Abstract This study reports optimization and simulation of biodiesel synthesis from waste cooking oil through supercritical transesterification reaction without the use of any catalyst. Although the catalyst enhances the reaction rate but due to the presence of water contents in waste cooking oil, the use of catalyst could cause a negative impact on the biodiesel yield. The transesterification reaction without catalyst also offers the advantage of the reduction of pretreatment cost. This study comprises of two steps; first step involves the development and validation of process simulation scheme. The second step involves the optimization using Response Surface Methodology. Face-centered central composite design of experiments is used for experimental matrix development and subsequent statistical analysis of the results. Analysis of variance (ANOVA) is employed for optimization purpose. In addition, a sensitivity study of the process parameters including pressure, temperature, and molar ration of oil-to-methanol was conducted. The statistical analysis reveals that temperature is the most influential process parameter as compared to pressure and oil-to-methanol molar ratio. The optimization study results in the maximum biodiesel yield (94.16%) at an optimum temperature of 274.8°C, 7.02 bar pressure, and an oil-to-methanol molar ratio of 12.43.