Determining the contribution of wood smoke to air pollution in large cities such as London is becoming increasingly important due to the changing nature of domestic heating in urban areas. During winter, biomass burning emissions can exceed the contributions from traffic emissions, and have been identified as a major cause of exceedences of European air quality limits. The aim of this work was to quantify the contribution of biomass burning in London to concentrations of PM2.5 and determine whether local emissions or regional contributions were the main source of biomass smoke. To achieve this, a number of biomass burning chemical tracers were analysed at a site within central London and two sites in surrounding rural areas. Concentrations of levoglucosan, elemental carbon (EC), organic carbon (OC) and K+ were generally well correlated across the three sites. At all the sites, biomass burning was found to be a source of OC and EC, with the largest contribution of EC from traffic emissions, while for OC the dominant fraction likely included contributions from secondary organic aerosols, primary biogenic and cooking sources. Source apportionment of the EC and OC using average source ratios from published data was found to give reasonable estimation of the total carbon from non-fossil and fossil fuel sources based upon comparison with estimates derived from 14C analysis. Black carbon (BC) data from 2 and 7 wavelength Aethalometers were also apportioned into the contributions from biomass burning and traffic, based upon the enhanced absorption of wood smoke at UV wavelengths compared to BC. While the source apportionment of BC using this approach found similar trends to that observed for EC, higher percentage contributions of wood burning to BC were estimated. Based on a wood smoke mass conversion factor for levoglucosan, mean wood smoke mass at the sites was found to range from 0.78–1.0 μg m−3 during the campaign in January–February 2012. Measurements on a 160 m tower in London suggested a similar ratio of brown to black carbon (reflecting wood burning and traffic respectively) in regional and London air. Peaks in the levoglucosan and K+ concentrations were observed to coincide with low ambient temperature, consistent with domestic heating as a major contributing local source in London. Overall, the source of biomass smoke in London was concluded to be a background regional source overlaid by contributions from local domestic burning emissions. This could have implications when considering future emission control strategies during winter and may be the focus of future work in order to better determine the contributing local sources.