Abstract Chalcopyrite (CuFeS2) is both the most economically important and the most difficult copper mineral to (bio)leach. The main reason for the slow rate of chalcopyrite dissolution is the formation of a layer on the surface of the mineral that hinders dissolution, termed “passivation”. The nature of this layer is still under debate. In this work, the role of bacterial activity was examined on the leaching efficiency of chalcopyrite by mimicking the redox potential conditions during moderately thermophilic bioleaching of a pure chalcopyrite concentrate in an abiotic experiment using chemical/electrochemical methods. The results showed that the copper recoveries were equal in the presence and absence of the mixed culture. It was found that the presence of bulk jarosite and elemental sulphur in the abiotic experiment did not hamper the copper dissolution compared to the bioleaching experiment. The leaching curves had no sign of passivation, rather that they indicated a hindered dissolution. XPS measurements carried out on massive chalcopyrite samples leached in the bioleaching and abiotic experiments revealed that common phases on the surface of the samples leached for different durations of time were elemental sulphur and iron-oxyhydroxides. The elemental sulphur on the surface of the samples was rigidly bound in a way that it did not sublimate in the ultra-high vacuum environment of the XPS spectrometer at room temperature. Jarosite was observed in only one sample from the abiotic experiment but no correlation between its presence and the hindered leaching behaviour could be made. In conclusion, a multi-component surface layer consisting of mainly elemental sulphur and iron-oxyhydroxides was considered to be responsible for the hindered dissolution.