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Objective： A method for simultaneous detection of 16 quinolones in food of animal origin was developed byclean-up column pretreatment combined with ultra high performance liquid chromatography-tandem mass spectrometry. Methods： Food samples of animal origin were extracted using 80% acetonitrile （containing 0.2% formic acid）. After purification on a Speedy Prep^®-Quino 1 column, 16 quinolone residues were detected by ultra-high performance liquid chromatography tandem mass spectrometry. Results： The results showed that the linear range of 16 quinolones was 1.6～40.0 μg/kg, the correlation coefficient<i> r</i>≥0.996 1, the limits of detection were 0.14～0.80 μg/kg, and the limits of quantification were 0.47～2.68 μg/kg. The recoveries of seven matrix after pretreatment were 62%~112%, and the relative standard deviations were 0.9%~18.7%. Conclusion： The method has the characteristics of fast detection speed and high sensitivity, and can be applied to animal derived food such as mutton, duck, beef, fish, eggs, pig kidney, duck skin.