Homalomena aromatica Schott is a valuable medicinal aromatic plant having wide range of applications in ethnobotany, pharmacology, perfumery and flavor industry. The present study is subjected to assess anti‐inflammatory, anti‐diabetic, tyrosinase inhibitory, acetylcholinesterase (AChE), antimicrobial, and genotoxic efficacies of H. aromatica rhizome essential oil (HAEO). To validate the findings of biological assays, in‐silico methods including PASS tool, target prediction and molecular docking were employed.The chemical profiling was performed using GC/MS where HAEO yield was 1.09±0.641% (v/w) with 12 compounds where linalool was identified as major compound (76.29%). Anti‐inflammatory activity showed IC50 value for protease inhibitor 19.59 µL/mL and albumin denaturation assay 32.16 µL/mL. Tyrosinase inhibitory activity showed 73.62 µL/mL. Acetylcholinesterase inhibitory (AChEase) activity revealed an IC50 value of 38.13 µL/mL. Antimicrobial activity showed MIC values of 10±0.47µg/mL, and 45±0.47µg/mL against S.aureus and C.albicans. Genotoxicity analysis showed HAEO has moderate toxic effect. In‐silico results identified six potential targets via swiss target prediction: (ACHE, CYP51, PPAR, COX‐2 and TYRP1). Additionally, molecular docking studies revealed Linalool ‐7.4 kcal/mol, Spathulenol ‐8.5 kcal/mol, Aromadendrene oxide‐(2) ‐8.3 kcal/mol better docking scores. For the first time, this study reports on anti‐diabetic, anti‐tyrosinase, acetylcholinesterase inhibitory activities and genotoxicity of HAEO.