|LIPPOLIS, VINCENZO - Institute Of Sciences Of Food Production|
|PORRICELLI, ANNA C - Institute Of Sciences Of Food Production|
|MANCINI, ERMINIA - Institute Of Sciences Of Food Production|
|LATTANZIO, VERONICA M - Institute Of Sciences Of Food Production|
|CIASCA, BIANCAMARIA - Institute Of Sciences Of Food Production|
|DE GIROLAMO, ANNALISA - Institute Of Sciences Of Food Production|
|DE SAEGER, SARAH - Ghent University|
|LI, PEIWU - Chinese Academy Of Agricultural Sciences|
|LOGRIECO, ANTONIO - Institute Of Sciences Of Food Production|
|PASCALE, MICHELANGELO - Institute Of Sciences Of Food Production|
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 10/16/2019
Publication Date: 10/16/2019
Citation: Lippolis, V., Porricelli, A.C.R., Mancini, E., Lattanzio, V.M.T., Ciasca, B., De Girolamo, A., De Saeger, S., Maragos, C.M., McCormick, S.P., Li, P., Logrieco, A.F., Pascale, M. 2019. Fluorescence polarization immunoassays for the determination of trichothecenes and their modified forms in wheat [abstract].
Technical Abstract: The trichothecenes of major concern for cereals and cereal-based products, due to their incidence and toxicity, are deoxynivalenol (DON), T-2 toxin (T-2) and HT-2 toxin (HT-2). These toxins can be harmful to both human and animal health. Moreover, a large number of modified forms of DON, T-2 and HT-2 generated by fungi, plants and mammals have been isolated and characterized. To date, natural occurrence of modified forms of DON, such as 3-acetyl-deoxynivalenol (3Ac-DON), 15-acetyl-deoxynivalenol (15Ac-DON), and deoxynivalenol-3-glucoside (DON3G), and of T-2 and HT-2, such as T-2 glucoside (T-2G) and HT-2 glucoside (HT-2G), have been reported by several authors. The development of analytical methods able to simultaneously detect mycotoxins and their modified forms, also expressed as the sum, has high impact because it could meet possible future requirements of European or international regulations. For this reason, within the MycoKey Project (EU, H2020), two FPIAs have been developed and validated for the rapid (<15 min) and simultaneous determination, expressed as sum, of: (i) DON, 3Ac-DON, 15Ac-DON and DON3G in wheat; (ii) T-2, HT-2, T-2G and HT-2G in wheat. Different extraction protocols, using organic and non-organic solvents, were tested for the developed FPIAs. All the developed FPIAs showed analytical performances, in terms of recovery (89-112%) and precision (=13%) that fulfilled the criteria for acceptability of an analytical method for the determination of relevant native forms established by the European Union. Furthermore, in line with harmonized guidelines for the validation of screening methods, an experimental protocol for single-laboratory validation has been applied to the determination of these toxins by FPIAs according to Regulation (EU) No 519/2014. The satisfactory analytical performances, in terms of precision (repeatability =9%, within laboratory reproducibility =13%), cut-off levels and rate of false positive results (<0.1%) confirmed the applicability of the proposed FPIAs as screening methods for assessing the content of DON, 3Ac-DON, 15Ac-DON and DON3G (expressed as sum) and T-2, HT-2, T-2G and HT-2G (expressed as sum) in wheat at regulatory levels. Moreover, the developed FPIAs are low-cost, portable, can be automated, and do not require a high level of technical skills. These findings indicate that the proposed FPIAs are appropriate for high-throughput screening of these trichothecenes and their modified forms in wheat.