Abstract: Studies of environmental chemistry and its related life science are essential to understand the exposure of environmental pollution to living species and the resulted health problems. During this process, mass spectrometry is an indispensably powerful tool thanks to its qualitative and quantitative abilities and remarkable mass resolution. In this review, the state-of-the art mass spectrometric methods were introduced, including inductively coupled plasma mass spectrometry (ICP-MS), electron capture negative ionization mass spectrometry (ECNI-MS), electrospray ionization mass spectrometry (ESI-MS), matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), high-resolution tandem mass spectrometry (HR-MS) and native mass spectrometry (nMS) as well as mass spectrometry imaging (MSI), which have been applied in environmental chemistry and the related life science research, with an emphasis on their role in the determination, characterization, discovery and biological impacts research of environmental pollutants in the environmental media, and the related metabolites during life process. Furthermore, the trend of MS methods from fundamental to applied perspectives was outlooked, in order to solve the emerging issues, such as the ambiguous interaction mechanisms of the environmental pollutants and/or their metabolites with biomolecules. It is very much expected that novel MS-based strategy like reactive proteomics approach together with the design of new molecular ruler will help to solve the “mystery” of the environmental pollutants and/or their biological metabolites impacted on a normal physiological process, deepening our understanding on the influence of environmental pollution on health.