Mechanical-biological pre-treatment (MBP) of municipal solid waste (MSW) has gained evidence as a practice capable of accomplishing the requirements for environmental sustainable landfilling. In particular, MBP is effective in reducing the ammoniacal nitrogen content in the leachate. However, few data are available on the modifications of the nitrogen forms occurring during MBP and on the role played by processes such as nitrification and generation of refractory organic compounds. The dynamic transformations of nitrogen were investigated during the MBP. MSW was mechanically and biologically pre-treated; samples were collected at different stages of the process and analysed to investigate the evolution of nitrogen forms; batch and column leaching tests were performed as well. The results indicate that nitrification is negligible and volatilization can only partially explain the low ammoniacal nitrogen content in the leachate. Incorporation of ammoniacal nitrogen into a refractory organic form was assessed and is likely to play an important role. The maximum content of refractory organic nitrogen in the solid waste was achieved after about 60 days of aerobic pre-treatment; therefore, the minimal duration of the MBP should be about 8-9 weeks in order to optimize the ammoniacal nitrogen incorporation, unless the waste is characterized by a low C/N ratio.