Relatively few radiometrically dated records are available for the central Mediterranean spanning the marine oxygen isotope stage 6–5 (MIS 6–5) transition and the first part of the Last Interglacial. Two flowstone cores from Tana che Urla Cave (TCU, central Italy), constrained by 19 U/Th ages, preserve an interval of continuous speleothem deposition between ca. 159 and 121 ka. A multiproxy record (δ18O, δ13C, growth rate and petrographic changes) obtained from this flowstone preserves significant regional-scale hydrological changes through the glacial/interglacial transition and multi-centennial variability (interpreted as alternations between wetter and drier periods) within both glacial and interglacial stages. The glacial stage shows a wetter period between ca. 154 and 152 ka, while the early to middle Last Interglacial period shows several drying events at ca. 129, 126 and 122 ka, which can be placed in the wider context of climatic instability emerging from North Atlantic marine and NW European terrestrial records. The TCU record also provides important insights into the evolution of local environmental conditions (i.e. soil development) in response to regional and global-scale climate events.