Disruption of the natural patterns of freshwater flow into estuarine ecosystems occurred in many locations around the world beginning in the twentieth century. To effectively restore these systems, establishing a pre-alteration perspective allows managers to develop science-based restoration targets for salinity and hydrology. This paper describes a process to develop targets based on natural hydrologic functions by coupling paleoecology and regression models using the subtropical Greater Everglades Ecosystem as an example. Paleoecological investigations characterize the circa 1900 CE (pre-alteration) salinity regime in Florida Bay based on molluscan remains in sediment cores. These paleosalinity estimates are converted into time series estimates of paleo-based salinity, stage, and flow using numeric and statistical models. Model outputs are weighted using the mean square error statistic and then combined. Results indicate that, in the absence of water management, salinity in Florida Bay would be about 3 to 9 salinity units lower than current conditions. To achieve this target, upstream freshwater levels must be about 0.25 m higher than indicated by recent observed data, with increased flow inputs to Florida Bay between 2.1 and 3.7 times existing flows. This flow deficit is comparable to the average volume of water currently being diverted from the Everglades ecosystem by water management. The products (paleo-based Florida Bay salinity and upstream hydrology) provide estimates of pre-alteration hydrology and salinity that represent target restoration conditions. This method can be applied to any estuarine ecosystem with available paleoecologic data and empirical and/or model-based hydrologic data.