A rapid fluorescence polarization immunoassay for the determination of T-2 and HT-2 toxins in wheat
| Autor: | Sabine Baumgartner, Michelangelo Pascale, Vincenzo Lippolis, Stefania Valenzano, Rudolf Krska, Valeria Pluchinotta, Angelo Visconti |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Time Factors
medicine.drug_class HT-2 toxin Monoclonal antibody medicine.disease_cause Biochemistry Analytical Chemistry chemistry.chemical_compound Fusarium Limit of Detection T-2 toxin Fluorescence polarization Fluorescence Polarization Immunoassay medicine Mycotoxin Triticum Detection limit Immunoassay Chromatography medicine.diagnostic_test Toxin Extraction (chemistry) Contamination chemistry Wheat Fluorescence anisotropy |
| Zdroj: | Analytical & bioanalytical chemistry 401 (2011): 2561–2571. doi:10.1007/s00216-011-5379-3 info:cnr-pdr/source/autori:Lippolis V., Pascale M., Valenzano S., Pluchinotta V., Baumgartner S., Krska R., Visconti A./titolo:A rapid fluorescence polarization immunoassay for the determination of T-2 and HT-2 toxins in wheat./doi:10.1007%2Fs00216-011-5379-3/rivista:Analytical & bioanalytical chemistry (Print)/anno:2011/pagina_da:2561/pagina_a:2571/intervallo_pagine:2561–2571/volume:401 |
| DOI: | 10.1007/s00216-011-5379-3 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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