Objective A sensitive and rapid method was developed for the determination of 17 β-adrenergic receptor blockers(β-blockers) by ultra performance liquid chromatography tandem mass spectrometry(UPLC-MS). Methods The β-blockers were extracted from porcine sample with acetonitrile, purified by solid phase extraction column Oasis Prime HLB. The analytes were then separated by the Endeavorsil C18 chromarographic column with methanol and 0.1% formic acid as gradient mobile phase. The ion fragment information analytes were obtained using ultra performance liquid chromatography-tandem mass spectrometry under multiple reaction monitoring mode and the isotope internal standards were used to quantify by calibration curves. Results The linear ranges of 17 β-blockers were from 0.2 to 20 μg/kg, with correlation coefficient(R)＞0.999. The limits of detection(LOD) were 0.05-0.07 μg/kg, and the limits of quantification(LOQ) were 0.15-0.2 μg/kg. The average recoveries of β-blockers(added into blank porcine samples at three concentration levels) ranged from 84.97% to 123.40%, and the relative standard deviation (RSD) was 0.41% to 13.94%. Conclusion The method is simple, sensitive, accurate and is suitable for the confirmation and quantification of β-blockers in porcine samples.