Integrated electron microprobe analyses (EMPAs) on glass and Sr–Nd isotope analyses have been performed on 17 tephras from the Middle Pleistocene Mercure lacustrine succession, southern Apennines. Two 40Ar/39Ar ages and the recognition of four relevant tephras from Colli Albani, Sabatini and possibly Roccamonfina volcanoes allowed us to ascribe the investigated succession to the late Marine Isotope Stage (MIS) 15–12 interval (560–440 ka). The Sr–Nd isotopes and major element glass compositions allowed us to attribute 10 out of the other 13 tephras to a poorly known activity of the Roccamofina volcano, whereas two layers were tentatively attributed to previously unknown Middle Pleistocene activity of Ponza Island or Campanian volcanoes, and one to Salina Island. The tephrostratigraphic correlation of the Mercure tephras with the Acerno lacustrine pollen record (Campania) also allowed us to evaluate the climatostratigraphic position of the tephras within the framework of the MIS 15–12 climatic variability. These results were obtained by combining the Sr–Nd isotope ratio with EMPA and 40Ar/39Ar geochronological data. This confirms the notable consistency of this approach for studying the Mediterranean Middle Pleistocene tephrostratigraphy, which, despite its great potential for both volcanology and Quaternary studies, has been hitherto barely explored.