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Oxygenated fuels are studied in spark combustion engines because of their potentially positive impact on greenhouse emissions, and as part of alternative renewable fuels. Furthermore, engine test results position them as a promising lever to reduce engine-out emissions, and most notably, particles. This study focuses on oxygenated fuel Butanol, which is a potential output of recent developments on Algae and Cyanobacteria harvest process. Its blending into gasoline and application into spark ignition engines is investigated. Blending levels of n-Butanol and iso-Butanol have been proposed based on standard gasoline’s octane number, RON, at two ethanol concentration levels, 10 and 25%. Fuel blend impact on combustion, and on regulated and non-regulated emissions is analysed. Fuel knock resistance properties, RON and MON, determine the knocking tendencies for ethanol and butanol at 2000 rpm. However, test results highlight different knocking sensibility behaviour at higher engine speed.