Eucommia ulmoides Oliver seed oil (E.u oil) as a functional oil is rich in many natural active components such as α-linolenic acid (56% to 63%), vitamin E, aucubin, and so on. In this study, water-in-oil (W/O) microemulsions composed of Eucommia ulmoides Oliver seed oil, distilled water, a blend of Sorbitan monooleate 80 (Span 80) and Polysorbate (20) sorbitan monooleate (Tween 80), and propylene glycol were prepared for improving the compatibility of Eucommia ulmoides Oliver seed oil. Pseudoternary phase diagrams were built to illustrate the phase behavior of the microemulsions, based on hydrophilic-lipophilic balance values, cosurfactant type, the proportion of cosurfactant, and the changing environmental stress. Dynamic light scattering, transmission electron microscopy, and electrical conductivity measurements were performed to characterize the microstructural aspects. The optimum process conditions at which the Eucommia ulmoides Oliver seed oil-loaded microemulsion had good tolerance to pH and salinity were: Propylene glycol served as cosurfactant, water-Propylene glycol, and Span 80-Tween 80 ratios separately kept constant at 1:1 and 6:4. These microemulsions with narrow size distribution, nanoscale particle size (below 60 nm), transparent appearance had a wide range of oil phase content and free-radical scavenging capacity toward DPPH and ABTS radicals with half-maximal inhibitory concentration (IC ) values of 49.20 and 33.43 mg/mL, respectively. PRACTICAL APPLICATION: This nanostructure, environmental stability, and antioxidant activity of microemulsions containing Eucommia ulmoides Oliver seed oil is a potential delivery system as an alternative to α-linolenic acid and can be used for the delivery of peptides, proteins, antioxidants, and water-soluble nutrients.