title: A rapid fluorescence polarization immunoassay for the determination of T-2 and HT-2 toxins in wheat.
abstract: A rapid fluorescence polarization (FP) immunoassay has been developed for the simultaneous determination of T-2 and HT-2 toxins in naturally contaminated wheat samples. Syntheses of four fluorescein-labelled T-2 or HT-2 toxin tracers were carried out and their binding response with seven monoclonal antibodies was evaluated. The most sensitive antibody-tracer combination was obtained by using an HT-2-specific antibody and a fluorescein-HT-2 tracer. The developed competitive FP immunoassay in solution showed high cross-reactivity for T-2 toxin (CR% = 100%) while a very low CR% for neosolaniol (0.12%) and no cross-reactivity with other mycotoxins frequently occurring in wheat. A rapid extraction procedure using 90% methanol was applied to wheat samples prior to FP immunoassay. The average recovery from spiked wheat samples (50 to 200 μg kg(-1)) was 96% with relative standard deviation generally lower than 8%. A limit of detection of 8 μg kg(-1) for the combined toxins was determined. Comparative analyses of 45 naturally contaminated and spiked wheat samples by both the FP immunoassay and high-performance liquid chromatography/immunoaffinity clean-up showed a good correlation (r = 0.964). These results, combined with the rapidity (10 min) and simplicity of the assay, show that this method is suitable for high throughput screening as well as for quantitative determination of T-2 and HT-2 toxins in wheat.
authors: Institute of Sciences of Food Production (ISPA), CNR-National Research Council of Italy, Via G. Amendola 122/O, 70126 Bari, Italy.
authors: Vincenzo
authors: V
authors: Lippolis
authors: Michelangelo
authors: M
authors: Pascale
authors: Stefania
authors: S
authors: Valenzano
authors: Valeria
authors: V
authors: Pluchinotta
authors: Sabine
authors: S
authors: Baumgartner
authors: Rudolf
authors: R
authors: Krska
authors: Angelo
authors: A
authors: Visconti
mesh_terms: D016231:Fluorescence Polarization Immunoassay / Q000191:economics / Q000379:methods*; D005670:Fusarium / Q000737:chemistry*; D057230:Limit of Detection; D013605:T-2 Toxin / Q000031:analogs & derivatives* / Q000032:analysis* / Q000302:isolation & purification; D013997:Time Factors; D014908:Triticum / Q000382:microbiology*
chemical_list: D013605:T-2 Toxin; C012351:HT-2 toxin