The main purpose of this study is to deepen our knowledge of the relevant phenomena surrounding the combustion process in a medium-duty diesel engine, using two types of biofuels derived from sugar cane. The analysis was made through a broad base of information obtained from experimental tests. Three variables were considered as fundamental aspects of engine operation: brake thermal efficiency, nitrogen oxide and particulate matter emissions. This work aims to study the trade-off effect for the same engine running under the same conditions for a fossil fuel and for two types of biofuels. The trade-off graph will represent how the change of fuel influences on the engine’s brake thermal efficiency and also nitrogen oxide and particulate matter emissions. The tests was carried out on the MAN D0834 LF05, a dual-turbocharged engine with intake air cooling (intercooler), 16.5 compression ratio and a high-pressure exhaust gas recirculation system with gas cooling. The results of this study were satisfactory, as the particulate matter and nitrogen oxide emissions met the standards of the Brazilian Association of Technical Standards. These biofuels achieved emissions below the standards while maintaining their performance, proving to be a more sustainable choice over fossil fuels.