Sr isotope data on groundwater samples from coal and overlying sandstone aquifers in the eastern Powder River Basin, Wyoming, demonstrate that the Sr isotope ratio effectively identifies groundwater from different aquifers where major ion geochemistry and O and H stable isotope data fail. Groundwaters from sandstone aquifers have a uniform 87Sr/86Sr ratio of 0.7126–0.7127. Waters from coal seams vary from 87Sr/86Sr ratio = 0.7127 near the recharge area to 0.7151 farther into the basin. The distinct Sr isotope signatures of sandstone and coal aquifers may reflect different sources of Sr in these two rock types: Sr in sandstones is held primarily in carbonate cement, whereas coals contain more radiogenic Sr in organic matter. The Sr isotope ratio is useful in identifying wells that contain mixed waters, whether due to well construction or to incomplete aquifer isolation. Measurement and continued monitoring of the Sr isotope ratio in groundwaters should provide a powerful tool for characterizing the impact of the burgeoning coal-bed methane industry on the hydrology of the Powder River Basin.