The use of ethanol in gasoline has become a worldwide tendency as an alternative to reduce net CO2 emissions to the atmosphere, increasing gasoline octane rating and reducing dependence on petroleum products. However, recently environmental authorities in large urban centers have expressed their concerns on the true effect of using ethanol blends of up to 20% v/v in in-use vehicles without any modification in the setup of the engine control unit (ECU), and on the variations of these effects along the years of operation of these vehicles. Their main concern is the potential increase in the emissions of volatile organic compounds with high ozone formation potential. To address these concerns, we developed analytical and experimental work testing engines under steady-conditions. We also tested carbureted and fuel-injected vehicles every 10,000 km during their first 100,000 km of operation. We measured the effect of using ethanol-gasoline blends on the power and torque generated, the fuel consumption and CO2, CO, NOx and unburned hydrocarbon emissions, including volatile organic compounds (VOCs) such as acetaldehyde, formaldehyde, benzene and 1,3-butadiene which are considered important ozone precursors. The obtained results showed statistically no significant differences in these variables when vehicles operate with a blend of 20% v/v ethanol and 80% v/v gasoline (E20) instead of gasoline. Those results remained unchanged during the first 100,000 km of operation of the vehicles. We also observed that when the vehicles operated with E20 at high engine loads, they showed a tendency to operate with greater values of λ (ratio of the actual air-fuel ratio to the stoichiometric air-fuel ratio) when compared to their operation with gasoline. According to the Eco-Indicator-99, these results represent a minor reduction (<1.3%) on the impact to human health, and on the deterioration of the ecosystem. However, it implies a 12.9% deterioration of the natural resources. Thermal equilibrium analysis, at the tailpipe conditions (~100 °C), showed that ethane, formaldehyde, ethylene and ethanol are the most relevant VOCs in terms of the amount of mass emitted. The use of ethanol in the gasoline reduced 20–40% of those emissions. These reductions implied an average reduction of 17% in the ozone formation potential.