To reduce CO₂ emissions from automobiles, it is essential to improve system efficiency through the electrification of vehicles with internal combustion engines (ICEs), such as hybrid electric vehicles (HEVs) and plug-in hybrid electric vehicles (PHEVs), as well as through enhancements in ICE thermal efficiency. Additionally, biofuels and synthetic fuels are gaining attention as promising options to reduce CO₂ emissions from existing vehicles. Among these alternative fuels, ethanol, a bio-derived fuel, is already used at varying concentrations in many countries, and its further adoption is expected. Expanding the fleet of flex-fuel vehicles (FFVs) capable of running on high ethanol blends is one approach; however, increasing ethanol content in conventional gasoline, which is more widely used, is considered to have a greater impact on CO₂ reduction. A key issue is how existing vehicles adapt to increased ethanol concentrations such as E20, E30, and E40. This study focuses on turbocharged engines typically found in heavier passenger vehicles, which are less likely to be electrified, to assess the effects of varying ethanol concentrations on performance, efficiency, emissions, and reliability. The evaluation showed that increasing ethanol concentration from E0 to E40 in a fixed Blend stock for Oxygenate Blending (BOB) resulted in comparable or slightly improved torque and thermal efficiency, with emissions remaining similar and abnormal combustion tendencies suppressed. No reliability issues were observed in endurance testing. Furthermore, advancing ignition timing to take advantage of ethanol’s knock resistance within the constraints of mass-produced engines revealed that medium ethanol concentrations are sufficient to realize these benefits. Therefore, to improve output and thermal efficiency across a wider range of vehicles using the same ethanol volume, medium concentrations may be more advantageous than higher concentrations.