1. Organic matter may sequester nutrients as it decomposes, increasing in total N and P mass via multiple uptake pathways. During leaf litter decomposition, microbial biomass and accumulated inorganic materials immobilize and retain nutrients, and therefore, both biotic and abiotic drivers may influence detrital nutrient content. We examined the relative importance of these types of nutrient immobilization and compared patterns of nutrient retention in recalcitrant and labile leaf litter. 2. Leaf packs of water oak (Quercus nigra), red maple (Acer rubrum) and Ogeechee tupelo (Nyssa ogeche) were incubated for 431 days in an intermittent blackwater stream and periodically analysed for mass loss, nutrient and metal content, and microbial biomass. These data informed regression models explaining temporal changes in detrital nutrient content. Informal exploratory models compared estimated biologically associated nutrient stocks (fungal, bacterial, leaf tissue) to observed total detrital nutrient stocks. We predicted that (i) labile and recalcitrant leaf litter would act as sinks at different points in the breakdown process, (ii) plant and microbial biomass would not account for the entire mass of retained nutrients, and (iii) total N content would be more closely approximated than total P content solely from nutrients stored in leaf tissue and microbial biomass, due to stronger binding of P to inorganic matter. 3. Labile litter had higher nutrient concentrations throughout the study. However, lower mass loss of recalcitrant litter facilitated greater nutrient retention over longer incubations, suggesting that it may be an important long-term sink. N and P content were significantly related to both microbial biomass and metal content, with slightly stronger correlation with metal content over longer incubations. 4. Exploratory models demonstrated that a substantial portion of detrital nutrients was not accounted for by living or dead plant and microbial biomass, especially in the case of N. This suggests increased importance of both N and P sorption to inorganic matter over time, with possible additional storage of N complexed with lignin. A better understanding of the influence of these mechanisms may improve our understanding of detrital nutrient uptake, basal resource quality and retention and transport of nutrients in aquatic ecosystems.