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

Toxins

Vincenzo Lippolis¹  VeronicaM. T. Lattanzio¹  Annalisa De Girolamo¹  Michelangelo Pascale¹  Biancamaria Ciasca¹  AntonioF. Logrieco¹  AnnaC. R. Porricelli¹  Erminia Mancini¹  Peiwu Li²  ChrisM. Maragos³  Susan McCormick³ 

[1] Institute of Sciences of Food Production (ISPA), National Research Council of Italy (CNR), 70126 Bari, Italy;Key Lab for Mycotoxins Detection, Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, China;Mycotoxin Prevention and Applied Microbiology Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Peoria, IL 61604, USA;

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

DOI  :  10.3390/toxins11070380

来源: DOAJ

【 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 different 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 µg/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-off level (80 and 96 µg/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.

【 授权许可】

Unknown