Concentrations less than 10 ng/L of 17 β-estradiol (E2), a natural estrogen, have been linked to adverse health effects in fish, including skewed sex distributions, reproductive failure, and organ impairment. The persistence of E2 in carbonate aquifer systems is not well documented. Water and sediment from cave streams within the Ozark Plateau of Missouri, USA, were collected and analyzed for E2. The persistence of E2 in the water was examined in two separate experiments, in which the holding temperatures (20°C vs. 4°C), bottle type, exposure to light, and filtration were varied. During two trials, no statistical difference was observed in the concentration of E2, suggesting that E2 is stable within the water. The fate of E2 was also examined in sediment-pore water collected from the cave streams in two independent trials. In trial 1, a significant decrease in E2 was noted over the 29 days of the experiment. However, in trial 2, no change in E2 concentration was observed. The results indicate that E2 is relatively stable in cave stream water and may persist in the sediment.