AbstractConverting biomass to charcoal produces physical and chemical changes greatly increasing environmental recalcitrance, leading to great interest in the potential of this carbon form as a long-term sequestration strategy for climate change mitigation. Uncertainty remains, however, over the timescale of charcoal’s environmental stability, with estimates varying from decadal to millennial scales. Uncertainty also remains over charcoal’s effect on other aspects of carbon biogeochemical cycling and allied nutrient cycles such as nitrogen. Radiocarbon is a powerful tool to investigate charcoal mineralization due to its sensitivity; here we report the results of a study using ¹⁴C-dead charcoal (pMC=0.137±0.002) in organic-rich soil (pMC=99.76±0.46), assessing charcoal degradation over 55 days of incubation. Using this method, we discriminated between decomposition of indigenous soil organic matter (SOM) and charcoal by microorganisms. SOM was the major source of carbon respired from the soil, but there was also a contribution from charcoal carbon mineralization. This contribution was 2.1 and 1.1% on days 27 and 55, respectively. We also observed a negative priming effect due to charcoal additions to soil, where SOM mineralization was repressed by up to 14.1%, presumably arising from physico-chemical interactions between soil and charcoal.