Luteolin was formulated into transfersome as lipid-based carrier. Development of transfersome was carried out using Central Composite design and the selected independent variables were total lipid (phospholipid-Tween 80) and luteolin concentration. Transfersome formulas were optimized by response surface methodology (RSM) for four responses of particle size (Z-average), polydispersity index, zeta potential and entrapment efficiency. Series of 14 formulas of luteolin transfersome were prepared by thin film hydration followed by sonication method. The effect of total lipid and luteolin interaction was found to be significant at entrapment efficiency but not at the other responses. Optimum formula of 4.88% total lipid and 0.5% luteolin with desirability value of 0.609 has been verified to ensure conformity with prediction responses. Vesicle image using transmission electron microscope (TEM) revealed spherical particles and occurrence of particle aggregation. Optimum formula of luteolin transfersome had particle size of 286.03±8.46 nm, polidispersity index of 0.480±0.013, zeta potential of -18.67±0.379 mV and entrapment efficiency of 94.97±0.28 %. Those responses values were not correspond to the predicted value. It was confirmed by the low value of adjusted and predicted R-squared. These regression models could not be used to predict the response value of a subsequent test.