Background: Xylopia aethiopica is well known to treat neurodegenerative diseases in traditional medicine and there is no scientific evidence for this claim. Objective: The current research aimed at investigating chemical characterization, antioxidant, and anticholine esterase activity of the essential oil from X. aethiopica. Materials and Methods: Essential oil extraction was carried out by the use of the steam distillation method in a modified Clevenger-type apparatus. The chemical composition of the essential oil from seeds of X. aethiopica (African pepper) was determined using gas chromatography coupled with mass spectrometry (GC-MS) and its potentials as antioxidant and anticholine esterase were evaluated for the first time. Results: The oil yield was 5.2% (v/w) in X. aethiopica. The GC-MS analysis identified a total of 52 compounds corresponding to 100% of the total oil in X. aethiopica. The major constituents of X. aethiopica essential oil are terpinen-4-ol (11.88%), α-terpineol (5.93%), cyclohexane methanol (4.79%), and β-copaene (4.74%). The most abundant classes of compounds from the essential oil were oxygenated monoterpenes (MT) which amounted to (37.6%), followed by oxygenated sesquiterpenes (29.61%), sesquiterpenes (14.67%), oxygenated diterpenes (9.05%), nonterpenoid aliphatic and aromatic hydrocarbons (4.81%), diterpenes (3.8%), and MT (0.47%) of all the identified constituents. A significant antioxidant (IC50value of 2,2-diphenyl-1-picrylhydrazyl = 2.19 ± 0.09 mg/mL) and anticholine esterase activity (IC50 =1.21 ± 0.06 mg/mL) was obtained for the essential oil of X. aethiopica. Conclusion: The study established the chemical composition, antioxidant, and anticholine esterase activities of the essential oil of the plant seeds.