This study demonstrates a simple method to estimate the extent to which the exposure of cyclists to NO₂ is reduced by using off-road cycle routes rather than on-road cycle routes. Diffusion tubes were used to measure monthly NO₂ concentrations on three paired sets of on- and off-road cycle journeys in and around the City of York in August and September 2008. These measurements were combined with estimates of journey times to calculate time-weighted average concentration and exposure on each route. The average concentration of NO₂ was significantly reduced by a mean of 37.5% when off-road routes were used in place of on-road routes and, despite the longer journey times for off-road routes, exposure was also significantly reduced by a mean of 25.5%. The method described in this study could be adopted more widely to provide a cost effective and simple means of assessing the benefits of alternative cycle routes that are provided by Local Authorities.