PurposeHypolimnetic hypoxia has become increasingly prevalent in stratified water bodies in recent decades due to climate change. One primary sink of dissolved oxygen (DO) is sediment oxygen uptake (JO2\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$${J}_{{O}_{2}}$$\\end{document}). On the water side of the sediment–water interface (SWI), JO2\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$${J}_{{O}_{2}}$$\\end{document} is controlled by a diffusive boundary layer (DBL), a millimeter-scale layer where molecular diffusion is the primary transport mechanism. In previous studies, the DBL was determined by visual inspection, which is subjective and time-consuming.Material and methodsIn this study, a computational procedure is proposed to determine the SWI and DBL objectively and automatically. The procedure was evaluated for more than 300 DO profiles in the sediment of three eutrophic water bodies spanning gradients of depth and surface area. Synthetic DO profiles were modeled based on sediment characteristics estimated by laboratory experiments. The procedure was further verified adopting the synthetic profiles.Results and discussionThe procedure, which was evaluated for both measured and synthetic DO profiles, determined the SWI and DBL well for both steady and non-steady state DO profiles. A negative relationship between DBL thickness and aeration rates was observed, which agrees with existing literatures.ConclusionsThe procedure is recommended for future studies involving characterizing DBL to improve efficiency and consistency.