The Southeast Coast Network assessed water-quality at Timucuan Ecological and Historical Preserve as part of the National Park Service Vital Signs Monitoring Program. Monitoring was conducted following methods developed by the Environmental Protection Agency (EPA) as part of the National Coastal Assessment Program. Laboratory analysis measured chlorophyll a and total and dissolved concentrations of nitrogen and phosphorous. Field measurements included water temperature, pH, dissolved oxygen, and salinity. Water clarity, which requires a Secchi depth measurement, was calculated when conditions allowed. All measured parameters were rated as good, fair, or poor based on thresholds set by the EPA’s National Coastal Assessment Program. The Southeast Coast Network's monitoring approach is extensively documented in the Protocol for Monitoring Estuarine Water and Sediment Quality in Selected Southeast Coast Network Parks: Protocol Narrative version 2.0 (Starkey et al. 2023) and associated Standard Operating Procedures (SOPs). Version 1.0 of the protocol (Gregory et al. 2013) was implemented between 2007 and 2018. Version 2.0 of the protocol was implemented 2019 to present and was updated to be aligned with new technology and techniques documented in revised Standard Operating Procedures. Monitoring objectives remain consistent across both versions of the protocol. The monitoring objective associated with this dataset is: Determine status and spatial variability of water and nutrient conditions in estuarine waters every five years near SECN parks.