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Development of a multiple immunoaffinity column for simultaneous determination of multiple mycotoxins in feeds using UPLC–MS/MS

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Abstract

A sensitive and specific immunoaffinity column to clean up and isolate multiple mycotoxins was developed along with a rapid one-step sample preparation procedure for ultra-performance liquid chromatography–tandem mass spectrometry analysis. Monoclonal antibodies against aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, zearalenone, ochratoxin A, sterigmatocystin, and T-2 toxin were coupled to microbeads for mycotoxin purification. We optimized a homogenization and extraction procedure as well as column loading and elution conditions to maximize recoveries from complex feed matrices. This method allowed rapid, simple, and simultaneous determination of mycotoxins in feeds with a single chromatographic run. Detection limits for these toxins ranged from 0.006 to 0.12 ng mL−1, and quantitation limits ranged from 0.06 to 0.75 ng mL−1. Concentration curves were linear from 0.12 to 40 μg kg−1 with correlation coefficients of R 2 > 0.99. Intra-assay and inter-assay comparisons indicated excellent repeatability and reproducibility of the multiple immunoaffinity columns. As a proof of principle, 80 feed samples were tested and several contained multiple mycotoxins. This method is sensitive, rapid, and durable enough for multiple mycotoxin determinations that fulfill European Union and Chinese testing criteria.

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Acknowledgments

This project was supported by the Outstanding Young Talents Plan of China, Special Fund for Grain-scientific Research in the Public Interest (201513006-2), Special Fund for Agro-scientific Research in the Public Interest (201203094-4).

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    Authors and Affiliations

    1. Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan, 430062, China

      Xiaofeng Hu, Zhaowei Zhang, Peiwu Li & Qi Zhang

    2. Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan, 430062, China

      Xiaofeng Hu, Zhaowei Zhang & Peiwu Li

    3. Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan, 430062, China

      Xiaofeng Hu, Zhaowei Zhang, Peiwu Li & Qi Zhang

    4. Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan, 430062, China

      Xiaofeng Hu & Peiwu Li

    5. State Key Laboratory of Magnetic Resonance and Atomic Molecular Physics, Wuhan Institute of Physics and Mathematics, Institute of Hydrobiology, Chinese Academy of Sciences, 7 Donghu Rd, Wuhan, 430072, China

      Rui Hu & Min Wang

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    1. Xiaofeng Hu
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    2. Rui Hu
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    Correspondence to Zhaowei Zhang or Peiwu Li.

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    Additional information

    Published in the topical collection Immunoanalysis for Environmental Monitoring and Human Health with guest editors Shirley J. Gee, Ivan R. Kennedy, Alice Lee, Hideo Ohkawa, Tippawan Prapamontol, and Ting Xu.

    Xiaofeng Hu and Rui Hu contributed equally to this work.

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    Hu, X., Hu, R., Zhang, Z. et al. Development of a multiple immunoaffinity column for simultaneous determination of multiple mycotoxins in feeds using UPLC–MS/MS. Anal Bioanal Chem 408, 6027–6036 (2016). https://doi.org/10.1007/s00216-016-9626-5

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    • DOI: https://doi.org/10.1007/s00216-016-9626-5

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