To systematically identify protein targets interacting with traditional Chinese medicine Zuogui Wan, limited proteolysis mass spectrometry (LiP-MS) was employed as a high-throughput screening platform. To investigate the molecular mechanisms underlying chronic unpredictable mild stress (CUMS)-induced Premature ovarian insufficiency (POI), we performed data-independent acquisition (DIA) quantitative proteomics on ovarian tissues from three generations of rats (Control vs. Model groups).