Dataset Information

Analysis of Aflatoxins, Fumonisins, Deoxynivalenol, Ochratoxin A, Zearalenone, HT-2, and T-2 Toxins in Animal Feed by LC-MS/MS Using Cleanup with a Multi-Antibody Immunoaffinity Column.

ABSTRACT:

Background

Regulations limiting aflatoxin levels in animal feed and guidance values for maximum levels for fumonisins (FB1 and FB2), deoxynivalenol (DON), ochratoxin A (OTA), zearalenone (ZON), HT-2, and T-2 toxins are in place both to protect animal health and to minimize potential transfer to animal products for human consumption. A multi-mycotoxin method which can handle complex feed matrices such as distillers dried grains with solubles (DDGS) is essential for analysis and accurate quantification without the need to revert to separately analyze individual mycotoxins.

Objective

The objective of this study is to generate single laboratory validation data for a method employing a multi-antibody immunoaffinity column (IAC) capable of providing cleanup for eleven mycotoxins, followed by LC-MS/MS quantification without the need for isotopic labelled and matrix-matched standards. The applicability of method is to be demonstrated for corn feed, pig feed, and DDGS by fortification and naturally occurring mycotoxins covering the range of regulated limits.

Methods

Feed sample (1 kg) ground by milling to approximately 1-2 mm particle size and sub-sample (5 g) extracted with acetonitrile-water-formic acid, passing through a multi-mycotoxin IAC, washing, and eluting prior to LC-MS/MS analysis monitoring selected ion transitions.

Results

Recoveries were in the range 74 to 117% (excluding five outliers) for aflatoxins, FB1, FB2, DON, OTA, ZON, HT-2, and T2- toxins spiked into three commercial animal feed matrixes (n = 84) and within-day RSDs averaged 1.7 to 10.3% (n = 99).

Conclusion

Single laboratory validation of a multi-antibody IAC method coupled with LC-MS/MS has shown the method to be suitable for accurate quantification of eleven regulated mycotoxins in DDGS, pig feed, and poultry feed.

Highlights

IAC method capable of accurately quantifying eleven regulated mycotoxins in complex feed matrices.

SUBMITTER: Mackay N

PROVIDER: S-EPMC9446684 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature

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Publications

Analysis of Aflatoxins, Fumonisins, Deoxynivalenol, Ochratoxin A, Zearalenone, HT-2, and T-2 Toxins in Animal Feed by LC-MS/MS Using Cleanup with a Multi-Antibody Immunoaffinity Column.

Mackay Naomi N Marley Elaine E Leeman Dave D Poplawski Cezary C Donnelly Carol C

Journal of AOAC International 20220901 5

<h4>Background</h4>Regulations limiting aflatoxin levels in animal feed and guidance values for maximum levels for fumonisins (FB1 and FB2), deoxynivalenol (DON), ochratoxin A (OTA), zearalenone (ZON), HT-2, and T-2 toxins are in place both to protect animal health and to minimize potential transfer to animal products for human consumption. A multi-mycotoxin method which can handle complex feed matrices such as distillers dried grains with solubles (DDGS) is essential for analysis and accurate q ...[more]

PMID: 35258598

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Project description:BackgroundThe 11+Myco MS-PREP® immunoaffinity column (IAC) contains a gel suspension of monoclonal antibodies specific to the toxins of interest. Following sample extraction, the IAC is used for cleanup and concentration of mycotoxins prior to analysis by LC with tandem mass spectrometry (LC-MS/MS).ObjectiveThis study evaluated the IAC with LC-MS/MS method for Performance Tested MethodSM certification for simultaneous determination and confirmation of aflatoxins (AF) B1, B2, G1, G2, and M1; deoxynivalenol (DON), fumonisins B1, B2, and B3; ochratoxin A (OTA); T-2; HT-2; and zearalenone (ZON) from corn, wheat, cereal-based baby food (with and without dairy ingredients), paprika, chili powder, and animal feed.MethodsA single extraction method using acetonitrile-water (1 + 1, by volume) was used for all matrixes. The method developer validated all matrixes and an independent laboratory verified method performance on corn and animal feed. Data were analyzed for recovery, repeatability precision, LOD, LOQ, confirmation of identity, and method selectivity.ResultsRecovery (72-138%) and repeatability (0.46-24%), with the exception of sporadic data points, were within acceptance criteria. LOQ was estimated as AFB1 0.018-0.32 μg/kg, AFB2 0.037-0.28 μg/kg, AFG1 0.019-0.14 μg/kg, AFG2 0.036-0.28 μg/kg, DON 4.0-75 μg/kg, fumonisin B1 4.9-37 μg/kg, fumonisin B2 4.0-32 μg/kg, fumonisin B3 2.0-16 μg/kg, OTA 0.15-4.4 μg/kg, T-2 0.5-7.5 μg/kg, HT-2 0.70-7.5 μg/kg, and ZON 1.3-7.2 μg/kg, depending on matrix. Method performance was verified with reference and QC materials. Selectivity and confirmation of identity were also demonstrated.ConclusionThe 11+Myco MS-PREP IAC with LC-MS/MS method demonstrated acceptable performance for simultaneous determination of 12 mycotoxins in seven matrixes.HighlightThe data were reviewed by the AOAC Performance Tested MethodsSM Program and approval was granted for certification of the 11+Myco MS-PREP Method as PTM 112401.

Project description:Fusarium mycotoxins often co-occur in broiler feed, and their presence negatively impacts health even at subclinical concentrations, so there is a need to identify the concentrations of these toxins that do not adversely affect chickens health and performance. The study was conducted to evaluate the least toxic effects of combined mycotoxins fumonisins (FUM), deoxynivalenol (DON), and zearalenone (ZEA) on the production performance, immune response, intestinal morphology, and nutrient digestibility of broiler chickens. A total of 960 one-day-old broilers were distributed into eight dietary treatments: T1 (Control); T2: 33.0 FUM + 3.0 DON + 0.8 ZEA; T3: 14.0 FUM + 3.5 DON + 0.7 ZEA; T4: 26.0 FUM + 1.0 DON + 0.2 ZEA; T5: 7.7 FUM + 0.4 DON + 0.1 ZEA; T6: 3.6 FUM + 2.5 DON + 0.9 ZEA; T7: 0.8 FUM + 1.0 DON + 0.3 ZEA; T8: 1.0 FUM + 0.5 DON + 0.1 ZEA, all in mg/kg diet. The results showed that exposure to higher mycotoxin concentrations, T2 and T3, had significantly reduced body weight gain (BWG) by 17% on d35 (p < 0.05). The T2, T3, and T4 groups had a significant decrease in villi length in the jejunum and ileum (p < 0.05) and disruption of tight junction proteins, occludin, and claudin-4 (p < 0.05). Higher mycotoxin groups T2 to T6 had a reduction in the digestibility of amino acids methionine (p < 0.05), aspartate (p < 0.05), and serine (p < 0.05); a reduction in CD4+, CD8+ T-cell populations (p < 0.05) and an increase in T regulatory cell percentages in the spleen (p < 0.05); a decrease in splenic macrophage nitric oxide production and total IgA production (p < 0.05); and upregulated cytochrome P450-1A1 and 1A4 gene expression (p < 0.05). Birds fed the lower mycotoxin concentration groups, T7 and T8, did not have a significant effect on performance, intestinal health, and immune responses, suggesting that these concentrations pose the least negative effects in broiler chickens. These findings are essential for developing acceptable thresholds for combined mycotoxin exposure and efficient feed management strategies to improve broiler performance.

Dataset Information

Analysis of Aflatoxins, Fumonisins, Deoxynivalenol, Ochratoxin A, Zearalenone, HT-2, and T-2 Toxins in Animal Feed by LC-MS/MS Using Cleanup with a Multi-Antibody Immunoaffinity Column.

Background

Objective

Methods

Results

Conclusion

Highlights

Publications

Analysis of Aflatoxins, Fumonisins, Deoxynivalenol, Ochratoxin A, Zearalenone, HT-2, and T-2 Toxins in Animal Feed by LC-MS/MS Using Cleanup with a Multi-Antibody Immunoaffinity Column.

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