The insufficient capabilities of current surface-enhanced Raman scattering (SERS) substrates in enriching dilute analytes from complex media severely restrict detection sensitivity, hampering practical applications. To meet this demand, in this study, a novel super hydrophobic membrane that can be directly prepared on a large scale based on the silver nanoparticles (AgNPs) functioning with perfluorodecanethiol (PFDT) is fabricated and evaluated as an SERS substrate. Firstly, polyester (PET) films modified with sodium chloride were proven to be capable of loading AgNPs, and the sizes of AgNPs were investigated. In addition, the PFDT concentration and reaction time for functionalizing the surface of AgNPs have been optimized. The relationship between the hydrophobic properties of the film and its SERS performance was then studied. The PET@Ag-PFDT film demonstrates two orders of magnitude superior SERS performance than the unmodified PET@Ag substrate, with a detection limit of folic acid approaching 5 × 10