Liquid Chromatographic Determination of Fumonisins B 1 , B 2 , and B 3 in Corn Silage (original) (raw)
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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.
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...
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....
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 protection
A specific sheep polyclonal antiserum was applied to the competitive direct enzyme-linked immunosorbent assay (CD-ELISA) of fumonisins in Fusarium cultures, fresh corn, animal feed, and human foods, and the results were related to fumonisin B1 (FB1) levels determined by liquid chromatography (LC). The limit of detection of FB1 for the CD-ELISA was 0.1 ng/ml. Cross reactivity (defined as [ng/ml FBI required for 50% inhibition]/[ng/ml of analogue required for 50% inhibition] × 100) for the antiserum in the ELISA was 100, 24, and 30%, for FB1 FB2, and FB3, respectively. FB1 was detected in F. moniliforme cultures grown on corn (≤760 μg/g) but not in those for F. graminearum. Fumonisin estimates were 2.8-fold higher by CD-ELISA than FB1 estimates by LC. Upon using the antiserum in a commercial CD-ELISA format (Veratox), mean recovery of FB1 from spiked corn (0.1 to 3 μg/g)was 85.2 ± 25.3%, whereas the mean recovery by LC was 74.1 ± 6.2%. Analyses of 43 fresh corn, 28 animal feed, and 14...
Estimation of dietary intakes of fumonisins B1 and B2 from conventional and organic corn
Food Control, 2007
The dietary intakes of fumonisins from 60 samples of conventional and organic corn were assessed. A 13.3% of the conventional corn samples contained fumonisin B 1 and B 2 at mean levels of 43 and 22 ng/g, respectively, while 10% of the organic corn samples contained fumonisins at somewhat lower levels of 35 ng/g (FB 1 ) and 19 ng/g (FB 2 ). Overall, the fumonisin levels in the corn samples were much lower than the maximum level of 2000 ng/g (as the sum of FB 1 and FB 2 ) proposed for unprocessed maize in a recent EU regulation. The fumonisins present in conventional and organic maize are estimated to contribute with very low percentages of 0.21% and 0.17%, respectively, to the level considered at risk for human health. Based on the data exposed in this paper, the farming system is probably not of decisive importance for the Wnal contamination of agricultural products with these mycotoxins.
J Agr Food Chem, 1994
The fumonisin mycotoxins are secondary metabolites of Fusarium moniliforme, which are common worldwide in maize. Thin-layer chromatography (TLC) and competitive indirect immunoassay (CI-ELISA) methods were compared for detection of fumonisin B1 in maize. Corn from the 1991 Missouri maize variety trials (322 samples) was collected, milled, subsampled, and analyzed independently by two laboratories using different screening methods. Fifty-two percent of the samples tested negative for fumonisin B1 a t 1 ppm by both methods. By TLC, an additional 14% of the samples had less than 1 ppm of fumonisin B1 and more than 1 ppm by CI-ELISA. TLC found 34% of the samples were from 1 to 10 ppm and only 1% of the samples were above 10 ppm of fumonisin B1. CI-ELISA found 28% of the samples contained fumonisin levels between 1 and 10 ppm and 20% had greater than 10 ppm of fumonisin. In the remaining samples, CI-ELISA consistently reported higher fumonisin levels than TLC (160/322), while TLC was higher in only 10 of 322 samples. The discrepancy may be due to fumonisin B1 alone being detected by TLC, while CI-ELISA measures total fumonisins. Both methods are well suited for rapid screening of maize samples for fumonisin contamination.
Toxins, 2015
Three compounds, hypothesized as fumonisin A1 (FA1), fumonisin A2 (FA2), and fumonisin A3 (FA3), were detected in a corn sample contaminated with mycotoxins by high-resolution liquid chromatography-Orbitrap mass spectrometry (LC-Orbitrap MS). One of them has been identified as FA1 synthesized by the acetylation of fumonisin B1 (FB1), and established a method for its quantification. Herein, we identified the two remaining compounds as FA2 and FA3, which were acetylated fumonisin B2 (FB2) and fumonisin B3 (FB3), respectively. Moreover, we examined a method for the simultaneous analysis of FA1, FA2, FA3, FB1, FB2, and FB3. The corn samples were prepared by extraction using a QuEChERS kit and purification using a multifunctional cartridge. The linearity, recovery, repeatability, limit of detection, and limit of quantification of the method were >0.99, 82.9%-104.6%, 3.7%-9.5%, 0.02-0.60 μg/kg, and 0.05-1.98 μg/kg, respectively. The simultaneous analysis of the six fumonisins revealed ...
Affinity column clean‐up for the analysis of fumonisins and their hydrolysis products in corn
Food and Agricultural Immunology, 1997
Fumonisins, toxic metabolites of certain Fusarium molds, can be found in corn and corn-based foods at levels sufficient to cause disease in livestock. Widely used methods of analysis involve organic extraction followed by isolation with either C 18 or strong anion exchange columns. An affinity column procedure that simultaneously isolated both intact and hydrolyzed fumonisins from corn is reported. The columns were prepared using two monoclonal antibodies with differing specificities, the first (P2A5-3-F3) bound intact fumonisins, while the second (P2F11-3-H7) bound their hydrolysis products. Corn samples were extracted with methanol/phosphate buffer (80:20, v:v) and the extract was diluted and applied to the affinity column. The recoveries of FB 1 from corn spiked with 0.5-8.0 ppm averaged 82%. The recoveries of HFB 1 over the range 0.25-1.25 ppm averaged 102%. Affinity columns capable of binding both intact and hydrolyzed fumonisins allow for the analysis of both types of toxins simultaneously from a single methanolic extract.
Food Additives and Contaminants, 2001
The determination of fumonisins in cornXakes is a challengin g matter as the actually available methods for the analysis of corn do not perform well when applied to this more complex matrix. After testing several factors that may aVect the analytical performance, an accurate method for the determination of fumonisin B 1 (FB 1 ) and B 2 (FB 2 ) in cornXakes has been developed. The method uses immunoaYnity chromatography for clean-up and high performance liquid chromatograph y (HPLC) for quantiWcation of the toxins. Samples were extracted twice with acetoni-trile± methanol± water (25:25:50) and the combined extracts were diluted with phosphate buVered saline (PBS) and applied to a FumoniTest TM immunoaYnity column. After washing with PBS, fumonisins were eluted from the column with methanol and reacted with o-phthaldialdehyde /2-mercaptoethanol to form Xuorescent derivatives. Fumonisin derivatives were analysed by reversed phase HPLC with Xuorometric detection using methanol± 0.1 M phosphat e buVer (77:23; pH adjusted at 3.35) as mobile phase. The average recoveries for FB 1 and FB 2 spiked in the ranges of 0.33± 2.80 ·g/g and 0.17± 1.40 ·g/g were 102.6% and 95.1% , respectively, with average relative standard deviations of 9% and 8% . The limit of quantiWcation for FB 1 and FB 2 was 0.005 ·g/g based on a signal-to-noise ratio of 6:1 by using a sensitive Xuorescence detector. The method was used to analyse 18 cornXakes and cornXake cereals samples for FB 1 and FB 2 contamination. All but one sample were found to be contaminated, with maxi-mum FB 1 and FB 2 concentrations of 1.092 ·g/g and 0.235 ·g/g, respectively. Mean FB 1 and FB 2 concentrations were 0.157 ·g/g and 0.036 ·g/g, respectively.