Two key challenges facing regulatory bodies are 1) detecting and monitoring the presence of per- and polyfluoroalkyl substances (PFAS) within the environment and, 2) effectively assessing their risk to organisms at field-realistic concentrations. Therefore, it is critical for regulators to examine the presence, movement and effects of PFAS within an Australian context. While levels in water and soil can provide baseline information on the scale of PFAS contamination, biomonitors can provide an evaluation of exposure. European honeybees (Apis mellifera) have unique characteristics that make them an ideal candidate as a biomonitoring species for terrestrial ecosystems. They are widely distributed, globally important pollinators that effectively sample the environment surrounding a colony. The accumulation of PFAS within these species could have sublethal effects, which leads to changes in biomolecular profiles that can be assessed via metabolomics (the study of small biological metabolites such as fats, sugars and amino acids). Non-traditional end-points such as metabolomics are often overlooked in risk assessments. In that context, this study aimed to assess the types and concentrations of PFAS found in honeybees from 21 Citizen Scientist beekeepers around Melbourne, and trial the use of metabolomics to understand if the concentrations of PFAS found are impacting metabolomic pathways. The results will improve our understanding of the presence, fate and chemical transport of PFAS within the environment, which will enhance environmental risk assessment and regulatory guidelines.