Fatty alcohols are naturally produced hydrocarbons present in all living organisms. They are also used in detergent and cosmetic formulations, may be sourced from either petroleum or biological materials, and are typically disposed of down the drain. This study was conducted on the Luray catchment, Virginia, USA, where sales data indicate that approximately 2 kg of fatty alcohols from detergent enter the wastewater every day. Reconstructing fatty alcohols in the influent on the basis of sales data indicated a mix of odd and even chain compounds, with C(12) being dominant. This profile was influenced strongly by liquid laundry detergents (69%). Sediment and soil samples from the catchment were analyzed by gas chromatography-mass spectrometry and by stable isotope ratio mass spectrometry to determine the δ(13)C and δ(2)H signatures. The long-chain components in agricultural soils and river sediments were distinguishable clearly from the algal fatty alcohols produced within the river system. The wastewater was a mixture of fecal and detergent sources of fatty alcohols in a ratio of 75:25%. The fatty alcohols in the effluent had different stable isotopic signatures and chain-length profiles from the influent, indicating that these compounds are not the same as those that entered the treatment plant. The total quantity of fatty alcohols leaving the treatment plant through the effluent pipe was low compared with the input. Analysis of the contributions based on the stable isotopes and profiles suggests that of the fatty alcohols present in the river system downstream of the treatment plant, 84% were derived from terrestrial plant production, 15% came from in situ algal synthesis, and 1% were derived from the effluent.