| Achoimre: | <i>Dryopteris</i> sp. is known for its various pharmacological effects and is used as a traditional medicine in Asia. The present study investigated the chemical composition and antimicrobial activity of <i>Dryopteris</i> sp. distributed in Korea. The chemical compounds in the ethanolic extracts of <i>Dryopteris lacera</i> and <i>Dryopteris bissetiana</i> were investigated by ultra-high performance liquid chromatography–quadrupole time-of-flight–mass spectrometry analysis and identified by exploring the UNIFI traditional medicine library. Flavonoids such as juglanin, 6-hydroxyluteolin 7-O-laminaribioside, peltatoside, kaempferitrin, hyperoside, and astragalin were identified in both <i>D. lacera</i> and <i>D. bissetiana</i>. Neochlorogenic acid was identified as a caffeoylquinic acid in <i>D. bissetiana</i>. Both extracts of <i>D. lacera</i> and <i>D. bissetiana</i> exhibited antibacterial activity against Gram-positive pathogens, <i>Staphylococcus aureus</i> and <i>Streptococcus mutans</i>. The minimum inhibitory concentration of <i>D. bissetiana</i> against <i>S. aureus</i> was less than 625 ppm. The antibacterial activity was attributed to the identified phenolic compounds, juglanin, 6-hydroxyluteolin 7-O-laminaribioside, kaempferitrin, astragalin, and neochlorogenic acid. Therefore, <i>D. lacera</i> and <i>D. bissetiana</i> can be used as Gram-positive selective antibiotics for further investigation.
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