The determinants of the broadband absorption spectrum of eumelanins are still largely unknown. Herein we report a novel approach to investigate eumelanin chromophore which is based on the biomimetic oxidation of the key monomer precursor, 5,6-dihydroxyindole (DHI, 1), with peroxidase/hydrogen peroxide in phosphate buffer, pH 7, containing 1–5% polyvinylalcohol (PVA, 27 000 Da). This approach relies on the discovery that as low as 1% PVA can prevent precipitation of the growing melanin polymer thus allowing investigation of the chromophoric phases accompanying oxidation of DHI without confounding scattering effects. Spectrophotometric monitoring showed the initial development of a band around 530 nm persisting for about 1 h before gradually changing into the typical broadband spectrum of eumelanin. Reductive treatment caused a significant absorbance decrease in the visible region without affecting an absorption band around 320 nm. Initial product analysis indicated an altered formation ratio of 2,4′-biindolyl (2) and 2,7′-biindolyl (3) relative to control experiments. Overall, these results demonstrate for the first time that the development in solution of visible chromophores since the early oligomer stages is independent of strong aggregation/precipitation phenomena.