Abstract: Modern mass spectrometry plays an important role in the study of metabolism and pharmacokinetics of traditional Chinese medicine due to its high sensitivity, high analysis speed and good stability. First, this article gives a brief introduction to the development of modern mass spectrometry techniques. Second, the application of modern mass spectrometry is summarized in the study of metabolism and pharmacokinetics of traditional Chinese medicine including in vitro and in vivo studies. In vitro research is reviewed with a focus on metabolism of intestinal bacteria and cytochrome P450 enzymes; in vivo research is covered mainly with a fo cuse on the application of microdialysis-MS, GC/LC-MS, new ambient mass spectrometry, and imaging mass spectrometry in metabolism and pharmacokinetics.