Eucommia ulmoides seed oil is the main byproduct of E. ulmoides cultivation. To better understand its functions, E. ulmoides seed oil is characterized comprehensively in this work. The composition of E. ulmoides seed, physicochemical properties, thermal properties, fatty acid composition, triacylglycerol (TAG) composition and Vitamin E composition of E. ulmoides seed oil were determined. The results show that the E. ulmoides seed contained about 34.63% oil. The excellent physicochemical properties of E. ulmoides seed oil ensured it has a potential to be developed as an edible oil. The main fatty acids in E. ulmoides seed oil were linolenic acid (61.36%), oleic acid (17.02%), and linoleic acid (12.04%). HPLC-ELSD method determined that LnLnLn (37.99%), LnLnO (22.62%), LnLnL (14.5%), and LnLnP (8.78%) were the oil's major TAG components. The oil exhibited a unique thermal curve which contained 2 melting peaks at -38.45 and -22.22 °C, respectively. The total content of vitamin E in E. ulmoides seed oil was 190.96 mg/100g, which exist mainly in γ-tocopherol and δ-tocopherol isomer. Overall, the results indicated that E. ulmoides seed oil is a promising oil in food, pharmaceutics, cosmetics and other nonfood industries.