This study aimed to assess the inclusion of antioxidant diphenylamine (DPA) to a Jatropha biodiesel (B30) blend to affects engine performance and exhaust emissions. In this research, three fuel blends has been utilized: diesel, B30, and 100 ppm of antioxidant diphenylamine is added to the B30 blend referred to as B30 + DPA100. A Response Surface Methodology (RSM) has been employed to analyze experimental data, resulting in the development of a regression equation. Additionally, an Adaptive Neuro-Fuzzy Inference System (ANFIS) has been employed to assess and verify the consistency of the developed model. The results from the experiments revealed that the inclusion of the antioxidant had a notable impact on reducing NOx emissions with only a slight effect on brake thermal efficiency (BTHE). Specifically, B30 + DPA100 showed a 7.48 and 15.53% decrease in average NOx emissions, while the average BTHE experienced a modest 0.78% decrease and a 2.67% increase compared to diesel and B30 biodiesel blend, respectively. The Brake Mean Effective Pressure (BMEP) of B30 + DPA100 has been found to be 3.91% and 1.3% higher than diesel and B30, respectively. The Brake Specific Fuel Consumption (BSFC) was 8.68% lower than B30 but 1.49% higher than diesel. Overall, these findings suggest that B30 + DPA100 can be used in diesel engines without modification, offering reduced NOx emissions.