This study assessed air quality indicators before and after enactment of the Spanish anti-smoking law. Mass and number concentrations and the chemical composition of particles were evaluated. Microscopy analyses were also conducted. Real time concentrations of PM10, PM2.5, PM1 and ultrafine particles were measured under ventilated and non-ventilated conditions and PM10 samples were collected for detailed inorganic and organic chemical characterization. Before enactment of the law in 2010, tobacco smoke produced significant indoor ambient particulate matter pollution, with elevated particulate matter mass concentrations (PM10 and PM1 concentrations of 122–220 and 48–85μg/m3, respectively) and ultrafine particle numbers (75,000 and 48,000cm–3 under ventilated and non-ventilated conditions, respectively). Typical tobacco smoke tracers including iso- and anteiso-alkanes and elements including La and Ce from the ignition of lighters were abundant. Additionally, several toxic substances derived from tobacco smoke, including Cd (3.1ng/m3) and benzo[a]pyrene (1.0ng/m3) were present at concentrations approximately 10 times greater than those measured after enactment of the anti-smoking law. The anti-smoking law significantly reduced exposure to potentially toxic compounds by approximately 90%. This law is expected to have a positive health impact, particularly for people who spend considerable time in affected environments, such as employees.