In this paper, the effects of biodiesel on performance and emission of the current and new-coming regulation cycles, namely the New European Driving Cycle (NEDC) and the Worldwide Harmonized Light Vehicles Test Cycle (WLTC), were investigated by conducting tests on a passenger car, a Euro-5 Ford Fiesta, equipped with a 1.5-L diesel engine. In a two-axle chassis dynamometer test bed, NEDC and WLTC were performed with pure diesel and biodiesel-to-diesel blend (30% biodiesel, 70% diesel in volume). A substantial reduction in CO (34%, 55%), HC (33%, 40%), and particulate number (PN) (22%, 31%) emissions was observed respectively for both the NEDC and WLTC when biodiesel was used. Besides, it was found that the WLTC has higher load and velocity profile compared to the NEDC. Moreover, lower CO, HC, and PN emissions were observed with B30 fuel under WLTC compared to the NEDC. Nevertheless, slightly higher CO2 and substantially higher NOx emissions were observed for the WLTC compared to the NEDC.