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Butanol has been proposed as a biologically derived fuel that has significant advantages over ethanol in terms of energy density and miscibility with diesel. This has generated the need to study separately and evaluate comparatively the in-engine combustion of the four isomers of butanol. Previous studies on the combustion of butanol isomers in laminar premixed flames have shown that while the isomers exhibit several similar combustion characteristics, including adiabatic flame temperature and flame speed, pollutant formation is highly dependent on the precise chemical structure of each isomer. The objective of this study is to build on these findings by investigating the effect of three of the four butanol isomers (n-butanol, isobutanol, and sec-butanol) on engine performance and emissions. The three isomers were blended as 30% butanol and 70% gasoline on a mass basis. These fuel blends were tested in a single-cylinder port-injection spark-ignition engine.