Resumo. Abstract Aim: In this research, we investigated the spectral variability of the specific phytoplankton absorption coefficient, a*φ, measured in a tropical meso-to-hypertrophic reservoir, aiming to find spectral features associated with the chlorophyll-a (chla) and other accessory pigments present in different phytoplankton species. Methods To accomplish this research, two fieldworks were carried out in different seasons in order to report a high bio-optical variation. Phytoplankton absorption coefficient, aφ, and chla concentration were measured in laboratory to estimate a*φ. Results The outcomes have indicated that there is a remarkable phytoplankton community dynamic as spatially as seasonally. Chla absorption features were well-defined at 440 nm and 675 nm. Conclusions All the a*φ spectra exhibited the absorption peak around 630 nm associated with phycocyanin pigment present in cyanobacteria. Some spectra have shown a peak at about 460 nm, which is related to chlorophyll b and chlorophyll c (chlb and chlc, respectively) found in different phytoplankton species. In turn, absorption features of carotenoids around 490 nm also were identified, however, well defined just in curves measured in austral autumn. Such spectral features are found in phytoplankton groups already identified in the study area such as Chlorophyceae, Bacillariophyceae, Cyanophyceae, Conjugatophyceae, Chrysophyceae, among others. We expect that the results are useful in researches about remote sensing of phytoplankton and eutrophication in reservoirs.