Abstract: A method has been developed for the determination of T-2 toxin in cereal grains by high performance liquid chromatography with fluorescence detection after immunoaffinity column clean-up and precolumn derivatization.The derivatization reaction was used to develop a sensitive, reproducible and accurate method for the determination of T-2 toxin in wheat, corn, barley and rice. T-2 toxin was extracted with methanol-water (80∶20, v/v), purified by immunoaffinity columns containing antibodies specific for T-2 toxin, and quantified by reversed-phase high performance liquid chromatography with fluorescence detection (excitation wavelength, 381 nm; emission wavelength, 470 nm) after derivatization with 1-anthroylnitrile （1-AN） and 4-dimethylaminopyridine (DMAP). ZORBAX Eclipse XDB-C18 column and mobile phase of acetonitrile-water (80∶20, v/v) were used for the analysis. Recoveries from the different cereals spiked with T-2 toxin at levels ranging from 0.01 to 1.5 μg/g were from 79.7% to 94.5%, the relative standard deviations were lower than 7% and the limit of detection was 0.01 μg/g based on a signal-to-noise ratio of 3∶1.

Key words: cereal grains , fluorescence detection, high performance liquid chromatography （HPLC）, precolumn derivatization, T-2 toxin, immunoaffinity column