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<p>Dissolved organic matter (DOM) connects aquatic and terrestrial ecosystems, plays an important role in carbon (C) and nitrogen (N) cycles, and supports aquatic food webs. Understanding DOM chemical composition and reactivity is key for predicting its ecological role, but characterization is difficult as natural DOM is comprised of a large but unknown number of distinct molecules. Photochemistry is one of the environmental processes responsible for changing the molecular composition of DOM, and DOM composition also defines its susceptibility to photochemical alteration. Reliably differentiating the photosensitivity of DOM from different sources can improve our knowledge of how DOM composition is shaped by photochemical alteration and aid research into photochemistry's role in various DOM transformation processes. Here we describe an approach for measuring and comparing DOM photosensitivity consistently, based on the kinetics of changes in DOM fluorescence during 20 h photodegradation experiments. We identify several methodological choices that affect photosensitivity measurements and offer guidelines for adopting our methods, including the use of reference material, precise control of conditions affecting photon dose, leveraging actinometry to estimate photon dose instead of expressing results as a function of exposure time, and frequent (every 20 min) fluorescence and absorbance measurements during exposure to artificial sunlight. We then show that our approach can generate photosensitivity metrics across several sources of DOM, including freshwater wetlands, a stream, an estuary, and <i>Sargassum</i> sp. leachate and observed differences in these metrics that may help identify or explain differences in their composition. Finally, we offer an example of applying our approach to compare DOM photosensitivity in two adjacent freshwater wetlands as seasonal hydrologic changes alter their DOM sources.</p>