Abstract Mitigation measures to reduce greenhouse gas emissions may have adverse effects on urban air quality and human exposure to harmful pollutants. The use of bioethanol fuelled vehicles is increasing worldwide and may create new undesired pollution effects. Different measurement campaigns were performed in a pilot study to contribute to the understanding of the consequences associated with the use of bioethanol blended fuel (E95) on a series of pollutants. Ambient screening measurements of NO2, O3, acetic acid, formaldehyde and acetaldehyde were performed at different urban locations, exposed and not exposed to the circulation of bioethanol buses. In addition, volatile organic compounds were measured at the exhaust pipe of a bioethanol fuelled bus, both under idling conditions (carbonyls; DNPH cartridge) and under on-road driving conditions applying online monitoring (PTR-TOF). Higher ambient acetaldehyde values were measured at locations exposed to bioethanol fuelled buses than at locations not exposed, and very high acetaldehyde and acetic acid values were measured from the exhaust pipe during driving conditions (acetaldehyde>150ppm; acetic acid≈20–30ppm) and modelled at close distance to the bioethanol bus. Human exposure to high concentration of acetaldehyde is expected, and it may involve a significantly increased chance in developing cancer. The high concentration of acetic acid will involve odour annoyance and significant material degradation or corrosion.