When aging, melanin in human retinal pigment epithelium (RPE) undergoes oxidative modifications, which increase its photoreactivity and reduce its antioxidant capacity, elevating the risk of chronic phototoxicity to the retina. The aim of this research was to examine the effect of iron on the degradation of melanin induced by hydrogen peroxide and light, and to elucidate the role of hydrogen peroxide and singlet oxygen in the photodegradation of melanin. A water-soluble synthetic model of eumelanin with and without iron ions was treated either with exogenous hydrogen peroxide or with intense violet light. Oxidative modifications of melanin were analyzed by electron paramagnetic resonance (EPR) spectroscopy, absorption spectrophotometry, dynamic light scattering (DLS) and by chemical analysis of melanin subunits. The results showed that although iron strongly accelerated melanin degradation induced by hydrogen peroxide, it had very little influence on the rate of photodegradation of melanin. On the other hand, the photodegradation of melanin was partly inhibited by NaN3. The determination of hydrogen peroxide together with oxygen uptake indicates that irradiated melanin generates similar amounts of singlet oxygen and hydrogen peroxide. Analysis of melanin samples exhibiting comparable reduction of their EPR signal revealed that the loss of the representative melanin subunits was much higher in irradiated samples than in those treated with hydrogen peroxide in the dark. In conclusion, hydrogen peroxide, formed during the aerobic photolysis of melanin, is not responsible for the accompanying oxidative modifications of melanin. On the other hand, singlet oxygen can be considered as a key oxidizing agent involved in the photodegradation of melanin.