Abstract: Standard analytical methods were used to determine the physicochemical characteristics and fatty acid profile of Polyalthia longifolia seed oil with the objective of determining its industrial applications. Physicochemical analysis of the seed oil produced the following results; color (reddish brown); specific gravity (0.875); pH (5.37); viscosity (1.026); acid value (13.46 mg KOH/g Oil); % free fatty acid (7.73); saponification value (120.00 mg KOH/g Oil); ester value (106.54 mg KOH/g Oil); % unsaponifiables (10.60); iodine value (95.0 wij’s) and peroxide value (730.0 mg eq/kg Oil). The fatty acids in the oil were determined using a Perkin Elmer Auto sampler XL Gas chromatograph. The results obtained showed that Polyalthia longifolia seed oil contained saturated fatty acids (9.04-10.79%), mono unsaturated fatty acids (16.64-20.29%) and poly unsaturated acids (11.55-12.78%). The average fatty acid profiles of the saturated fatty acids showed hexadecanoic acid to be the major saturated fatty acid. The mono unsaturated fatty acids present are C18:1cis (17.07%); C20:1cis (0.29%) C22:1cis (1.26%); cis-9-octadecenoic acid being the most abundant. The polyunsaturated acids included C18:2cis (10.08%) and C18:3cis (2.09%), giving ω6/3 value of 4.82~5. In conclusion Polyalthia longifolia seed oil may likely be useful in the Food, Pharmaceutical and Cosmetics industries.