Radiometals are increasingly used in nuclear medicine for both diagnostic and therapeutic purposes. The DOTA ligand (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) is widely used as a chelating agent for various radionuclides, including 89Zr, with high thermodynamic stability constants and great in vivo stability. However, in contact with radioisotopes, chelating molecules are subjected to the effects of radiation, which can lead to structural degradation and induce alteration of their complexing properties. For the first time, the radiolytic stability of the Zr–DOTA complex in aqueous solution was studied and compared to the stability of the DOTA ligand. The identification of the major degradation products allows us to propose two different degradation schemes for the DOTA ligand and Zr–DOTA complex. DOTA is degraded preferentially by decarboxylation and cleavage of an acetate arm CH2–COOH, whereas in Zr–DOTA, DOTA tends to oxidize by the addition of the OH group in its structure. In addition, the degradation of the ligand, when involved in a Zr complex, is significantly less than when the ligand is free in solution, indicating that the metal protects the ligand from degradation. DFT calculations were performed to supplement the experimental data and give an improved understanding of the behaviour of DOTA and Zr–DOTA solutions after irradiation: the increase in stability upon complexation is attributed to the strengthening of the bonds in the presence of metal cations, which become less vulnerable to radical attack. Bond dissociation energies and Fukui indices are shown to be useful descriptors to estimate the most vulnerable sites of the ligand and to predict the protective effect of the complexation.