Montezuma Well is a unique natural spring located in a sinkhole surrounded by travertine. Montezuma Well is managed by the National Park Service, and groundwater development in the area is a potential threat to the water source for Montezuma Well. This research was undertaken to better understand the sources of groundwater to Montezuma Well. Strontium isotopes (87Sr/86Sr) indicate that groundwater in the recharge area has flowed through surficial basalts with subsequent contact with the underlying Permian aged sandstones and the deeper, karstic, Mississippian Redwall Limestone. The distinctive geochemistry in Montezuma Well and nearby Soda Springs (higher concentrations of alkalinity, As, B, Cl, and Li) is coincident with added carbon dioxide and mantle-sourced He. The geochemistry and isotopic data from Montezuma Well and Soda Springs allow for the separation of groundwater samples into four categories: (1) upgradient, (2) deep groundwater with carbon dioxide, (3) shallow Verde Formation, and (4) mixing zone. δ18O and δD values, along with noble gas recharge elevation data, indicate that the higher elevation areas to the north and east of Montezuma Well are the groundwater recharge zones for Montezuma Well and most of the groundwater in this portion of the Verde Valley. Adjusted groundwater age dating using likely 14C and δ13C sources indicate an age for Montezuma Well and Soda Springs groundwaters at 5,400–13,300 years, while shallow groundwater in the Verde Formation appears to be older (18,900). Based on water chemistry and isotopic evidence, groundwater flow to Montezuma Well is consistent with a hydrogeologic framework that indicates groundwater flow by (1) recharge in higher elevation basalts to the north and east of Montezuma Well, (2) movement through the upgradient Permian and Mississippian units, especially the Redwall Limestone, and (3) contact with a basalt dike/fracture system that provides a mechanism for groundwater to flow to the surface. While the exact nature of the groundwater flow connections is still uncertain, the available data indicate that flow to Montezuma Well may be more susceptible to future groundwater development in the Redwall Limestone than from any other geologic unit. Overall, the shallow groundwater in the surrounding Verde Formation appears to be largely disconnected from deeper groundwater flowing to Montezuma Well.