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Plants possess a wide range of cellular mechanisms that help them tolerate potentially toxic substances. Several studies that were carried out under laboratory conditions have demonstrated that the antioxidant compound glutathione plays a role in enabling mosses to tolerate high levels of heavy metals without toxic effects. However, until now, the antioxidant levels have not been studied in mosses under field conditions. In this study, we aimed to clarify the antioxidant concentrations of glutathione in the terrestrial moss Pseudoscleropodium purum growing in industrial environments characterised by high atmospheric levels of Cd, Ni, and Pb, and to evaluate the potential use of the compound as a biomarker. The results indicated the existence of a glutathione threshold response, which was significantly correlated with the metal toxicity and which may be influenced by metal bioavailability. Although future studies are needed to strengthen the findings, the present study suggests that total concentration of glutathione in P. purum could therefore be used as a biomarker in air pollution biomonitoring studies, provided that metal bioavailability is taken into account.