Anthropogenic alteration of the soil phosphorus (P) cycle leads to subsequent water quality issues in agricultural dominated watersheds. In the humid Northeastern United States (NE US), variably saturated areas can generate surface runoff that transports P and stimulates biogeochemical processes; these hydrologically dynamic locations are often called biogeochemical ‘hotspots’. Many studies have evaluated nitrogen and carbon cycling in biogeochemical hot spots but few have focused on P. We hypothesized seasonally wet parts of the landscape (i.e., hotspots) have smaller biologically available P pools because runoff events frequently carry away nutrients like P. To test this hypothesis, we generated soil wetness index (SWI) maps from soil (SURRGO) and elevation (LiDAR rescaled to 3 m) data and used these maps to direct seasonal soil sampling near Klingerstown, Pennsylvania (PA) and Ithaca, New York (NY). We collected 5cm deep soil samples in PA (bimonthly) and NY (monthly) along soil moisture gradients for a range of land cover types (forest, fallow, and cropped) from May through October. We measured soil moisture in the field and percent organic matter (OM), pH, and three increasingly strong soil P extractions (dilute-salt-extractable P, oxalate-extractable P, and total-extractable P) in the laboratory. Our results indicated a negative relationship between dilute-salt-extractable P concentrations and SWI in PA and no relationship between these same variables in NY. We also found positive relationships between each of the three P extractions in PA but only a positive relationship between oxalate-extractable P and total-extractable P in NY. Our findings in PA support our hypothesis; namely, less biologically available P (i.e. dilute-salt-extractable P) is found in wetter areas of the landscape. However, divergent P availability patterns in NY point to further complexities and confounding variables in our understanding in soil P processes. Further studies will look into the importance of environmental variables such as OM and pH on P patterns under changing soil moisture regimes. The knowledge gained from this study will improve our understanding of P cycling in biogeochemical hotspots and can be used to improve the effectiveness of agricultural management practices in the NE US.