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Optimization and Validation of a Fluorescence Polarization Immunoassay for Rapid Detection of T-2 and HT-2 Toxins in Cereals and Cereal-Based Products

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Abstract

A fluorescence polarization (FP) immunoassay has been optimized and validated for rapid quantification of T-2 and HT-2 toxins in both unprocessed cereals, including oats, barley and rye, and cereal-based products for direct human consumption, such as oat flakes, oats crispbread and pasta. Samples were extracted with 90 % methanol, and the extract was filtered and diluted with water or sodium chloride solution prior to the FP immunoassay. Overall mean recoveries from spiked oats, rye, barley, oat flakes, oats crispbread and pasta ranged from 101 to 107 %, with relative standard deviations lower than 7 %. Limits of detection (LODs) of the FP immunoassay were 70 μg/kg for oats, 40 μg/kg for oat flakes and barley, 25 μg/kg for pasta and 20 μg/kg for rye and oats crispbread. The trueness of the immunoassay was assessed by using two oat and oat flake reference materials for T-2 and HT-2 toxins, showing good accuracy and precision. Good correlations (r > 0.953) were observed between T-2 and HT-2 toxin contents in naturally and artificially contaminated samples determined by both FP immunoassay and ultra-high-performance liquid chromatography (UHPLC) with immunoaffinity column cleanup used as reference method. These results, combined with rapidity and simplicity of the assay, show that the optimized assay is suitable for high-throughput screening, as well as for reliable quantitative determination of T-2 and HT-2 toxins in cereals and cereal-based products.

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Acknowledgments

This work has been supported by the Italian Ministry of Education, University and Research, MIUR (P.O.N. “Ricerca & Competitività” 2007–2013), project no. 02_00186_341751212792-S.I.Mi.S.A. “New Strategies for Improvement of Food Safety: Prevention, Control, Correction” and project no. CTN01_00230 CL.A.N. Cluster Tecnologici Nazionali—SAFE&SMART project “New enabling technologies for food safety and food chain integrity within a global scenario”. The authors would like to thank Prof. Simon G. Edwards of Harper Adams University College (Newport, UK) for providing naturally contaminated oat samples. The authors would like to thank Giuseppe Panzarini (ISPA-CNR) for his technical help in the HPLC analysis.

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Authors and Affiliations

  1. Institute of Sciences of Food Production (ISPA), National Research Council of Italy (CNR), via G. Amendola 122/O, 70126, Bari, Italy

    Anna Chiara R. Porricelli, Vincenzo Lippolis, Stefania Valenzano, Marina Cortese & Michelangelo Pascale

  2. Barilla Food Research Labs, via Mantova 166, 43122, Parma, Italy

    Michele Suman & Sandro Zanardi

Authors
  1. Anna Chiara R. Porricelli
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  2. Vincenzo Lippolis
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  3. Stefania Valenzano
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  4. Marina Cortese
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  5. Michele Suman
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  6. Sandro Zanardi
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  7. Michelangelo Pascale
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Correspondence to Vincenzo Lippolis.

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Conflict of Interest

Anna Chiara R. Porricelli declares that she has no conflict of interest. Vincenzo Lippolis declares that he has no conflict of interest. Stefania Valenzano declares that she has no conflict of interest. Marina Cortese declares that she has no conflict of interest. Michele Suman declares that he has no conflict of interest. Sandro Zanardi declares that he has no conflict of interest. Michelangelo Pascale declares that he has no conflict of interest.

Ethical Approval

This article does not contain any studies with human or animals performed by any of the authors.

Informed Consent

Informed consent was obtained from all individual participants included in the study.

Funding

This research was funded by the Project S.I.Mi.S.A. “New strategies for improvement of food safety: prevention, control, correction” Italian Ministry of Education, University and Research, P.O.N. “Ricerca & Competitività” 2007–2013, project no. 02_00186_341751212792. The work was also funded by Project SAFE&SMART “New enabling technologies for food safety and food chain integrity within a global scenario”, CL.A.N. Cluster Tecnologici Nazionali, project no. CTN01_00230.

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Porricelli, A.C.R., Lippolis, V., Valenzano, S. et al. Optimization and Validation of a Fluorescence Polarization Immunoassay for Rapid Detection of T-2 and HT-2 Toxins in Cereals and Cereal-Based Products. Food Anal. Methods 9, 3310–3318 (2016). https://doi.org/10.1007/s12161-016-0527-1

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  • DOI: https://doi.org/10.1007/s12161-016-0527-1

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