Stable isotopes of water were applied to estimate water yield to fifty lakes in northeastern Alberta as part of an acid sensitivity study underway since 2002 in the Athabasca Oil Sands Region (AOSR). Herein, we apply site-specific water yields for each lake to calculate critical loads of acidity using water chemistry data and a steady-state water chemistry model. The main goal of this research was to improve site-specific critical load estimates and to understand the sensitivity to hydrologic variability across a Boreal Plains region under significant oil sands development pressure. Overall, catchment water yields were found to vary significantly over the seven year monitoring period, with distinct variations among lakes and between different regions, overprinted by inter-annual climate-driven shifts. Analysis of critical load estimates based on site-specific water yields suggests that caution must be applied to establish hydrologic conditions and define extremes at specific sites in order to protect more sensitive ecosystems. In general, lakes with low (high) water yield tended to be more (less) acid sensitive but were typically less (more) affected by interannual hydrological variations. While it has been customary to use long-term water yields to define a static critical load for lakes, we find that spatial and temporal variability in water yield may limit effectiveness of this type of assessment in areas of the Boreal Plain characterized by heterogeneous runoff and without a long-term lake-gauging network. Implications for predicting acidification risk are discussed for the AOSR.