Fluorescence polarization as a means for determination of fumonisins in maize.
Fumonisins, mycotoxins produced by certain species of Fusaria, are commonly found worldwide as contaminants in maize. This paper reports the development of a rapid, portable fluorescence polarization-based assay for fumonisins in maize. The assay was based on the competition of unlabeled fumonisin, from a sample, with a fluorescently tagged fumonisin (FB(1)-FL) for a fumonisin-specific monoclonal antibody in solution. The fluorescence polarization (FP) of the tagged fumonisin was increased upon binding with the antibody. In the presence of free toxin, less of the FB(1)-FL was bound and the polarization signal was decreased. The assays were very simple to perform, requiring only mixing of an aqueous extract of maize with the tagged fumonisin and antibody, and required <2 min per sample, excluding extraction time. Two permutations of the assay were tested, one with each sample matrix serving as its own blank, and the other with all of the samples compared relative to a PBS blank with normalization of the data similar to an ELISA. The limit of detection, defined as the toxin content associated with a fluorescence polarization signal 5 standard deviations from that of a fumonisin-free control, was 0.5 microg of FB(1)/g in spiked maize. Recoveries from spiked maize over the range of 0.5-20 ppm averaged 94.3 +/- 13.8%. Forty-eight samples of field-contaminated maize were tested by the FP and an established HPLC method, with a good correlation between the two (r(2) = 0.85-0.88). For these samples, the two variations of the FP assay also compared well to one another (r(2) = 0.97), suggesting the assay principle is very robust. The results, combined with the speed and ease of use for the assay, suggest that this technology has substantial potential as a screening tool for mycotoxins in foods.
Mycotoxin Research Unit, NCAUR, Agricultural Research Service, U.S. Department of Agriculture, Peoria, Illinois 61604, USA. maragocm@mail.ncaur.usda.gov
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Maragos
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Jolley
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Plattner
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Nasir
D000911:Antibodies, Monoclonal; D002264:Carboxylic Acids / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005454:Fluorescence Polarization / Q000379:methods; D005456:Fluorescent Dyes; D037341:Fumonisins; D005670:Fusarium; D009183:Mycotoxins / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D003313:Zea mays / Q000737:chemistry* / Q000382:microbiology*
D000911:Antibodies, Monoclonal; D002264:Carboxylic Acids; D005456:Fluorescent Dyes; D037341:Fumonisins; D009183:Mycotoxins; C056933:fumonisin B1