Objective To establish a method for the simultaneous determination of ofloxacin, norfloxacin, ciprofloxacin, pefloxacin, danofloxacin, lomefloxacin, enrofloxacin, ciprofloxacin, pefloxacin, oxolinic acid and flumequine,tetracycline, chlortetracycline, oxytetracycline and doxycycline in animal tissue by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) with automatic solid phase extraction. Methods The samples were extracted with 0.1 mol/L EDTA-Mcllcaine buffer solution (pH=4.0), and the resulting extracts were cleaned-up on HLB solid phase extraction column, and then eluted with 100% methanol. The target components were separated on a UPLC C₁₈ column (100 mm×2.1 mm, 1.6 μm), with 0.1% formic acid water and methanol-acetonitrile (40∶60, V/V) solution as mobile phase in gradient elution. Multiple reaction monitoring (MRM) in positive was used, qualitative confirmation was performed from retention time and secondary mass characteristic ions, and the matrix-matched external standard calibration curves were used for quantitative analysis. Results The target components and impurity could be well separated, it was showed good linearity in the range of 1.25-50.0 μg/kg (r=0.998 5 above). The detection limits and quantitation limits of 11 kinds of quinolones were 0.013-0.069 and 0.043-0.23 μg/kg, the limits of 4 kinds of tetracyclines were 0.033-0.093 and 0.11-0.31 μg/kg. The recovery tests at high, medium, and low spiked levels:the recoveries were 70.8%-105.4%, and relative standard deviation (RSDs) were 0.5%-7.4% in chicken; the recoveries were 75.6%-115.2%, and RSDs were 0.8%-8.9% in pork; the recoveries were 73.7%-117.5%, and RSDs were 0.5%-14.8% in shrimp. The method was applied to the analysis of 60 samples from Guangdong, and the result showed that ofloxacin, ciprofloxacin, enrofloxacin and doxycycline were detected in 4 samples, the maximum content of enrofloxacin and doxycycline in shrimp was 400 μg/kg, the content of doxycycline in chicken was 220 μg/kg, which exceeded the limit value of 100 μg/kg. Conclusion The method was accurate, efficient and simple, synchronous confirmation and quantitative analysis of the residues for chicken, pork and shrimp meat and other musculature of quinolones and tetracyclines, and was suitable for analysis and confirmation in food safety risk monitoring.