Abstract:

A rapid analytic method using high performance liquid chromatography (HPLC) was established for
the simultaneous determination of aflatoxins, including AFB1, AFB2, AFG1 and AFG2, ochratoxin A, zearalenone,
deoxynivalenol and T-2 toxin in grains. The samples were extracted with acetonitrile-water, and cleared up with an immunoaffinity
column (IAC). The separation of the targeted compounds was performed on an Agilent Elipse Plus C18 column
(100 mm×4 mm, 3.5 μm) using a mobile phase consisting of a mixture of acetonitrile : methanol : water : acetic acid with
a flow rate of 1 mL/min at 35 ℃. The injection volume was 20 μL. The detection system was variable wavelength detector
(VWD)-PHRED system-fluorescence detector (FLD). The limits of detection (LODs) of AFB1, AFB2, AFG1, AFG2, ZEN,
OTA, DON, and T-2 toxin were 0.446, 0.152, 0.523, 0.334, 7, 0.7, 200, and 100 ng/mL, respectively. The recoveries of
samples spiked with these mycotoxins were in the ranges of 80.0% - 104.5% for corn and 83.2% - 102.8% for wheat. The
precision values associated with the method, expressed as relative standard deviation (RSD) values, were between 2.6% and
10.2%. It took only 2 hours to complete an analysis. The method was suitable for the determination of common 8 mycotoxins
in raw grains, with the advantages of simplicity, rapidness, sensitivity, and good reproducibility.

Key words: aflatoxins (AFB1, AFB2, AFG1, AFG2), zearalenone, deoxynivalenol, T-2, ochratoxin A, grain, HPLC, immunoaffinity columns (IAC)