Fluorescence Polarization Immunoassay for the Determination of T-2 and HT-2 Toxins and Their Glucosides in Wheat

文献类型： 外文期刊

第一作者： Lippolis, Vincenzo

作者： Lippolis, Vincenzo;Porricelli, Anna C. R.;Mancini, Erminia;Ciasca, Biancamaria;Lattanzio, Veronica M. T.;De Girolamo, Annalisa;Logrieco, Antonio F.;Pascale, Michelangelo;Maragos, Chris M.;McCormick, Susan;Li, Peiwu

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关键词： fluorescence polarization immunoassay; T-2 toxin; HT-2 toxin; T-2 glucoside; HT-2 glucoside; wheat; validation study; screening method

期刊名称：TOXINS （ 影响因子：4.546； 五年影响因子：4.8 ）

ISSN：

年卷期： 2019 年 11 卷 7 期

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收录情况： SCI

摘要： T-2 and HT-2 toxins and their main modified forms (T-2 glucoside and HT-2 glucoside) may co-occur in cereals and cereal-based products. A fluorescence polarization immunoassay (FPIA) was developed for the simultaneous determination of T-2 toxin, HT-2 toxin and relevant glucosides, expressed as sum. The developed FPIA, using a HT-2-specific antibody, showed high sensitivity (IC50 = 2.0 ng /mL) and high cross-reactivity (100% for T-2 toxin and 80% for T-2 and HT-2 glucosides). The FPIA has been used to develop two rapid and easy-to-use methods using two di ff erent extraction protocols, based on the use of organic (methanol /water, 90: 10, v /v) and non-organic (water) solvents, for the determination of these toxins in wheat. The two proposed methods showed analytical performances in terms of sensitivity (LOD 10 g /kg) recovery (92-97%) and precision (relative standard deviations 13%), fulfilling the criteria for acceptability of an analytical method for the quantitative determination of T-2 and HT-2 toxins established by the European Union. Furthermore, the methods were then validated in accordance with the harmonized guidelines for the validation of screening methods included in the Regulation (EU) No. 519 /2014. The satisfactory analytical performances, in terms of intermediate precision ( 25%), cut-o ff level (80 and 96 g /kg for the two methods) and rate of false positives (< 0.1%) confirmed the applicability of the proposed methods as screening method for assessing the content of these toxins in wheat at the EU indicative levels reported for T-2 and HT-2 toxins.

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