Determination of the Fusarium mycotoxins nivalenol, deoxynivalenol, 3-acetyldeoxynivalenol, and 15-O-acetyl-4-deoxynivalenol in contaminated whole wheat flour by liquid chromatography with diode array detection and gas chromatography with electron capture detection.

A rapid and sensitive method was developed for simultaneous detection of nivalenol (NIV), deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-A-DON), and 15-O-acetyl-4-deoxynivalenol (15-A-DON) in wheat flour. Samples were extracted with acetonitrile-water (84 + 16), and the extract was filtered and purified by a column containing a combination of charcoal, celite, and other adsorbents. For screening analysis, the column eluate was only extracted with ethyl acetate. After evaporation of the solvent, the dried residue was redissolved in acetonitrile-water (2 + 8) and then analyzed by reversed-phase liquid chromatography (LC) with diode array detection. Recoveries of NIV, DON, 3-A-DON, and 15-A-DON from whole wheat flour spiked at 2 levels were 49-55, 92-97, 98-100, and 100-105%, respectively. To quantitate mycotoxin amounts lower than 1 ppm, purified column extracts were evaporated to dryness, derivatized with heptafluorobutyric anhydride, and analyzed by gas chromatography with electron capture detection (GC-ECD). Average recoveries of NIV, DON, 3-A-DON, and 15-A-DON from whole wheat flour spiked at 2 levels, were 45-52, 91-103, 81-85, and 84-92%, respectively. GC-ECD detection limits for all mycotoxins tested at a signal-to-noise ratio of 4:1 were < 30 ng/g. Results of GC-ECD analysis for whole wheat flour samples spiked with mycotoxins at 3 and 10 ppm compared well with results (2.8 and 9.9 ppm) for the same samples analyzed by LC.