Objective To explore the progress on the application of in vivo drug analysis techniques in clinical pharmacy work. Methods Relevant literature was reviewed to provide an overview of the characteristics of clinical samples, common in vivo drug analysis methods used in the clinic, the application and existing problems of in vivo drug analysis in clinical pharmacy. Results and Conclusion In recent years, with the increasing demand for individualized and precise treatment in clinical practice and the continuous development of analytical techniques, in vivo drug analysis techniques have been widely used in clinical pharmacy work, which have become one of the important auxiliary techniques to promote rational clinical drug use, improve individualized treatment and reduce the occurrence of adverse reactions. However, in the actual application, there were still problems such as the invasive blood sampling that hinders sampling, the weak ability to interpret drug monitoring results and clinical testing methods that still need to be improved. These problems should be taken seriously and continuously improved and solved in the subsequent research and application.