A drug target is a specific structure in a cell, tissue or organ that interacts with a drug to make it work. Comprehensive identification of drug targets is essential to understanding the mechanism of action of drugs and their potential side effects. Currently, widely used drug target identification and validation tech-niques, such as activity-based protein profiling (ABPP), compound-centric chemical proteomics (CCCP), re-quire the modification of small-molecule compounds, which may reduce or alter the activity of drug mole-cules. Therefore, advanced techniques that do not require chemical modification of drug molecules, such as drug affinity responsive target stability (DARTS), cellular thermal shift assay (CETSA), and thermal proteome profiling (TPP), are becoming important tools for drug target research. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is a powerful tool for identification of drug target proteins. This paper reviewed drug target research techniques based on LC-MS/MS and future development of drug target identification.