Perfluorooctanesulfonic acid (PFOS) is a persistent organic pollutant (POP) with non-biodegradable and bioaccumulation. Recent studies identified it as a risk factor for cardiovascular disease. However, research on the specific toxic mechanisms of PFOS in platelets, particularly its effects on blood coagulation, remains limited, particularly in relation to thrombosis, related to cardiovascular disease. Therefore, this study investigated whether PFOS exposure induces phosphatidylserine (PS) exposure in human platelets, thereby increasing blood coagulation. Furthermore, we identified the effect of PFOS to mitochondrial dysfunction and oxidative stress as molecular mechanisms. PFOS significantly increased platelet membrane PS exposure in a concentration-dependent manner, this attributed to the inhibition of flippase activity and the increase of scramblase activity. It also increased intraplatelet calcium (Ca²⁺) concentration, depleted ATP levels, and induced a collapse of the mitochondrial membrane potential. PFOS also promoted intracellular ROS production and depleted the antioxidant glutathione (GSH). Importantly, pretreatment with exogenous antioxidants (N-acetylcysteine, Catalase) and a calcium chelator (BAPTA-AM) effectively inhibited PFOS-induced PS exposure, membrane potential collapse, and thrombin generation. This shows that exposure PFOS promotion of blood coagulation activity, and that PFOS exposure may affect platelet-mediated thrombosis and cardiovascular disease.