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Automated regenerable microarray-based immunoassay for rapid parallel quantification of mycotoxins in cereals

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Abstract

An automated flow-through multi-mycotoxin immunoassay using the stand-alone Munich Chip Reader 3 platform and reusable biochips was developed and evaluated. This technology combines a unique microarray, prepared by covalent immobilization of target analytes or derivatives on diamino-poly(ethylene glycol) functionalized glass slides, with a dedicated chemiluminescence readout by a CCD camera. In a first stage, we aimed for the parallel detection of aflatoxins, ochratoxin A, deoxynivalenol, and fumonisins in cereal samples in a competitive indirect immunoassay format. The method combines sample extraction with methanol/water (80:20, v/v), extract filtration and dilution, and immunodetection using horseradish peroxidase-labeled anti-mouse IgG antibodies. The total analysis time, including extraction, extract dilution, measurement, and surface regeneration, was 19 min. The prepared microarray chip was reusable for at least 50 times. Oat extract revealed itself as a representative sample matrix for preparation of mycotoxin standards and determination of different types of cereals such as oat, wheat, rye, and maize polenta at relevant concentrations according to the European Commission regulation. The recovery rates of fortified samples in different matrices, with 55–80 and 58–79 %, were lower for the better water-soluble fumonisin B1 and deoxynivalenol and with 127–132 and 82–120 % higher for the more unpolar aflatoxins and ochratoxin A, respectively. Finally, the results of wheat samples which were naturally contaminated with deoxynivalenol were critically compared in an interlaboratory comparison with data obtained from microtiter plate ELISA, aokinmycontrol® method, and liquid chromatography–mass spectrometry and found to be in good agreement.

Principle of the competitive chemiluminescence ELISA using the microarray chip

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Acknowledgments

This research project was supported by the German Ministry of Economics and Technology (via AiF) and the FEI (Forschungskreis der Ernährungsindustrie e.V. Bonn); project AiF 381 ZN. Further, the authors thank Dr. T. Westermair and Dr. P. Walser (muva Kempten) for LC-MS analysis of deoxynivalenol in grain samples and Martina Kirsch, Rosenmühle Landshut (Germany) for providing naturally contaminated grain samples and performing analysis of deoxynivalenol by the aokinmycontrol® system. We are also grateful to Dr. G. Lystik (Soft Flow Biotechnology, Gödöllo, Hungary) for providing the anti-OTA monoclonal antibody and Huntsman Corporation for the free samples of DAPEG.

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Authors and Affiliations

  1. Institute of Hydrochemistry and Chair of Analytical Chemistry, Technische Universität München, Marchioninistrasse 17, 81377, Munich, Germany

    S. Oswald, X. Y. Z. Karsunke, R. Niessner & D. Knopp

  2. Hygiene and Technology of Milk, Ludwig-Maximilians Universität München, Schönleutnerstrasse 8/219, 85764, Oberschleissheim, Germany

    R. Dietrich & E. Märtlbauer

Authors
  1. S. Oswald
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  2. X. Y. Z. Karsunke
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  3. R. Dietrich
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  4. E. Märtlbauer
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  5. R. Niessner
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  6. D. Knopp
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Corresponding author

Correspondence to D. Knopp.

Additional information

This paper is dedicated to Professor Franz Dickert on the occasion of his 70th birthday.

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Oswald, S., Karsunke, X.Y.Z., Dietrich, R. et al. Automated regenerable microarray-based immunoassay for rapid parallel quantification of mycotoxins in cereals. Anal Bioanal Chem 405, 6405–6415 (2013). https://doi.org/10.1007/s00216-013-6920-3

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  • DOI: https://doi.org/10.1007/s00216-013-6920-3

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