Determination of fumonisins B1 and B2 in corn by liquid chromatography/mass spectrometry with immunoaffinity column cleanup: single-laboratory method validation (original) (raw)
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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 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.
Liquid Chromatographic Determination of Fumonisins B 1 , B 2 , and B 3 in Corn Silage
Journal of Agricultural and Food Chemistry, 2004
A liquid chromatographic (LC) method for simultaneous determination of fumonisins B1 (FB1), B2 (FB2), and B3 (FB3) in corn was subjected to a collaborative study involving 12 participants from 10 countries, in which the accuracy and reproducibility characteristics of the method were established. Mean analyte recoveries from corn ranged from 81.1 to 84.2% for FB1 (at a spiking range of 500 to 8000 ng/g), from 75.9 to 81.9% for FB2 (at a spiking range of 200 to 3200 ng/g), and from 75.8 to 86.8% for FB3 (at a spiking range of 100 to 1600 ng/g). The valid data were statistically evaluated after exclusion of outliers. Relative standard deviations for within-laboratory repeatability ranged from 5.8 to 13.2% for FB1, from 7.2 to 17.5% for FB2, and from 8.0 to 17.2% for FB3. Relative standard deviations for between-laboratory reproducibility varied from 13.9 to 22.2% for FB1, from 15.8 to 26.7% for FB2, and from 19.5 to 24.9% for FB3. HORRAT ratios, calculated for the individual toxin analogues, ranged from 0.75 to 1.73. The LC method for determination of fumonisins B1, B2, and B3 in corn (at concentrations of 800-12800 ng total fumonisins/g) has been adopted by AOAC INTERNATIONAL.
Food chemistry, 2017
This work involved the optimization and validation of a method, according to Directive 2002/657/EC and the Analytical Quality Assurance Manual of Ministério da Agricultura, Pecuária e Abastecimento, Brazil, for simultaneous extraction and determination of fumonisins B1 and B2 in maize. The extraction procedure was based on a matrix solid phase dispersion approach, the optimization of which employed a sequence of different factorial designs. A liquid chromatography-tandem mass spectrometry method was developed for determining these analytes using the selected reaction monitoring mode. The optimized method employed only 1g of silica gel for dispersion and elution with 70% ammonium formate aqueous buffer (50mmolL(-1), pH 9), representing a simple, cheap and chemically friendly sample preparation method. Trueness (recoveries: 86-106%), precision (RSD ≤19%), decision limits, detection capabilities and measurement uncertainties were calculated for the validated method. The method scope wa...
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.
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.
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 ...
A Rapid, Sensitive Thin Layer Chromatography Procedure for the Detection of Fumonisin B1 and B2
Journal of Veterinary Diagnostic Investigation, 1992
A thin layer chromatography (TLC) method was developed for the detection of fumonisin B1 and B2 in corn and corn-based feedstuffs. Finely ground samples were extracted with acetonitrile: water (1: 1), filtered, and applied to C18 cleanup columns. The columns were washed with 1% aqueous KCl followed by acetonitrile: 1% aqueous KC1 (1:9), and the fumonisins were eluted with acetonitrile: water (7:3). The eluants were concentrated and spotted on reverse-phase C18 TLC plates along with fumonisin B1 and B2 standards, and the plates were developed in methanol: 4% aqueous KCl (3:2). The fumonisins were visualized by spraying the TLC plates successively with 0.1 M sodium borate buffer, fluorescamine, and 0.01 M boric acid. The plates were then dried and examined under longwave ultraviolet light. Fumonisin B1 and B2 appeared as bright yellowish-green fluorescent bands at Rfs of 0.5 and 0.1, respectively. The detection limit for the fumonisins on the TLC plate was 0.1 ppm in corn. Recoveries ...
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.