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&NA; A novel strategy was developed for dual‐target screening of bioactive components from traditional Chinese medicines (TCMs). This strategy was based on the use of low‐cost microporous hollow fibers filled with target enzymes as baits to “fish out” the ligands in TCM extracts, followed by identification of the ligands dissociated from the target‐ligand complexes by liquid chromatography–mass spectrometry. Ganjiang Huangqin Huanglian Renshen Decoction (GHHRD), a classical TCM prescription for diabetes treatment, was chosen as a model sample to evaluate the feasibility of the proposed strategy. Three bioactive components were successfully screened out from GHHRD. Coptisine was identified as the ligand of &agr;‐glucosidase and baicalin as the ligand of angiotensin‐converting enzyme (ACE). Berberine was found to be a dual inhibitor of &agr;‐glucosidase and ACE. The results were further verified by enzyme inhibitory assay and molecular docking simulation. The study suggested that our developed strategy would be a powerful tool for screening bioactive components from multi‐component and multi‐target TCMs. Graphical abstract Figure. No caption available. HighlightsA novel ligand fishing strategy based on dual‐target screening for TCM was developed.Hollow fiber‐based ligand fishing in combination with LC–MS was devised.Ligands of &agr;‐glucosidase and ACE were identified from complex matrix simultaneously.Screening results were verified by enzyme inhibitory assay and molecular docking.