Rapid and High-Throughput Determination of Sixteen <i>β</i>-agonists in Livestock Meat Using One-Step Solid-Phase Extraction Coupled with UHPLC-MS/MS

β-agonists are illegally added to animal feed because they can greatly increase carcasses’ leanness, which impairs the safety of animal-derived foods and indirectly endangers human health. This study aimed to develop an ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for determining sixteen β-agonists in livestock meat. The homogenized samples were subjected to enzymatic hydrolysis using β-glucuronidase/sulfatase at 40 °C for 2 h, extracted with acetonitrile containing 1% acetic acid (v/v), and purified by the one-step Qvet-AG extraction column. The residue was redissolved by 0.1% aqueous formic acid/methanol (9:1, v/v) after blow-drying by nitrogen, and then determined by UHPLC-MS/MS. The results demonstrated that the well linearity was in the range of 0.1–50 μg/L with the correlation coefficient (R2) ≥0.9928, and the limits of detection (LOD) and quantification (LOQ) were 0.01–0.11 μg/kg and 0.04–0.38 μg/kg, respectively. With intraday and interday relative standard deviations (RSDs) being less than 10%, the average recoveries of pork, beef, and lamb at various spiked levels ranged from 62.62–115.93%, 61.35–106.34%, and 62.00–111.83%, respectively. In conclusion, the established method is simple, efficient, sensitive, and suitable for the simultaneous detection of several β-agonist residues in livestock meat