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A very competitive alcohol for use in diesel engines is butanol. Butanol is of particular interest as a renewable bio-fuel, as it is less hydrophilic and it possesses higher heating value, higher cetane number, lower vapor pressure, and higher miscibility than ethanol or methanol. These properties make butanol preferable to ethanol or methanol for blending with conventional diesel or gasoline fuel. In this paper, the spray and combustion characteristics of pure n-butanol fuel was experimentally investigated in a constant volume combustion chamber. The ambient temperatures were set to 1000 K, and three different oxygen concentrations were set to 21%, 16%, and 10.5%. The results indicate that the penetration length reduces with the increase of ambient oxygen concentration. The combustion pressure and heat release rate demonstrate the auto-ignition delay becomes longer with decreasing of oxygen concentrations.