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 orHT-2 toxin tracers were carried out and their binding response with sevenmonoclonal 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 crossreactivity 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 ?g kg-1) was 96% with relative standard deviation generally lower than 8%. A limit of detection of 8 ?g 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/immunoaffinityclean-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.

Autore Pugliese

• Pascale Michelangelo , Lippolis Vincenzo

Tutti gli autori

• Lippolis V.; Pascale M.; Valenzano S.; Pluchinotta V.; Baumgartner S.; Krska R.; Visconti A.

Titolo volume/Rivista

Analytical & bioanalytical chemistry

Anno di pubblicazione

2011

ISSN

1618-2642

ISBN

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Numero di citazioni Wos

Nessuna citazione

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Numero di citazioni Scopus

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Settori ERC

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Codici ASJC

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