Multiple stable isotopes were used to determine the effectiveness of distinguishing among several dominant riparian species and aquatic macrophytes both spatially (three sites) and temporally (three seasons) along an 8-km reach of a blackwater stream. The differences in isotopic composition were used to assess contributions of various organic matter sources to the detrital pool of the stream. Samples of riparian and aquatic macrophyte vegetation and detritus were collected at three times to represent early leaf-out (April), mid-summer (August), and just prior to abscission (October). Each sample was analyzed for stable isotopes of carbon ฮดC, nitrogen ฮดN, and sulfur ฮดS Within a site and sampling date, ฮดC-values were significantly different among certain riparian species and detritus samples. Species differences persisted between seasons. ฮดS values were the most variable of the three elements examined although they remained fairly constant through time within each species and site. The results suggest that temporal changes in isotopic compositions of riparian species and aquatic macrophytes are site-specific. Discriminant analysis dissimilarity plots (based on all three isotopes) demonstrated that the contribution of species to the detrital pool depended on the site and season. At the upper site, detritus was isotopically most similar to Quercus laurifolia and Sparganium americanum in April, and the aquatic macrophytes (S. americanum and Potamogeton spp.) in August and October. At the middle site, detritus was most similar to Carpinus caroliniana and Q. nigra in April but no single source was similar to detritus in August or October. At the lower site, detritus was most similar to Taxodium distichum for all three seasons.