Determination of Deoxynivalenol in Wheat Bran and Whole-Wheat Flour by Fluorescence Polarization Immunoassay

Abstract

A rapid and accurate fluorescence polarization (FP) immunoassay has been optimized for the determination of deoxynivalenol (DON) in wheat bran and whole-wheat flour. A preliminary treatment with activated charcoal was used to eliminate the strong matrix effect due to highly colored interfering compounds present in raw wheat bran extracts. In particular, matrix effect was removed by adding activated charcoal to the wheat bran extract (3.5 mg/mL) and mixing for 3 min of incubation time prior to the FP immunoassay analysis. No preliminary treatment was necessary for whole-wheat flour. Average recoveries from samples spiked with DON at levels of 500, 1,000, and 1,500 μg/kg were 95 % for wheat bran and 94 % for whole-wheat flour, with relative standard deviation generally lower than 13 %. Limits of quantification of the optimized FP immunoassay were 120 μg/kg for both matrices. The overall time of analysis was lower than 15 min for wheat bran and 10 min for whole-wheat flour. Good correlations (r > 0.971) were observed between DON contents obtained by both FP immunoassay and high-performance liquid chromatography with immunoaffinity cleanup for 37 and 23 samples of naturally contaminated wheat bran and whole-wheat flour, respectively. These results show that the FP immunoassay is suitable for high-throughput screening as well as for quantitative determination of DON in wheat bran and whole-wheat flour.