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In recent years, due to the favorable climate conditions of Iran, the cultivation of rapeseed has increased significantly. The aim of this study was to investigate the possibility of biodiesel production from one genotype of rapeseed (TERI (OE) R-983). An ultrasonic approach was used in order to intensify the reaction. Response surface methodology (RSM) was applied to identify the optimum conditions of the process. The results of this research showed that the conversion of biodiesel was found to be 87.175% under the optimized conditions of a 4.63:1 molar ratio (methanol to oil), 56.50% amplitude, and 0.4 s pulses for a reaction time of 5.22 min. Increasing the operating conditions, such as the molar ratio from 4:1 to 5.5:1, amplitude from 50% to 72.5%, reaction time from 3 min to 7 min, and pulse from 0.4 s to 1 s, increased the FAME (fatty acid methyl esters) yield by approximately 4.5%, 2.3%, 1.2%, and 0.5%, respectively. The properties of the TERI (OE) R-983 methyl ester met the requirements of the biodiesel standard (ASTM D6751), indicating the potential of the produced biodiesel as an alternative fuel.