Abstract Background/Objective: Per- and polyfluoroalkyl substances (PFASs) are hazardous synthetic chemicals that have been linked to adverse human health effects such as cancer and endocrine disruption. Firefighters have been shown to be occupationally exposed to PFAS through aqueous film-forming foam (AFFF) used during difficult-to-suppress fires, and in the textile liners of their safety turnout gear. PFASs have both hydrophobic and hydrophilic chemical structures, giving them strong thermal stability and making them resistant to environmental and physiological degradation. Studies of human exposures to PFAS have observed associations between some PFAS and increased risk of obesity or elevated body mass index (BMI). Given the unique occupational exposures of firefighters to PFAS, we examine the association between obesity and PFAS in a sample of Florida firefighters. Methods: A cross-sectional study design was used to collect a health survey and a blood sample from a non-probabilistic sample of career Florida fighters between June and October 2021. The health surveys, administered on REDCap using tablet devices, assessed information on firefighter socio-demographic and work characteristics. We quantified 7 distinct PFASs using a solid-phase extraction-high-performance liquid chromatography-isotope dilution-tandem mass spectrometry approach; the limit of detection was between 0.5 and 0.05 ng/mL, depending on the analyte. All 7 PFASs, including Perfluorobutanesulfonic Acid (PFBS), Perfluoroheptanoic Acid (PFHpA), Perfluorohexanesulfonic Acid (PFHxS), Perfluorooctanoic Acid (PFNA), Perfluorononanoic Acid (PFNA), Perfluorooctanesulfonic Acid (PFOS), and Perfluoroctanesulfonamide (PFOSA), were detected in the blood samples; concentrations below the limit of detection were replaced by the limit of detection/√2.35. Results: Among the 208 firefighters who completed the survey and provided a blood sample, 92.8% were male, 16.2% Hispanic/Latinx, 81.7% White, 89.8% used AFFF at their fire department, with a group mean age 40.2±10.3 standard deviation. Approximately 15.8% of firefighters were of healthy weight, 51.5% overweight, and 32.7% obese. Firefighter body mass index was positively correlated with serum concentrations of PFHxS (rs=.17;p=.020) and PFBS (rs=.60;p=.26). Group geometric mean of each PFAS congener (in ng/mL): PFBS (0.09), PFHpA (0.08), PFHxS (2.17), PFOA (1.38), PFNA (0.42), PFOS (2.69), and PFOSA (0.10). Total serum PFAS concentrations was highest among obese firefighters (7.62 ng/mL) and lowest in healthy weight firefighters (6.55 ng/mL). Conclusions: Firefighters who were overweight and obese, based on body mass index, had higher total serum PFAS concentrations than those firefighters of healthy weight. Both long- and short-chain PFASs, PFHxS and PFBS respectively, were positively correlated with firefighter body mass index. Citation Format: Alberto J. Caban-Martinez, Paola Louzado Feliciano, Simi Oduwole, Natasha Schaefer Solle, Catalina Gonzalez-Umana, Thomas Stone, Erin N. Kobetz. Per- and polyfluoroalkyl substances and obesity in Florida firefighters [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 25.