To develop the nano-liposomal system of turmeric essential oil for inhibiting Aspergillus flavus growth and aflatoxin generation. Methods: The stabilized and bio-safe liposomes loading turmeric essential oil were prepared by the ethanol injection-ultrasonic emulsification method with lecithin and cholesterol as excipients. Based on the combination of single factor screening and orthogonal design, liposomes were obtained by means of the optimized technology. Subsequently, following with the physicochemical characterization, the anti-Aspergillus flavus activity of liposomes were evaluated. Results: With the ratio of lecithin and cholesterol(10∶1, weight ratio), ratio of feeding drug and excipient(1:5, weight ratio), 1 mL/min of the ethyl alcohol adding speed and along with ultrasonic emulsification for 5 min, liposomes were prepared, exhibiting high encapsulation efficiency, suitable size distribution, and good storage stability. Particularly, after incubation with 2 mL essential oil liposome per 25 mL culture medium for 10d, almost 100% of Aspergillus flavus growth could be suppressed, without aflatoxin production. Conclusion: The preparation of turmeric essential oil based on nano-system not only increases the stability and homogeneity of the system but guarantees the efficacy of anti-Aspergillus flavus.