Radiosensitizers are used in radiotherapy to enhance tumour control of radioresistant hypoxic tumours. While the detailed mechanism of radiosensitization is still unknown, the formation of radical anions is believed to be a key step. Thus understanding the ionization reactions of radiosensitizers is crucial in evaluating the radiosensitization potential and in developing new and more effective drugs. The present work investigates the negative and positive electrospray ionization and subsequent collision-induced dissociation and electron-induced dissociation reactions of ions derived from nimorazole, misonidazole and related compounds using a hybrid linear ion trap – Fourier Transform Ion Cyclotron Resonance mass spectrometer (Finnigan-LTQ-FT). A key finding is that negative electrospray ionization of these radiosensitizers leads to the formation of radical anions, allowing their fragmentation reactions to be probed for the first time.