Per- and polyfluoroalkyl substances (PFAS) have become a subject of growing global concern due to their persistence, toxicity, and environmental bioaccumulation. This comprehensive review surveys the latest methodologies for PFAS detection across food, environmental, and biological domains, encompassing mass spectrometry, fluorescence detection, electrochemical sensing, and more. The review explores innovative sample pretreatment techniques, newly developed materials, and advanced methods, sensors, and platforms, while analyzing the strengths and limitations of diverse detection approaches. The pursuit of tailored detection mechanisms aligned with PFAS structure and physicochemical properties, coupled with the integration of emerging functional materials for sensors, platforms, and sample pretreatment, has yielded exceptional detection performances. These achievements include time-efficient processes, high sensitivity, and remarkable precision and accuracy in PFAS analysis. The review concludes by outlining current trends and presenting future perspectives for advancing PFAS analysis methodologies.