Given the importance of dissolved organic matter (DOM) in the carbon cycling of aquatic ecosystems, information on its seasonal variability is crucial. In this study we assess the use of optical absorption indices available in the literature based on in situ data to both characterize the seasonal variability of DOM in a highly complex environment and for application in large-scale studies using remote sensing data. The study area comprises four lakes located in the Mamirauá Sustainable Development Reserve (MSDR). Samples for the determination of colored dissolved organic matter (CDOM) and measurements of remote sensing reflectance (Rrs) were acquired in situ. The Rrs was used to simulate the response of the visible bands of the Sentinel-2 MultiSpectral Instrument (MSI), which was used in the proposed models. Differences between lakes were tested using the CDOM indices. The results highlight the role of the flood pulse in the DOM dynamics at the floodplain lakes. The validation results show that the use of the absorption coefficient of CDOM (aCDOM) as a proxy of the spectral slope between 275 and 295 nm (S275295) during rising water is worthwhile, demonstrating its potential application to Sentinel-2 MSI imagery data for studying DOM dynamics on the large scale.