Sea level rise has increased the frequency of tidal flooding even without accompanying precipitation in many coastal areas worldwide. As the tide rises, inundates the landscape, and then recedes, it can transport organic and inorganic matter between terrestrial systems and adjacent aquatic environments. However, the chemical and biological effects of tidal flooding on urban estuarine systems remain poorly constrained. Here, we provide the first extensive quantification of floodwater nutrient concentrations during a tidal flooding event and estimate the nitrogen (N) loading to the Lafayette River, an urban tidal sub-tributary of the lower Chesapeake Bay (USA). To enable the scale of synoptic sampling necessary to accomplish this, we trained citizen-scientist volunteers to collect 190 flood water samples during a perigean spring tide to measure total dissolved N (TDN), dissolved inorganic N (DIN) and phosphate concentrations, and Enterococcus abundance from the retreating ebb tide while using a phone application to measure the extent of tidal inundation. Almost 95% of Enterococcus results had concentrations that exceeded the standard established for recreational waters (104 MPN 100 mL(-1)). Floodwater dissolved nutrient concentrations were higher than concentrations measured in natural estuarine waters, suggesting floodwater as a source of dissolved nutrients to the estuary. However, only DIN concentrations were statistically higher in floodwater samples than in the estuary. Using a hydrodynamic model to calculate the volume of water inundating the landscape, and the differences between the median DIN concentrations in floodwaters and the estuary, we estimate that 1,145 kg of DIN entered the Lafayette River during this single, blue sky, tidal flooding event. This amount exceeds the annual N load allocation for overland flow established by federal regulations for this segment of the Chesapeake Bay by 30%. Because tidal flooding is projected to increase in the future as sea levels continue to rise, it is crucial we quantify nutrient loading from tidal flooding in order to set realistic water quality restoration targets for tidally influenced water bodies.