Objective:This study aimed to develop a method of rapid detection of tetracyclines including tetracycline, oxytetracycline, aureomycin and doxycyclin in aquatic products with deep eutectic solvents (DES) extraction and ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).Methods:Different parameters of DES, such as the types of solvents, molar ratio of solvents, water content, and pH value were optimized.Results:The optimum deep eutectic solvent conditions for the medicament analysis of four tetracyclines in aquatic products were choline chloride-glycerol combination(molar ratio 1∶4) with 90% moisture content and 0.1% formic acid. Under the control of these condition, the four types of tetracyclines could be significantly separated, with a linear correlation of 0.999 4 in the mass concentration range of 5.0~200.0 ng/mL and LOD of 20.0 μg/kg. The recovery of each target component ranged from 82.2% to 102.8%, and the RSD was less than 10%. The analytical method developed was used to detect four kinds of tetracyclines in different samples including 13 batches of commercially available aquatic products, one quality control (QC) sample, and one international proficiency testing sample. The result indicated that oxytetracycline and doxycycline could be detected in one batch of shrimp sample from the market with a concentration of 64.8 μg/kg, and 25.4 μg/kg, respectively, while the content of tetracyclines in QC sample and international proficiency testing sample were all within the limits of given characteristic values.Conclusion:The novel DES-UPLC-MS/MS method developed is feasible to both the qualitative and quantitative analysis of the tetracyclines in aquatic products, which an advantage of simple pre-processing and high accuracy and efficiency.