The Amazon River plume (ARP) has been shown to support high rates of nitrogen fixation and primary production. However, nitrogen fixation alone cannot account for total primary production determined in the region, hinting that other nitrogen uptake processes might play a role. For the first time, we measured nitrate uptake rates in the ARP during three cruises in May 2018, June 2019 and April/May 2021, along with primary production rates and an analysis of phytoplankton community composition via high performance liquid chromatography. Based on a classification according to the salt content the region was divided into estuarine (ES), mesohaline (MH) and oceanic (OC) stations. Primary production was light limited near the river mouth at ES stations and was maximal off the coasts of French Guiana and Suriname, where also nitrate uptake was highest with rates of 11.4 mmol m−2 d−1. The role of eddies pinching off a deflecting plume are discussed as possible reason for higher nutrient concentrations at the MH stations. Surprisingly, at most MH stations north of 5°N, nitrate uptake rates were low despite the presence of sufficient substrate concentration (up to 1.44 μM nitrate). Diatoms, dinoflagellates or Synechococcus sp. dominated phytoplankton communities. OC stations showed lowest productivity rates in accordance with oligotrophic conditions. However, rates seem to be sufficient to completely deplete the remaining riverine nitrate, preventing its export to the open ocean.