Fe2+/H2O2-mediated oxidation of homogentisic acid indicates the production of ochronotic and non-ochronotic pigments. Implications in Alkaptonuria and beyond.

Abstract

Homogentisic acid (HGA) can be oxidized by a combination of Fe2+ and H2O2 into a darkly colored high molecular mass pigment. Increasing the concentration of H2O2 can lead to the formation of a high molecular mass material that exhibits less absorbance in the visible range of the electromagnetic spectrum, while maintaining a strong absorbance in the UV range of the electromagnetic spectrum. FT-IR spectroscopy did indicate the presence of a chemical feature in the pigments generated through H2O2-mediated oxidation that is absent in pigments generated through air-mediated oxidation. Our observations could have implications in the pathophysiology of alkaptonuria. In alkaptonuria, patients suffer from homogentisic aciduria from birth, but develop ochronosis (darkening) of collagenous tissues much later in life due to the formation of a melanin-like pigment. Three major questions regarding ochronosis remain to be answered: 1) how is the pigment formed, 2) why does it appear by the third decade of life and 3) why is it sporadic in appearance? Our results suggest that ochronotic or non-ochronotic pigments can be generated from HGA depending on the oxidation reaction conditions. Thus, the absence of any visible pigment, as for younger alkaptonuria patients, could not necessarily mean the absence of HGA-derived melanin-like molecules. We compare our results and their potential implications for ochronosis to the changes in pigmentation observed in vitiligo or aging (greying) hair.