Fossil fuel vehicles, emitting air toxics into the atmosphere, impose a heavy burden on the economy through additional health care expenses and ecological degradation. Air pollution is responsible for millions of deaths and chronic and acute health problems every year, such as asthma and chronic obstructive pulmonary disease. The fossil-fuel-based transportation system releases tons of toxic gases into the atmosphere putting human health at risk, especially in urban areas. This analysis aims to determine the economic burden of environmental and health impacts caused by Highway 401 traffic. Due to the high volume of vehicles driving on the Toronto Highway 401 corridor, there is an annual release of 3771 tonnes of carbon dioxide equivalent (CO2e). These emissions are mainly emitted onsite through the combustion of gasoline and diesel fuel. The integration of electric and hydrogen vehicles shows maximum reductions of 405−476 g CO2e per vehicle-kilometer. Besides these carbon dioxide emissions, there is also a large amount of hazardous air pollutants. To examine the impact of air pollution on human health, the mass and concentrations of criteria pollutants of PM2.5 and NOx emitted by passenger vehicles and commercial trucks on Highway 401 were determined using the MOVES2014b software. Then, an air dispersion model (AERMOD) was used to find the concentration of different pollutants at the receptor’s location. The increased risk of health issues was calculated using hazard ratios from literature. Finally, the health cost of air pollution from Highway 401 traffic was estimated to be CAD 416 million per year using the value of statistical life, which is significantly higher than the climate change costs of CAD 55 million per year due to air pollution.