Simultaneous determination of Deoxynivalenol, Deoxynivalenol-3-Glucoside and Nivalenol in wheat grains by HPLC-PDA with immunoaffinity column cleanup

Deoxynivalenol-3-glucoside (D3G) is a modified mycotoxin formed by the metabolism of plants through the conjugation of deoxynivalenol (DON) with glucose. Toxicology studies of D3G for human and animal health are still under investigation, and the development of practical and reliable methods for its direct determination, especially in cereal matrices, is of great importance. In the present study, a methodology for simultaneous determination of D3G, DON, and nivalenol (NIV) in wheat grains, using immunoaffinity column (IAC) cleanup, separation by C18 column and detection by ultraviolet (UV) absorption, was optimized and in-house validated. The results demonstrated adequate values of D3G recovery from IAC and spiked samples. Intraday precision, linearity, limit of detection and limit of quantification (LOQ) were also adequate for the determination of these mycotoxins. Range of applicability varied from 47.1 to 1000 g/kg for D3G and from 31.3 to 1000 g/kg for DON and NIV, with recovery ranging from 84.7±7.2 % to 112.3±8.1Felipe Trombete is grateful for a doctoral fellowship provided by the Brazilian Federal Agency for Support and Evaluation of Graduate Education (CAPES)

Occurrence of Fusarium head blight and mycotoxins as well as morphological identification of Fusarium species in winter wheat in Kosovo

Wheat (Triticum aestivum L.) is an important cereal crop in Kosovo and a major component of population food. Fusarium head blight (FHB) of wheat is an economically very significant disease. FHB leads to various losses in quality like reduced germination of seeds, reduced baking quality and reduced nutritional quality through mycotoxin contamination. In 2010 and 2011 the incidence and identity of the Fusarium spp. infecting wheat in Kosovo as well as mycotoxin contamination was investigated. The results of two years research work show that the incidence of FHB on winter wheat in 2010 was low (<6%). In the year 2011 the disease incidence was clearly higher (up to 31%). Based on morphological characters, F. graminearum was the most frequently Fusarium sp. identified on wheat kernels in the year 2010 (100%) and 2011 (98%). Less frequently isolated species included F. cerealis (<1%) and F. avenaceum (<1%). Wheat flour samples were analyzed by liquid chromatography coupled to mass spectrometry and found to be contaminated with a variety of mycotoxins, most importantly deoxynivalenol and zearalenone. This is the first report on the incidence as well as on the identification of Fusarium species isolated from naturally infected winter wheat in Kosovo

Colour-encoded paramagnetic microbead-based direct inhibition triplex flow cytometric immunoassay for ochratoxin A, fumonisins and zearalenone in cereals and cereal-based feed

A combined (triplex) immunoassay for the simultaneous detection of three mycotoxins in grains was developed with superparamagnetic colour-encoded microbeads, in combination with two bead-dedicated flow cytometers. Monoclonal antibodies were coupled to the beads, and the amounts of bound mycotoxins were inversely related to the amounts of bound fluorescent labelled mycotoxins (inhibition immunoassay format). The selected monoclonal antibodies were tested for their target mycotoxins and for cross-reactivity with relevant metabolites and masked mycotoxins. In the triplex format, low levels of cross-interactions between the assays occurred at irrelevant high levels only. All three assays were influenced by the sample matrix of cereal extracts to some extent, and matrix-matched calibrations are recommended for quantitative screening purposes. In a preliminary in-house validation, the triplex assay was found to be reproducible, sensitive and sufficiently accurate for the quantitative screening at ML level. The triplex assay was critically compared to liquid chromatography–tandem mass spectrometry using reference materials and fortified blank material. Results for the quantification of ochratoxin A and zearalenone were in good agreement. However, the fumonisin assay was, due to overestimation, only suitable for qualitative judgements. Both flow cytometer platforms (Luminex 100 and FLEXMAP 3D) performed similar with respect to sensitivity with the advantages of a higher sample throughput and response range of the FLEXMAP 3D and lower cost of the Luminex 100

Safe food and feed through an integrated toolbox for mycotoxin management: the MyToolBox approach

There is a pressing need to mobilise the wealth of knowledge from the international mycotoxin research conducted over the past 25-30 years, and to perform cutting-edge research where knowledge gaps still exist. This knowledge needs to be integrated into affordable and practical tools for farmers and food processors along the chain in order to reduce the risk of mycotoxin contamination of crops, feed and food. This is the mission of MyToolBox – a four-year project which has received funding from the European Commission. It mobilises a multi-actor partnership (academia, farmers, technology small and medium sized enterprises, food industry and policy stakeholders) to develop novel interventions aimed at achieving a significant reduction in crop losses due to mycotoxin contamination. Besides a field-to-fork approach, MyToolBox also considers safe use options of contaminated batches, such as the efficient production of biofuels. Compared to previous efforts of mycotoxin reduction strategies, the distinguishing feature of MyToolBox is to provide the recommended measures to the end users along the food and feed chain in a web-based MyToolBox platform (e-toolbox). The project focuses on small grain cereals, maize, peanuts and dried figs, applicable to agricultural conditions in the EU and China. Crop losses using existing practices are being compared with crop losses after novel pre-harvest interventions including investigation of genetic resistance to fungal infection, cultural control (e.g. minimum tillage or crop debris treatment), the use of novel biopesticides suitable for organic farming, competitive biocontrol treatment and development of novel modelling approaches to predict mycotoxin contamination. Research into post-harvest measures includes real-time monitoring during storage, innovative sorting of crops using vision-technology, novel milling technology and studying the effects of baking on mycotoxins at an industrial scale

Determination of zearalenone and its metabolites in endometrial cancer by coupled separation techniques

This study presents a selective method of isolation of zearalenone (ZON) and its metabolite, α-zearalenol (α-ZOL), in neoplastically changed human tissue by accelerated solvent and ultrasonic extractions using a mixture of acetonitrile/water (84/16% v/v) as the extraction solvent. Extraction effectiveness was determined through the selection of parameters (composition of the solvent mixture, temperature, pressure, number of cycles) with tissue contamination at the level of nanograms per gram. The produced acetonitrile/water extracts were purified, and analytes were enriched in columns packed with homemade molecularly imprinted polymers. Purified extracts were determined by liquid chromatography (LC) coupled with different detection systems (diode array detection - DAD and mass spectrometry - MS) involving the Ascentis RP-Amide as a stationary phase and gradient elution. The combination of UE-MISPE-LC (ultrasonic extraction - molecularly imprinted solid-phase extraction - liquid chromatography) produced high (R ≈ 95–98%) and repeatable (RSD < 3%) recovery values for ZON and α-ZOL

Alcohol Drinking in Never Users of Tobacco, Cigarette Smoking in Never Drinkers, and the Risk of Head and Neck Cancer: Pooled Analysis in the International Head and Neck Cancer Epidemiology Consortium

Background At least 75% of head and neck cancers are attributable to a combination of cigarette smoking and alcohol drinking. A precise understanding of the independent association of each of these factors in the absence of the other with the risk of head and neck cancer is needed to elucidate mechanisms of head and neck carcinogenesis and to assess the efficacy of interventions aimed at controlling either risk factor. Methods We examined the extent to which head and neck cancer is associated with cigarette smoking among never drinkers and with alcohol drinking among never users of tobacco. We pooled individual-level data from 15 case-control studies that included 10244 head and neck cancer case subjects and 15227 control subjects, of whom 1072 case subjects and 5775 control subjects were never users of tobacco and 1598 case subjects and 4051 control subjects were never drinkers of alcohol. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using unconditional logistic regression models. All statistical tests were two-sided. Results Among never drinkers, cigarette smoking was associated with an increased risk of head and neck cancer (OR for ever versus never smoking = 2.13, 95% CI = 1.52 to 2.98), and there were clear dose-response relationships for the frequency, duration, and number of pack-years of cigarette smoking. Approximately 24% (95% CI = 16% to 31%) of head and neck cancer cases among nondrinkers in this study would have been prevented if these individuals had not smoked cigarettes. Among never users of tobacco, alcohol consumption was associated with an increased risk of head and neck cancer only when alcohol was consumed at high frequency (OR for three or more drinks per day versus never drinking = 2.04, 95% CI = 1.29 to 3.21). The association with high-frequency alcohol intake was limited to cancers of the oropharynx/hypopharynx and larynx. Conclusions Our results represent the most precise estimates available of the independent association of each of the two main risk factors of head and neck cancer, and they exemplify the strengths of large-scale consortia in cancer epidemiolog