<p>Connecting tropical rainforests to larger rivers, tropical headwaters export large quantities of carbon and nutrients as dissolved organic matter (DOM), and are thus a key component of the global carbon cycle. This DOM transport is not passive, however; sunlight and microbial activity alter DOM concentrations and compositions, affecting riverine greenhouse gas emissions and downstream ecosystems. The effects of sunlight and microbial turnover/activity on DOM concentrations and compositions in tropical headwaters are currently poorly understood, but novel Size Exclusion Chromatography (SEC) techniques coupled to suitable detectors can for the first time quantify their influences. Here, we present in-situ incubation experiments from from headwaters of the Essequibo River, in the Iwokrama Rainforest, Guyana, where sunlight oxidised up to 9% of dissolved organic carbon (DOC) over 12 hours, at higher rates than in larger tropical rivers. DOM transformations occurred in both photo-sensitive and supposedly photo-resistant pools. Microbial activity had varying, less clear influences on DOC concentrations over the same time span; compositionally, this appeared to extend beyond known bio-labile components. Biopolymers were particularly reactive to both processes. We show sunlight has the greater potential to mineralise headwater DOM and thus potentially influence degassing. Our approach provides a future template to constrain DOM transformations along river networks, identify biogeochemical activity hotspots, and improve greenhouse gas emissions estimations under changing environmental conditions.</p>