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The Amazon Basin is the largest on the planet, and its aquatic ecosystems affect and are affected by the Earth’s processes. Specifically, Amazon aquatic ecosystems have been subjected to severe anthropogenic impacts due to deforestation, mining, dam construction, and widespread agribusiness expansion. Therefore, the monitoring of these impacts has become crucial for conservation plans and environmental legislation enforcement. However, its continental dimensions, the high variability of Amazonian water mass constituents, and cloud cover frequency impose a challenge for developing accurate satellite algorithms for water quality retrieval such as chlorophyll-a concentration (Chl-a), which is a proxy for the trophic state. This study presents the first application of the hybrid semi-analytical algorithm (HSAA) for Chl-a retrieval using a Sentinel-3 OLCI sensor over five Amazonian floodplain lakes. Inherent and apparent optical properties (IOPs and AOPs), as well as limnological data, were collected at 94 sampling stations during four field campaigns along hydrological years spanning from 2015 to 2017 and used to parameterize the hybrid SAA to retrieve Chl-a in highly turbid Amazonian waters. We implemented a re-parametrizing approach, called the generalized stacked constraints model to the Amazonian waters (GSCMLAFW), and used it to decompose the total absorption αt(λ) into the absorption coefficients of detritus, CDOM, and phytoplankton (αphy(λ)). The estimated GSCMLAFWαphy(λ) achieved errors lower than 24% at the visible bands and 70% at NIR. The performance of HSAA-based Chl-a retrieval was validated with in situ measurements of Chl-a concentration, and then it was compared to literature Chl-a algorithms. The results showed a smaller mean absolute percentage error (MAPE) for HSAA Chl-a retrieval (36.93%) than empirical Rrs models (73.39%) using a 3-band algorithm, which confirms the better performance of the semi-analytical approach. Last, the calibrated HSAA model was used to estimate the Chl-a concentration in OLCI images acquired during 2017 and 2019 field campaigns, and the results demonstrated reasonable errors (MAPE = 57%) and indicated the potential of OLCI bands for Chl-a estimation. Therefore, the outcomes of this study support the advance of semi-analytical models in highly turbid waters and highlight the importance of re-parameterization with GSCM and the applicability of HSAA in Sentinel-3 OLCI data.