Abstract: [Objective] The aims of the present study were to establish an ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry （UHPLC-MS/MS） method for simultaneous detection of 64 drugs in compound feed, and to improve the detection efficiency of illegal additives. [Method] Waters HSS T3 column （2.1 mm×100 mm, 1.8 μm） was used for separation. Gradient elution was performed using 0.1% formic acid aqueous solution as mobile phase A and acetonitrile containing 0.1% formic acid as mobile phase B, respectively, with a flow rate of 0.40 mL/min and sample size of 2 μL. Positive-ion electrospray ionization （ESI） source scanning mode was used for detection, and multiple reaction monitoring （MRM） mode was used for signal acquisition. The recovery rates of the target drugs obtained with 4 kinds of sample extraction solvents and 2 types of solid-phase extraction column were compared to determine the optimal method of sample pretreatment. Detection of the 64 drugs in 100 batches of compound feed samples of different sources collected in Ningxia Hui Autonomous Region was carried out with the established UHPLC-MS/MS method. [Result] The compound feed samples were extracted with acetonitrile aqueous solution （acetonitrile∶water=8∶2, V/V） containing 0.2% formic acid after homogenization. Oasis PRiME HLB solid-phase extraction column was used to purify the samples. The recovery rates of most target drugs was above 60%. In the concentration range of 5.0-200.0 μg/L, a good linear relationship of the 64 drugs was observed, and their correlation coefficients （R） were all greater than 0.99. The limit of quantitation of different drugs was 5.0-10.0 μg/kg. The average recovery rates of 3 spiking levels at 5.0, 20.0 and 50.0 μg/kg ranged from 41.00% to 120.49%. The intra- and inter-assay relative standard deviations （RSDs） were 0.54%-15.94% and 1.25%-13.64%, respectively. No target drug was detected in the 100 batches of compound feed samples. [Conclusion] The established UHPLC-MS/MS method has good linear relationship, high recovery rate, good precision, high reproducibility and good operability, and can be used for screening the 64 drugs illegally added in compound feed.

Key words: compound feed, ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry method, illegal addition, veterinary drug