The use of biofuels in vehicles becomes more advantageous than the consumption of fossil fuels, mainly because it uses renewable sources of energy. Recently there are some concerns about biodiesel sources, and hydrotreated vegetable oils (HVO) appear as a possible advanced solution. To understand the effect that the implementation of the new and old European type-approval test cycles (NEDC e WLTP) has on the results of these fuels considering pollutant emissions and fuel consumption results, a EURO V vehicle was subject to these cycles and also to engine performance evaluation tests. For this analysis, the fuels considered were: B0 (pure diesel), B7 (7% of biodiesel), B15 (15% of biodiesel), B100 (pure biodiesel), and HVO15 (15% of HVO). The findings lead to the conclusion that completely replacing fossil fuels with biofuels is not the most cost-effective approach. No significant differences were observed considering the two homologation cycles, the oldest (NEDC) and the actual (WLTP) and the use of HVO also does not present any relevant differences concerning the fuel consumption differences to B0 (+0.58% NEDC and +0.05%WLTP), comparing well with biodiesel behavior (−1.74% NEDC and −0.69%WLTP for B7 and +1.48% NEDC and 1.89% WLTP for B15). Considering the power of the engine obtained with the fuels, the differences are almost negligible, revealing variations smaller than 2% for B7, B15, and HVO15.