B-vitamins are recognized as essential organic growth factors for many organisms, although little is known about their abundance and distribution in marine ecosystems. Despite their metabolic functions regulating important enzymatic reactions, the methodology to directly measure different B-vitamins in aquatic environments has only recently been developed. Here, we present the first direct measurements of two B-vitamins, thiamin (B-1), and pyridoxine (B-6), in the Amazon River plume-influenced western tropical North Atlantic (WTNA) Ocean, an area known to have high productivity, carbon (C) and dinitrogen (N-2) fixation, and C sequestration. The vitamins B-1 and B-6 ranged in concentrations from undetectable to 230 and 40 pM, respectively. Significantly higher concentrations were measured in the surface plume water at some stations and variation with salinity was observed, suggesting a possible riverine influence on those B-vitamins. The influences of vitamins B-1 and B-6 on biogeochemical processes such as C and N-2 fixation were investigated using a linear regression model that indicated the availability of those organic factors could affect these rates in the WTNA. In fact, significant increases in C fixation and N-2 fixation were observed with increasing vitamin B-1 concentrations at some low and mesohaline stations (stations 9.1 and 1; p value <0.017 and <0.03, respectively). N2 fixation was also found to have a significant positive correlation with B-1 concentrations at station 1 (p value 0.029), as well as vitamin B-6 at station 9.1 (p value <0.017). This work suggests that there can be a dynamic interplay between essential biogeochemical rates (C and N-2 fixation) and B-vitamins, drawing attention to potential roles of B-vitamins in ecosystem dynamics, community structure, and global biogeochemistry.