Abstract: To prepare the β-cyclodextrin inclusion of Adlay seed oil with a saturated aqueous solution.The optimum by BoxBehnken response surface design were as follows-the ratio of β-CD to Adlay seed oil 9 ∶ 1 (g/g) , stirring temperature 45 ℃, inclusion time 6 h. The inclusion was characterized by SEM, DSC, FTIR and XRD. Entrapment efficiency of the inclusion determined by HPLC was 63%.In the acute toxicity test, the mice which were fed with the highest concentration (63 mg/m L) and maximum dosage (0.04 m L/g) , no death was found in the experimental mice after 14 days. No organ abnormalities were found and no organ pathology in sliced observation.The prepared inclusion complexes were safety for oral administration to be better for further study of efficacy.