While fossil fuel resources are limited, the increasing demand for energy in the industrial and transportation sectors has led to using renewable fuels. This study aims to improve biodiesel production from sunflower oil by combining cold plasma and oscillatory systems through a transesterification process. The four main operational variables are examined, such as molar ratio (MR) (1:6–1:12), catalyst concentration (CC) (0.5–1.25 wt%), plasma exposure time (PET) (25–75 s), and residence time (RT) (30–90 s). The smooth periodic constriction reactor's physical attributes, including the diameter of the tube (D), the diameter of the constricted region (d0), and the distance between baffles (l), as well as the properties of the cold plasma system, are determined. The data analysis presents that PET, RT, CC, and MR have the greatest effect on efficiency. This optimization indicates that by exposing the reaction mixture to plasma for 59.99 s, adding 1.09% by weight of the catalyst, and with a MR of 7.17 in 86.43 s, biodiesel with a conversion percentage of 94.75% is produced. The results show that the optimum point matches with EN 14214 standard.