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

使用多抗体免疫亲和柱净化,通过 LC-MS/MS 分析动物饲料中的黄曲霉毒素、伏马毒素、脱氧雪腐镰刀菌烯醇、赭曲霉毒素 A、玉米赤霉烯酮、HT-2 和 T-2 毒素

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作者:Naomi Mackay, Elaine Marley, Dave Leeman, Cezary Poplawski, Carol Donnelly

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.

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.

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.

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.

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).

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