Abstract: OBJECTIVE To apply the Box-Behnken experimental design to optimize the formulation of oxaliplatin liposome. METHODS The independent variables were investigated by using a three-factor, three-level Box-Behnken experimental design with the concentration of phospholipid(X₁), the mass ratio of phospholipids and Ox(X₂) and the mass ratio of phospholipid and cholesterol(X₃) as the research object and the response variable was encapsulation efficiency. RESULTS The optimized formulation was as follows: the concentration of phospholipid is 10.20 g·L^-1, the mass ratio of phospholipid and drug was 31∶1 and the mass ratio of phospholipid and cholesterol was 3.8∶1. The deviation between the predicted values of entrapment efficiency and its theoretical values were small. CONCLUSION The Box-Behnken experimental design method can be used to optimize the formulation of oxaliplatin liposome.