Natural fibrous epsomite crystals containing high amounts of toxic and radioactive elements have recently been discovered in the abandoned sulphur mine of Perticara (Italy). In the present study, all the different matrices that characterize the Perticara mine (air, water, host-rock, minerals, and bitumen) were sampled and investigated employing a multi-analytical approach to define the distribution and origin of these hazardous elements. Water samples showed high Al, Fe, Pb, Mg, and Mn content but not radioactive elements. The bitumen sample showed a higher amount of 210Po and 210Pb (0.12 Bq/g and 0.11 Bq/g, respectively), compared to the host-rock and fibrous sericolite samples, but lower than fibrous epsomite crystals (210Po 5.59 Bq/g; 210Pb 5.93 Bq/g). A high 222Rn concentration was also detected in the tunnels and it is likely the source of both 210Po and 210Pb, which are in radioactive equilibrium, especially in epsomite. This latter mineral, being capable of capturing 210Po and 210Pb, can be used as a helpful mineral indicator for the presence of radioactive elements in similar environmental conditions. Moreover, our results also highlight a slight anomaly in the 40K and 226Ra content of the host-rock, probably derived from the evaporitic sediments of the rock succession. The current risk for humans linked to the mine environment is minor since the activity of the mine ended several decades ago. However, the results of this study must be taken into serious consideration for any future development of this area (e.g., industrial archaeology, farming, educational, scientific and touristic purposes), and represent the basis for the calculation of radiotoxicity and dose assessment, which is fundamental for the correct and safe management of the mining environment and neighbouring areas.