Urine patches within pasture soils are hotspots for nitrogen (N) cycling and losses, where nitrification inhibitors (NI) offer a means of reducing such losses. Within urine influenced soil, more research has been conducted for dicyandiamide (DCD) than 3,4-dimethylpyrazole phosphate (DMPP). Differences in the efficacy of these NI are often ascribed to a greater mobility of DCD, which may lead to spatial separation from NH4+ and nitrifying microorganisms. We tested the mobility of 14C-labelled DCD and DMPP relative to sheep urine-derived NH4+ in soil columns of contrasting texture and organic matter content, following simulated rainfall. We also assessed factors influencing the vertical mobility of these NI in soils, including solubility, sorption/desorption processes and microbial degradation and uptake. Following 40-mm rainfall, without the presence of sheep urine, the distribution of both NI were similar in the soil columns; however, there was a greater retention of DCD compared to DMPP in the top 1 cm. Both NI appeared to co-locate well with urine-derived NH4+, and the presence of sheep urine altered the leaching profile of the NI (compared to rainfall application alone), but this effect was inhibitor and soil-type dependent. A greater sorption to the soil matrix was observed for DCD in comparison to DMPP in all three studied soils, and the presence of urine generally increased desorption processes. Of the NI applied to the soil columns, 18–66 % was taken up within 30 min by the microbial community. However, only small amounts (<1 %) were mineralized during this period. In conclusion, due to the greater adsorption of DCD as opposed to DMPP and similarity in the degree of co-location of both NI with urine NH4+, the results of this study suggest that differences in microbial uptake and degradation may be more important parameters for explaining differences in the efficacy of reducing nitrification. Further work is required to determine the comparative efficacy of both NI in reducing nitrification rates under field conditions in a range of soil types and environmental conditions.