Pharmaceuticals have been increasingly detected globally in a variety of environmental matrices, including surface water and sediments. The presence of these compounds in the environment presents a risk to aquatic organisms. Bioaccumulation of commonly detected anti-inflammatory pharmaceuticals (AIPs) such as ibuprofen, acetaminophen, and aspirin are of particular interest in filter-feeding organisms such as freshwater mussels. They are highly biodiverse and valuable to aquatic ecosystems; however, the decline in their biodiversity and populations emphasizes the importance of investigating the potential impacts of pharmaceuticals found within their habitats. To understand how different habitat characteristics (e.g. sediment particle size, water chemistry etc.) influence uptake of pharmaceuticals, we exposed Fatmucket mussels (Lampsilis siliquoidea) to relevant and above relevant concentrations of AIPs in mixture for 21 days with different sediment types, and used LCMS analysis to investigate concentrations across time from different compartments (i.e. water, sediment, and tissue). Our surface water data revealed differences in concentration that follows a dose-dependent relationship. Acetaminophen and ibuprofen were present in water at ranges of 0 to 34.8 ug/L and 0.51 to 75.6 ug/L respectively, while aspirin was found to be at a range of 0 to 24.2 ug/L. Aspirin was not detectable in tissue (suspected to be from possible biotransformation and excretion). However, both acetaminophen and ibuprofen were detected at a maximum of 10 ng/g and 203 ng/g of wet weight, respectively. We found that site had a significant effect on water and tissue concentrations (p = <0.001 for both compartments), suggesting an environmental influence on AIP partitioning. Understanding these relationships can provide researchers with greater insight into how the environment can influence uptake in freshwater systems. Our findings also warrant further investigation into sublethal responses that may impose different physiological responses for freshwater mussels, gaining a more holistic view into factors influencing their decline.