This study investigated the physicochemical properties and antioxidant activity of Citrillus lanatus Seed Oil (CLSO). The oil's density, refractive index, acid value, peroxide value, iodine value, and saponification value were determined. CLSO exhibited a density of 0.953, slightly lower than water, and a refractive index of 1.46. The acid value (12.366 mg KOH/g/oil) and acidity (28.683%) were relatively high, indicating the presence of free fatty acids. The peroxide value (10 meq O2/kg oil) suggested moderate oxidation. The iodine value (1.455 g iodine/g) and saponification value (26.84 mg/KOH/g) were lower than previously reported values for similar oils. Antioxidant activity was assessed using the DPPH radical scavenging assay, with CLSO demonstrating superior antioxidant capacity compared to ascorbic acid, exhibiting an IC50 of 1.15 µg/ml at 200 µg/ml concentration. These findings suggest that CLSO possesses significant antioxidant properties and unique physicochemical characteristics, warranting further investigation for potential applications in various industries.