Bryophytes, which include mosses, hornworts and liverworts, are early land plants that lack vascular tissues. Due to their small size and difficulties in identification, they have been neglected as a study material for a significant period of time. However, recent phytochemical studies focusing on mosses have revealed the presence of unique and biologically active substances. The objective of this research was to examine the volatile bioactive phytochemicals present in the ethanol extract of Hyophila involuta (Hook.) A. Jaeger using GC-MS and docking of a potential compound against the target proteins of Staphylococcus aureus. Through GC-MS analysis, a total of thirty-seven phytochemicals were identified, including beneficial substances like Neophytadiene, Squalene, Phytol, Lupan-3-ol acetate, Campesterol, Stigmasterol, Gamma-sitosterol, dl-alpha-tocopherol and more. Upon undergoing ADMET analysis with SwissADME, Squalene, a
triterpenoid, has been identified as an orally active compound that adheres to the Lipinski rule and displays drug-like characteristics, with biological activities suited best as a ligand molecule. The docking results of Squalene against the target proteins of S. aureus revealed the highest docking score when interacting with the active site of Undecaprenyl diphosphate synthase (4H8E), followed by Dihydrofolate reductase (3FYV), Sortase A (1T2P) and Dehydrosqualene synthase (2ZCO) with binding energies (Kcal/mole) of -9.99, -9.04, -7.68 and -7.35 respectively. Integrating high-throughput screening methods with the docking results can expedite the validation of potential antibacterial compounds, enabling researchers to identify novel treatments more efficiently.