The objective of the study was to determine the physicochemical characterization and analysis of fatty acids composition present in Citrullus vulgaris seed oil. The extraction was carried out by solvent extraction method using petroleum ether (40-600C) as solvent. The physicochemical properties (iodine, saponification, acid, free fatty acid, peroxide, specific gravity and refractive index) of the extracted oil were assayed using standard methods. The fatty acid profile was analyzed by gas chromatography with FID detector (GC-FID) was used. The seed oil had an golden yellow color and a very characteristic flavour. The oil was also stable at the room temperature. The physical and chemical characteristic shows that the iodine and saponification values are especially high. The specific gravity and refractive index of the oil are also relatively high. GC-MS confirms the presence of fatty acids (palmitic, stearic, oleic, linoleic and linolenic acids). Linoleic acid was the most abundant of the five fatty acids. Fatty acids content in Citrullus vulgaris has great applications as vegetable oils and could be utilized for therapeutic benefit.