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As a result of the increase of ecological awareness and restrictions for fumes emission, numerous studies have been conducted which aim at improving power supply systems and the structure of diesel engine. Another course of action taken to adjust to rigorous fumes emission norms is the replacement of hydrocarbon fuels with alternative ones. The research on the engine dynamometer, conducted by the authors, showed that the application of the mixture of canola oil with n-hexane positively influenced certain ecological parameters of the engine, particularly the nitric oxide (NOx) emission. Due to law regulations concerning the level of emission of toxic substances of exhaust gases, which require monitoring in real road traffic conditions, the attempts have been made to assess ecological parameters of diesel engine supplied with mixtures of canola oil with n-hexane. This article presents the research results for diesel engine in real operating conditions. The exhaust gas tester - Herman HGA 400 - was used for measuring ecological parameters. The level of the main toxic substances was measured during the research (CO, HC, NOx) and CO2, O2. The ecological parameters for diesel engine supplied with the mixture of canola oil and n-hexane were compared to those for diesel engine supplied with diesel fuel.