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This paper reports the results obtained in the ECOBUS Project. This project has been supported by the LIFE Program, one of the spearheads of European Union's environmental policy. The main aim for this project was to convert used cooking oil into biodiesel and then use this biodiesel in urban buses, contributing to reducing in one hand waste cooking oil that can create problems in the drainage city system and water contamination and on the other hand reducing engine polluting emissions. Departamento de Máquinas y Motores Térmicos (DMMT) facilities at the Universidad Politécnica de Valencia (Spain) were used to conduct detailed performance and emission measurements in a Diesel engine similar to those used by the urban transport operator in Valencia (Empresa Municipal de Transporte, EMT). The test engines were operated with conventional Diesel fuel (EN 590) and three different biodiesel blends: 30, 50 and 70%, using the pure biodiesel obtained from used cooking oil (EN 14214).