In the Argentinean Pampas, where Phaeozems soils are predominant, the conversion of native grasslands into croplands has negatively affected soil fertility and health. Nitrogen (N) mineralized during an anaerobic incubation (Nan), an estimator of N potential mineralization, is frequently used to evaluate soil health. The objectives of our study were to determine the magnitude of the Nan depletion caused by agriculture (dNan), and to assess the relative contribution of land use, climate, and soil properties on dNan in the Argentinean Pampas. We conducted a soil survey at 409 sites distributed throughout the Argentinean Pampas region. At each site, pairwise samples (0–20 cm) were collected from uncultivated and agricultural soils. At each site, different edaphic, climatic, and productive variables were evaluated. The direct and indirect effect of different variables over dNan was evaluated through a path analysis. The conversion of native grasslands from the Argentinean Pampas into croplands generally decreased Nan values, with Nan reductions ranging from 0 to 216 mg kg−1 (representing 0 to 79 % of the Nan value at uncultivated soils). The magnitude of the Nan depletion was influenced by edaphic variables [i.e. original Nan value (Nan at uncropped soils), soil organic matter depletion, clay content] and climatic variables (i.e. precipitation). These four variables explained 85 % of the dNan variability. From the path analysis, it was determined that soils with greater original Nan values (r = 0.89) and greater soil organic matter depletion (r = 0.65) presented larger dNan, while increases in clay content (r = 0.30) or mean annual precipitation (r = 0.25) resulted in lower dNan values. The identification of the drivers of Nan depletion can aid in the classification of areas with greater vulnerability to soil degradation and in the development of models to predict Nan depletion in soils. Such models would hold significant potential as tools for policymakers and program developers engaged in the formulation and implementation of targeted interventions aimed at halting or reversing this degradation process.