Geothermal energy exploration is based in part on interpretation of the chemistry, temperature, and discharge rate of thermal springs. Here we present the major element chemistry and the δD, δ¹⁸O, ⁸⁷Sr/⁸⁶Sr and δ¹¹B isotopic ratio of groundwater from the low-enthalpy geothermal system near the city of Viterbo in the Cimino-Vico volcanic district of west-Central Italy. The geothermal system hosts many thermal springs and gas vents, but the resource is still unexploited. Water chemistry is controlled by mixing between low salinity,HCO₃-rich fresh waters (<24.2°C) flowing in shallow volcanic rocks and SO₄-rich thermal waters (25.3°C to 62.2°C) ascending from deep, high permeability Mesozoic limestones. The (equivalent) SO₄/Cl (0.01–0.02), Na/Cl (2.82–5.83) and B/Cl ratios (0.02–0.38) of thermal waters differs from the ratios in other geothermal systems from Central Italy, probably implying a lack of hydraulic continuity across the region. The δ¹⁸O (−6.6‰ to −5.9‰) and δD (−40.60‰ to −36.30‰) isotopic composition of spring water suggest that the recharge area for the geothermal system is the summit region of Mount Cimino. The strontium isotope ratios (⁸⁷Sr/⁸⁶Sr) of thermal waters (0.70797–0.70805) are consistent with dissolution of the Mesozoic evaporite-carbonate units that constitute the reservoir, and the ratios of cold fresh waters mainly reflect shallow circulation through the volcanic cover and some minor admixture (<10%) of thermal water as well. The boron isotopic composition (δ¹¹B) of fresh waters (−5.00 and 6.12‰) is similar to that of the volcanic cover, but the δ¹¹B of thermal waters (−8.37‰ to −4.12‰) is a mismatch for the Mesozoic reservoir rocks and instead reflects dissolution of secondary boron minerals during fluid ascent through flysch units that overlie the reservoir. A slow and tortuous ascent enhances extraction of boron but also promotes conductive cooling, partially masking the heat present in the reservoir. Overall data from this study is consistent with previous studies that concluded that the geothermal system has a large energy potential.