Estuaries of the southeastern United States not only serve an important nursery function but also are common repositories of polycyclic aromatic hydrocarbons (PAHs) derived from upland activities. Thus, these habitats may be at risk for PAH phototoxicity. To better characterize this risk, a daily survey of ultraviolet-A (UV-A; 320–400 nm) irradiance was performed at Leadenwah Creek (Wadmalaw Island, SC, USA) on June 27 and August 1, 2003. In addition, laboratory assays were completed using two light exposure regimes: One that was typical of historical phototoxicity assessments (continuous light [C-UV]), and a more environmentally realistic regime (ER-UV). On both survey days, irradiance at a depth of 10 cm exhibited a pattern generally similar to that observed at the surface, whereas irradiance at the bottom of the creek was a function of both tidal height and time of day. Total UV-A dose at a 10-cm depth on June 27 and August 1, 2003 was 4.37 and 4.78 J/cm2, respectively. Attenuation coefficients on both days varied as a function of tidal height. In the laboratory, larval grass shrimp (Palaemonetes pugio) exposed to an ER-UV regime for these habitats (photoperiod, 12: 12-h light: dark; total daily UV-A dose, 4.40 J/cm2) exhibited a 2.5-fold decrease in toxicity compared with those exposed to the C-UV regime (photoperiod, 24: 0-h light: dark; total daily UV dose, 1.50 J/cm2), despite a threefold higher UV dose in the ER-UV regime. The lower potency under the ER-UV regime likely is attributable to the presence of a 12-h dark period allowing for recovery. The consequences of these results are discussed in the context of habitat-specific UV-A dose and its relevance to future laboratory assessments of PAH phototoxicity.