Investigation of a new type of fuel for the internal combustion engine, which can be successfully used in both the power generation and the automotive industries, is presented in this article. The proposed fuel is a blend of 75% n-butanol and 25% glycerol. The engine tests conducted with this glycerol−butanol blend were focused on the performance, combustion thermodynamics, and exhaust emissions of a spark-ignition engine. A comparative analysis was performed to find potential similarities and differences in the engine fueled with gasoline 95 and the proposed glycerol−butanol blend. As measured, CO exhaust emissions increased, NOx emissions decreased, and UHC emissions were unchanged for the glycerol−butanol blend when compared to the test with sole gasoline. As regards the engine performance and combustion progress, no significant differences were observed. Exhaust temperature remarkably decreased by 3.4%, which contributed to an increase in the indicated mean effective pressure by approximately 4% compared to gasoline 95. To summarize, the proposed glycerol−butanol blend can be directly used as a replacement for gasoline in internal combustion spark-ignition engines.