Abstract: Objective To develop a method for simultaneous determination of sixteen antibiotics in human urine by ultra performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS）. Methods With Piperacillin as an internal standard, the target antibiotics in urine samples were enriched and purified by Oasis HLB solid phase extraction（SPE） cartridges, then separated in a ZORBAX SB-C18 column with a gradient elution of mobile phase of 0.1% formic acid water and acetonitrile, finally analyzed with multiple reaction monitoring（MRM） mode. Results The limits of detection（LOD） for these sixteen antibiotics were in the range of 0.05-10.0 ng/mL and the limits of quantification（LOQ） in the range of 0.25-20.0 ng/mL. Within the related linear range, the related coefficient （r） of sixteen antibiotics were all more than 0.995. Accuracies for these antibiotics were ranged from 82.0％-119.3%, the within-day precision were less than 13.9％. Conclusion The developed method is sensitive, specific and appropriate for the analysis of antibiotics in forensic toxicology and therapeutic drugs monitoring.

Key words: forensic toxicology, antibiotics, urine, ultra performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS）