It is well documented that the bioavailability of hydrophobic organic chemicals (HOCs) can vary substantially among sediments. This makes risk assessments based on total sediment concentrations problematic. The present study investigates the application of thin-film solid-phase extraction to measure bioavailable concentrations of phenanthrene in estuarine sediment by comparing concentrations of phenanthrene in the amphipod Corophium colo and in thin ethylene/vinyl acetate films at different concentrations in three geochemically different sediments. For all sediment types, concentrations of phenanthrene in sediments and thin films followed linear relationships, indicating first-order exchange kinetics. Organism/thin-film concentration ratios did not vary systematically among sediment types but dropped significantly with increasing phenanthrene concentration in the sediments. While at low phenanthrene concentrations in the sediment fugacities of phenanthrene in the amphipods approached the fugacities in the thin films, they were significantly lower than those in the sediments at higher concentrations. While phenanthrene concentrations in the three sediment types were identical, biota sediment accumulation factors and concentrations in amphipods and thin films were consistently lower in sediments enriched with black carbon than in sediments with sedimentary organic matter bearing a more diagenetic organic signature. It is concluded that, for the range of concentrations tested, thin-film solid-phase extraction can be a useful tool in the characterization of differences in bioavailability of HOCs among sediment types.