Molecular characterization of dissolved organic nitrogen and phosphorus in agricultural runoff and surface waters

•FT-ICR-MS revealed DON and DOP molecular characteristics in runoff and surface waters.•Less-bioavailable DON and DOP compounds were abundant in all waters.•Within less-available DON and DOP compounds, lignin-like compounds dominant in all waters.•Photochemical and/or microbial processes altered DON and DOP signatures in all waters. Agricultural runoff is a significant contributor to nitrogen (N) and phosphorus (P) pollution in water bodies. Limited information is available about the molecular characteristics of the dissolved organic N (DON) and P (DOP) species in the agricultural runoff and surface waters. We employed Fourier Transform-Ion Cyclotron Resonance-Mass Spectrometry (FT-ICR-MS) to investigate the changes in the molecular characteristics of DON and DOP at three watershed positions (upstream water, runoff from agricultural fields, and downstream waters). Across three watershed locations, more-bioavailable compounds (such as amino sugars, carbohydrates, lipids, and proteins) accounted for 95% of DON and 69–96% of DOP. Of the dissolved organic matter, runoff waters from agricultural fields contained the greatest proportion of DON formulas (20–25%) than upstream (18%) and downstream (13–14%) waters, indicating the presence of a greater diversity of DON species in the runoff. Various nutrient sources present in agricultural fields such as crop residues, soil organic matter, and transformed fertilizers likely contributed to the diverse composition of DON and DOP in the runoff, which were likely altered as the surface water traversed along the flow pathways in the watershed. The presence of more-bioavailable molecules detected in upstream compared to agricultural runoff and downstream waters suggests that photochemical and/or microbial processes likely altered the characteristics of DON and DOP compounds. The findings of this study increase our understanding of DON and DOP compounds lability and transformations in runoff and surface waters , which may be useful in quantifying the contribution of organic N and P sources to water quality impairment in aquatic ecosystems. [Display omitted]