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United States Department of Agriculture

Agricultural Research Service

Title: Fluorescence Polarization Based Assays for the Determination of Don (Deoxynivalenol) and Aflatoxins in Grains

Authors
item Nasir, Mohammad - DIACHEMIX, GRAYSLAKE, IL
item Maragos, Chris
item Plattner, Ronald
item Jolley, Michael - DIACHEMIX, GRAYSLAKE, IL

Submitted to: American Chemical Society National Meeting
Publication Type: Abstract Only
Publication Acceptance Date: August 30, 2001
Publication Date: N/A

Technical Abstract: Fluorescence polarization (FP) has been used for the homogeneous analysis of molecular interactions of small molecules with proteins, antigen- antibody interactions, hormone-receptor interactions, monitoring therapeutic drug levels and the detection of substances of abuse. Recently we published a fluorescence polarization based assay for quantitative determination of fumonisin in corn. The assay is rapid, inexpensive and can be used in the field. As compared to the normal use of organic solvents for fumonisin extraction, we used water as the extraction solvent, avoiding problems associated with the solvents in the extracts. The extraction is simple and is virtually complete in 5 min. The assay is very fast (complete in 1 min) and requires minimal training to perform. DON (deoxynivalenol, vomitoxin) is produced in various grains such as wheat, corn and barley. It causes various diseases in animals, humans and laboratory animals. Various methods for the quantitative analysis of DON and aflatoxins are in use (TLC, GC, HPLC and immunochemical methods, such as ELISA). Most of these methods require extensive cleanup procedures after extraction and are not suitable for field-testing. Here we report two similar assays for the quantitative determination of DON and aflatoxins in various grain samples. Analysis of naturally-contaminated wheat samples by FP correlated very well with the analysis independently carried out using HPLC. The cross-reactivity of the DON tracer and one of the antibodies to 21 related trichothecene mycotoxins was tested. Of these 21 compounds, only three (DON, 15-acetyl-DON and HT-2 toxin) were active with a cross reactivity relative to DON of 100%, 357% and 9%. These tests could be a major contribution to the grain community and growers.

Last Modified: 9/2/2014
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