientific Committee dietary recommendations. A total of twelve c…

Occurrence of Fusarium mycotoxins and their dietary intake through beer consumption by the European population

Abstract Since cereals are raw materials for production of beer and beer-based drinks, the occurrence mycotoxins in 154 beer samples was topic of investigation in this study. The analyses were conducted using QuEChERS extraction and gas chromatography–tandem mass spectrometry determination. The analytical method showed recoveries for vast majority of analytes ranged from 70% to 110%, relative standard deviations lower than 15% and limits of detection from 0.05 to 8 μg/L. A significant incidence of HT-2 toxin and deoxynivalenol (DON) were found in 9.1% and 59.7% of total samples, respectively. The exposure of European population to mycotoxins through beer consumption was assessed. No toxicol…

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…

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

Translation and adaptation to spanish language of the quality of life questionnaire for celiac people called Canadian Celiac Health Survey

Introducción: Adaptar y valorar el cuestionario de calidad de vida denominado Canadian Celiac Health Survey (CCHS). Objetivo: Traducir y adaptar en castellano el cuestionario CCHS para poder ser utilizado por la población de habla hispana puesto que se trata de un cuestionario específico para la celiaquía. Método: La adaptación del CCHS, que consta de 76 ítems distribuidos en 11 secciones diferentes, se realizó mediante el método de traducción-retrotraducción y tras ser revisado y consensuado se procedió a realizar una prueba piloto con 25 personas celíacas, de forma individual y por un miembro del grupo de investigación, para valorar la comprensión de los ítems y sus secciones. Las aportac…

Rapid whole protein quantification of staphylococcal enterotoxin B by liquid chromatography

Abstract Food poisoning caused by Staphylococcus aureus is one of the most important foodborne diseases in the world. The ability of these bacteria to produce one or more enterotoxins in milk and dairy products is linked to staphylococcal food poisoning. Enterotoxin B (SEB) is an exotoxin produced by S. aureus and is one of the compounds most frequently involved in staphylococcal food poisoning worldwide. In this work, 20 samples of milk collected from restaurants have been studied for the presence of S. aureus enterotoxigenic strains. All the isolates from milk samples have been analysed by liquid chromatography-coupled with diode array detector for the rapid identification and quantificat…

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…

Evaluation of Organophosphorus Pesticide Residues in Citrus Fruits from the Valencian Community (Spain)

Abstract Approximately 200 citrus samples from markets of the Valencian Community (Spain) were analyzed to establish their residue levels in 12 organophosphorus pesticide residues during the 1994-1995 campaign. The organophosphorus pesticides carbophenothion, chlorpyriphos, chlorfenvinphos, diazinon, ethion, fenitrothion, malathion, methidation, methylparathion, phosmet, quinalphos, and tetradifon were simultaneously extracted by matrix solidphase dispersion and determined by gas chromatography- mass spectrometry using selected ion monitoring mode. A total of 32.25% contained pesticide residues and 6.9% exceeded the European Union Maximum Residue Levels (MRLs). The pesticides found in the s…

Multi-mycotoxin contamination of green tea infusion and dietary exposure assessment in Moroccan population

Green tea infusion is one of the most widely drunk beverages worldwide due to its health benefits associated with microelements, essential oils, and polyphenols, etc. Several studies have reported that green tea is subjected to contamination by various toxigenic fungi. Thus, this work aims to investigate the co-occurrence of 15 mycotoxins [four aflatoxins (AFB1, AFB2, AFG1, AFG2), ochratoxin A (OTA), beauvericin (BEA), four enniatins (ENA, ENA1, ENB, ENB1), zearalenone (ZEN), alternariol (AOH), tentoxin (TENT), T-2 and HT-2 toxins] in green tea samples available in Morocco by liquid chromatography tandem mass spectrometry method. Analytical and consumption data were then used to assess the …

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…

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…

Incidence of microbial flora in lettuce, meat and Spanish potato omelette from restaurants

A total of 370 samples including lettuce, meats (beef, pork and chicken) and Spanish potato omelette from restaurants were studied to evaluate the incidence of Escherichia coli, E. coli O157:H7,Staphylococcus aureus, Salmonella spp.,Yersinia enterocolitica, enterococci and some micro-organisms that can cause spoilage orcan be usedas indicators for food safety. Escherichia coliand enterococci were harboured with the highest incidence in lettuce, whereas incidence of Staphylococcus aureus was higherin meat than in the other foods studied. Enterobacter cloacae and Klebsiella pneumoniaewere isolated from the three food groups. Chryseomonas luteola, Enterobacter sakazakii, Klebsiella ozaenae, Mo…

In vitro biosynthesis of beauvericin and enniatins by Fusarium avenaceum and Fusarium poae strains isolated from Italian durum wheat grains

Assessment of the microbiological quality and wash treatments of lettuce served in University restaurants

Abstract One hundred and forty-four samples of lettuce from 16 University restaurants were analyzed. The mesophilic aerobic counts of all samples ranged from 3.01 to 7.81 log 10 CFU g −1 . Results of total coliforms ranged from 3.38 log 10 most probable number (MPN) g −1 . Of the lettuce samples, 25.7% harbored Escherichia coli , 22.9% Staphylococcus aureus and 84% group D streptococci. Similarly, 10.4% of the samples harbored Aeromonas hydrophila , 2.8% Pseudomonas aeruginosa , and coliforms such as 14.6% Citrobacter freundii , 8.3% Klebsiella pneumoniae , 4.2% Enterobacter cloacae and 1.4% Providencia spp. Salmonella , Shigella and E. coli O157:H7 were not detected. When sodium hypochlori…

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…

Identification and Quantification of Enniatins and Beauvericin in Animal Feeds and Their Ingredients by LC-QTRAP/MS/MS

Emerging fusariotoxins, mainly enniatins (ENNs) and beauvericin (BEA), are secondary toxic metabolites produced by Fusarium spp. and are widely distributed contaminants of cereals and by-products. Mycotoxin contamination in these products supposes an important risk to feed supply security in the feed industry due to the common use of cereals in feed formulations. Hence, continuous monitoring of both raw materials and feed mixtures is highly recommended as stated by sanitary authorities. Therefore, an analytical procedure based on liquid chromatography tandem mass spectrometry and an acetonitrile-based extraction followed by a d-SPE (QuEChERS) step for the simultaneous determination of emerg…

Biocatalyzed acidolysis of soybean oil triacylglycerols to increase oleic acid content

Lipase catalyzed acidolysis of triacylglycerols (TAG) of soybean oil with oleic acid in organic solvent was studied; immobilized lipase from Rhizomucor miehei was used and the effects of reaction time, incubation temperature and enzyme load on TAG total and positional fatty acid (FA) percentage compositions were investigated. The results show that oleic acid incorporation was high after 24 and 48 h, while after 72 h a lower level of oleic acid in TAG was observed. Moreover, for the reactions carried out at 30 and 40 degrees C, it was observed that the oleic acid level was about 46.5% while ligtly higher values (about 49%) were observed at 50 degrees C; however, under this last condition, th…

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.

Application of hybrid linear ion trap-high resolution mass spectrometry to the analysis of mycotoxins in beer

This paper reports the application of liquid chromatography electrospray ionization ion trap-orbitrap mass spectrometry for the determination of 18 mycotoxins (aflatoxins, fumonisins, trichothecenes, ochratoxin A, sterogmatocystin, beauvaricin, zearalenone and zearalenol) in beer. The extraction procedure was carried out by solid phase extraction (SPE): SPE columns were conditioned with acetonitrile/methanol and water. Beer was loaded onto the column which was washed with water. In these conditions, the recoveries were more than 65% and the relative standard deviation (RSD) were below 18%. The lowest limits of quantification (LLOQ) ranged from 9 to 155 ng ml(-1). Matrix-matched calibration …

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…

Isolation, purification, LC–MS/MS characterization and reactive oxygen species induced by fumonisin B1 in VERO cells

Fumonisins are mycotoxins produced by Fusarium verticillioides that commonly contaminate maize and maize products. The present work shows the results of a comparative study of three different fermentation's techniques (solid and liquid medium of corn and a solid agarized medium) for the production of fumonisins B-1, B-2 and B-3 with strains of F. verticillioides. The solid medium of corn was the most effective in the production of fumonisins, being Fumonisin B-1 the one produced with higher concentration, so the extract obtained by solid fermentation process was used for FB1 purification. Fumonisins characterization and quantification were performed with reversed-phase high-performance liqu…

Presence of Fusarium emerging mycotoxins in tiger-nuts commercialized in Spain

Abstract Tiger-nuts are tubers from the plant Cyperus esculentus which are commercialized in some regions of West and Central Africa, USA and Spain. These tubers have a high microbial charge such as bacteria, moulds and yeasts which could remain until the finished product. In this study we examined 47 samples of tiger-nut tubers purchased from Spanish supermarkets for contamination with the emerging Fusarium mycotoxins: Enniatins ENs (EN A, EN A1, EN B and EN B1), beauvericin (BEA) and fusaproliferin (FUS). The extraction of the samples was carried out with methanol using an Ultraturrax homogenizer. Mycotoxins were analyzed with a liquid chromatography (LC) coupled to a diode array detector…

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, …

On-line determination of bipyridylium herbicides in water by HPLC

Selective on-line solid phase extraction (SPE) and liquid chromatography determination (HPLC) of diquat, paraquat and difenzoquat from environmental water samples has been accomplished with Graphitized Carbon Black (GCB) as both extraction and analytical columns. The method involved passing of 50 mL of water through a cartridge filled with Carbograph. In the elution step, the herbicides were transferred from the cartridge to the analytical column (Hypercarb) by mean of a gradient of pH 3 aqueous solution of tetramethylammonium hydroxide (TMAOH) and ammonium sulphate and methanol. Hypercarb columns were found to give a low probability of false positives for bypiridylium herbicides and are ve…

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…

Ciclohexadespipeptide beauvericin degradation by different strains of Saccharomyces cerevisiae

Abstract The interaction between the mycotoxin beauvericin (BEA) and 9 yeast strains of Saccharomyces cerevisiae named LO9, YE-2, YE5, YE-6, YE-4, A34, A17, A42 and A08 was studied. The biological degradations were carried out under aerobic conditions in the liquid medium of Potato Dextrose Broth (PDB) at 25 °C for 48 h and in a food/feed system composed of corn flour at 37 °C for 3 days, respectively. BEA present in fermented medium and corn flour was determined using liquid chromatography coupled to the mass spectrometry detector in tandem (LC–MS/MS) and the BEA degradation products produced during the fermentations were determined using the technique of the liquid chromatography coupled …

Occurrence and co-occurrence of Fusarium mycotoxins in wheat grains and wheat flour from Romania

Abstract In this study, the presence of fourteen Fusarium mycotoxins, legislated by the European Union – deoxynivalenol, zearalenone, HT-2 and T-2 toxins (EC/1881/2006; 2013/165/EU), or non-legislated (five trichothecens and five “emerging” mycotoxins), was evaluated in 31 whole unprocessed wheat samples and 35 white wheat flour samples from different areas of Romania. For this purpose, a validated multi-mycotoxins liquid chromatography tandem mass spectrometry method was applied. Seventy three percent of the analyzed samples contained at least one mycotoxin. The highest occurrence was for enniatin B, 71% of the analyzed samples being positive (21–407 μg kg−1). Regarding the legislated myco…

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

Biosynthesis of beauvericin and enniatins in vitro by wheat Fusarium species and natural grain contamination in an area of central Italy

Contamination of wheat grain by beauvericin (BEA) and enniatins (ENs) is a global emerging mycotoxicological food problem. In this study, strains of Fusarium avenaceum (FA), Fusarium poae (FP), Fusarium equiseti and Fusarium sporotrichioides, all potential BEA and EN producers, isolated from 162 grain samples of durum and soft wheat harvested in 2009 and 2010 collected in an area of central Italy, were preliminarily screened for the presence of the esyn1 gene, encoding the multifunctional enzyme enniatin-synthetase for the detection of potential hexadepsipeptide-producing isolates. All positive isolates were tested for their ability to biosynthesize BEA and ENs in vitro. In addition, all wh…

Ochratoxin A in the morning and afternoon portions of urine from Coimbra and Valencian populations.

Abstract The widespread contamination of foodstuffs and beverages by mycotoxins, such as ochratoxin A (OTA), has made the monitoring of human contamination levels essential. By using a sensitive, accurate and speedy method that combines extraction with 5% NaHCO 3 , immunoaffinity column clean-up and HPLC with fluorescence detection, the human exposure to OTA through urine analysis can be monitored. This method is less invasive than blood monitoring and has the potential to be a good marker of human exposure. The limit of quantification of the method was 0.007 ng/mL of urine, with recoveries of OTA, from urine samples spiked at levels between 0.02 and 0.1 ng/mL, higher than 91% with RSD lowe…

Evaluation of enniatins A, A1, B, B1 and beauvericin in Portuguese cereal-based foods.

Sixty-one samples of Portuguese cereal-based foods were analysed for the occurrence of emerging mycotoxins called enniatins (A, A1, B and B1) and beauvericin. Samples were extracted with a mixture of acetonitrile/water (85/15, v/v) using an Ultra-Turrax homogeniser, and mycotoxins were detected with liquid chromatography coupled to a mass spectrometer. This method was validated and adequate values of recovery (70-103%) and relative standard deviation (15%) were obtained. Signal suppression/enhancement was studied and matrix-matched calibration used to minimise this effect, but no additional clean-up step was necessary. The mass spectrometer was operated in selected reaction monitoring (SRM)…

Determination of aflatoxins in peanuts by matrix solid-phase dispersion and liquid chromatography

A new method based on matrix solid-phase dispersion (MSPD) extraction was studied to determine aflatoxin B1, B2, G1 and G2 from peanuts. Optimization of different parameters, such as type of solid supports for matrix dispersion and elution solvents were carried out. The method used 2 g of peanut sample, 2 g of C18 bonded silica as MSPD sorbent and acetonitrile as eluting solvent. Recoveries of each aflatoxin spiked to peanut samples at 2.5 ng/g (5 ng/g for aflatoxin G2) level were between 78 and 86% with relative standard deviations ranging from 4 to 7%. The limits of quantification ranged from 0.125 to 2.5 ng/g for the four studied aflatoxins using liquid chromatography (LC) with fluoresce…

The importance of a registered dietitian in restaurants: a pilot study in Valencia (Spain)

University students have the option to purchase their meals at university restaurants; in general, food eaten away from home has high contents of saturated fat and energy. The aim of this work was to perform a pilot study of the nutritional adequacy of meals served at two university restaurants from Valencia. Energy, macronutrient and micronutrient contents were calculated and compared against the Spanish Recommended Dietary Intake (SRDI). Daily menus provided a high amount of energy; lunch from restaurant 1 provides 60 and 78% of the energetic SRDI for male and female, respectively. In restaurant 2 these values are 49.5 and 64.6%, respectively. Menus from one university restaurant are plan…

Presence of mycotoxin in commercial infant formulas and baby foods from Italian market

In this study a total of 75 commercially Italian samples of baby foods, including 13 infant formula milks (infant formula powders, ready-to-use preparation), 11 dairy products (cheese and yogurt), 25 cereal-based baby foods, 16 fruit and vegetables compotes, and 10 fruit and vegetables purees (composed of pear, peach, banana and for apple), were analyzed to provide an overview on mycotoxin presence. The presence was carried out by evaluating of 23 mycotoxins: ochratoxin-A (OTA), patulin (PAT), two aflatoxins (AFM1, AFB1), three zearalenones (ZONs), which include zearalenone (ZON) and its metabolites (α-zearalenol (α-ZOL) and β-zearalenol (β-ZOL)), nine trichothecenes: deoxynivalenol (DON), …

Liquid chromatography–electrospray quadrupole ion-trap mass spectrometry of nine pesticides in fruits

A liquid chromatographic method, with electrospray ionization tandem mass spectrometry (LC-ESI-MS-MS), has been developed for determining acrinathrin, carbosulfan, cyproconazole, lambda-cyhalothrin, kresoxim methyl, pyrifenox, pyriproxyfen, propanil, and tebufenpyrad in fruits. The ions prominent in ESI spectra were [M + H]+ and [M + Na]+. In the mass analyzer, collision-induced dissociation fragmentation involved common pathways, for example, product ions of [M + H]+ resulted from the cleavage of the carbamic group or an oxygen bound. The utility of the method is demonstrated by the analysis of crude extracts obtained by matrix solid-phase dispersion (MSPD) using C18 as dispersant and dich…

Antibacterial effects of enniatins J1 and J3 on pathogenic and lactic acid bacteria

Abstract Enniatins (ENs) are N -methylated cyclohexadepsipeptides, secondary metabolites produced by various species of the genus Fusarium . They are known to act as antifungal, antiyeast and antibacterial and to possess antiinsecticidal and phytotoxic properties. In this study we evaluated for the first time the antibiotic effect of pure fractions of EN J 1 and J 3 on several pathogenic strains and lactic acid bacteria. The ENs J 1 and J 3 were purified from the fermentation extract of Fusarium solani growth on solid medium of wheat kamut, using the technique of the low pressure liquid chromatography (LPLC) followed by a semipreparative liquid chromatography (LC). The purity and the struct…

Multi‐Mycotoxin Analysis in Durum Wheat Pasta by Liquid Chromatography Coupled to Quadrupole Orbitrap Mass Spectrometry

A simple and rapid multi‐mycotoxin method for the determination of 17 mycotoxins simultaneously is described in the present survey on durum and soft wheat pasta samples. Mycotoxins included in the study were those mainly reported in cereal samples: ochratoxin‐A (OTA), aflatoxin B1 (AFB1), zearalenone (ZON), deoxynivalenol (DON), 3‐and 15‐acetyl‐deoxynivalenol (3‐AcDON and 15‐AcDON), nivalenol (NIV), neosolaniol (NEO), fusarenon‐X, (FUS‐X), T‐2 toxin (T‐2) and HT‐2 toxin (HT‐2), fumonisin B1 and B2 (FB1 and FB2), and four emerging mycotoxins: three enniatins (ENA, ENA1, and ENB), and beauvericin (BEA). Twenty‐nine samples were analyzed to provide an overview on mycotoxin presence: 27 samples…

APPLICATION OF REAL-TIME POLYMERASE CHAIN REACTION FOR RAPID DETERMINATION OFSALMONELLAIN RESTAURANT FOODS

Several foods have been identified as frequently or occasionally contaminated with Salmonella, such as eggs, raw milk and all raw foods of animal origin that may carry salmonella and other bacteria. Salmonellosis is the most common food-borne illness, but it is usually preventable according to the implementation of a food safety system that includes Hazard Analysis and Critical Control Point. In this study, 96 food samples of the University restaurant of Valencia (Spain) were divided into four groups: first course; main course; accompaniment; and dessert, and were analyzed simultaneously with classical and fluorogenic polymerase chain reaction microbiological analysis. Neither of the proced…

Comparison of four mass analyzers for determining carbosulfan and its metabolites in citrus by liquid chromatography/mass spectrometry

Four liquid chromatography/mass spectrometry (LC/MS) systems, equipped with single quadrupole, triple quadrupole (QqQ), quadrupole ion trap (QIT) and quadrupole time-of-flight (QqTOF) mass analyzers, were evaluated for the analysis of carbosulfan and its main transformation products. The comparison of quantitative aspects (sensitivity, precision and accuracy) was emphasized. Results showed that the triple quadrupole instrument reaches at least 20-fold higher sensitivity (LOD from 0.04 to 0.4 microg kg(-1)) compared to the single quadrupole (4-70 microg kg(-1)), the QIT (4-25 microg kg(-1)) and the QqTOF (4-23 microg kg(-1)) instruments. Recoveries were over 70% for all the analytes, except …

Occurrence of fourteen mycotoxins in tiger-nuts

Abstract A previous developed matrix solid-phase dispersion (MSPD) extraction method was applied for the routine analysis of aflatoxins (AFs), ochratoxin A (OTA), fumonisins (FB 1 and FB 2 ), beauvericin (BEA), nivalenol (NIV), deoxynivalenol (DON), the toxin T-2 (T-2), toxin HT-2 (HT-2), diacetoxyscirpenol (DAS) and zearalenone (ZEN) in tiger-nuts by liquid chromatography–triple-quadrupole linear ion trap (HPLC–QTRAP ® ). The extraction solid support used was C 18 , while the elution solvent was acetonitrile/methanol (50/50, v/v) 1 mM ammonium formate. Using matrix-matched calibration, recoveries and repeatabilities were in the range 67–89% and 2–11% relative standard deviation (RSD), resp…

Determination of lead on the airborne particulates of urban Valencia city

A method for the determination of lead in airborne particulates by graphite furnace atomic absorption spectrometry after extraction with HCl-HNO3 (120–150°C) is described. Particulate lead levels in the atmosphere have been measured in samples collected throughout a year, sampling has been made at eleven sites within the city of Valencia. The results obtained in this study show that annual means of lead levels are within the range of 0.526 to 7.033 μg/m3 depending upon the sampling site. Lead levels are higher in winter and autumn. Lead and nitrogen oxide levels are well correlated.

Determination of Fusarium mycotoxins enniatins, beauvericin and fusaproliferin in cereals and derived products from Tunisia

Abstract In this study, 51 samples of cereals (wheat, Barley, maize and Sorghum) and by-products (mainly pasta and couscous) purchased from Tunisian supermarkets were examined for contamination with the emerging Fusarium mycotoxins: Enniatins ENs (EN A, EN A 1 , EN B and EN B 1 ), beauvericin (BEA) and fusaproliferin (FUS). The extraction of the samples was performed with methanol using an Ultra-turrax homogenizer. Mycotoxins were analyzed with a liquid chromatography (LC) coupled to a diode array detector (DAD). The frequencies of contamination of total samples with ENs were 96%. EN A 1 was the most common EN found with the highest prevalence of 92.1%, levels ranged between 11.1 and 480 mg…

Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry Analysis of Carbosulfan, Carbofuran, 3-Hydroxycarbofuran, and Other Metabolites in Food

The potential of liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QqTOF-MS) to identify and confirm carbosulfan and seven of its main metabolites (carbofuran, 3-hydroxycarbofuran, 3-ketocarbofuran, 3-hydroxy-7-phenol carbofuran, 3-keto-7-phenolcarbofuran, 7-phenolcarbofuran, dibutylamine) at trace levels from food is explored for the first time. The analytical method developed consists of pressurized liquid extraction (PLE) and LC-QqTOF-MS in positive ion mode, which attains unequivocal identification and quantification of the studied compounds in food, at levels well below of those of concern (0.05 mg/kg for the sum of carbosulfan, carbofuran, and 3-hydroxycarbofuran).…

Applicability of hybrid linear ion trap-high resolution mass spectrometry and quadrupole-linear ion trap-mass spectrometry for mycotoxin analysis in baby food.

Abstract Recent developments in mass spectrometers have created a paradoxical situation; different mass spectrometers are available, each of them with their specific strengths and drawbacks. Hybrid instruments try to unify several advantages in one instrument. In this study two of wide-used hybrid instruments were compared: hybrid quadrupole-linear ion trap-mass spectrometry (QTRAP ® ) and the hybrid linear ion trap-high resolution mass spectrometry (LTQ-Orbitrap ® ). Both instruments were applied to detect the presence of 18 selected mycotoxins in baby food. Analytical parameters were validated according to 2002/657/CE. Limits of quantification (LOQs) obtained by QTRAP ® instrument ranged …

Amylase–Trypsin Inhibitors in Wheat and Other Cereals as Potential Activators of the Effects of Nonceliac Gluten Sensitivity

Nonceliac gluten sensitivity (NCGS) is a gluten-related gastrointestinal disorder distinct from celiac disease (CD) and gluten allergy that is not easy to diagnose due to the lack of biomarkers. It is characterized by intestinal symptoms and extraintestinal manifestations with the consumption of gluten-containing foods. In contrast to CD, NCGS patients do not present a genetic predisposition or intestinal villi atrophy. Recent studies question the proinflammatory triggering activity of α-gliadin fraction contained in wheat, since it has been demonstrated that the amylase-trypsin inhibitors (ATIs) exert a strong activating effect on the innate immune response. We aimed to analyze the role of…

Influence of prebiotics, probiotics and protein ingredients on mycotoxin bioaccessibility

The aim of this study was to investigate the influence of prebiotic compounds (cellulose and inulin), food ingredients (milk whey, β-lactoglobulin and calcium caseinate) and several probiotic microorganisms on the bioaccessibility of beauvericin (BEA), enniatins (ENs A, A1, B, B1), deoxynivalenol (DON) and zearalenone (ZEA) present in wheat crispy bread produced with wheat flour previously fermented with F. tricinctum, F. culmorum and G. zeae. The bioaccessibility of mycotoxins was determined by a dynamic simulated gastrointestinal digestion system, imitating the human digestive physiological conditions of the gastrointestinal tract. Mycotoxins were determined in the simulated intestinal fl…

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

Incidence of ochratoxin A in rice and dried fruits from Rabat and Salé area, Morocco.

One hundred samples of dried fruits (20 dried raisins, 20 walnuts, 20 peanuts, 20 dried figs and 20 pistachios) and 20 samples of rice purchased from retail shops in the Rabat and Salé area in Morocco were analysed for ochratoxin A (OTA) by immunoaffinity clean-up (IAC) and liquid chromatography (LC) with fluorescence detection. The limit of quantification (LOQ) (S/N = 10:1) of OTA was 0.02 ng g(-1) in rice, 0.03 ng g(-1) in pistachio, peanut and walnut, and 0.03 ng g(-1) in dried raisins and dried figs. The incidences of occurrence of OTA in dried raisins, walnuts, peanuts, dried figs and rice were 30, 35, 25, 65 and 90%, respectively. Analytical results showed that pistachio samples conta…

Multimycotoxin Determination in Tunisian Farm Animal Feed

Mycotoxins presence was evaluated in animal feed marketed in Tunisia for the first time ever. A QuEChERS method was performed to analyze the natural copresence of 22 mycotoxins (enniatins, beauvericin, ochratoxin A, aflatoxins, alternariol monomethyl ether, alternariol, tentoxin, zearalenone, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, nivalenol, neosolaniol, diacetoxyscirpenol, T-2 toxin, and HT-2 toxin) in 122 Tunisian marketed feed samples, intended for poultry (n = 43), cattle (n = 35), rabbit (n = 12), sheep (n = 16), and horse (n = 16). Analytes detection and quantification were done using both liquid chromatography and gas chromatography coupled to tandem mass sp…

Survey of microbial quality of plant-based foods served in restaurants

This study was carried out to evaluate the microbiological quality of plant-based foods obtained from foodservice establishments. The samples included cereals, legumes, fruits and vegetables. According to the European Commission Regulation (No. 2073/2005 and No. 1441/2007) and Spanish microbiological criteria (No. 3484/2000), vegetables were the plant-based dishes where more samples exceed the adopted limits of mesophilic aerobic counts and Enterobacteriaceae. Furthermore, Staphylococcus aureus and Escherichia coli were also found in several vegetable dishes. E. coli and Salmonella spp. were detected in 6.6% and 0.7% of lettuce samples, respectively. However, all the samples were negative f…

Microbial evaluation of Spanish potato omelette and cooked meat samples in University restaurants.

The focus of this study was to evaluate the microbial quality of Spanish potato omelette and cooked meat samples including pork loin, chicken croquettes, long pork sausage, chicken breast, and meatballs from University restaurants. Microbiological analyses of Spanish potato omelette and cooked meat samples resulted in aerobic plate counts from1.00 to 2.90 and from1.00 to 6.04 log10 CFU g(-1), respectively. Total coliforms ranged from3 to 43 most probable number (MPN) g(-1) and from3 to2,400 MPN g(-1) for Spanish potato omelette and meat products, respectively. Escherichia coli, coagulase-positive staphylococci, and Lancefield group-D streptococci were detected in 1.7%, 3.5%, and 12.9% of Sp…

Exposure assessment of fruits contaminated with pesticide residues from Valencia, 2001– 03

A total of 634 samples of oranges, tangerines, peaches, nectarines, khakis and watermelons were collected from an Agricultural Valencia Community Cooperative during the May 2001 to April 2003 campaigns and they were analysed for 15 pesticides among those recommended for pest treatment. A conventional multiresidue analytical procedure based on ethyl acetate extraction was used followed by gas chromatography coupled to a nitrogen phosphorus detector for routine analysis; and mass spectrometry was performed for confirmation. Recovery studies with spiked samples at 0.5 mg kg-1 for each pesticide ranged from 52% for acephate to 87% for fenthion with a standard deviation20%. Limits of quantificat…

Incidence of microorganisms from fresh orange juice processed by squeezing machines

Abstract This study was carried out to evaluate the microbiological quality of orange juice obtained from squeezing machines in foodservice establishments. The samples included fresh squeezed orange juice and juice which is maintained in metal jugs until consumption. According to the European Commission Regulation (No. 2073/2005 and No. 1441/2007) and Spanish microbiological criteria (No. 3484/2000), 12% and 43% of the total examined lots exceed the adopted limits of mesophilic aerobic counts and Enterobacteriaceae, respectively. Possibly, this contamination is caused by incorrect handling of oranges and juices and also by inadequate cleaning and sanitization of squeezing machine and metal …

Chemoprotective effect of carotenoids from Lycium barbarum L. on SH-SY5Y neuroblastoma cells treated with beauvericin

Goji berry has recently been introduced in Mediterranean diet and its consumption is increasing. This study aims to determine cytoprotection of lutein (LUT), zeaxanthin (ZEAX) and goji berry extract (GBE) rich in carotenoids against Beauvericin (BEA)-induced cytotoxicity on SH-SY5Y neuroblastoma cells. Both carotenoids and GBE showed cytoprotective effects. Cytoprotection was evaluated by simultaneous combination of the two xanthophylls LUT and ZEAX with BEA, as well as using pre-treatment assays. The highest protective effect occurred in 16%, 24% and 12% respectively for LUT, ZEAX and LUT + ZEAX incubating simultaneously with BEA, while by pre-treatment assay LUT showed a cytoprotection ef…

Application of an HPLC-MS/MS method for mycotoxin analysis in commercial baby foods

Abstract This article describes the validation of an analytical method for the detection of 21 mycotoxins in baby food. The analytical method is based on the simultaneous extraction of selected mycotoxins by matrix solid-phase dispersion (MSPD) followed by liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS) using a hybrid triple quadrupole-linear ion trap mass spectrometer (QTRAP®). Information Dependent Acquisition (IDA), an extra confirmation tool for samples that contain the selected mycotoxins, was used. The matrix effects were evaluated, and the corrections for the matrix effects were performed using two calibration approaches: external matrix-matched calibration and…

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…

A review of the application of the hazard analysis and critical control point system to salads served in the restaurant of Valencia University

Development of an Antifungal and Antimycotoxigenic Device Containing Allyl Isothiocyanate for Silo Fumigation

The aims of this study were to evaluate the antifungal activity of the bioactive compound allyl isothiocyanate (AITC) against Aspergillus flavus (8111 ISPA) aflatoxins (AFs) producer and Penicillium verrucosum (D-01847 VTT) ochratoxin A (OTA) producer on corn, barley, and wheat. The experiments were carried out initially in a simulated silo system for laboratory scale composed of glass jars (1 L). Barley and wheat were contaminated with P. verrucosum and corn with A. flavus. The cereals were treated with a hydroxyethylcellulose gel disk to which 500 &micro

Occurrence of Fusarium mycotoxins in Italian cereal and cereal products from organic farming.

In the present study, the occurrence of eighteen mycotoxins, nine trichothecenes (deoxynivalenol, 3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, nivalenol, neosolaniol, diacetoxyscirpenol, fusarenon-X, T-2 toxin and HT-2 toxin), three zearalenones (zearalenone, α-zearalenol and β-zearalenol), and six emergent mycotoxins, beauvericin and five enniatins (A, A1, B, B1 and B4), was monitored in different Italian organic cereals and cereal products by using a liquid chromatography coupled to triple quadrupole mass spectrometry method. A total of 93 organic cereal samples (wheat, barley, rye and oat) were collected from Italy. Limits of quantification ranged from 5 to 15 μg/kg. 80% of analyze…

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,…

Levels of ochratoxin A in wheat and maize bread from the central zone of Portugal.

Ochratoxigenic fungi are natural contaminants of cereal and the produced toxins are harmful to humans and animals. Ochratoxin A (OTA) is among the most important mycotoxins, and the International Agency for Research on Cancer (IARC) classifies it as possibly carcinogenic to humans (group 2B). A total of 61 samples of bread from the central zone of Portugal were analysed for OTA by liquid chromatography (LC) with fluorescence detection (FD). For confirmation two procedures were applied, methyl ester derivatization with boron trifluoride-methanol and liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI/MS/MS). As far as we know, this is the first report where on-line…

Ochratoxin A in rice on the Moroccan retail market

One hundred (100) samples of rice purchased from retail markets in five different cities (Rabat, Témara, Salé, Casablanca and Méknès) in Morocco from January to October 2006 were surveyed for the presence of ochratoxin A (OTA) using Accelerated Solvent Extraction (ASE) coupled to liquid chromatography with fluorescence detection. The identification of OTA in positive rice samples was confirmed by methyl ester derivatization. Analytical results showed a frequency of contamination of 26% of total analyzed rice samples. The percentage of contamination of samples was 24, 26.6, 16.6, 27.7 and 30% in Rabat, Témara, Méknès, Salé and Casablanca respectively. Levels of OTA in positive samples ranged…

Presence of mycotoxins in sorghum and intake estimation in Tunisia

Sorghum samples (n = 60) from Tunisian markets were analysed for the occurrence of 22 of both traditional and emerging mycotoxins. Samples were extracted with a QuEChERS-like method and mycotoxins were detected by LC-MS/MS. This method was validated and adequate analytical parameters were obtained. All samples had contamination with mycotoxins and several samples had higher contamination levels than European Union legislative limits (MLs). The most frequently found mycotoxins were ENB (100%), OTA (98%), ENA₁ (63%), ENB₁ (56%), BEA (48%), AFB1 (38%) and STG (33%). Mean contaminations were 30.7, 1.93, 33.2, 51.0, 15.4, 1.49 and 20.5 µg kg(-1), respectively. While two samples were contaminated…

Isolation, purification and antibacterial effects of fusaproliferin produced by Fusariumsubglutinans in submerged culture.

To evaluate the fusaproliferin (FUS) production, Fusariumsubglutinans ITEM 2404 was grown in a liquid medium of potato being this mycotoxin purified by high-performance liquid chromatography (HPLC) with a C18 semipreparative column using a mobile phase of acetonitrile/H(2)O using gradient conditions. The purity of the fusaproliferin was verified by analytical HPLC, ultraviolet absorbance measurements, LC/MS-MS, (1)H NMR spectroscopy. The isolated FUS was shown to be free of impurities and can be used as a standard for routine analysis. The pure fusaproliferin was utilized to study the biological activity on Escherichiacoli and Staphylococcusaureus. This study demostred that FUS not showed s…

First report on the presence of emerging Fusarium mycotoxins enniatins (A, A1, B, B1), beauvericin and fusaproliferin in rice on the Moroccan retail markets

Seventy samples of rice purchased from local markets in six cities from Morocco (Rabat, Casablanca, Kenitra, Mohammadia, Tanger and Errachidia) were analyzed for the presence 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) by using an ultra-turrax homogenizer. Mycotoxins were then identified and quantified with liquid chromatography (LC) coupled to diode array detector (DAD). Positive samples were confirmed with an LC-MS/MS. Analytical results showed that BEA was present in 75.7% of total analyzed samples. BEA levels varied between 3.8 and 26.3 mg/kg…

Isolation, Identification and Investigation of Fermentative Bacteria from Sea Bass (Dicentrarchus labrax): Evaluation of Antifungal Activity of Fermented Fish Meat and By-Products Broths

During fish production processes, great amounts of by-products are generated, representing &asymp

Simultaneous analysis of twenty-six mycotoxins in durum wheat grain from Italy

Abstract A multi-mycotoxin analysis method based on liquid chromatography coupled to triple quadrupole mass spectrometry was validated and applied for the determination of twenty-six mycotoxins, including eight trichothecenes, three fumonisins, sterigmatocystin, ochratoxin A, four aflatoxins, zearalenone, five “emerging” mycotoxins and three Alternaria mycotoxins in 74 durum wheat samples from central Italy. Eighty-eight percent of the analysed samples contained one or more mycotoxins, with enniatin B showing the highest contamination levels (78% of samples with levels ranging from 23 to 1826 ng/g), followed by enniatin B1 (10–1384 ng/g) and deoxynivalenol (48–2267 ng/g). This work gives an…

Analysis of pesticide residues in fruit and vegetables by matrix solid phase dispersion (MSPD) and different gas chromatography element-selective detectors

A comparison between different element selective detectors for the determination of organophosphorus and organochlorine pesticide residues, from fruit and vegetables, was performed by capillary GC with electron capture detector (ECD), nitrogen phosphorus detector (NPD), flame photometric detector (FPD) in the sulphur and phosphorus modes, and mass spectrometry detector (MSD) in selected ion monitoring (SIM) mode. Pesticides were extracted from the different foodstuffs by Matrix Solid Phase Dispersion (MSPD). Recoveries of 41–108 % with relative standard deviation of 2–14 % in the concentration range 0.5–10 μg L−1 were obtained in oranges, lemons, grapefruit, pears, plums, lettuces and tomat…

Listeria species in raw and ready-to-eat foods from restaurants.

From September 1999 to March 2000, meat (pork, beef, and chicken), fish (salmon, hake, and sole), vegetable (lettuce and spinach), and Spanish potato omelette samples obtained at restaurants were collected and tested for the occurrence of Listeria spp. Listeria monocytogenes was isolated from 3 (2.9%) out of 103 studied samples. Other species isolated were Listeria grayi (13.6%), Listeria innocua (1.9%), Listeria ivanovii (5.8%), Listeria seeligeri (3.9%), and Listeria welshimeri (1.9%). Listeria was neither isolated from beef nor any type of fish.

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…

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…

Simultaneous determination of Fusarium mycotoxins in wheat grain from Morocco by liquid chromatography coupled to triple quadrupole mass spectrometry

Abstract In the present study, eighteen (18) mycotoxin produced by the genus Fusarium (fumonisins (FB1, FB2 and FB3), type-B trichothecenes (NIV, DON, FUS-X, 3Ac-DON and 15Ac-DON), type-A trichothecenes (NEO, DAS, T2 and HT2), zearalenone (ZEA), beauvericin (BEA), and enniatins (ENA, ENA1, ENB and ENB1)) were monitored in different samples of wheat grain commercialized in Morocco. A liquid chromatography coupled to triple quadrupole mass spectrometry method previous matrix solid phase dispersion extraction was used for sample analysis. A total of eighty (80) samples of durum wheat were collected in different local markets from several areas in Morocco. Analytical results showed that 54 out …

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…

Glucose influence on the production of T-2 toxin by Fusarium sporotrichioides

Toxigenic isolate of Fusarium sporotrichioides was tested for the T-2 toxin production on PDA plates during 10 days under various glucose concentrations. T-2 toxin was determined by LC-MS and confirmed with LC-MS/MS. This analytical method has been applied, for the first time, to an extensive study of T-2 accumulation. Results showed that the production of this mycotoxin is directly correlated to the concentration of glucose present in the medium. Concentrations of T-2 toxin produced by the strain of F. sporotrichioides ranged from 0 to 1.45 mg/kg. The better T-2 production was evidenced in the fermentation operated with 20% of glucose.

Application of White Mustard Bran and Flour on Bread as Natural Preservative Agents.

In this study, the antifungal activity of white mustard bran (MB), a by-product of mustard (Sinapis alba) milling, and white mustard seed flour (MF) was tested against mycotoxigenic fungi in the agar diffusion method. The results obtained were posteriorly confirmed in a quantitative test, determining the minimum concentration of extract that inhibits the fungal growth (MIC) and the minimum concentration with fungicidal activity (MFC). Since MF demonstrated no antifungal activity, the MB was stored under different temperature conditions and storage time to determine its antifungal stability. Finally, an in situ assay was carried out, applying the MB as a natural ingredient into the dough to …

Rapid whole protein quantitation of staphylococcal enterotoxins A and B by liquid chromatography/mass spectrometry

Abstract Staphylococcus aureus is an important pathogen and has been indicated as the fifth causative agent of food-borne human illness throughout the world. Staphylococcal enterotoxins (SEs) are toxic compounds excreted mainly by strains of S. aureus. Among these toxins, enterotoxins A (SEA) and B (SEB) are both of the most prevalent compounds in staphylococcal food poisoning. In this work, reverse phase liquid chromatography coupled to ESI mass spectrometry (LC–ESI/MS) has been applied for its rapid identification and quantification. Limit of detection (LOD) values were 0.5 and 0.2 ng for SEA and SEB, respectively and limit of quantification (LOQ) value was 1 ng for both enterotoxins. SEA…

Occurrence of Mycotoxins in Botanical Dietary Supplement Infusion Beverages.

The aim of the present work was to study the occurrence of mycotoxins [aflatoxins (1-4), 3-acetyldeoxyniavlenol (5), 15-acetyldeoxynivalenol (6), nivalenol (7), HT-2 (8), T-2 (9), ochratoxin A (10), zearalenone (11), enniatin A (12), enniatin A1 (13), enniatin B (14), enniatin B1 (15), and beauvericin (16)] present in potable products derived from herbal teas. Analysis was carried out by liquid chromatography coupled to ion-trap tandem mass spectrometry (LC-MS/MS-IT) after a dispersive liquid-liquid microextraction procedure (DLLME) was conducted. The DLLME method was applied to 52 commercial samples of chamomile, chamomile with anise, chamomile with honey, linden, pennyroyal mint, thyme, v…

A chemical approach for the reduction of beauvericin in a solution model and in food systems.

Abstract Beauvericin (BEA) is a bioactive compound produced by the secondary metabolism of several Fusarium strains with a strong antibacterial, antifungal, and insecticidal activities. This study evaluated the reduction of BEA added at 25 mg/kg in phosphate buffer saline (PBS) solutions at pH of 4, 7 and 10, or to different cereal products (kernels and flours) by the bioactive compounds phenyl isothiocyanate (PITC) and benzyl isothiocyanate (BITC). The concentration of the mycotoxin was evaluated using liquid chromatography coupled to the diode array detector (LC-DAD). In solution, BEA reduction ranged from 9% to 94% on a time-dependent fashion and lower pH levels resulted in higher BEA re…

Use of the modified quick easy cheap effective rugged and safe sample preparation approach for the simultaneous analysis of type A- and B-trichothecenes in wheat flour

A suitable extraction and purification method for the simultaneous liquid chromatography-mass spectrometry (LC-MS) determination of five mycotoxins, three type A, diacetoxyscirpenol (DAS), T-2 toxin (T-2) and HT-2 toxin (HT-2), and two type B-trichothecenes, deoxynivalenol (DON) and nivalenol (NIV), has been optimised using a modified "Quick Easy Cheap Effective Rugged and Safe" (QuEChERS) method. Different solvents were studied in the extraction procedure to obtain better recoveries, which ranged from 86 to 108%, using a 85/15 (v/v) mixture of methanol/acetonitrile. The values obtained for recovery, repeatability and reproducibility of the optimized method are in agreement with Commission …

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…

Transfer of Fusarium mycotoxins from malt to boiled wort

Malting was simulated using two different batches of barley as raw material: a naturally contaminated batch and laboratory inoculated (with a deoxynivalenol (DON) and zearalenone (ZEA) producing Fusarium graminearum strain) one. Up to three contamination levels were prepared, every process being carried out in triplicate. A significant washout effect on DON was observed by the end of the first water phase (between 22.4 and 34 % reduction) with an even more pronounced reduction (up to 75 % decrease) by the end of the steeping process. ZEA content remained almost unchanged (no significant difference between the initial and the final concentration). Germination was characterized by an increase…

Influence of different coffee drinks preparations on ochratoxin A content and evaluation of the antioxidant activity and caffeine variations.

Coffee is one of the most frequently consumed beverages in North America and Europe. It is well known that coffee contains caffeine and that coffee beans can be contaminated by Ochratoxin A (OTA). The operating conditions however affect OTA and caffeine extraction from the roasted coffee. OTA content found in the beverages can be greater than that found in the roasted coffee used to prepare it, representing a potential OTA related risk factor for the human health. Moreover the coffee beans and coffee based beverages have an anti oxidant activity. This study investigates the OTA content, the amount of caffeine, and the antioxidant activity in five different preparations: American coffee, Mok…

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…

Listeria spp. in Street-Vended Ready-to-Eat Foods.

Street-vended ready-to-eat food sold in Egypt, including sandwiches and dishes of traditional food, was examined for the presence ofListeriaspecies. Out of 576 samples, 24% were found to containListeriaspecies.L. monocytogenesandL. innocuawere isolated from 57% and 39% of the contaminated samples, respectively. OtherListeriaspp. were detected with lower frequency.L. monocytogenesof ≥103 CFU/g were detected in 7% of the total examined samples, which represent 49% of the contaminated food samples (meat, poultry, seafood, dairy products, and products of plant origin). Most of the samples contaminated byL. monocytogeneshad high levels of total viable bacterial counts. The results obtained may h…

Comparación del somatotipo, evaluación nutricional e ingesta alimentaria entre estudiantes universitarios deportistas y sedentarios

Resumen Fundamento y objetivo El estudio del somatotipo y de la ingesta alimentaria entre la poblacion universitaria es importante para implantar politicas internas de mejora de la salud y de la prevencion de enfermedades. En este estudio se ha realizado la comparacion del somatotipo, evaluacion energetica-nutricional e ingesta alimentaria entre estudiantes universitarios deportistas frente a los sedentarios. Sujetos y metodo La muestra esta formada por 1.299 jovenes universitarios (420 varones y 879 mujeres) de la Universitat de Valencia, durante 2008-2009 y 2009-2010, evaluandose en ellos diversos parametros antropometricos, el somatotipo y la ingesta nutricional y alimentaria mediante un…

Occurrence of Free and Conjugated Mycotoxins in Aromatic and Medicinal Plants and Dietary Exposure Assessment in the Moroccan Population.

Aromatic and medicinal plants (AMPs), as herbal material, are subjected to contamination by various mycotoxin-producing fungi, either free and conjugated. Such a problem is associated with poor storage practices, and lack of adopting good agricultural practices and good harvesting practices. Nevertheless, AMPs are poorly investigated. The purpose of this study was to investigate the co-occurrence of 15 mycotoxins (four aflatoxins (AFB1, AFB2, AFG1, and AFG2), ochratoxin A (OTA), beauvericin (BEA), four enniatins (ENA, ENA1, ENB, and ENB1), zearalenone (ZEN), alternariol (AOH), tentoxin (TENT), T-2, and HT-2 toxins) in 40 samples of AMPs frequently consumed in Morocco by using liquid chromat…

Development of a Bioactive Sauce Based on Oriental Mustard Flour with Antifungal Properties for Pita Bread Shelf Life Improvement

Ochratoxin A (OTA) is a mycotoxin produced in the secondary metabolism of fungus belonging to the genus Aspergillus and Penicillium. In this study, the employment of oriental mustard flour (OMF) as an ingredient in a packaged sauce was evaluated for the generation in situ of the antimicrobial compound allyl isothiocyanate (AITC) in order to preserve pita bread contaminated with Penicillium verrucosum VTT D-01847, an OTA producer, in an active packaging system. Four different concentrations (8, 16, 33 and 50 mg/g) were tested. Mycelium formation, mycotoxin production, AITC absorbed by the food matrix, and volatilization kinetics were studied for each concentration. The results obtained were …

Incidence of Staphylococcus aureus in meals from cafeterias

A study to determine the incidence of Staphylococccus aureus in 705 meals from several cafeterias in Valencia, Spain was undertaken. Out of the 705 studied meals, 81 samples (I I. I %) had coagulase positive staphylococci with a range of S. aureus from 100 to 53000 cfu/g. me four highest values (> l0,ooO S. aureus &/g) were obtained with Russian type salad, meatballs, chicken croquettes and salad. These products involved more handling stages. These data show that food handlers may contribute to food poisoning and that there are some handling practices that require more attention.

Occurrence of fumonisins in organic and conventional cereal-based products commercialized in France, Germany and Spain

A fast and effective method for quantitative determination of fumonisins B1 and B2 (FB1 and FB2) in different cereal-based products was developed. Fumonisins were extracted combining a solid liquid extraction (SLE) pre-treatment and immunoaffinity columns (IACs), which were applied for an effective clean-up. Consecutively, high pressure liquid chromatography (HPLC) coupled to hybrid triple quadrupole-linear ion trap mass spectrometer (QTrap®) was used for the separation, detection and quantification of targeted mycotoxins. The analytical method showed acceptable recoveries ranged from 89.7% to 99.1%, as well as a relative standard deviation (%, RSD) lower than 12%. A total of 1250 samples f…

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…

Aflatoxins and A. flavus Reduction in Loaf Bread through the Use of Natural Ingredients

In this study, the antifungal activity of yellow mustard (YMF) and oriental mustard (OMF) meal extracts against 14 strains of fungi was tested on a solid medium. The results obtained with the YMF were next confirmed in liquid medium determining the minimum inhibitory concentration (MIC) and the minimum fungicide concentration (MFC). Finally, the use of YMF as a natural preservative to extend the useful life of bread was evaluated. Breads with different concentrations of YMF (2, 4, 6 and 8 g/kg) were prepared and contaminated with Aspergillus flavus ISPA 8111 and Penicillium nordicum CECT 2320. For 10 days the formation of mycelium was observed, and after that the fungal growth and the mycot…

In vitro antifungal activity of allyl isothiocyanate (AITC) against Aspergillus parasiticus and Penicillium expansum and evaluation of the AITC estimated daily intake

Abstract Isothiocyanates (ITCs) are natural compounds derived from cruciferous vegetables produced by enzymatic conversion of metabolites called glucosinolates. They are potentially useful antimicrobial compounds for food applications have been shown to be promising agents against cancer in human cell culture, animal models, and in epidemiological studies. In this study, the antifungal activity of the allyl isothiocyanate (AITC) was evaluated on two mycotoxigenic fungi as Aspergillus parasiticus and Penicillium expnsum , aflatoxins (AFs) and patulin (PAT) producers, employing an assay on solid medium. Also an approximation of the risk evaluation associated to the intake of food treated with…

Coffee Silverskin and Spent Coffee Suitable as Neuroprotectors against Cell Death by Beauvericin and α-Zearalenol: Evaluating Strategies of Treatment

Coffee silverskin and spent coffee have been evaluated in a neuroblastoma cell line (SH-SY5Y cells) against beauvericin (BEA) and α-zearalenol (α-ZEL)-induced cytotoxicity with different strategies of treatment. First, the direct treatment of mycotoxins and coffee by-products extracts in SH-SY5Y cells was assayed. IC50 values for α-ZEL were 20.8 and 14.0 µM for 48 h and 72 h, respectively and, for BEA only at 72 h, it was 2.5 µM. Afterwards, the pre-treatment with spent coffee obtained by boiling water increased cell viability for α-ZEL at 24 h and 48 h from 10% to 16% and from 25% to 30%, respectively

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…

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…

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, …

Enterotoxinomics: The omic sciences in the study of staphylococcal toxins analyzed in food matrices

Abstract Foodomics covers all several disciplines related with emerging working areas and relies on them to get a broader knowledge about food quality and food safety to improve consumer's well-being, health and confidence. Enterotoxin-producing S. aureus is one of the most common causes of food-borne human illness throughout the world. Because of the possible significance of these enterotoxins for public health and food safety, greater knowledge of their occurrence and an efficient means of screening at the protein level are needed. It is under these circumstances where Enterotoxinomic science has an important role. The term Enterotoxinomics refers to the omic sciences applied to enterotox…

Probiotic characterization of Lactobacillus strains isolated from breast milk and employment for the elaboration of a fermented milk product

Abstract A total of seven strains of lactic acid bacteria (LAB) isolated from breast milk, were investigated for their probiotic properties. Lactobacillus plantarum 5H1 and 5L1 were selected for their beneficial properties and used for probiotic fermented milk manufacture. The fermentation parameters (pH and lactic acid) and LAB viability during fermented milk processing and storage were evaluated. The results showed that the strains had a broad spectrum of antimicrobial activity against pathogenic bacteria and toxigenic fungi. These also evidenced capacity of adhesion and reduction of Salmonella adhesion to Caco-2 cells. During milk fermentation process, an increase in lactic acid content,…

Reduction of the aflatoxins B1, B2, G1 and G2 in Italian piadina by isothiocyanates

Abstract Aflatoxins (AFs) are mycotoxins produced mainly by the molds Aspergillus flavus , Aspergillus parasiticus and A . nomius . These mycotoxins are contaminants of cereals. AFB 1 , the most abundant and toxic metabolite, is known to cause several toxic responses, such as hepatotoxicity, teratogenicity and mutagenicity. Isothiocyanates (ITCs) are natural compounds produced by the enzymatic hydrolysis of glucosinolates (GLs), which have shown potent antimicrobial activity in food applications. In this study, ITCs derived from oriental and yellow mustard (0.1, 0.5 and 1 g of flour) were used to avoid the production of AFs in piadina (a typical Italian flatbread) contaminated with A . para…

Reduction in vitro of the minor Fusarium mycotoxin beauvericin employing different strains of probiotic bacteria

Abstract The interaction between the minor Fusarium mycotoxins BEA and 13 bacterial strains characteristic of the gastrointestinal tract as Bifidobacterium longum , Bifidobacterium bifidum , Bifidobacterium breve , Bifidobacterium adolescentes , Lactobacillus rhamnosus , Lactobacillus casei-casei , Lactobacillus plantarum , Eubacterium crispatus , Salmonella fecalis , Salmonella termofilus , Lactobacillus ruminis , Lactobacillus casei and Lactobacillus animalis was studied. The fermentations were carried out in the liquid medium of MRS during 4, 12, 16, 24 and 48 h at 37 °C, under anaerobic conditions. Levels of BEA in the fermentation liquid, on the cell walls and on the internal part of t…

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…

Microbial contamination of milk and dairy products from restaurants in Spain

This study was carried out to evaluate the microbiological quality of milk and some dairy products, including 95 lots of warm milk, 95 lots of milk conserved at room temperature, and 75 lots of dairy products collected from restaurants in Spain. According to the European microbiological criteria (92/46/EEC, 93/43/EEC, and Commission Regulation No. 2073/2005), 31% and 35% of the total examined lots exceed the adopted limits of mesophilic aerobic counts and Enterobacteriaceae, respectively. This may be due to incorrect handling of milk and dairy products and inadequate cleaning and sanitization of milk container. Further, 2% of all the lots examined were positive for the presence of Escherich…

Analytical Methods for Pesticide Residue Determination in Bee Products

Monitoring pesticide residues in honey, wax, and bees helps to assess the potential risk of these products to consumer health and gives information on the pesticide treatments that have been used on the field crops surrounding the hives. The present review seeks to discuss the basic principles and recent developments in pesticide analysis in bee products and their application in monitoring programs. Consideration is given to extraction, cleanup, chromatographic separation, and detection techniques.

Presence of Enniatins and Beauvericin in Romanian Wheat Samples: From Raw Material to Products for Direct Human Consumption

In this study, a total of 244 wheat and wheat-based products collected from Romania were analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS) in order to evaluate the presence of four enniatins (ENs; i.e., ENA, ENA1, ENB, and ENB1) and beauvericin (BEA). For the wheat samples, the influence of agricultural practices was assessed, whereas the results for the wheat-based products were used to calculate the estimated daily intake of emerging mycotoxins through wheat consumption for the Romanian population. ENB presented the highest incidence (41% in wheat and 32% in wheat-based products), with its maximum levels of 815 μg kg−1 and 170 μg kg−1 in wheat and wheat-based products,…

A survey of mycotoxins in random street-vended snacks from Lagos, Nigeria, using QuEChERS-HPLC-MS/MS

Abstract A survey in African snacks was carried out in order to evaluate the intake of 23 mycotoxins. The African snack samples were purchased from street vendors within Lagos metropolis (Nigeria) and evaluated for the presence of 23 mycotoxins using a modified QuEChERS procedure coupled with liquid chromatography-triple quadrupole linear ion trap mass spectrometer. The snacks included akara, baked coconut, coconut candy, donkwa, groundnut cake (kulikuli), lafun, milk curd (wara), fresh and dried tiger-nuts, and yam flour. Only three mycotoxins were detected in 23.8% of the studied snacks, and at concentrations ranging from 6 to 54 μg kg−1. The concentrations of aflatoxin B1 (AFB1) and AFB2…

Occurrence of mycotoxins in refrigerated pizza dough and risk assessment of exposure for the Spanish population.

Mycotoxins are toxic metabolites produced by filamentous fungi, as Aspergillus, Penicillium and Fusarium. The first objective of this research was to study the presence of mycotoxins in 60 samples of refrigerated pizza dough, by extraction with methanol and determination by liquid chromatography associated with tandem mass spectrometry (LC-MS/MS). Then, the estimated dietary intakes (EDIs) of these mycotoxins, among the Spanish population, was calculated and the health risk assessment was performed, comparing the EDIs data with the tolerable daily intake values (TDIs). The mycotoxins detected in the analyzed samples were aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), zearale…

Multi-mycotoxin contamination of couscous semolina commercialized in Morocco.

The multi-mycotoxin contamination of ninety-eight (98) couscous semolina samples collected from various areas in Morocco was investigated in this study. Samples were surveyed for the presence of 22 mycotoxins (four aflatoxins, ochratoxin A, diacetoxiscyrpenol (DAS), three fumonisins, beauvericin (BEA), deoxynivalenol (DON), 15-acetyl-deoxynivalenol (15-ADON), 3-acetyl-deoxynivalenol (3-ADON), nivalenol (NIV), sterigmatocystin (STG), zearalenone (ZEA), four enniatins, T-2 and HT-2 toxins). Results showed that 96 out of 98 total couscous samples (98%) were contaminated by at least one mycotoxin. Enniatin B (ENB), Enniatin B1 (ENB1), Enniatin A1 (ENA1) and zearalenone (ZEA) have shown the high…

Development of an Antifungal Device Based on Oriental Mustard Flour to Prevent Fungal Growth and Aflatoxin B1 Production in Almonds

The present study describes the manufacture of an antifungal device composed of oriental mustard flour and hydroxyethyl-cellulose (H-OMF) and evaluates its efficacity in inhibiting Aspergillus flavus growth and aflatoxin B1 (AFB1) production in almonds. Additionally, it compares the H-OMF with allyl isothiocyanate (AITC) and a freeze-dried extract of yellow mustard flour (YMF-E); such substances were previously described as antifungal. Minimum inhibitory concentration (MIC), Minimum fungicidal concentration (MFC), the H-OMF in vitro antifungal activity, and the residual fungal population, as well as the production of AFB1 in almonds were determined. AITC and YMF-E showed significant antifun…

Natural co-occurrence of mycotoxins in wheat grains from Italy and Syria

This article describes the application of an analytical method for the detection of 25 mycotoxins in wheat grain based on simultaneous extraction using matrix solid-phase dispersion (MSPD) followed by liquid chromatography coupled to tandem mass spectrometry, a hybrid triple quadrupole-linear ion trap mass spectrometer (QTrapÒ). Information Dependent Acquisition (IDA), an extra confirmation tool for samples that contain the target mycotoxins, was used. The analysis of 40 Syrian and 46 Italian wheat grain sam- ples interestingly showed that Syrian samples were mainly contaminated with ochratoxin A and aflatox- ins, whereas Italian samples with deoxynivalenol and 15-acetyldeoxynivalenol. Emer…

Occurrence and daily intake of ochratoxin A of organic and non-organic rice and rice products

Abstract Ochratoxin A (OTA) was extracted from 84 rice samples and rice products by using accelerated solvent extraction (ASE) and analysed with liquid chromatography coupled with fluorescence detection. Samples were collected from rice cultivars, local markets and supermarkets; 64 were of non-organic and 20 of organic production. 7.8% of non-organic samples had OTA levels from 4.3 to 27.3 μg/kg and in 30% of organic samples was detected the presence of this mycotoxin varying from 1.0 to 7.1 μg/kg. OTA presence was confirmed by methyl-ester derivatization. Rice and rice products labelled with denomination of origin (DO) were not detected OTA due to the fact that its production has implement…

Mass spectrometry applications

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

Presence of aflatoxin M1 in pasteurized milk from Morocco

Fifty four samples of pasteurized milk produced by five different dairies from Morocco were surveyed for the presence of aflatoxin M1 (AFM1) using immunoaffinity columns and liquid chromatography coupled to fluorescence detection. Confirmation of AFM1 identity in positive samples was based on the formation of AFM1 hemi-acetal derivative (AFM2a) after derivatization with trifluoracetic acid. Analytical results showed that 88.8% of the samples were contaminated with AFM1; 7.4% being above the maximum level of 0.05 microg/L set by the Moroccan and European regulations for AFM1 in liquid milk. The incidence of AFM1 in milk from these dairies was 100, 92.3, 90, 83.3 and 77.7% respectively, with …

One-year monitoring of aflatoxins and ochratoxin A in tiger-nuts and their beverages.

A sensitive and selective liquid chromatography-triple quadrupole-tandem mass spectrometry (LC-ESI-MS-MS) method was developed for the routine analysis of aflatoxins (AFB(1), AFB(2), AFG(1) and AFG(2)) and ochratoxin A (OTA) in tiger nuts and tiger-nut beverage (horchata). A matrix solid phase dispersion was adapted to eliminate lipidic interferences. The solid support was C(18), while the elution solvent was acetonitrile. Mean recoveries obtained at two fortification levels were 72-83% and 71-81% for horchata and tiger nut respectively with relative standard deviations (RSDs) <13% and 15% respectively. The LC-MS-MS method allowed quantification and identification at low levels in two matri…

Simple liquid chromatography assay for analyzing ochratoxin A in bovine milk

Abstract Ochratoxin A (OTA) is a mycotoxin with teratogenic and carcinogenic properties. Animal intake of feedstuffs contaminated with OTA may cause that some residues may be found in bovine milk, therefore, its analysis requires a highly sensitive, simple and precise technique. This method is based on a liquid–liquid extraction with methanol, followed by filtration and extract concentration. Liquid chromatography coupled to fluorescence detection was used for OTA analysis. In this way, several impurities are filtered off and OTA is quantified with a mean recovery of 93.0 ± 7.4% and a limit of detection of 0.01 ng mL −1 . Therefore, this methodology allows a simple quantitative extraction o…

Evaluation of matrix solid-phase dispersion (MSPD) extraction for multi-mycotoxin determination in different flours using LC-MS/MS

An existing matrix solid-phase dispersion (MSPD) method for aflatoxins (AFs) and ochratoxin A (OTA) extraction was extended by further 14 mycotoxins. After it careful optimization, this method was applied to determine the occurrence of these mycotoxins on commercial flour samples (with different cereals composition) collected from local markets. In a total of 49 samples investigated, 9 mycotoxins were identified. Nivalenol (NIV) and Beauvericin (BEA) were the mycotoxins found most frequently. The samples that presented major contamination were wheat flours and bakery preparations. Despite of the great number of positives finding, only one wheat flour sample exceeded the maximum limits (ML) …

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…

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…

Cytoprotective effects of carotenoids-rich extract from Lycium barbarum L. on the beauvericin-induced cytotoxicity on Caco-2 cells.

Abstract In this work, the cytotoxicity of Beauvericin (BEA), lutein (LUT), zeaxanthin (ZEAX) and goji berries extract (GBE) rich in carotenoids, was investigated, as well as cytoprotective effects of these carotenoids against BEA induced-cytotoxicity on Caco-2 cells. Cytotoxicity was carried out using MTT and protein content (PC) assays during 24 and 48 h of exposure. Only BEA showed cytotoxic effect obtaining a reduction in cell proliferation range from 6.5 to 92.8%. Simultaneous combination of LUT and ZEAX with BEA slightly increased cell proliferation compared to BEA tested alone. LUT, ZEAX and GBE showed cytoprotective effects against cytotoxicity induced by BEA on Caco-2 cells. Pre-tr…

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…

A preliminary study of presence of resveratrol in skins and pulps of European and Japanese plum cultivars

BACKGROUND: Resveratrol is a polyphenol with health properties being mainly present in the skins of several foods. However, any study has been carried out to analyze the presence of this stilbene in the plum fruit from the genus Prunus in European and Japanese cultivars. RESULTS: The analysis of resveratrol from the skin in different cultivars of plums from Spanish markets with liquid chromatography coupled to ultraviolet (LC-UV) detector with subsequent confirmation by LC-MS/MS has been demonstrated that contents of this compound in plums ranged from 0.1 to 6.2 µg/g. CONCLUSIONS: Values of resveratrol in European plum cultivars is higher than in Japanese cultivars. Copyright © 2012 Society…

Rapid determination of ochratoxin A in cereals and cereal products by liquid chromatography.

A new method based on extraction with octylsilica (C8) followed by liquid chromatography coupled with fluorescence detection (LC-FLD) was studied to determine ochratoxin A (OTA) from cereals and cereal products. Optimization of different parameters, such as type and amount of solid phase, type and volume of eluent and amount of sample were carried out. Recovery of OTA from rice samples spiked at 10 ng/g level was of 86% with relative standard deviation of 5%. The limits of detection and quantification of the proposed method were 0.25 and 0.75 ng/g, respectively. Furthermore, LC-FLD after of OTA methylation and liquid chromatography coupled to mass spectrometry with an electrospray interface…

Dietary administration of high doses of pterostilbene and quercetin to mice is not toxic.

The aim of this study is to evaluate possible harmful effects of high doses of t-pterostilbene (t-PTER) and quercetin (QUER) in Swiss mice. Mice were fed during 28 days at doses of 0, 30, 300, and 3000 mg/kg body weight/day of t-PTER, QUER, or a mixture of both, t-PTER + QUER, which are equivalent to 5, 50, and 500 times, respectively, the estimated mean human intake of these polyphenols (25 mg/day). Daily oral administration of QUER, t-PTER, or a mixture of both of them did not cause mortality during the experimental period. There were no differences in food and water consumption on sex. No significant body weight gain in the male or female groups was observed. Red blood cell number and th…

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…

Stinging nettle (Urtica dioica L.) as a functional food additive in egg pasta: Enrichment and bioaccessibility of Lutein and β-carotene

Abstract The use of stinging nettles as an ingredient in egg pasta has been evaluated with respect to food enrichment with carotenoids. Bioaccessibility of lutein and β-carotene has been estimated by dynamic simulation of the digestion process, with particular attention to duodenum and colon stages. Higher bioaccessibility for the two carotenoids occurs between 2 and 24 h of colonic fermentation and it is around 35% for lutein and 10% for β-carotene. However, the results reveal that the food matrix has a significant role in carotenoid release during the digestion process. In general, nettle enriched pasta has a lower carotenoid bioaccessibility than dietary supplement at duodenum and after …

Rapid screening of organophosphorus pesticides in honey and bees by liquid chromatography—Mass spectrometry

Twenty-two organophosphorus pesticide residues were simultaneously determined in honey and honeybee samples by extraction procedures based on solid-phase and liquid chromatography-atmospheric pressure chemical ionization-mass in positive and negative ionization modes. Pesticides in honeybee samples were extracted by matrix solid phase dispersion with C18 silica and subsequently eluted with a mixture of dichloromethane-methanol (85∶15, v/v). The average recoveries from spiked honeybees (0.5–10 μg g−1) ranged from 76% for coumaphos to 100% for omethoate, with relative standard deviations between 3 and 17%. Limits of detection were ≤0.13 μg g−1 and limits of quantification were ≤0.43 μg g−1. H…

Characterization of heat-labile toxin-subunit B from Escherichia coli by liquid chromatography-electrospray ionization-mass spectrometry and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

The possibilities of characterizing the heat-labile enterotoxin (LT) of enterotoxigenic Escherichia coli (ETEC) by liquid chromatography electrospray mass spectrometry (LC/ESI-MS) and matrix-assisted laser desorption with time-of-flight mass spectrometry (MALDI-TOF-MS) were investigated. The B subunit from recombinant E. coli (expression in Pichia pastoris) can be detected by LC/ESI-MS expressed in P. pastoris and the charge envelope signals can be observed; LC/ESI-MS and MALDI-TOF-MS analysis allowed the acquisition of labile toxin subunit B (LTB) molecular weight and preliminary structural characterization of LTB toxin. MALDI-TOF analysis after reduction and alkylation of the protein evid…

Reduction of earwax with hypoacusia in Saharawi children

Summary We report case reports of earwax with hypoacusia (1.8%) in 270 Saharawi children, aged from 6 to 13 years, for a two-month period during the summer according to the “Holidays in Peace” program developed in collaboration with a local non-governmental organization (NGO). Our values are lower than those of other authors due to an initiative of systematic checks carried out by trained general doctors and nursing on children's hearing in Saharawi refugee camps in the last years.

Determination of trichothecenes and zearalenones in grain cereal, flour and bread by liquid chromatography tandem mass spectrometry

Although analytical methods have been already reported for legislated mycotoxins as trichothecenes and zearalenone (ZON) separately, we describe the optimization of a simple and rapid multimycotoxin method for the determination of a total of 12 mycotoxins simultaneously, nine trichothecenes (NIV, DON, FUS-X, DAS, 15-AcDON, 3-AcDON, NEO, HT-2, T-2 T2), and zearalenone and its metabolites (ZON, a-ZOL, b-ZOL), of different origin (wheat, oat, barley and spelt) and in three different products where these substance can be present (grain, flour and bread) reach the food chain and cause toxic effect either in humans or animals. The extraction procedure was based on a mixture of acetonitrile/water …

Biopreservation potential of lactic acid bacteria from Andean fermented food of vegetal origin

Abstract Microbial fermentations have long represented a way of natural biopreservation of raw materials, which frequently originated new food products. Among them, traditionally fermented products still manufactured by native populations all around the world are source of lactic acid bacteria (LAB) strains with high biotechnological potential. LAB are food grade microorganisms and therefore a good alternative to chemicals to be applied in food preservation. A total of 130 LAB isolates recovered from “chicha” and “tocosh”, traditional fermented Andean products of vegetal origin, were screened for antimicrobial activities against spoiler fungi Meyerozyma guilliermondii CECT 1021 (synonym Pic…

Development of a nutritional HACCP plan.

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…

Analysis of enniatins and beauvericin by LC-MS/MS in wheat-based products

Due to the matrix complexity for wheat-based products, a comparative study of different rapid extraction procedures was performed for the extraction of enniatins (ENA, ENA1, ENB, ENB1) and beauvericin in flour, pasta, breakfast cereals, and biscuits. Three different approaches were studied during the extraction and purification steps (shaker, Ultra-Turrax, and QuEChERS) for each matrix. Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) with electrospray source working in a positive mode was used. For the analysis of the five mycotoxins, the three methods were tested in terms of recovery, matrix effect, and sensibility, concluding that Ultra-Turrax extraction was the most …

Thermal and non-thermal preservation techniques of tiger nuts' beverage "horchata de chufa". Implications for food safety, nutritional and quality properties.

"Horchata de chufa" is a traditional Spanish beverage produced from tiger nuts (Cyperus esculentus L.). Due to its richness in nutritional compounds, it is highly perishable and its conservation by pasteurization and/or adding preservatives is required. Although efficient, conventional thermal treatment for pasteurization induces changes in the nutritional and sensory properties. Replacing conventional pasteurization by non-thermal technologies such as pulsed electric fields, ultraviolet, and high pressure, combined with moderate temperatures (<40°C) allows a reduction of energy consumption, along with the preservation of the most thermo-sensitive molecules. Accordingly, this review deals w…

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 …

Dietary intake of ochratoxin A from conventional and organic bread

Ochratoxin A (OTA) was extracted from 100 bread samples by using accelerated solvent extraction (ASE) and analyzed with liquid chromatography coupled with fluorescence detection. The presence of OTA was confirmed by methyl-ester derivatization. Bread samples were bought from different bakeries and supermarkets, 74 of non-organic and 26 of organic bread. The incidence of OTA varied between 20.3% and 23.0% for non-organic and organic bread, respectively. The highest values were obtained with non-organic versus organic products, five samples exceeded the European maximum permitted limit of OTA (3 ng/g) for this product. Estimated daily intake of OTA in this study was 1.6 ng/kg b.w./day. This v…

Matrix solid-phase dispersion extraction procedure for multiresidue pesticide analysis in oranges.

A multiresidue extraction method based on matrix solid-phase dispersion (MSPD) is optimized for the extraction and gas chromatographic screening of eighteen insecticides (aldrin, carbophenothion, captafol, chlorpyriphos, chlorfenvinphos, diazinon, dicofol, alpha-endosulfan, beta-endosulfan, ethion, fenitrothion, folpet, methidathion, malathion, methyl-azinphos, methyl-parathion, phosmet, and tetradifon) from oranges. After optimization of different parameters, such as type of solid phase used and the amount of solid phase or eluent, recoveries ranged from 67 to 102% with relative standard deviations ranging from 2 to 10%. The limits of detection, calculated as 3 times the baseline noise ran…

Potential Application of Lactic Acid Bacteria to Reduce Aflatoxin B1 and Fumonisin B1 Occurrence on Corn Kernels and Corn Ears

Fungal spoilage is an important issue for the food industry, leading to food sensory defects, food waste, economic losses and public health concern through the production of mycotoxins. Concomitantly, the search for safer natural products has gained importance since consumers began to look for less processed and chemically treated foods. In this context, the aim of this study was to evaluate the antifungal and antimycotoxigenic effect of seven strains of Lactobacillus plantarum. Lactic acid bacteria (LAB) were grown on Man Rogosa Sharpe (MRS) broth at 37 &ordm

Antifungal activity of peracetic acid against toxigenic fungal contaminants of maize and barley at the postharvest stage

Abstract Contamination of maize and barley grain during storage by the toxigenic fungi Aspergillus flavus (A. flavus) and Penicillium Verrucosum (P. verrucosum) is both an economic and a public health problem, especially in less industrialized countries. Peracetic acid (PA) is a compound used for the disinfection of food and food contact surfaces. Unlike other disinfectants, it leaves no toxic residues and its decomposition products (CH3COOH, O2 and H2O) are environmentally friendly. In order to apply PA to preserve maize and barley grain during storage, first, the Minimum Inhibitory Concentration (MIC) and Minimum Fungicidal Concentration (MFC) against both fungi were determined in a 96-we…

Gas chromatographic evaluation of pesticide residue contents in nectarines after non-toxic washing treatments

Washing with aqueous solutions of citric acid, ethanol, glycerol, hydrogen peroxide, potassium permanganate, sodium metabisulfite, sodium laurylsulfate (SLS), sodium hypochlorite, and urea is evaluated for pesticide residue reduction in nectarines and compared with simple tap water washing. Residues of pesticides commonly utilized in nectarines (chlorpyrifos, fenarimol, iprodione, malathion, methidathion, myclobutanil, parathion and pirimicarb) are extracted with ethyl acetate and anhydrous sodium sulfate, extract is concentred and analyzed by GC with nitrogen-phosphorus detection. The formation of possible toxic by-products (chlorpyrifos oxon, malaoxon, methidaoxon and paraoxon methyl) is …

Determination of carbamate residues in fruits and vegetables by matrix solid-phase dispersion and liquid chromatography-mass spectrometry.

Abstract Thirteen carbamates were analysed in orange, grape, onion and tomatoes by matrix solid-phase dispersion (MSPD) followed by liquid chromatography–mass spectrometry (LC–MS). Electrospray (ES) and atmospheric pressure chemical ionisation (APCI) were compared and both gave similar results in terms of sensitivity and structural information because at 20 V fragmentor voltages the fragmentation is minimal. The efficiency of different solid-phases (C18, C8, cyano, amine and phenyl) for the MSPD was compared. Mean recoveries using C8 varied from 64 to 106% with relative standard deviations of 5–15% in the concentration range of 0.01–10 mg kg−1. Matrix constituents did not interfere signific…

Bioactive compounds from mustard flours for the control of patulin production in wheat tortillas

Abstract Patulin (PAT) is a toxic fungal metabolite produced by Penicillium, Aspergillus and Byssochlamys growing especially in fruit and cereals. PAT exhibits a number of toxic effects in animals and its presence in food is undesirable. In this study the reduction of the mycotoxin PAT produced by a strain of Penicillium expansum, on wheat tortillas was studied using volatile bioactive compounds present in the oriental and yellow mustard flour and also using the standard solution of the antifungal compound allyl isothiocyanate (AIT), developing an active packaging with two different systems of release of those bioactive compounds. Also the kinetic of volatilization of the compounds used in …

Chemical reduction of the mycotoxin beauvericin using allyl isothiocyanate.

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 investigated the reduction of BEA present in the concentration of 25 mg/kg on a solution model (phosphate buffer saline at pH 4 and 7) and in wheat flour using allyl isothiocyanate (AITC) as a reactant. The concentration of the mycotoxin studied was evaluated using liquid chromatography coupled to the diode array detector (LC-DAD), whereas adducts formed between the BEA and AITC were examined by liquid chromatography coupled to mass spectrometry-linear ion trap (LC-MS-LIT). In solution, BEA reduction ranged from 20% t…

Antifungal and antimycotoxigenic activity of hydrolyzed goat whey on Penicillium spp: An application as biopreservation agent in pita bread

Abstract Whey is a by-product of the cheese industry, yet it contains proteins that have a high nutritional value and are an important source of antifungal peptides. Food deterioration caused by toxigenic fungi is one of the challenges of food safety. In this context, trypsin was used to hydrolyse goat milk whey at 37. The resultant peptides were characterised by LC–ESI–TOF-MS. Antifungal activity of the goat milk whey hydrolysate (HGW) was determined against 10 toxigenic fungi from the genus Penicillium, in solid and liquid media. Furthermore, HGW was used as an ingredient for bread elaboration. Bread elaborated with HGW and inoculated with toxigenic fungi was included in a shelf-life stud…

Bioaccessibility of Deoxynivalenol and its natural co-occurrence with Ochratoxin A and Aflatoxin B1 in Italian commercial pasta

Abstract Cereals products for direct human consumption are rarely contaminated by moulds, unlike raw materials, which are often infected, either in the field or during storage. In this study, 27 samples of dried pasta characterised by size, packaging and marketing intended for young children consumption were collected and analysed by liquid chromatography (LC) and liquid chromatography–tandem mass spectrometry (LC–MS/MS) for Deoxynivalenol (DON), Ochratoxin A (OTA) and Aflatoxin B 1 (AFB 1 ) determination. The samples that showed the highest amounts of one of the mycotoxins were cooked for 10 min, digested with an in vitro gastrointestinal protocol and bioaccessibility values were calculate…

Development of microextraction techniques in combination with GC-MS/MS for the determination of mycotoxins and metabolites in human urine.

Simple and highly efficient sample preparation procedures, namely, dispersive liquid–liquid microextraction and salting-out liquid–liquid extraction for the analysis of ten Fusarium mycotoxins and metabolites in human urine were compared. Various parameters affecting extraction efficiency were carefully evaluated. Under optimal extraction conditions, salting-out liquid–liquid extraction showed a better accuracy (84–96%) and precision (<14%) than dispersive liquid–liquid microextraction. Hence, a multibiomarker method based on salting-out liquid–liquid extraction followed by gas chromatography with tandem mass spectrometry was proposed. Satisfactory results in terms of validation were achiev…

Propuesta de valores de referencia ambientales microbiológicos en los servicios de restauración

Introducción: Proponer una nueva referencia microbiológica ambiental en los servicios de restauración. Objetivo: El presente trabajo muestra la determinación y evaluación de la contaminación microbiológica generada en un establecimiento de servicio de alimentos. Método: Se basa en el muestreo microbiológico de superficie utilizando filtros de membrana de ésteres mixtos de celulosa y en el muestreo de aire utilizando placas de Petri. Resultados: Los límites de contaminación se establecieron antes y durante la elaboración de alimentos, mediante el análisis microbiológico de las superficies y del medio ambiente, sistemas de equipos, hasta límites confiables y estableciendo niveles de aceptació…

Simple liquid chromatography assay for analyzing ochratoxin a in bovine milk

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…

Rapid mycotoxin analysis in human urine: A pilot study

A simple and rapid method effective for quantitative determination of deoxynivalenol (DON), T-2 toxin (T-2), HT-2 toxin (HT-2), zearalenone (ZEN), ochratoxin A (OTA), aflatoxins (AFs) B(1), B(2), G(1) and G(2) and fumonisins FB(1) and FB(2) in urine was developed. The urine was diluted with phosphate buffer solution (PBS) and thoroughly mixed. For clean-up and extraction, the mixture was loaded on a MYCO 6in1 IAC. Hybrid triple quadrupole-linear ion trap mass spectrometer (QTrap) was used for the detection. Extra tools for confirmation of selected mycotoxins in positive samples, Information Dependent Acquisition (IDA) experiments, were also developed. The use of immunoaffinity columns follo…

The Role of the Liquid Chromatography-Mass Spectrometry in Pesticide Residue Determination in Food

The use of liquid chromatography (LC) in pesticide residue determination was usually limited to groups of compounds or single compounds for which no suitable gas chromatographic (GC) conditions were available. However, recent developments have significantly enlarged the LC scope in this field of analysis. One of the most important advances was the on-line coupling of efficient LC separation with mass spectrometry detectors (LC-MS and LC-MS/MS) that makes this technique an excellent method for the determination of pesticides and their transformation products in complex matrices such as food. This review considers the application of LC-MS/MS in this field. Emphasis is placed on the tandem MS …

Limited survey for the occurrence of aflatoxins in cereals and poultry feeds from Rabat, Morocco

Fifty eight (58) samples of cereals for human consumption (20 corn flour, 17 wheat flour) and poultry feeds (n=21) purchased from popular markets of Rabat in Morocco were analyzed for aflatoxins (AF) by HPLC with immunoaffinity column (IAC) clean-up and fluorimetric detection. The incidence of AF in corn, wheat flour and poultry feeds was about 80, 17.6 and 66.6% respectively. High contamination levels were found in corn and poultry feeds samples. Levels of contamination of analyzed samples with AFB1 ranged from 0.23 to 11.2, 0.03 to 0.15 and 0.05 to 5.38 ng/g for corn, wheat and poultry feeds respectively. Results showed also that the contamination of 10% of samples of corn was higher than…

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…

Degradation of the minor Fusarium mycotoxin beauvericin by intracellular enzymes of Saccharomyces cerevisiae

Abstract Beauvericin (BEA) is a cyclic depsipeptide with antibiotic and insecticidal effects. It was discovered for the first time from the fungus Beauveria bassiana , but more significantly, is produced by several Fusarium strains, and considered a contaminant of several cereals like corn, wheat and barley. This study investigated the degradation of BEA by intracellular raw enzymes of four strains of Saccharomyces cerevisiae , named LO9, YE5, A34, and A17. The BEA at 25 mg/kg in a model solution and in corn flour was co-incubated with the raw enzymes from the four yeast strains, respectively. The reduction of BEA was evaluated using liquid chromatography coupled to a diode array detector (…

A survey of trichothecenes, zearalenone and patulin in milled grain-based products using GC-MS/MS.

An analytical protocol based on QuEChERS and gas chromatography-tandem mass spectrometry (GC-MS/MS) was successfully applied for the determination of trichothecenes, patulin and zearalenone in 182 milled grain-based samples. The analytical method was validated following the SANCO 1495/2011 document. LOQs were lower than 10μgkg(-1) for the selected mycotoxins. Recoveries of fortified cereals ranged between 76-108% and 77-114% at 20 and 80μgkg(-1), respectively, with relative standard deviation lower than 9%. More than 60% of the samples analysed showed deoxynivalenol contamination, followed by HT-2 toxin and nivalenol with frequencies of 12.1% and 10.4%, respectively. Co-occurrence of mycoto…

Dietary exposure to mycotoxins through the consumption of commercial bread loaf in Valencia, Spain

Abstract In this study, 80 commercial samples of bread loaves were purchased from different supermarkets located in Valencia (Spain). These samples were investigated for the presence of legislated and non-legislated mycotoxins. Results showed that samples were contaminated with Aflatoxins (AFs), Zearalenone (ZEA) and Enniatins (ENs) with a frequency of 20, 96, and 65% respectively. Aflatoxin B1 (AFB1), Aflatoxin B2 (AFB2) and Aflatoxin G1 (AFG1) were detected with concentrations ranged from 0.5 to 7.1 μg/kg. The samples contaminated with AFB1 showed values exceeding the maximum limit allowed in the EU. The sum of AFs also exceeded the maximum limit allowed in 6 samples. ENs contamination da…

Real-time quantitative PCR of Staphylococcus aureus and application in restaurant meals.

Staphylococcus aureus is considered the second most common pathogen to cause outbreaks of food poisoning, exceeded only by Campylobacter. Consumption of foods containing this microorganism is often identified as the cause of illness. In this study, a rapid, reliable, and sensitive real-time quantitative PCR was developed and compared with conventional culture methods. Real-time quantitative PCR was carried out by purifying DNA extracts of S. aureus with a Staphylococcus sample preparation kit and quantifying it in the LightCycler system with hybridization probes. The assay was linear from a range of 10 to 10(6) S. aureus cells (r2 > 0.997). The PCR reaction presented an efficiency of >85%. …

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.

Improvement of accessibility for the disabled in university restaurants

AMELIORATION DE L’ACCESSIBILITE DE RESTAURANTS UNIVERSITAIRES A DES HANDICAPES Une etude a ete realisee en vue d’ameliorer l’acces des handicapes aux 19 restraurants universitaires de Valence en Espagne, puisque le taux des etudiants handicapes, ces trois dernieres annees universitaires, est de l’ordre de 0,12 a 0,13 %. Au debut de cette etude, 16 etablissements disposaient d’un acces correct avec un pourcentage de 87,5, 25 et 6,3 concernant respectivement les rampes, les ascenseurs et les balustrades. En outre, toutes les portes d´acces de ces etablissements etaient equipees des poignees a leviers, ou etaient concues de sorte qu’une prise serree et qu’une torsion du poignet ne soient pas n…

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

Multi-mycotoxin determination in barley and derived products from Tunisia and estimation of their dietary intake.

A study on raw barley and derived products (barley soup and beers) was carried out to determine the natural presence of twenty-four mycotoxins by both liquid chromatography and gas chromatography coupled to tandem mass spectrometry (MS/MS). The developed multi-mycotoxin procedure was based on both SLE and QuEChERS extraction steps. 66% of analyzed samples presented mycotoxin contamination and only one sample, which was soup of barley (6 ng/g), exceeded the maximum level (ML) established by EU for OTA (5 ng/g). Raw barley was the most contaminated matrix (62%), which concentrations ranged from 1.70 to 287.13 ng/g) and type of detected mycotoxins (DON, 15AcDON, NEO, NIV, HT2, FB1, OTA, ENA, E…

Tiger nut and its by-products valorization: From extraction of oil and valuable compounds to development of new healthy products

Abstract Consumer's growing demand for consumption of “Horchata de chufa”, a Spanish beverage produced from tiger nut tubers, has led to large-scale production of tiger nuts, and its subsequent processing for the food industry. Recent investigations clearly show that tiger nut is a valuable source of stable vegetable oils, rich in monounsaturated fatty acids, and phytosterols as well as high-added value compounds (proteins, carbohydrates, vitamins, minerals and bioactive compounds). Several conventional (Soxhlet) and alternative innovative (SC-CO2, enzyme, high pressure, etc.) extraction methods have been developed for the efficient recovery of tiger nut oil and high-added value compounds. …

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…

Determination of carbosulfan and its metabolites in oranges by liquid chromatography ion-trap triple-stage mass spectrometry.

Liquid chromatography ion-trap mass spectrometry LC-MSn has been successfully applied to identify and confirm carbosulfan and seven of its metabolites in oranges after pressurized liquid extraction (PLE) with dichloromethane. Mass spectra of carbosulfan and its metabolites were investigated using multiple stages of mass spectrometry. Although interpretation of the fragmentation pathways, based on mass spectra, enables structural elucidation and identification of these compounds, the proposed fragmentation pathways and ion structures need verification by exact mass measurements. The analytical method--PLE and LC-MS3 --was validated: limits of quantification (LOQ) ranged from 0.01 to 0.07 mg …

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…

Reducing the effect of beauvericin on neuroblastoma SH-SY5Y cell line by natural products

Abstract In the present work, different natural compounds from coffee by-product extracts (coffee silverskin and spent coffee) rich in polyphenols, was investigated against beauvericin (BEA) induced-cytotoxicity on SH-SY5Y cells. Spent coffee arise as waste products through the production of instant coffee and coffee brewing; while the silverskin is a tegument which is removed and eliminated with toasting coffee grains. First of all, polyphenol extraction methods, measurement of total polyphenols content and its identification were carried out. Afterwards evaluating in vitro effects with MTT assay on SH-SY5Y cells of coffee by-product extracts and mycotoxins at different concentrations and …

Prevalence of bacteria and absence of anisakid parasites in raw and prepared fish and seafood dishes in Spanish restaurants

This study evaluated the presence of bacteria and anisakid parasites in 45 samples of raw anchovies in vinegar, a dish widely eaten in Spain, and in 227 samples of cooked fish and cephalopods served in Spanish food service establishments. Our analysis showed that, according to European and Spanish regulation, 14 to 30% of the prepared fish and cephalopod dishes exceeded the maximum allowable level for mesophilic aerobic counts, and 10 to 40% of these samples exceeded the allowable levels for Enterobacteriaceae. None of the studied samples showed evidence of anisakid parasites, Escherichia coli, Staphylococcus aureus, Salmonella, or Listeria monocyto genes. These results indicate that applic…

Fusarium species, chemotype characterisation and trichothecene contamination of durum and soft wheat in an area of central Italy

BACKGROUND Fusarium head blight (FHB) of wheat is an important disease causing yield losses and mycotoxin contamination. The aim of the work was to detect and characterise trichothecene producing Fusarium species in durum and soft wheat cultivated in an area of central Italy in 2009 and 2010 and to determine trichothecene contamination by LC-MS/MS in the grain. RESULTS F. graminearum s. str. was the most frequent species. In 2009, the occurrence of F. avenaceum and F. poae was higher than in 2010. Among F. graminearum strains, the 15-acetyl deoxynivalenol (15-ADON) chemotype could be found more frequently, followed by nivalenol (NIV) and 3-ADON chemotypes, while all F. culmorum isolates bel…

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…

Determination of Alternaria mycotoxins in strawberries and storage conditions

Pressurized liquid extraction coupled to liquid chromatography for the analysis of ochratoxin A in breakfast and infants cereals from Morocco

Abstract A sensitive and reliable method using pressurized liquid extraction (PLE) and liquid chromatography (LC) has been developed for the analysis of ochratoxin A (OTA) in breakfast and infants cereals. Influence of several extraction solvents that affect PLE efficiency was studied. The selected PLE operating method was: 10 g of sample was packed into 22 ml stainless-steel cell and OTA was extracted with acetonitrile/water (80:20) at 40 °C, 34 atm in one cycle of 5 min at 60% flush. The mean recovery of OTA was 82 ± 4 at fortification level of 3 ng/g OTA. The limit of quantification (LOQ) of OTA was 0.25 ng/g. The proposed method was successfully applied to the analysis of 68 samples of …

Disorders of malnutrition among the Saharawi children

It is first article that reviews the literature on the problem of disorders of malnutrition among Saharawi children. Most of the studies reveal problems mainly of celiac disease, goiter, geophagia, undernutrition, wasting, stunting, overweight, scurvy, rickets and iron-deficiency anaemia.

Effect of introduction of HACCP on the microbiological quality of some restaurant meals

The microbial quality of Spanish potato omelette and pork loin before and after implementation of the HACCP system in University restaurants was used as indicator of food safety in this work. The prevalence of aerobic plate counts and incidences of Staphylococcus aureus, Escherichia coli, E. coli O157:H7, Listeria monocytogenes, Salmonella spp. and Clostridium perfringens were analysed. Results of implementation of the HACCP system show a lower incidence of studied microorganisms. On the other hand, a documented training in personal hygiene, good manufacturing practices (GMPs), cleaning and sanitation procedures and personal safety in addition to the rearrangement in the infrastructure of t…

Shelf life improvement of the loaf bread using allyl, phenyl and benzyl isothiocyanates against Aspergillus parasiticus

Abstract Fungal growth inhibition and aflatoxins (AFs) reduction using allyl (AITC), benzyl (BITC) and phenyl (PITC) isothiocyanates were studied in loaf bread contaminated with Aspergillus parasiticus . Two inoculated loaf bread slices were introduced into a plastic tray together with paper filters or small plastic bags paper filters soaked with AITC, BITC or PITC, the final concentration inside the package was of 0.5, 1 or 5 μL/L. The plastic trays, incubated at room temperature, were visual examined for the shelf life evaluation during 8 days. The quantification of the AFs was carried out using liquid chromatography coupled to mass spectrometry (LC-MS/MS). Shelf life increase of three an…

Development and Validation of a LC-ESI-MS/MS Method for the Determination of Alternaria Toxins Alternariol, Alternariol Methyl-Ether and Tentoxin in Tomato and Tomato-Based Products

Alternaria species are capable of producing several secondary toxic metabolites in infected plants and in agricultural commodities, which play important roles in food safety. Alternaria alternata turn out to be the most frequent fungal species invading tomatoes. Alternariol (AOH), alternariol monomethyl ether (AME), and tentoxin (TEN) are some of the main Alternaria mycotoxins that can be found as contaminants in food. In this work, an analytical method based on liquid chromatography (LC) tandem mass spectrometry (MS/MS) detection for the simultaneous quantification of AOH, AME, and TEN in tomato and tomato-based products was developed. Mycotoxin analysis was performed by dispersive liquid-…

Liquid chromatography-ultraviolet detection and quantification of heat-labile toxin produced by enterotoxigenic E. coli cultured under different conditions

Abstract Enterotoxigenic Escherichia coli (ETEC) is the main bacterial cause of dehydrating infant diarrhoea in less-developed countries. Labile toxin (LT) is the major virulent factor of ETEC. Easy diagnostic tests are necessary to reduce the number of cases. Immunological methods have some drawbacks and also have important limitations. For that reason, a Liquid Chromatography coupled to UV detector technique (LC-UV) has been optimize to a rapid identification and quantification of LT from bacteria cultures. It is also important to know optimal conditions for LT and with this purpose several enterotoxigenic E. coli strains have been studied to determine the influence of glucose concentrati…

Evaluation of Alternaria mycotoxins in strawberries: quantification and storage condition

Alternariol (AOH), alternariol methyl ether (AME) and tentoxin (TEN) are Alternaria mycotoxins produced by the most common post-harvest pathogens of fruits. The production of these metabolites depends on several environmental factors, mainly temperature, water activity, pH and the technological treatments that have been applied to the product. In this study, the occurrence of AOH, AME and TEN was evaluated in strawberries samples stored at different temperatures ranges (at 22 ± 2 or 6 ± 2°C) and different periods (up to 1 month) simulating the current practice of consumer's storage conditions. Sample extraction was performed using a liquid-liquid extraction method prior to LC-MS/MS analysis…

Phenol profiling and nutraceutical potential of Lycium spp. Leaf extracts obtained with ultrasound and microwave assisted techniques

In recent years, agricultural and industrial residues have attracted a lot of interest in the recovery of phytochemicals used in the food, pharmaceutical, and cosmetic industries. In this paper, a study on the recovery of phenol compounds from Lycium spp. leaves is presented. Ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) have been used with alcoholic and hydroalcoholic solvents. Methanolic UAE was the most successful technique for extracting phenols from Lycium leaves, and we used on leaves from L. barbarum and L. chinense cultivated in Italy. The extracts were then characterized as regards to the antioxidant properties by in vitro assays and the phenol profil…

Development of a GC–MS/MS strategy to determine 15 mycotoxins and metabolites in human urine

The widespread mycotoxins contamination of food commodities has made the monitoring of their levels essential. To overcome the disadvantages of the indirect approach by food analysis, detection of mycotoxin as biomarkers in urine provides a useful and specific data for exposure assessment to these food contaminants. In this work, a sensitive, rapid and accurate method based on gas chromatography-tandem mass spectrometry procedure to determine 15 mycotoxins and metabolites in human urine was optimized and validated taking into consideration the guidelines specified in Commission Decision 2002/657/EC and 401/2006/EC. A salting-out assisted acetonitrile-based extraction was used for sample pre…

Comparison of various liquid chromatographic methods for the analysis of avermectin residues in citrus fruits.

Abstract Various liquid chromatographic (LC) techniques for analyzing avermectin (Abamectin) were compared after extraction of residues from citrus fruit samples by matrix solid-phase dispersion (MSPD). LC with UV and fluorescence detection were used as also was LC coupled to the mass spectrometer by an electrospray interface. The results obtained by the three methods were compared in terms of sensitivity and selectivity. The combination of MSPD extraction and LC with fluorescence detection have made it possible to quantify 0.5 μg kg −1 of Abamectin in 0.5 g of orange sample, with an overall average recovery of 94%. The procedure provides a simple and sensitive method for monitoring Abamect…

Key analytical features of mycotoxins

Evaluation of Mycotoxins in Infant Breast Milk and Infant Food, Reviewing the Literature Data

In this review, an analysis focusing on mycotoxin determination in infant breast milk and infant food has been summarised for the last fifteen years of research focused on the intended population group of 1–9 months. The objective was to know the level of exposure of the child population to an estimated daily intake (EDI) of mycotoxins from the consumption of habitual foods. The EDI was compared with the tolerable daily intake (TDI) established by EFSA to estimate risk. In breast milk, the high prevalence and levels were for samples from Africa (Egypt and Tanzania) with aflatoxin M1 (1.9 μg/L and 10%), and Asia (Iran) with ochratoxin-A (7.3 μg/L and 100%). In infant formulas, high incidence…

Antifungal and antimycotoxigenic activity of allyl isothiocyanate on barley under different storage conditions

Abstract The present study evaluated the efficacy of allyl isothiocyanate (AITC) in avoiding the growth of Penicillium verrucosum and consequent ochratoxin A (OTA) production in barley during storage for 90 d. Environmental humidity was controlled using saturated salt solutions and moisture content (MC) of the grain was analyzed. Moreover, the residual concentration of AITC on stored barley was also examined. Samples with 20.6% of MC presented the highest absorption of AITC with levels ranging from 75 to 4 mg/kg at day 1 and 90, respectively. The population of P. verrucosum was significantly reduced after 24 h of AITC exposure. After 90 d, the non-treated control group reached a fungal popu…

Review on the toxicity, occurrence, metabolism, detoxification, regulations and intake of zearalenone: An oestrogenic mycotoxin

Zearalenone (ZEA) is a mycotoxin produced mainly by fungi belonging to the genus Fusarium in foods and feeds. It is frequently implicated in reproductive disorders of farm animals and occasionally in hyperoestrogenic syndromes in humans. There is evidence that ZEA and its metabolites possess oestrogenic activity in pigs, cattle and sheep. However, ZEA is of a relatively low acute toxicity after oral or interperitoneal administration in mice, rat and pig. The biotransformation for ZEA in animals involves the formation of two metabolites alpha-zearalenol (alpha-ZEA) and beta-zearalenol (beta-ZEA) which are subsequently conjugated with glucuronic acid. Moreover, ZEA has also been shown to be h…

Apple-products phytochemicals and processing: a review.

Epidemiological studies have linked the consumption of apples with reduced risk of some cancers, cardiovascular disease, asthma, and diabetes. Extensive research exists on apples and the health benefits of their beverages and phytochemicals. The purpose of this paper is to review the most recent literature in this area focusing on phytochemicals, phytochemical bioavailability and antioxidant behavior.

Beauvericin and ochratoxin A mycotoxins individually and combined in HepG2 cells alter lipid peroxidation, levels of reactive oxygen species and glutathione

The co-presence of more than one mycotoxin in food is being evidenced in last food surveys as reported in the literature. Beauvericin (BEA) is a non-legislated emergent mycotoxin while Ochratoxin A (OTA) has been widely studied and legislated. Concentration range individually studied was from 2.5 to 0.3 μM for BEA and from 25 to 3.1 μM for OTA; binary mixture [BEA + OTA] comprised concentrations of 1:10 ratio from [2.5 + 25] to [3.1 + 0.3] μM. Potential of toxicity of BEA in HepG2 cells was the highest at all times assayed (24, 48 and 72h). LPO was performed through malondyaldehyde (MDA) detection denoting in the binary mixture for [1.25 + 12.5] μM and at 24 and 72h the highest disturbance …

Report of toxic shock syndrome toxin 1 (TSST-1) from Staphylococcus aureus isolated in food handlers and surfaces from foodservice establishments.

A set of 53 Staphylococcus aureus isolates collected from food handlers and foodservice establishments in Spain was analyzed for toxic shock syndrome toxin (TSST-1) production. S. aureus strains were isolated from 908 samples collected from different surfaces such as dish towels, workers’ hands, cutting boards, stainless steel tables and slicers, but they were not detected neither in clean plates nor in kitchen knives. Only one food worker hand has been reported to be contaminated by TSST-1 in a restaurant. Despite this, proper hygiene practices should be respected for the surfaces of contact with food, as well as for the hands of the manipulators This is the first article, in Spain, that r…

Gaseous allyl isothiocyanate to inhibit the production of aflatoxins, beauvericin and enniatins by Aspergillus parasiticus and Fusarium poae in wheat flour

Abstract There is a growing concern about the presence of mycotoxins in foods, since up to 25% of cereals and cereal foods are contaminated with these compounds. Moreover, the general public is against the use of synthetic preservatives in foods and the use of natural antimicrobials in foods is a current trend. Allyl isothiocyanate (AITC) is a volatile antimicrobial derived from oriental and black mustard. The objective of this work was to evaluate the capacity of gaseous AITC in inhibiting the production of mycotoxins by Aspergillus parasiticus (aflatoxin producer) and Fusarium poae (beauvercin and enniatin producer) in wheat flour. Petri dish lids filled with 2 g of wheat flour were inocu…

Impact of ultrasound extraction parameters on the antioxidant properties of Moringa oleifera leaves

Recently, much interest has been focused on Moringa oleifera L., a highly versatile and sustainable plant. In addition to its nutritional properties, numerous bioactive compounds have been identified in M. oleifera leaves, for which healthy properties have been reported. In the present research, the impact of ultrasound-assisted extraction (UAE) on the recovery of the bioactive compounds from leaves was investigated. Firstly, an experimental design approach has been used to highlight the influence of some extraction parameters (solvent, solvent/dry leaves ratio, temperature, time) on phenol compound recovery and antioxidant activity. Solvent composition was the most influential factor

Concentration of ochratoxin A in wines from supermarkets and stores of Valencian Community (Spain)

Ochratoxin A (OTA) is a mycotoxin produced by fungi species belonging to the genera Aspergillus and Penicillium being isolated in alcoholic beverages. The aim of this work is developed and applied a procedure for the analysis of OTA in wines. An analytical method based on immunoaffinity column (IAC) for clean-up, liquid chromatography with fluorescence detection (LC-FD), and LC-FD after of OTA methylation was used to determine the occurrence of OTA in wines. Recoveries of this mycotoxin spiked to red wines at 0.5 ng/ml level were >90% with an average of relative standards deviations of 4%. Furthermore, 116 wine samples from designation of origin (DO) and three samples from food stores of Va…

Proposal of reference values of microbiological environment monitoring in foodservice establishments

Introducción: Proponer una nueva referencia microbiológica ambiental en los servicios de restauración. Objetivo: El presente trabajo muestra la determinación y evaluación de la contaminación microbiológica generada en un establecimiento de servicio de alimentos. Método: Se basa en el muestreo microbiológico de superficie utilizando filtros de membrana de ésteres mixtos de celulosa y en el muestreo de aire utilizando placas de Petri. Resultados: Los límites de contaminación se establecieron antes y durante la elaboración de alimentos, mediante el análisis microbiológico de las superficies y del medio ambiente, sistemas de equipos, hasta límites confiables y estableciendo niveles de aceptació…

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 …

Determination of ochratoxin A in maize bread samples by LC with fluorescence detection.

Ochratoxin A (OTA) is a secondary fungal metabolite produced by several moulds, mainly by Aspergillus ochraceus, A. carbonarius, A. niger and by Penicillium verrucosum. The present work shows the results of comparative studies using different procedures for the analysis of OTA in maize bread samples. The studied analytical methods involved extraction with different volumes of PBS/methanol, different extraction apparatus, and clean-up through immunoaffinity columns. The separation and identification were carried out by high-performance liquid chromatography with fluorescence detection. The optimized method for analysis of OTA in maize bread involved extraction with PBS:methanol (50:50), and …

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…

Reduction of beauvericin and enniatins bioaccessibility by prebiotic compounds, evaluated in static and dynamic simulated gastrointestinal digestion

Abstract The aim of this study was to investigate the bioaccessibility of beauvericin (BEA) and enniatins (ENs) present in wheat crispy breads. A microbial fermentation was performed by a BEA- and ENs-producer Fusarium strain, adding inulin and fructooligosaccharides (FOS) at two concentrations (1% and 5%). The bioaccessibility of mycotoxins was determined by static and dynamic simulated gastrointestinal digestion systems, imitating the digestive physiological conditions until the colonic compartment. BEA and ENs were determined in the intestinal fluids by liquid chromatography-tandem mass spectrometry (LC-MS/MS). BEA and ENs bioaccessibilities in the static model (46.7–61.1% and 6.2–44.9%,…

Bio-Preservative Potential of Microorganisms Isolated from Red Grape against Food Contaminant Fungi

Fungal spoilage is one of the main reasons of economic losses in the food industry, especially in the wine sector. Consequently, the search for safer and new preservation techniques has gained importance in recent years. The objective of this study was to investigate the antifungal and anti-mycotoxigenic activity from 28 microorganisms (MO) isolated from red grape. The antifungal activity of a cell free supernatant of fermented medium by the isolated MO (CFS) was tested with the agar diffusion method and the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) assay. Additionally, different antifungal compounds from the CFS were identified and quantified (organi…

Extraction of Phenolic Compounds from Fresh Apple Pomace by Different Non-Conventional Techniques

Red Delicious apple pomace was produced at laboratory scale with a domestic blender and different non-conventional extraction techniques were performed to isolate phenolic compounds, such as ultrasound-assisted extraction (UAE), ultraturrax extraction (UTE), accelerated solvent extraction (ASE) and pulsed electric field (PEF) extraction pre-treatment. Total phenolic content (TPC) was determined by Folin–Ciocalteu assay. Phloridzin, the main phenolic compound in apples, was determined by chromatographic analysis Q-TOF-LC/MS. The results obtained with these techniques were compared in order to identify the most efficient method to recover polyphenols. The highest value of TPC (1062.92 ± 59.80…

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…

Quantification of Listeria monocytogenes in salads by real time quantitative PCR

Abstract A real time quantitative PCR (RTQ-PCR) was carried out purifying DNA extracts of Listeria monocytogenes using a High Pure Listeria Sample Preparation Kit and quantifying in a LightCycler system with hybridisation probes. A standard curve was constructed with serial dilutions. A range linear relationship, from 10 to 10 5 L. monocytogenes colony forming units (CFU), was observed between threshold cycle ( C t ) and logarithmic concentration of the serial dilutions. The assay was linear in a range from 10 to 10 5 L. monocytogenes CFU and the coefficient of determination ( r 2 ) was > 0.98. RTQ-PCR presented an efficiency of > 85%. The accuracy of the PCR-based assay, expressed as % bia…

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.

Limited survey for the presence of aflatoxins in foods from local markets and supermarkets in Valencia, Spain.

Aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2) were extracted by matrix solid-phase dispersion with C18 silica and acetonitrile as the eluting solvent, analysed by liquid chromatography with fluorescence detection and confirmed by liquid chromatography with mass spectrometry using an electrospray interface in 58 samples grouped as cereals, dried fruits, herbs and spices, pulses, snacks, and nuts and nut products collected from local markets and supermarkets in Valencia, Spain. All samples analysed by the proposed method were previously studied with an enzyme-linked immunosorbent assay as a screening protocol for the fast detection of mycotoxins. The samples containing residues (3/5…

Study of mycotoxin calibration approaches on the example of trichothecenes analysis from flour

The aim of this study was to evaluate the applicability of different calibration approaches in trichothecenes analysis from wheat flour. Concretely eight trichothecenes (five type B trichothecenes and three type A trichothecenes) were analyzed by matrix solid-phase dispersion (MSPD) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). In the first set of experiments the presence of matrix effects was evaluated; values ranged between 59% and 79%. In the second set of experiments, solutions to compensate these signal suppressions were examined. Different calibration methods showed to tackle matrix effects obtaining values between 69% and 85% for external matrix matched calibration a…

Bioaccessibility Study of Aflatoxin B1 and Ochratoxin A in Bread Enriched with Fermented Milk Whey and/or Pumpkin

The presence of mycotoxins in cereals and cereal products remains a significant issue. The use of natural ingredients such as pumpkin and whey, which contain bioactive compounds, could be a strategy to reduce the use of conventional chemical preservatives. The aim of the present work was to study the bioaccessibility of aflatoxin B1 (AFB1) and ochratoxin (OTA) in bread, as well as to evaluate the effect of milk whey (with and without lactic acid bacteria fermentation) and pumpkin on reducing mycotoxins bioaccessibility. Different bread typologies were prepared and subjected to an in vitro digestion model. Gastric and intestinal extracts were analyzed by HPLC&ndash;MS/qTOF and mycotoxins bio…

Assessment of microbial quality of commercial and home-made tiger-nut beverages

Aims:  We aimed to assess the microbiological quality of Spanish commercial tiger-nut beverages as well as home-made samples collected from supermarkets, street vendors, juice bars and ice-cream parlours located in Valencia. Methods and Results:  Microbiological analysis of 44 tiger-nut beverages samples were carried out according to International Standard Organization (ISO) norms and published works which included the total viable count, Enterobacteriaceae, Escherichia coli, Staphylococcus aureus, Salmonella, Bacillus cereus, yeasts, moulds, Yersinia enterocolitca, Clostridium perfringens, Vibrio spp. and Listeria monocytogenes. The obtained results indicated that all the commercial sample…

Multi-occurrence of twenty mycotoxinsin pasta and a risk assessment in the moroccan population

In the present study, the multi-occurrence of twenty (20) mycotoxins in pasta samples consumed in Morocco was assessed. For this, a modified Quick, Easy, Cheap Effective, Rugged, and Safe method was validated. The mycotoxins studied were identified and quantified by liquid chromatography&ndash

First study on trichothecene and zearalenone exposure of the Romanian population through wheat-based products consumption

Abstract In this study, a dietary exposure assessment of mycotoxins was conducted for the Romanian population using the contamination data of a various categories of wheat-based products for direct human consumption. Wheat-based foods (n = 181) commercialized in Romania, including flour, bread, biscuits, breakfast cereals and pasta, were evaluated by GC-QqQ-MS/MS for the occurrence of deoxynivalenol (DON), 3-acetyldeoxynivalenol (3AcDON), 15-acetyldeoxynivalenol (15AcDON), fusarenon-X, nivalenol, HT-2 and T-2 toxins, diacetoxyscirpenol, neosolaniol and zearalenone (ZEA). DON and 15AcDON were detected in 63 and 5% of all the analyzed samples, whereas 13-AcDON, HT-2, T-2, NIV and ZEA were not…

Study on Trichothecene and Zearalenone Presence in Romanian Wheat Relative to Weather Conditions

To evaluate the influence of weather conditions on mycotoxin presence in wheat, deoxynivalenol (DON), 3-acetyldeoxynivalenol (3AcDON), 15-acetyldeoxynivalenol (15AcDON), fusarenon-X (FUS-X), nivalenol (NIV), HT-2 toxin (HT-2), T-2 toxin (T-2), diacetoxyscirpenol (DAS), neosolaniol (NEO) and zearalenone (ZEN) were evaluated in 102 Romanian wheat samples coming from five wheat growing areas during 2015. Only six mycotoxins were detected, while FUS-X, DAS, NEO and NIV were not present in the wheat samples. Mycotoxin concentrations were correlated with precipitation and temperature values during anthesis and the preharvest period. Overall, the highest frequency was registered for DON, while the…

Occurrence of deoxynivalenol and T-2 toxin in bread and pasta commercialised in Spain

Abstract Deoxynivalenol and T-2 toxin were extracted from wheat-based bread ( n  = 75) and pasta ( n  = 75) samples using a mixture of acetonitrile:water (86:14 v/v); for analysis, gas chromatography/mass spectrometry after derivatisation with trifluoroacetic anhydride was utilised. The recovery of deoxynivalenol and T-2 toxin from both food matrixes ranged from 90.1 to 94.0%. The occurrence of these mycotoxins in bread was 28.0% and 2.6% for deoxynivalenol and T-2 toxin, respectively, whereas in pasta, the occurrence of both mycotoxins was higher, varying from 9.3 to 62.7%. The mean content of deoxynivalenol (42.5 μg/kg) in bread was lower than the content of T-2 toxin (68.37 μg/kg), while…

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…

Traducción y adaptación al castellano del cuestionario de salud para celíacos Canadian Celiac Health Survey

Introducción: Adaptar y valorar el cuestionario de calidad de vida denominado Canadian Celiac Health Survey (CCHS). Objetivo: Traducir y adaptar en castellano el cuestionario CCHS para poder ser utilizado por la población de habla hispana puesto que se trata de un cuestionario específico para la celiaquía. Método: La adaptación del CCHS, que consta de 76 ítems distribuidos en 11 secciones diferentes, se realizó mediante el método de traducción-retrotraducción y tras ser revisado y consensuado se procedió a realizar una prueba piloto con 25 personas celíacas, de forma individual y por un miembro del grupo de investigación, para valorar la comprensión de los ítems y sus secciones. Las aportac…

Application of matrix solid-phase dispersion to the determination of a new generation of fungicides in fruits and vegetables.

A method based on matrix solid-phase dispersion (MSPD) and gas chromatography to determine eight fungicides in fruits and vegetables is described. Fungicide residues were identified and quantified using nitrogen-phosphorus detection and electron-capture detection connected in parallel and confirmed by mass spectrometric detection. The method required 0.5 g of sample, C18 bonded silica as dispersant sorbent, silica as clean-up sorbent and ethyl acetate as eluting solvent. Recoveries from spiked orange, apple, tomato, artichoke, carrot and courgette samples ranged from 62 to 102% and relative standard deviations were less than 15% in the concentration range 0.05-10 mg kg(-1). Detection and qu…

Mycotoxin Occurrence and Risk Assessment in Gluten-Free Pasta through UHPLC-Q-Exactive Orbitrap MS

Celiac disease (CD) is a genetic-based autoimmune disorder which is characterized by inflammation in the small intestinal mucosa due to the intolerance to gluten. Celiac people should consume products without gluten, which are elaborated mainly with maize or other cereals. Contamination of cereals with mycotoxins, such as fumonisins (FBs) and aflatoxins (AFs) is frequently reported worldwide. Therefore, food ingestion is the main source of mycotoxin exposure. A new analytical method was developed and validated for simultaneous analysis of 21 mycotoxins in gluten-free pasta, commonly consumed by celiac population as an alternative to conventional pasta. Ultrahigh-performance liquid chromatog…

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…

Optimization of Matrix Solid-Phase Dispersion method for simultaneous extraction of aflatoxins and OTA in cereals and its application to commercial samples

Abstract A method based on Matrix Solid-Phase Dispersion (MSPD) has been developed for the determination of 5 mycotoxins (ochratoxin A and aflatoxins B and G) in different cereals. Several dispersants, eluents and ratios were tested during the optimization of the process in order to obtain the best results. Finally, samples were blended with C 18 and the mycotoxins were extracted with acetonitrile. Regarding to matrix effects, the results clearly demonstrated the necessity to use a matrix-matched calibration to validate the method. Analyses were performed by liquid chromatography–triple quadrupole-tandem mass spectrometry (LC–QqQ-MS/MS). The recoveries of the extraction process ranged from …

Potential application of lactic acid bacteria in the biopreservation of red grape from mycotoxigenic fungi

Background Filamentous fungi are the main contamination agent in the viticultural sector. Use of synthetic fungicides is the regular answer to these contaminations. Nevertheless, due to several problems associated with the use of synthetic compounds the industry demands new and safer methods. In the present work, the biopreservation potential of four lactic acid bacteria (LAB) strains was studied against the principal grape contaminant fungi. Results Agar diffusion test evidenced that all four culture-free supernatant (CFS) had antifungal properties against all tested fungi. The Minimum Inhibitory Concentration (MIC) and Minimum Fungicidal Concentration (MFC) test values evidenced that medi…

Evaluation of mycotoxins and their metabolites in human breast milk using liquid chromatography coupled to high resolution mass spectrometry

Humans can be exposed to mycotoxins through the food chain. Mycotoxins are mainly found as contaminants in food and could be subsequently excreted via biological fluids such as urine or human breast milk in native or metabolised form. Since breast milk is usually supposed as the only food for new-borns, the occurrence of mycotoxins in thirty-five human milk samples was evaluated by a newly developed method based on QuEChERS extraction and UHPLC-HRMS detection. The method described here allows the detection of target mycotoxins in order to determine the quality of this initial feeding. The method has been fully validated, with recoveries ranging from 64% to 93% and relative standard deviatio…

Climatic conditions influence emerging mycotoxin presence in wheat grown in Romania – A 2-year survey

Abstract The correlation between the occurrence of four enniatins (ENA, ENA1, ENB, and ENB1) and beauvericin (BEA) and the weather parameters during anthesis and preharvest period was studied in 97 wheat samples collected in 2014 and 2015 across three counties from central and south Romania (Brasov, Dâmboviţa, and Teleorman). The highest mean values of ENA (16.1 μg kg −1 ) and ENB (147.1 μg kg −1 ) were measured in the samples from Brasov county in the harvest year 2015, whereas for ENA1 and ENB1 the highest means (55.2 μg kg −1 , and 108.0 μg kg −1 , respectively) were noted in samples from Teleorman county in 2014. Statistically significant differences ( P r s  = 0.8745 and r s  = 0.9326;…

Factors affecting the presence of ochratoxin A in wines.

Ochratoxin A (OTA) are synthesized mainly by different species of Aspergillus and Penicillium being its human toxicological effects reflected in different countries due to the consumption of different foods and beverages such as red, white, rose, and special wines. This review presents an overview of the direct (meteorological conditions, grape cultivation, and wine-making techniques) and indirect (latitude, year of production, use of pesticides, presence of spoilage microorganisms, conditions of storage of the harvested grapes, type of maceration, and conditions of fermentation), factors affecting the presence of OTA in wines.

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.

Antifungal effect of phenolic extract of fermented rice bran with Rhizopus oryzae and its potential use in loaf bread shelf life extension

In this study the antifungal potential of a phenolic extract obtained from rice bran fermented with Rhizopus oryzae CECT 7560 and its application in the elaboration of bread was assessed.; Results: Eighteen compounds with antifungal potential were identified by LC-ESI-qTOF-MS in the extract: organic acids, gallates and gallotannins, flavonoids, ellagic acid and benzophenone derivatives. The extract was active against strains of Fusarium, Aspergillus and Penicillium, with minimum inhibitory concentration ranging from 390 to 3100 µg mL-1 and minimum fungicidal concentration variable from 780 to 6300 µg mL-1 . The strains that were most sensitive to the phenolic extract were F. graminearum, F.…

Urinary levels of enniatin B and its phase I metabolites: First human pilot biomonitoring study

Abstract Enniatins (Enns) are mycotoxins produced by Fusarium spp. and are widely distributed contaminants of cereals and derivate products. Among the different identified enniatins, Enn B is the most relevant analogue in cereals in Europe. Therefore, the aim of this study was to investigate for the first time the occurrence of Enn B and Enn B phase I metabolites in 300 human urine samples throughout an ultrahigh-performance liquid chromatography-high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) methodology. Three different sample preparation procedures were evaluated and salting-out liquid-liquid extraction showed satisfactory validation results. Enn B was quantified in 83.7% of sa…

Antifungal effects of the bioactive compounds enniatins A, A1, B, B1

To produce enniatin (ENs), Fusarium tricinctum CECT 20150 was grown in a liquid medium of potato (PDB), being mycotoxin purified by high performance liquid chromatography (HPLC) with a reverse phase semipreparative column using a mobile phase of acetonitrile/water using gradient condition. The purity of the ENs fractions was verified by analytical HPLC and LC/MS-MS. The pure fractions of ENs were utilized to study the biological activity on several mycotoxigenic moulds as Fusarium verticilloides, Fusarium sporotrichioides, Fusarium tricinctum, Fusarium poae, Fusarium oxysporum, Fusarium proliferatum, Beauveria bassiana, Trichoderma harzianum, Aspergillus flavus, Aspergillus parasiticus, Asp…

Evaluating the impact of supercritical-CO2 pressure on the recovery and quality of oil from “horchata” by-products: Fatty acid profile, α-tocopherol, phenolic compounds, and lipid oxidation parameters

Abstract The effect of supercritical carbon dioxide (SC-CO2) (10–40 MPa) and conventional extraction (CE) to recover oil from by-products obtained during “horchata” production was assessed. To evaluate both extraction techniques, the fatty acid composition, polyphenols, α-tocopherol, antioxidant capacity and lipid oxidation parameters of the extracts obtained were analysed. A linear relationship between extraction pressure and oil yield was observed. However, the highest oil yield was obtained under conventional extraction. The by-products from “horchata” presented a profile where monounsaturated fatty acids (MUFA) were the predominant, representing ≈ 70% of total fatty acids. The amount of…

Bioaccessibility of glucoraphanin from broccoli using anin vitrogastrointestinal digestion model

This study investigated the effect of cooking methods on the degradation of glucoraphanin (GR) in broccoli and the bioaccessibility of this compound through simulated gastrointestinal digestion. Broccoli was cooked using three different techniques: boiling, steaming and microwave cooking. Then, GR was extracted and quantified by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). In addition, the cooked samples were added to a system that simulates the digestion characteristics of the mouth, stomach and duodenum. Samples were drawn before and after the digestion, and GR bioaccessibility was calculated. GR losses were higher when broccoli was boiled (47.03%), followed by …

Determination of pesticide residues in fruit and vegetables.

A review concerning the determination of pesticide residues in fruit and vegetables is presented. The basic principles and recent developments in the extraction and quantitation of pesticides are discussed. Consideration is given to solid phase and supercritical extraction techniques, automation and robotic systems, and immunoassay procedures.

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%…

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.

Exposure assessment approach through mycotoxin/creatinine ratio evaluation in urine by GC–MS/MS

Abstract In this pilot survey human urine samples were analyzed for presence of 15 mycotoxins and some of their metabolites using a novel urinary multi-mycotoxin GC–MS/MS method following salting-out liquid–liquid extraction. Fifty-four urine samples from children and adults residents in Valencia were analyzed for presence of urinary mycotoxin and expressed in gram of creatinine. Three out of 15 mycotoxins were detected namely, HT-2 toxin, nivalenol and deoxynivalenol (DON). 37 samples showed quantifiable values of mycotoxins. Co-occurrence of these contaminants was also observed in 20.4% of assayed samples. DON was the most frequently detected mycotoxin (68.5%) with mean levels of 23.3 μg/…

Analysis of organophosphorus pesticides in honeybee by liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry.

Pesticides applied in extended agricultural fields may be controlled by means of bioindicators, such as honeybees, in which are the pesticides bioaccumulate. Liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (LC-APCI-MS) experiments with positive (PI) and negative (NI) ion modes were optimized for the analysis of 22 organophosphorus pesticides in honeybee samples. The extraction required 3 g of sample, which was extracted with acetone. The extract was purified with coagulating solution and reextracted with Cl(2)CH(2). Pesticides studied could be detected by both ionization modes except for parathion, parathion-methyl, and bromophos, which did not give signals …

Quantification of Imidacloprid in Honeybees: Development of a Chemiluminescent ELISA

A Chemiluminescente Enzyme-Linked Immuno-Sorbent Assay (CL-ELISA) for determination and quantification of the fungicide imidacloprid in honeybees was developed in an indirect competitive format. The assay was optimized by determining: the optimal coating conjugate concentration and anti-imidacloprid antiserum dilution, the effect of the incubation time on the competitive step, the tolerance to organic solvents. The IC50 and the limit of detection (LOD) values were 14.8 ng mL-1 and 0.11 ng mL -1, respectively, similar to those of colorimetric ELISA with a calibration range of 0.1 – 2600 ng mL-1. Cross reactivity of some related compounds such as three imidacloprid metabolites, 6-chloro nicot…

Determination of ochratoxin A in organic and non-organic cereals and cereal products from Spain and Portugal

The objective of this work was to know the occurrence of OTA in organic and non-organic cereals and cereal products from Spain and Portugal. A method based on extraction with matrix solid phase dispersion (MSPD) using octylsilica (C8) followed by liquid chromatography coupled with fluorescence detection (LC-FD) was used to determine OTA from the selected samples. Recoveries of OTA from the studied samples spiked at 10 ng/g level ranged from 78% to 89% with a standard deviation of 3.66. The limits of detection and quantification of this method were 0.05 and 0.19 ng/g, respectively. Furthermore, LC-FD after OTA methylation was used to confirm the identity of OTA in all positive samples. This …

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…

Accelerated solvent extraction of ochratoxin a from rice samples.

In this paper, accelerated solvent extraction (ASE) for the analysis of ochratoxin A (OTA) is applied for the first time. Optimization of the method is given for the extraction of OTA from rice samples. Several parameters such as type of solvent, temperature, pressure, static time, and cell size were investigated thoroughly to find the optimal extraction conditions. The optimum ASE operating conditions were methanol as extraction solvent, 1500 psi, 40 degrees C, 5 min of static time, 50% flush volume, 60 s of purge, 1 cycle, and 11 mL cell size. The total extraction time was approximately 15 min. OTA was determined by liquid chromatography coupled with a fluorescence detector and confirmed …

Pesticide residues in oranges from Valencia (Spain)

One hundred and fifty citrus samples from an agricultural co-operative of the Valencian Community (Spain) were analysed for pre- and post-harvest pesticide residues using high performance liquid chromatography and gas chromatography. Among the residues from post-harvest treatments, imazalil was detected in 112 (74.7%) samples at a mean level of 1.2 mg/kg, thiabendazole in 21 (14.0%) samples at a mean level of 0.47 mg/kg and carbendazim in 5 (3.3%) samples at a mean level of 1.05 mg/kg. Among the residues from pre-harvest treatment, dicofol was detected in 28 (18.7%) samples at a mean level of .28 mg/kg chlorpyriphos in 19 (12.7% samples at a mean level of 0.16 mg/kg and endosulfan in 11 (7.…

Routine application using single quadrupole liquid chromatography-mass spectrometry to pesticides analysis in citrus fruits.

Abstract A rapid and sensitive liquid chromatography–electrospray ionization–mass spectrometry method has been developed for the routine analysis of buprofezin, bupirimate, hexaflumuron, tebufenpyrad, fluvalinate and pyriproxyfen in citrus fruits. Extracts were obtained by matrix solid-phase dispersion (MSPD) using C 18 as dispersant and dichloromethane-methanol (80:20, v/v) as eluent. Matrix effects were tested for all matrices by addition of standard to sample blank extracts (samples containing no detectable residues). Mean recoveries obtained at fortification levels between 0.01 and 5 mg kg −1 were 57–97% with relative standard deviations (RSDs) from 5 to 19%. The limits of quantificatio…

Dietary intake and food pattern among university students

Dietary intake and food pattern were determined among university students. Nine hundred-eighteen students (384 males and 534 females) completed a 24 hour recall and a questionnaire. This study showed an average energy intake of 10.3 and 8.0 MJ among male and female university students. The mean intakes of energy and magnesium for males and energy, iron, magnesium and zinc for females were lower to the Spanish Recommended Intakes. For males, breakfast, lunch, dinner and snacks bring about 18, 33, 25 and 27% of total dairy energy, respectively, however, for females a 15, 34, 23 and 27% of total dairy energy were obtained for breakfast, lunch, dinner and snacks, respectively. The university po…

Absence Ochratoxin A in soy sauce

A method is described for the determination of ochratoxin A (OTA) in soy sauce using phosphate-buffered saline (PBS) extraction, an immunoaffinity clean-up, a liquid chromatographic determination with fluorescence detection (LC-FD) and confirmation with LC-FD after methylation of OTA. Recoveries of OTA spiked to soy sauce samples at 0.25 ng/ml level were 90% with relative standard deviations of 4%. The limit of detection was 0.01 ng/ml for OTA using the proposed method. Furthermore, the proposed method was applied to 60 soy sauce samples from China and Japan and none of them were found to contain OTA.

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…

Mass spectrometry strategies for mycotoxins analysis in European beers

In this work, an existent solid-phase extraction (SPE) procedure was used to study the occurrence of mycotoxins in different European beers. HPLC-QqQ-MS/MS and ultra high resolution mass analyser have been optimized for the analysis of 18 mycotoxins: the methods were validated according to the EU Commission Decision 2002/657/EC guidelines. In this sense, matrix-matched calibration was performed for each type of beer, obtaining an effective quantification. The recoveries ranged from 63 to 91% and repeatability and reproducibility expressed as relative standard deviation (RSD%) were lower than 17%. On one hand, HPLC-LTQ-Orbitrap ® was used for unambiguous identification of target mycotoxins, …

Influence of the antimicrobial compound allyl isothiocyanate against the Aspergillus parasiticus growth and its aflatoxins production in pizza crust.

Abstract Aflatoxins (AFs) are secondary metabolites produced by different species of Aspergillus, such as Aspergillus flavus and Aspergillus parasiticus, which possess mutagenic, teratogenic and carcinogenic activities in humans. In this study, active packaging devices containing allyl isothiocyanate (AITC) or oriental mustard flour (OMF) + water were tested to inhibit the growth of A. parasiticus and AFs production in fresh pizza crust after 30 d. The antimicrobial and anti-aflatoxin activities were compared to a control group (no antimicrobial treatment) and to a group added with commercial preservatives (sorbic acid + sodium propionate). A. parasiticus growth was only inhibited after 30 d…

Effect of allyl isothiocyanate on transcriptional profile, aflatoxin synthesis, and Aspergillus flavus growth.

Abstract The goals of this study were to determine the efficacy of allyl isothiocyanate (AITC) against the growth of A. flavus and Aflatoxin B1 (AFB1) production as well as to evaluate changes in the transcriptome profile when colonizing maize. A. flavus was inoculated in potato dextrose agar (PDA), the plates were placed inside glass jars and the mycelial growth (MG) was monitored for 7 d. Likewise, maize grains were contaminated with A. flavus in glass jars of 1 L and treated with 0.125, 0.25, 0.5, 1 and 5 µL of AITC. The moisture content (MC) of grains was 15 and 21%. After 7 days of storage, the MG was significantly reduced in doses higher than 0.125 µL/L of AITC. All doses of AITC redu…

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

Evaluation of beauvericin and enniatins in Italian cereal products and multicereal food by liquid chromatography coupled to triple quadrupole mass spectrometry

In this study, 48 multicereal baby foods samples including 25 of pasta and 23 of multicereal baby foods from supermarkets of Campania region (Italy) were analysed for evaluating the presence of beauvericin (BEA) and enniatins (ENs) A, A1, B, B1 and B4. Subsequently to evaluate the risk exposure of Italian population and infant population over the consumption of pasta or multicereal baby food, was, respectively, evaluated. For the above mentioned evaluation, a method developed in our laboratory by liquid chromatography tandem mass spectrometry was used. A liquid phase dispersion procedure was optimised for the simultaneous extraction of BEA and the five ENs from multicereal baby food samples…

Occurrence and legislation of mycotoxins in food and feed from Morocco

Mycotoxins are natural food and feed contaminants, mainly produced by moulds of genera Aspergillus, Penicillium and Fusarium. The number of mycotoxins known to exert toxic effect on human and animal health is constantly increasing as well as the legislative provisions taken to control their presence in food and feed. Morocco, a North African country, surrounded by the Mediterranean Sea and Atlantic Ocean, has a climate characterized by high humidity and high temperature which favor growth of moulds. This paper gives an overview about the contamination levels and the occurrence of some mycotoxins (e.g. aflatoxins, ochratoxin A, and Fusarium toxins) in cereals, bread, milk, spices, wine, oliv…

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.

Analysis of carbamate and phenylurea pesticide residues in fruit juices by solid-phase microextraction and liquid chromatography–mass spectrometry

A new analysis method to detect carbamates and phenylurea pesticide residues in fruit juices was developed using solid-phase microextraction (SPME) coupled with liquid chromatography-single quadrupole mass spectrometry (LC/MS) and liquid chromatography-quadrupole ion trap mass spectrometry (LC/QIT-MS). The pesticide residues present in watery matrices as fruit juices were extracted using three types of fibers: 50-microm Carbowax/templated resin (CW/TPR), 60-mum poly(dimethylsiloxane)/divinylbenzene (PDMS/DVB) and 85-microm polyacrylate. The different extraction conditions were evaluated choosing as the best parameters 90 min (time), 20 degrees C (temperature) and 1 ml (volume). After extrac…

Exposure to patulin from consumption of apple-based products.

Patulin is a mycotoxin produced by species of Penicillium, Aspergillus and Byssochylamys. Several Scientific Committees classify patulin as mutagenic, embryotoxic and immunotoxic. It has been found as a natural contaminant of processed apple products and its presence may be indicative of the quality of fruit used in production. In this work, a method for the analysis of patulin is described, based on a simple liquid-liquid extraction with acetonitrile; patulin is analyzed using liquid chromatography with UV detection. Patulin identity was confirmed by GC-MS after its reaction with N-methyl-N-(trimethylsilyl)trifluoroacetamide. Fifty-three apple-containing products were analyzed and patulin …

Comparison of liquid chromatography using triple quadrupole and quadrupole ion trap mass analyzers to determine pesticide residues in oranges.

Liquid chromatography-triple quadrupole/mass spectrometry (LC-TQ/MS) and liquid chromatography-quadrupole ion trap/mass spectrometry (LC-QIT/MS) for determining bupirimate, hexaflumuron, tebufenpyrad, buprofezin, pyriproxyfen, and fluvalinate in fruits have been compared. The differences in the mass spectra obtained by triple and ion trap quadrupoles are discussed, showing how both of them provide interesting features. The evaluation of the two instruments was carried out by ethyl acetate extraction of oranges spiked with the studied pesticides at LOQ and 10 times the LOQ. Results obtained by LC-TQ/MS correlated well with those obtained by LC-QIT/MS. Recoveries were 70-94% by LC-TQ/MS and 7…

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…