Abstract:A method for the determination of cannabinol，cannabidiol，and tetrahydrocannabinol in biscuits using ultra-high performance liquid chromatography tandem mass spectrometry technology was established.The sample was extracted with acetonitrile-acetic acid（99+1），added to the acetic acid system for salt package ultrasonic centrifugation，and then added to the adsorbent for centrifugation. Using a 0.1% formic acid aqueous solution and methanol as the mobile phase，gradient elution was performed，separated by C18 chromatographic column，and monitored by atmospheric pressure chemical ionization（APCI）positive ion multiple reaction mode.By examining different types of extraction solvents，ultrasonic time，salt package system selection，and target compound recovery rate of adsorbent composition，the optimal pre-treatment method was determined. The results showed that cannabinol，cannabidiol，and tetrahydrocannabinol exhibited a good linear relationship in the range of 2-100 ng/mL，with correlation coefficients（R2）all greater than 0.999. The limit of detection of the method was 0.01 mg/kg，and the limit of quantitation was 0.02 mg/kg. In the low，medium，and high concentration spiked recovery experiment，the recovery rate of cannabinol ranged from 80.3% to 91.1%，with a relative standard deviation（RSD）of 2.4%-6.5%.The recovery rate of cannabidiol ranged from 78.4% to 88.5%，with an RSD of 2.9%-7.5%.The recovery rate of tetrahydrocannabinol ranged from 81.5% to 91.3%，with an RSD of 2.6%-6.8%. This method is fast，simple，highly accurate，has good reproducibility，and is suitable for the detection of cannabidiol，cannabidiol，and tetrahydrocannabidiol in biscuits.