Objective To develop an analytical method for determination of 5 kinds of poppy shell alkaloids in hot pot condiment by ultra-high performance liquid chromatography-high resolution mass spectrometry. Methods The samples were extracted and cleaned up based on dispersive micro solid-phase extraction (DMSPE) using strong cationic exchange adsorbent (PCX). The chromatographic separation of alkaloids was performed on an HSS T3 column (2.1 mm×100 mm, 1.8 μm) using 0.1% formic acid aqueous solution and acetonitrile with 0.1% formic acid as the mobile phase with gradient elution. The alkaloids were analyzed by high resolution mass spectrometry using parallel reaction monitoring mode and the external standard method was selected for quantitative analysis. Results Five kinds of alkaloids showed a good linearity in a certain concentration range with relative coefficient of R²>0.999. Limits of detections (LODs) were in the range of 0.1-0.3 μg/kg, and limits of quantitations (LOQs) were in the range of 0.4-1.0 μg/kg. At the spiked levels of 1,10, 100 μg/kg, the recoveries of alkaloids in hot pot condiment were from 78.5% to 104.2%, with the intra-day precision (RSD_r) of 1.8%-6.8% and the inter-day precision (RSDR) of 4.9%-13.7%. Conclusion Using DMSPE cleanup technique with PCX as adsorbent, this method was simple, rapid, low-cost, high sensitivity, and had good reproducibility on the basis of ensuring effective cleanup effect. It could be applied to the qualitative and quantitative determination of five poppy shell alkaloids in hot pot condiment.