Method For Determination Of Deoxynivalenol Research Articles

The Validation of Neogen Reveal® Q+ for DON.

Reveal® Q+ for DON is an immunochromatographic test for quantitative determination of deoxynivalenol (DON) in grains. A study was conducted to validate performance of this method for determination of DON in naturally contaminated corn and wheat and in DON-spiked corn/soy blend, soybeans, barley, malted barley, buckwheat, brown rice, sorghum, and distillers dried grain. In addition to matrix testing, LOD, linearity, selectivity, robustness, and stability/lot-to-lot consistency testing were performed. The LOD was determined to be 0.014 ppm in corn and 0.037 ppm in wheat, and the LOQ 0.042 ppm in corn and 0.11 ppm in wheat. Recovery ranged from 90 to 104% across a range of reference values of 0.5 to 34.5 ppm. Linearity calculation comparing test results with reference values produced R2 values of 0.999 in both matrixes. Internal results with corn and wheat were corroborated in independent laboratory testing. For DON-spiked commodities, mean recovery across a range of DON concentration from 0.5 to 30 ppm ranged from 90 to 109%. Results of selectivity testing showed no cross-reactivity with other mycotoxins and no interference in detection of DON. Reagent lot-to-lot consistency and stability studies showed consistent results across a range of DON levels and established expiration dating for the test of 18 months after manufacture when stored under specified conditions. Conclusions and Highlights: The Reveal Q+ for DON test offers reliable performance as well as the advantages of aqueous sample extraction, procedural simplicity, minimal labor and equipment requirements, and rapid results. The Reveal Q+ for DON test is validated as a Performace Tested Method in Corn, Wheat, and a variety of other grains. The test provides rapid results from a simple aqueous extraction and requires minimal labor and equipment.

Simple Validated Method for Determination of Deoxynivalenol and Zearalenone in Some Cereals Using High Performance Liquid Chromatography

Simple Validated Method for Determination of Deoxynivalenol and Zearalenone in Some Cereals Using High Performance Liquid Chromatography

Development and validation of a gas chromatography–mass spectrometry method for determination of deoxynivalenol and its metabolites in human urine

The determination of deoxynivalenol (DON) and its metabolites such as deepoxy-deoxynivalenol (DOM-1) in human urine is complicated due its low levels (ng/mL) and the complexity of the matrix. A gas chromatography–mass spectrometry method was optimized and validated for the confirmation analysis of DON and its metabolites in urine samples using 13C isotopic-labeled DON as internal standard. In the sample preparation the type and amount of β-glucuronidase for enzymatic hydrolysis was investigated as well as the cleanup procedure, being compared the immunoaffinity column with solid-phase extraction (SPE). As far as we know, SPE C18 cleanup procedure was applied for the first time in the analysis of DON and its metabolites in human urine. Using this analytical methodology the detection and quantification limits achieved ranged from 0.06 to 0.30ng/mL and from 0.2 to 1.0ng/mL, respectively. Recoveries were higher than 73% for fortification levels between 25 and 100ng/mL and repeatability were lower than 13%. The natural occurrence of DON and its metabolites in human urine samples from the north zone of Portugal was studied. Free DON was detected in 15% of the samples whereas total (free+conjugated) DON was detected in 69% of the samples. Deepoxy-deoxynivalenol, 3-acetyldeoxynivalenol and 15-acetyldeoxynivalenol were not detected in any of the samples analyzed.

Simultaneous Determination of Deoxynivalenol and Nivalenol in Traditional Chinese Medicine by SPE and LC

A liquid chromatography method for simultaneous determination of deoxynivalenol (DON) and nivalenol (NIV) in traditional Chinese medicine (TCM) is described for the first time. Different clean-up columns and extraction solvents have been optimized. The DON purification column was chosen as the best. The detection limits for DON and NIV were 62.5 and 50.0 μg kg−1, respectively. Recoveries from different TCMs spiked with DON and NIV at levels ranging from 0.5 to 10 mg kg−1 were 82.8–99.9% and 80.8–100.3%, respectively. None of the 30 commercially available TCM samples analyzed, were found to contain any detectable amounts of DON and NIV.

Sensitive method for determination of DON in cocoa by means of HPLC-techniques

The mycotoxin deoxynivalenol (DON) is one of a group of mycotoxins known as type B trichothecenes and is particularly formed by the mould speciesFusarium graminearum andFusarium culmorum. The frequency of the occurrence of DON in certain raw materials and the concentrations found make it one of the world's most significant mycotoxin contaminants. Positive findings of the toxin especially have been established in cereal-based foods, as well as in oilseeds.The main objective of this study was to set up a current situation assessment of the possible occurrence of deoxynivalenol in cocoa and cocoa products. As there was no analytical method for determining DON in cocoa and cocoa products, a special method was developed. The applicability and consistency of the method was confirmed by performing recovery assays on various cocoa products. A special post-column derivatisation procedure was developed to increase selectivity and raise sensitivity by a factor of 80.The method was used to test 230 samples for possible DON content, ranging from cocoa beans to cocoa bean shells, nibs, cocoa liquor and cocoa powders through to finished cocoa-based products. The results suggest that DON may occasionally occur in cocoa beans in very low concentrations.

Determination of deoxynivalenol in cereals by HPLC-UV

A simple method for determination of deoxynivalenol (DON) in cereal samples is described. DON was extracted with methanol, the solvent evaporated, and the residue redissolved with water. This extract was purified on immunoaffinity columns. DON was determined by HPLC with UV-detection. The limits of detection (LOD) and quantification (LOQ) were 10 and 50 µg/kg, respectively.

A simple method for determination of deoxynivalenol in cereals and flours

An effective and fast method for determination of deoxynivalenol (DON) in cereals and flours has been developed. The immunoaffinity column was used for the isolation of DON from wheat, corn, rice and flour extract. The determination was carried out by using the HPLC/UV method. The limit of detection was 0.02 mg/kg. The recoveries for the assay range 0.1 to 2 mg/kg were generally higher than 80%, ranging from 83 to 96% with an average relative standard deviation of 3.8%. The trueness of the method using the DON test – HPLC column was established by use of certified reference material CRM 379. The certified value was 0.67 mg/kg. The result obtained from three replicates was 0.68 ± 0.05 mg/kg. The corresponding confidence interval at 95% probability ranged from 0.63 to 0.73 mg/kg. A comparative study of the DON testTM – HPLC/UV and the Mycosep 225 – GC/ECD methods was carried out. Six naturally contaminated wheat samples were analysed by both methods. Linear regression analysis demonstrates that DON testTM – HPLC is a statistically significant predictor of the GC/ECD method using the Romer Mycosep 225 column.  

NON–DESTRUCTIVE DETERMINATION OF DEOXYNIVALENOL LEVELS IN BARLEY USING NEAR–INFRARED SPECTROSCOPY

The objective of this research was to develop a neural network based method for determination of Deoxynivalenol (DON) levels in barley using near–infrared (NIR) spectroscopy. The NIR spectra of 188 barley samples with DON level between 0.3 to 50.8 ppm were collected using the FOSS NIR Systems 6500 Near Infrared Spectrometer. The DON levels were measured with GC/mass spectrometry (GC/MS). With the NIR spectra as input variable and GC/mass measured DON as output variable, neural networks were developed and trained to predict DON levels. The prediction accuracy of the models was tested using randomly selected production sets, which had not been seen by the models. The effects of wavelength interval and ranges on the prediction accuracy of models were also examined. The results demonstrate that the combination of neural networks and NIR spectra can be conveniently used to determine DON levels in barley.

ガスクロマトグラフィーによる穀類中のデオキシニバレノール,ニバレノールの効率的な定量分析法

ガスクロマトグラフィーによる穀類中のデオキシニバレノール,ニバレノールの効率的な定量分析法

Efficient Method for Determination of Deoxynivalenol and Nivalenol in Wheat and Barley by Gas Chromatography

We have developed an analytical method for rapidly determining deoxynivalenol (DON) and nivalenol (NIV), typical mycotoxins in cereals infected by Fusarium spp., in wheat and barley. Since we used a solvent of high polarity and water to elute columns of mixed alumina-charcoal-celite and Amberlite for extraction and clean-up, we were able to omit partition and dehydration processes. The separated mycotoxin was determined by ECD-gas chromatography after trimethylsilylation. The limits of determination of DON and NIV are 12ng/g, and that of fusarenon-X (F-X) is 24ng/g. These limits are adequate for economic screening analysis. When DON and NIV were added to wheat and barley at the 120ng/g level and F-X at the 240ng/g level, the recoveries were 101 and 95%, 93 and 92%, and 96 and 96%, and the coefficients of variation were 2.9 and 3.4%, 3.7 and 2.4%, and 3.5 and 3.4%, respectively. When F-X was detected by this method, its identity was confirmed by mass chromatography. So far we have not detected F-X in cereals. Further studies are in progress. This method was designed for rapid determination of wheat and barley, but should also be applicable to the analysis of mixed feed. It is easy to change the limits of determination by changing the quantities of sample and solvent. One analyst can analyze 8 samples in a day, and two analysts, 20 samples.

Thin Layer Chromatographic Method for Determination of Deoxynivalenol in Wheat: Collaborative Study

A collaborative study of a rapid method for the determination of deoxynivalenol (DON) in winter wheat was successfully completed. The method involves sample extraction with acetonitrile-water (84 + 16), cleanup using a disposable column of charcoal, Celite, and alumina, and detection by thin layer chromatography after spraying with an aluminum chloride solution. Each of the 15 collaborators analyzed 12 samples, 2 of which were naturally contaminated, and 10 to which DON was added, in duplicate, at levels of 0, 50, 100, 300, and 1000 ng/g. Average recoveries of DON ranged from 78 to 96% with repeatabilities of 30-64% and reproducibilities of 33-87%. The results of the study show that false positives were not a problem and that all of the analysts could detect DON at the 300 ng/g level or higher. The method has been adopted official first action.