To establish an analytical method for the determination of zearalenone (ZEN) in Triticum aestivum L powder by molecularly imprinted solid-phase column purification-ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).

The ZEN from Triticum aestivum L powder samples were extracted with acetonitrile/water containing acetic acid, purified by a ZEN-specific molecularly imprinted solid-phase extraction column method, eluted with the mobile phase of 5 mmol/L ammonium acetate solution and acetonitrile, separated on the Luna®Omega Polar C₁₈ column (100 mm×2.1 mm, 3 μm) at a flow rate of 0.5 mL/min. The analysis was conducted using UPLC-MS/MS and then analyzed in negative ion mode of electrospray ionization source with multiple reaction monitoring mode for qualitative and quantitative analysis.

In the range of 1.0–50.0 μg/L, the peak area of ZEN was linearly correlated with the content, and the determinationn coefficient (r²) was 0.9997. Meanwhile, the average recoveries of ZEN in Triticum aestivum L powder were 80.2%–95.8% with the maximum relative standard deviations of 7.6% at the spiked concentration of 3.00, 6.00 and 30.00 μg/kg. The limit of detection was 3.0 μg/kg.

This study establishes an efficient detection method for ZEN in Triticum aestivum L powder using molecularly imprinted solid-phase extraction purification. This method effectively removes matrix interference and enriches ZEN in the sample, demonstrating high precision and accuracy. The research results provide reliable technical support and experimental data for establishing detection standards and quality control guidelines for ZEN in Triticum aestivum L powder.