We report the use of 25,27-diethyl-dithiocarbamic-26,28-dihydroxy-p-tert-butylcalix[4]arene in the functionalization of Ag nanoparticles for pyrene detection by surface-enhanced Raman scattering (SERS). SERS spectra provided information about the calixarene orientation on the metal surface and the interaction mechanism with pyrene. Thus, in this work, we have combined a powerful spectroscopy technique such as SERS, the electronic plasmon-based properties of nanostructured metals, the molecular size-selective recognition of calixarene, and the strong chelating properties of the dithiocarbamate group toward the metal surface in the detection of polycyclic aromatic hydrocarbons.