|Lippolis, V - National Research Council - Italy|
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 11/12/2014
Publication Date: 11/12/2014
Citation: Lippolis, V., Maragos, C.M. 2014. Fluorescence polarization immunoassays for rapid, accurate, and sensitive determination of mycotoxins [abstract].
Technical Abstract: Analytical methods for the determination of mycotoxins in foods are commonly based on chromatographic techniques (GC, HPLC or LC-MS). Although these methods permit a sensitive and accurate determination of the analyte, they require skilled personnel and are time-consuming, expensive, and unsuitable for screening purposes. Simple, rapid, and more effective screening methods for mycotoxins determination are highly demanded. Fluorescence polarization immunoassay (FPIA) is a type of homogeneous assay. For low molecular weight antigens, such as mycotoxins, it is based on the competition between an unlabeled antigen and its fluorescent-labelled derivative (tracer) for an antigen-specific antibody. The antigen content is determined by measuring the reduction of fluorescence polarization signal, which in turn is determined by the reduction of tracer molecules able to bind antibody in solution. The development of a competitive FPIA for mycotoxin analysis has three key elements: an antibody specific for the mycotoxin, the mycotoxin of interest conjugated with a fluorophore as tracer and an instrument able to measure the polarization. The selection of an appropriate antibody-tracer combination is the most important parameter in the development of FPIA because it determines the sensitivity, speed, and accuracy of the assay. Several FPIA methods for the detection of the major mycotoxins, including aflatoxins, fumonisins, ochratoxin A, deoxynivalenol, T-2 and HT-2 toxins and zearalenone in food and beverages have been developed in the last decade. Basic principles, advantages and limitations of these methods will be described. Their applications vary substantially in sensitivity, the time of analysis, and the degree of sample preparation required. The accuracy of many of the FPIA for mycotoxins has been supported through comparisons between FPIA and HPLC with spiked and naturally contaminated samples. The trueness of FPIA has also been established through the use of certified reference materials. These FPIA methods are simple, readily automated, rapid, and suitable for high-throughput screening, as well as for the reliable quantitative determination of mycotoxins in foods and commodities.