INTRODUCTION: Per- and polyfluoroalkyl substances (PFAS) are synthetic chemicals linked with adverse pregnancy outcomes, yet they remain understudied in Puerto Rico. Moreover, underlying biological mechanisms of PFAS are not fully understood, although oxidative stress and inflammation are suspected pathways. OBJECTIVES: We aimed to characterize PFAS exposure among pregnant women in Puerto Rico and examine associations between early to mid-pregnancy PFAS concentrations and repeated measures of oxidative stress biomarkers. METHODS: This study included 434 participants enrolled in the PROTECT birth cohort. We analyzed serum samples (∼18 weeks of gestation) for nine PFAS, while urinary oxidative stress biomarkers [8-isoprostaglandin F2α (8-IsoP), its metabolite (IsoP-M), and prostaglandin F₂α (PGF2α)] were measured up to three times during pregnancy. We examined associations between each PFAS and oxidative stress biomarker using linear mixed effects regression models and multivariable regression analyses, adjusting for maternal demographic, socioeconomic, and study-related factors. RESULTS: PFOS, PFNA, PFHxS, and PFOA were detected in over 50 % of participants, with PFOS being the most dominant. The majority of participants had total PFAS levels above 2 ng/mL. In repeated measures analyses, an interquartile range increase in PFHxS was associated with a 5.35 % (95 % CI: 0.12, 10.86) rise in IsoP-M levels. In categorical analyses, moderate levels of PFOS and PFNA were positively associated with PGF2α, while higher PFDeA was suggestively linked to 8-IsoP and IsoP-M. CONCLUSIONS: Despite relatively modest levels compared to the U.S. NHANES, certain PFAS were positively linked with oxidative stress or inflammation, highlighting the need for broader investigations to examine PFAS-related alteration of inflammatory processes during pregnancy.