In this paper, UPLC-MS/MS content determination method and beta-lactam drug resistance gene detection method for simultaneous detection of five beta-LAD in water were used to systematically study the environmental behaviors and drug resistance gene emission characteristics of five common beta-LADs (ie amoxicillin, ampicillin, ceftazidime, cefotaxime, cefepime) in drug production process. The results showed that the bacteria selected from the system not only had a high proportion of resistant bacteria, but also had strong drug resistance, and the drug resistance genes had emission patterns. Finally, the research method established in this study was successfully applied to the detection of beta-lactam concentration and drug resistance genes in the factory drainage system.