Abstract: Application of high-throughput drug screening, screening anti-epidermal growth factor receptor(EGFR)activity of Chinese herb extracts, and the constituents in Lonicera macranthoides Hand. -Mazz were determined by UPLC-Q-TOF/MS. Herbs were percolated through petroleum ether, ethanol extract, ethyl acetate extraction and boiling water, obtaining four corresponding parts. The activity of EGFR was detected based on HTRF. The variation of fluorescence signal and signal value at 620 nm and 665 nm were determined; the inhibition rate of extracts were calculated. Chromatographic separation was performed on a C₁₈ column(2. 1 mm×50 mm, 1. 7 μm)with a gradient elution of 0. 1% formic acid aqueous solution- acetonitrile. The mass spectrometer equipped with electrospray ionization source was used as defector and data were collected under the positive ion mode. Strong inhibitory activity of EGFR was found for the ethyl acetate part of L. macranthoides Hand. -Mazz, with IC₅₀ value of 2. 027 μg/mL. The information of positive ion and the relative molecular mass were distinguished by Q-TOF/MS, which identified the 7 corresponding parts of L. macranthoides Hand. -Mazz. : caffeic acid, esculetin, quercetin, 3, 5- O-dicaf feoylquinic acid, hinoki flavone, chlorogenic acid and β-sitosterol as well as 4 unknown substances. 3, 5-O-dicaf feoylquinic acid, caffeic acid and hinoki flavone are the main components. The results showed that L. macranthoides Hand. -Mazz contained phenolic acids and flavonoids inhibiting EGFR, which provided the basis for the research and development of antitumor drugs in the future.