Melanins are natural pigments with promising bioelectronic applications. Among their unique properties, the existence of a persistent paramagnetic signal can be considered one of the most intriguing and controversial features. Additionally, the possible influence of such centers on charge transport accentuates the need for a better understanding of their origin and specific characteristics. In this report, electron paramagnetic resonance spectra of melanin samples obtained at different experimental conditions were systematically studied. From the fitting procedure, three distinct resonant lines are proposed, two associated with carbon-centered radicals and one with semiquinone free radicals.