Essential oil has unstable properties and low bioavailability. In order to improve these problems, with the help of ultrasonic wave, stable nano-emulsions were prepared with different emulsifiers. In this experiment, the formation of nano-emulsions with different emulsifiers at different concentrations was studied, including sodium caseinate (SC), whey glycoprotein (WGP), soy protein isolate (SPI) and Tween 80. Meanwhile, we studied the effect of ultrasonic cell disruption apparatus and homogenizer on the particle size of nano-emulsions. Cinnamon essential oil nano-emulsion prepared under the optimum emulsifier and its concentration has good microstructure and rheological properties. The stability of cinnamon essential oil nano-emulsion in different environments was studied, including centrifugation, freeze-thaw, heating-freezing and long-term (30 d) stability. The nano-emulsion of cinnamon essential oil is stable in emulsifiers. The nano-emulsions prepared by different emulsifiers has different particle sizes in different environments and their advantages and disadvantages are compared. Finally, how different nano-emulsion were digested in vitro was also studied. The in vitro digestion of nano-emulsion by different emulsifiers was simulated. Its properties include particle size, zeta potential and polymer dispersion index (PDI).