Recent studies recognize the ability of chemical techniques such as solid phase microextraction (SPME) fibers and Tenax extraction to predict bioavailability more effectively than exhaustive chemical extractions for sediment-associated organic contaminants. While the majority of research using these techniques studied legacy compounds such as PCBs and PAHs, there is great potential for these methods to work well for highly toxic, rapidly biotransformed compounds such as pyrethroid insecticides. The current study compared the ability of the two techniques to predict the bioavailability of permethrin and bifenthrin to two benthic invertebrates (Lumbriculus variegatus and Hexagenia sp.). In addition, variations in the application of the two techniques, specifically duration and conditions of exposure of the SPME fibers and duration of extraction with Tenax, were explored. The SPME fiber concentrations correlated strongly to both 6 and 24 h Tenax concentrations. The SPME fiber concentrations and 6 h and 24 h Tenax extractable concentrations correlated with both the parent permethrin and bifenthrin concentrations in the tissues of both species at steady state. Parent compound tissue concentrations for both species could be predicted with a single relationship for individual pyrethroids. This demonstrated the potential value of these methods to predict the bioavailability of compounds subject to biotransformation and application to multiple species.