Response Surface Methodology for the Optimization of Lactoferrin Nano-Liposomes

The purpose of this study was to optimize the formulation of lactoferrin nano-liposomes using response surface methodology. Response surface methodology based on central composite rotatable design has been successfully used to model and optimize biochemical and biotechnological processes. The mass ratio of phosphatidylcholine and cholesterol (1-8), lactoferrin concentration (2-15 mg/mL), tween80 (0.5-2.5 mg/mL) and the volume ratio of organic phase and aqueous phase (2-10) were selected as independent variables with encapsulation efficiency and particle size as dependent variables. For each response, a 2-order polynomial model was developed using multiple linear regression analysis. Applying a desirability function method the optimum parameters were: phosphatidylcholine to cholesterol mass ratio of 5.77, lactoferrin concentration of 11.71 mg/mL, tween80 of 1.18 mg/mL and organic phase to aqueous phase volume ratio of 4.59. At this optimum point, particle size and encapsulation efficiency were found to be 214 nm and 60.21%, respectively. Furthermore, leakage ratio of nano-liposomes was used to determine the influence of storage period.