The behavior of perfluorinated persistent organic pollutants (POPs), especially perfluoroalkyl carboxylic and sulfonic acids, at aqueous interfaces is crucial for their transport and speciation in the environment and subsequent immunotoxicity. Here, we investigate the surface prevalence and interfacial interaction of a prototype perfluorinated-POP, perfluoroheptanoic acid (PFHA), with environmentally relevant amphiphiles of varying hydrophobicity and head groups (CnH2n+1-X; n: 8 vs 16; -X: -OH vs -COOH) using interface-selective vibrational sum frequency generation (VSFG) spectroscopy. SFG intensity spectra in the CH- and OH-stretch regions reveal that PFHA prevails at aqueous interfaces that contain amphiphiles of intermediate chain length such as 1-octanol (n = 8) and heptanoic acid (n = 6). PFHA partially expels as well as increases the alkyl chain order of octanol on the water surface. Whereas heptanoic acid, though less hydrophobic than octanol, is retained at the water surface through hydrogen-bonding with the PFHA head group ((COO)-C-(PFHA)-center dot center dot center dot HOOC(heptanoic-acid)). Long chain amphiphiles (n = 16) such as hexadecanol and palmitic acid expel PFHA from the water surface regardless of the difference in their head groups. Interestingly, the dangling OH (3710 cm(-1)) which is diminished at the hydrogenated-amphiphile-water interface is preserved at the perfluorinated-POP-water interface.