ABSTRACT As the world transitions away from fossil fuels, there is a growing need to balance energy security with energy demand. Several energy sources are being investigated and tested, primarily for driving automobiles. Biodiesel, a common vegetable oil derivative, is growing in popularity in several countries due to its substantial similarity to conventional diesel. Biodiesel can be promissorily used as a replacement liquid fuel for petrol-based diesel for compression ignition engines. At present, biodiesel production primarily relies on edible vegetable oils as its main source. This research explores the potential of a non-edible blend of Argemone Mexicana, Azadirachta Indica, and Melia Azedarach Linn seeds for biodiesel production. In the study, the raw oil was extracted from a novel ternary mixture of three non-edible oil seeds together. The raw oil was further processed for methyl ester production in two step process as the FFA was achieved more than 6%. Moreover, the biodiesel yield percentage was optimized by adjusting the parameters; catalyst concentration, molar ratio and heating time. The findings show that a 6:1 molar ratio, 60 min heating time and 0.6% by weight catalyst concentration yielded the highest percentage of biodiesel at 93%. The physio-chemical characteristics of the produced biodiesel were evaluated, and the outcomes satisfied the ASTM standards. The novel ternary mixture biodiesel exhibited equivalent chemical properties such as calorific value, viscosity and density as 39 MJ/kg, 7.2 mm2/s and 0.860 g/cm3 respectively. These findings demonstrate the feasibility of using non-edible oil from the seeds together for biodiesel production and the potential of this novel ternary mixture as an alternative to conventional diesel fuel.