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Abstract: Dissolved organic matter (DOM) represents the largest organic matter pool in freshwater systems, but much of it remains molecularly uncharacterized. Although freshwater systems cover only a small area of the earth’s surface, inland waters are an important component of the global carbon cycle. The traditional idea that rivers are simply conduits for refractory carbon delivery to coastal areas is inconsistent with carbon flux estimates, and streams have been shown to serve as reactors for DOM cycling. The overall quality of DOM, and its associated reactivity, can be related to its chemical composition and molecular structure. However, the variability of DOM composition in freshwater ecosystems, particularly in headwater streams, is poorly characterized. Detailed molecular studies of DOM from small streams across climatic regions, which could provide critical information regarding carbon dynamics on a more global scale, have not been performed. To address these issues, this study applies a multi-method analytical approach in an attempt to assess molecular characteristics of DOM and ultrafiltered DOM (UDOM) in headwater streams from different climatic regions in North America. In general terms the chemical and molecular characteristics of UDOM from six different biomes were determined in unsurpassed detail to feature some clear general similarities but also specific differences. While the degree of similarity is remarkable, and suggests similar source strengths, such as soil-derived organic matter and/or similar diagenetic degradation processes for DOM from vastly different environments, each sample was clearly unique in its overall composition, featuring some distinct molecular patterns for at least one or more of the analytical determinations. Molecular and compositional differences of DOM from headwater streams should result from variations in DOM sources and localized environmental conditions, and consequently feature different photo- and bio-reactivity and associated re-mineralization potentials during fluvial transport. Such knowledge could assist in predicting the consequences of global change and its relationship to global carbon cycling. [Copyright &y& Elsevier]