Biodegradable chemicals may become persistent due to reductions in their bioavailability thereby impacting on the rate and extent of biodegradation in soils and sediments. This chapter examines this – commonly neglected – contradictory face of persistence assessments from the light of the latest advancements in bioavailability science. They include the microbial influences on bioavailability, the different sorption capacities of carbonaceous components of soils and sediments, and the dissimilar bioavailability shown by chemicals when they are present as non-extractable residues. We also discuss possible pathways to improve the realism in persistence assessments from standardized biodegradation tests by incorporating new bioavailability-based approaches. Innovations of the standard tests are possible through the modified chemical application of enhanced dispersion and passive dosing. In addition, we offer a proposal for integrating bioavailability measurements into standard simulation tests with soils and sediments, by using desorption extraction and passive sampling methods to assess the removal of the bioavailable fractions, in addition to the total extractable concentration of the chemical. © 2020, Springer Nature Switzerland AG.