Determination of fumonisins B 1 and B 2 in cornflakes by high performance liquid chromatography and immunoaffinity clean-up (original) (raw)
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Journal of AOAC International
A liquid chromatographic (LC) method for the determination of fumonisins B1 (FB1) and B2 (FB2) in corn and corn flakes was collaboratively studied by 23 laboratories, which analyzed 5 blind duplicate pairs of each matrix to establish the accuracy, repeatability, and reproducibility characteristics of the method. Fumonisin levels in the corn ranged from <0.05 (blank) to 1.41 microg/g for FB1 and from <0.05 to 0.56 microg/g for FB2, whereas in the corn flakes they ranged from <0.05 to 1.05 microg/g for FB1 and from <0.05 to 0.46 microg/g for FB2. The method involved double extraction with acetonitrile-methanol-water (25 + 25 + 50), cleanup through an immunoaffinity column, and LC determination of the fumonisins after derivatization with o-phthaldialdehyde. Relative standard deviations for the within-laboratory repeatability (RSDr) of the corn analyses ranged from 19 to 24% for FB1 and from 19 to 27% for FB2; for the corn flakes analyses, RSDr ranged from 9 to 21 % for FB1 ...
Journal of Food Science, 2011
The occurrence of the fumonisins B 1 and B 2 in maize-based food products marketed in Italy was examined. A simply and reliable chromatographic method with fluorimetric detection and postcolumn o-phtalaldehyde derivatization was used for a monitoring of 100 samples (8 flours, 21 corn-meal, 16 snacks, 7 maize samples, 13 gluten-free products, and 35 corn-flakes) bought in local supermarkets during the years 2008 and 2009. The presence of both fumonisins B 1 and B 2 , at a concentration higher than 15 μg/kg, was observed in all samples of corn-meal and maize-flour, in 75% of snacks, in 57% of maize samples, in 54% of gluten-free products, and in 29% of corn-flakes. A total of 7 samples including 4 corn-meals, 2 maize-flours, and 1 maize showed a value exceeding the maximum level fixed in the Regulation 1126/2007/EC; no positive sample was observed in corn-flakes, snacks, and gluten-free foods. Fumonisins contamination, on the whole range of maize-based food products analyzed, emphasizes the need of improve agricultural practices, and increase official control and monitoring studies.
Journal of Chromatography A, 2008
A sensitive and selective analytical method was developed for the quantitative determination of fumonisins B 1 and B 2 in maize-based foods for direct human consumption. The method, based on highperformance liquid chromatography and fluorescence detection, presents a rapid and automated on-line post-column derivatization, performed with o-phtalaldehyde and N,N-dimethyl-2-mercaptoethylamine. Several factors affecting the separation and detection of fumonisins were investigated, including mobile phase composition, column features, derivatization agent flow-rate and both the excitation and the emission wavelengths. Optimal fluorescence detection was obtained by using a exc of 343 nm and a em of 445 nm. Under the optimized experimental conditions, a complete separation of fumonisins was obtained in less than 13 min by using a C 18 column and a gradient elution at 0.8 mL/min with methanol and 0.1 M phosphate buffer at pH 3.15. The limits of detection for FB 1 and FB 2 were 4 and 5 g/L corresponding to 5 and 6 g/kg in matrix. Each fumonisin was determined in the range 40-320 g/L that correspond to 50-400 g/kg in matrix. The necessary requirements for accuracy, reproducibility and sensitivity were fulfilled and recovery values ranged from 87 to 94% for FB 1 and from 70 to 75% for FB 2 in cornflake samples at three fortification levels in the range 100-300 g/kg. The potential of this method, combined with a simple clean-up procedure, was assessed by the measurements of FB 1 and FB 2 in maize-based products, such as maize flour, "polenta", tortillas and cookies.
Journal of AOAC International
A single-laboratory method validation was conducted to establish the effectiveness of an immunoaffinity column cleanup procedure followed by liquid chromatography/mass spectrometry (LCIMS) for the determination of fumonisins B1 and B2 (FBI + FB2) in corn. The test portion is extracted with acetonitrile-methanol-water (25 + 25 + 50). The extract is filtered, diluted with phosphate-buffered saline solution, and applied to an immunoaffinity column. FB1 + FB2 are removed with methanol and directly determined by reversed-phase LC with MS detection using selected-ion monitoring of 2 characteristic ions in each case. Test portions of blank corn samples were spiked with a mixture of FB1 + FB2 to give total levels of 200 and 500 ng/g, respectively. Recoveries of both FB1 and FB2 from spiked samples averaged 90.4-101%. Based on results for spiked raw corn (triplicates at 2 levels), the relative standard deviation for repeatability ranged from 2.8 to 7.1%. The accuracy of the method was demons...
Bioaccessibility of total bound fumonisin from corn flakes
Mycotoxin Research, 2009
Fumonisin B 1 (FB 1) is often found as a natural contaminant of corn and corn-based food. Several publications have demonstrated the presence of fumonisin bound to proteins and to other compounds of the matrix. In spite of the low oral bioavailability of FB 1 in rats, pigs, chickens, cows, and monkeys, FB 1 can cause agriculturally significant disease and possibly human cancer. The aim of this work was to determine the bioaccessibility of total bound FB 1 (TB FB 1) (percentage of TB FB 1 , released from corn flakes to the chyme) after in vitro digestion. Two samples of corn flakes washed with solvents were incubated with gastrointestinal tract solutions simulating saliva plus stomach and duodenal juices. After hydrolysis of the chyme with KOH, TB FB 1 was determined as hydrolyzed FB 1 (HFB 1). The bioaccessibility of TB FB 1 in chyme from corn flakes was 37-64%, indicating that these derivatives should be considered in evaluation of exposure to fumonisin.
Effects of muffin processing on fumonisins from 14C-labeled toxins produced in cultured corn kernels
Journal of food protection, 2003
The persistence of fumonisins during cooking is known to be affected by several factors, including thermal degradation and the presence of various ingredients in corn-based food recipes that can react with the toxin. A method for the production of corn kernels containing 14C-fumonisins was developed. The corn kernels were colonized by Fusarium verticillioides MRC 826 and supplemented with 1,2-14C-sodium acetate. The specific activity of 14C-FB1 produced made the study of its fate in cornmeal muffins possible. The double-extraction acetonitrile-water-methanol/immunoaffinity column/o-phthaldialdehyde high-performance liquid chromatography (HPLC) method was used to determine FB1 levels in cornmeal muffins. Reductions in FB1 levels in muffins spiked with 14C-labeled and unlabeled FB1 (43 and 48%, respectively) were similar, indicating that the extraction method was efficient and consistent with previous reports. However, with the labeled corn kernel material, recovery levels based on th...
Biointerface Research in Applied Chemistry, 2021
Fumonisins B1 and B2 are carcinogenic and commonly contaminate corn and corn-based products. Analysis of such toxins using C18 HPLC column is officially accredited but still unknown if all column types can effectively separate FB1 and FB2 or not. The present study evaluated the efficiency of 5 analytical columns with different dimensions, particle sizes, and porosities to determine these toxins in both agar cultures of Fusarium verticillioides and cornflakes. Interestingly, the traditional column 150mm of length with 5µm porous particles had close retention times to those of the short-fused core column 75mm of length with 2.7 µm reflecting in time and solvents saving. Using Sep-Pack C18 for clean-up played an important role in enhancement the limit of quantification (LOQ) for cornflake samples (5-13.7 and 16.1-39 µg kg-1 for FB1 and FB2, respectively). However, it was relatively higher for fungal culture samples that were not passed through the cleaning-up step (11.5-16 and 28.1-46....
Journal of The Serbian Chemical Society, 2005
Corn silage was dried, ground, and then extracted with 0.1 M ethylenediaminetetraacetic acid. The filtrate was applied to a FumoniTest immunoaffinity column. Fumonisins were derivatized with naphthalene-2,3-dicarboxaldehyde, separated on a C 18 liquid chromatographic column, and detected by fluorescence. The detection limits for fumonisin B 1 , fumonisin B 2 , and fumonisin B 3 were 50, 25, and 25 ng/g of dried silage, respectively. Recoveries of fumonisin B 1 , fumonisin B 2 , and fumonisin B 3 from wet and dried corn silage spiked over the range of 100-5000 ng/g averaged 91-106%. The method was applied to corn silage samples collected from the midwestern area of the United States during 2001-2002. Of 89 corn silage samples, fumonisin B 1 , fumonisin B 2 , and fumonisin B 3 were found in 86 (97%), 64 (72%), and 51 (57%) of the samples. The mean positive levels of fumonisin B 1 , fumonisin B 2 , and fumonisin B 3 were 615, 93, and 51 ng/g, respectively, in dried silage. This suggests that fumonisins may be frequent low level contaminants in corn silage.
Fumonisins are one of the most agriculturally significant environmental toxins produced by Fusarium and Aspergillus species that grow on agricultural commodities in the field or during storage. Cereals contaminated with fumonisins causes serious loss to agricultural produce leads to health problems in humans and other farm animals. In the present study, polyclonal hyperimmune sera was raised against FB1 in rabbits immunized with FB1–keyhole limpet haemocyanin (KLH). Purified antibodies were used to establish a sensitive gold nanoparticle based immunochromatographic strip (ICG) for detecting FB1 levels in cereal grains. Effective on-site detection of FB1 was achieved by developing a rapid and sensitive pAb based ICG strip. This strip had a detection limit of 5 ng mL−1 for FB1 in cereal samples and it could be completed within 3 min. Close examination of 150 cereal samples by ICG strip method revealed that 77 were fumonisin-positive. Results obtained by the developed method was further validated with well standardized HPLC method and results of strip method was correlated well with those obtained by HPLC method. In conclusion, the developed method was a better alternative for onsite detection of FB1 in cereal samples intended for human consumption to reduce risk of humans and other farm animals. The high level of FB1 concentrations recorded in present study warrants the need to develop an awareness creation programme to the farmers of India for safe handling of cereal grains at the time of harvesting and storage of grains.
Journal of AOAC International
An interlaboratory validation study was conducted to establish the method performance characteristics of an immunoaffinity column (IAC) cleanup procedure followed by LC/MS for the determination of fumonisins B1 (FB1) and B2 (FB2) and combined FB1 + FB2 in corn. The test portion is extracted with acetonitrile-methanol-water (25 + 25 + 50). The extract is filtered, diluted with phosphate-buffered saline solution, and applied to an IAC. FB1 and FB2 are removed with methanol, followed by water, then directly determined by RPLC with MS detection using selected-ion monitoring of two characteristic ions in each case. Naturally contaminated corn samples were milled to a fine powder and mixed to produce three samples with target levels of combined FB1 + FB2 ranging from 350 to 4000 microg/kg. Of 15 initially participating laboratories, two failed to report results and another did not follow the prescribed method. Thus, valid results were obtained from 12 participants located in 11 countries....