The present study aims to explore the relationship between perfluoroalkyl substances (PFAS) exposure and hyperlipidemia using data from the National Health and Nutrition Examination Survey. A total of 1600 subjects were included in the analysis, and nine kinds of PFAS were measured. Multivariate logistic regression analysis was performed to explore the association between serum PFAS and hyperlipidemia. Compared with the lowest quartile of perfluoromethylheptane sulfonic acid isomers (Sm-PFOS), the percentage change for hyperlipidemia was 57% and 41% in the third and highest quartile of PFOS. The positive association between Sm-PFOS and hyperlipidemia remained significant in population younger than 60 years, and the odds ratio for hyperlipidemia in fourth quartile of Sm-PFOS was 1.81. These findings indicated that serum Sm-PFOS was independently associated with a higher risk for hyperlipidemia. The epidemiological study warrants further study to elucidate the causal relationship between them.