This work determined the association between several parameters of biodiesel production from waste cooking oil (WCO) using waste bovine bone (WBB) as catalyst to achieve a high conversion to fatty acid methyl ester (%FAME). The effect of three independent variables was used as the optimum condition using response surface methodology (RSM) for maximizing the %FAME. The RSM analysis showed that the ratio of MeOH to oil (mol/mol), catalyst amount (%wt), and time of reaction have the maximum effects on the transform to FAME. Moreover, the coefficient of determination (R-2) for regression equations was 99.19%. Probability value (P < 0.05) demonstrated a very good significance for the regression model. The optimal values of variables were MeOH/WCO ratio of 15.49:1 mol/mol, weight of catalyst as 6.42 wt%, and reaction time of 128.67 min. Under the optimum conditions, %FAME reached 97.59%. RSM was confirmed to sufficiently describe the range of the transesterification parameters studied and provide a statistically accurate estimate of the best transform to FAME using WBB as the catalyst.