We measured the concentrations of 11 polyfluoroalkyl compounds (PFCs), including perfluorooctane sulfonic acid (PFOS), perfluorooctanoic acid (PFOA), and perfluorohexane sulfonic acid (PFHxS) in 1562 serum samples collected from a representative U.S. population 12 years of age and older in the 1999-2000 National Health and Nutrition Examination Survey. Participants represented both sexes, three race/ethnicities (non-Hispanic blacks, non-Hispanic whites, and Mexican-Americans), and four age categories (12-19 years, 20-39 years, 40-59 years, and 60 years and older). PFCs were extracted from 100 microL of serum using on-line solid-phase extraction coupled to isotope dilution-high performance liquid chromatography-tandem mass spectrometry; limits of detection ranged from 0.05 to 0.2 ng/ mL. PFOS, PFOA, PFHxS, and perfluorooctane sulfonamide were detected in all samples analyzed; 2-(N-ethyl-perfluorooctane sulfonamido) acetic acid, 2-(N-methyl-perfluorooctane sulfonamido) acetic acid, and perfluorononanoic acid were detected in more than 90% of samples, which suggests prevalent exposures to several PFCs in the U.S. population. The concentrations of most PFCs were similar regardless of the participants' ages but were higher in males than in females. Mexican Americans had lower concentrations than non-Hispanic blacks and non-Hispanic whites, whose concentrations were similar. Higher education was associated with higher concentrations of PFOS and PFOA. These data will serve as a nationally representative baseline of the U.S. population's exposure to PFCs to which other populations can be compared, and will play an important role in public health by helping set research priorities, ranging from health effects studies to defining sources and pathways of exposure.