A method is described for the analysis of 16 β-lactams in chicken muscle by UPLC-quadrupole(Q)-Orbitrap-MS with parallel reaction monitoring (PRM). QuEChERS approach includes clean-up step by sorbent of primary-secondary amine (PSA) and C18 was adopted for sample preparation. Q-Orbitrap with PRM showed high sensitivity with limits of detection (LODs) ranged from 0.01 μg kg−1 to 0.35 μg kg−1. The method was further validated by intra- and inter-day test with spiking levels less than MRLs (maximum residue limits, the European Union). Recovery (83–112%) and precision values (RSDs <15%) for all studied analytes were obtained. The result indicates that UPLC-Q-Orbitrap coupled with QuEChERS preparation can serve as a routine quantification method for β-lactam residues in chicken muscles.