Melanoidins, the brown-colored compounds formed through the Maillard reaction, are responsible for color development in dulce de leche (DL), a popular confectionary dairy product in the Río de la Plata region, particularly in Uruguay and Argentina. Color is a critical quality parameter that strongly influences consumer preference. This work aimed to develop a method to perform preliminary structural characterization of the chromophores produced by the Maillard reaction. Melanoidins are present in a water-insoluble fraction, linked to a protein backbone, conforming melanoproteins of high molecular weight. The insoluble melanoprotein fraction (10% total solids) was isolated, and the chromophores released by proteolysis and isolated by gel-permeation chromatography. The analysis of the products revealed that they present a high degree of molecular weight (MW) polydispersity, in a range of 300 to 2000 Da, where the compounds of higher molecular weight contributed the most to the color of the product. The isolated fractions were further analyzed by RP-HPLC using a diode array detector (DAD) detector. These results, together with H-NMR data, suggested that the chromophores isolated belonged to a relatively simple mixture of aromatic products with higher hydrophobic character relative to other products of the melanoprotein digestion. View Full-Text