Per- and polyfluoroalkyl substances (PFAS) comprise a large group of synthetic chemicals with a long history of use in industrial and consumer products. Regulatory and public health agencies have recognized that exposure to high levels of some PFAS may cause adverse health effects including reduced antibody responses to vaccines, increased cholesterol levels, low infant birth weight, and increased risk of high blood pressure. Although considerable effort has been devoted to the study of PFAS in the environment, there are significant gaps in our understanding of the potential human exposure to PFAS from food and food packaging. In 2020, a two-session symposium titled Identifying Science Gaps for Risk Assessment of Per- and Polyfluoroalkyl Substances (PFAS) in Food was held by ILSI North America (in 2021, ILSI North America has evolved to become the Institute for the Advancement of Food and Nutrition Sciences [IAFNS]). Recognizing the importance of measurement systems in PFAS risk assessment, the first session focused on analytical methods and science gaps for detecting and quantifying PFAS in various foods and packaging materials. The second session addressed exposure routes into foods, including an overview of the United States Department of Agriculture Food Safety Inspection Service work on PFAS and recent toxicological studies by the Food and Drug Administration on biopersistence and potential human effects of short-chain PFAS used as replacement for longer-chain biopersistent PFAS. Expert presentations encompassed US regulatory, academic, industry, and non-profit perspectives and were followed by panel discussions.