The antimalarial agent mefloquine was investigated using Fourier transform near-infrared (FT NIR) Raman and FT IR spectroscopy. The IR and Raman spectra were calculated with the help of density functional theory (DFT) and a very good agreement with the experimental spectra was achieved. These DFT calculations were applied to unambiguously assign the prominent features in the experimental vibrational spectra. The calculation of the potential energy distribution (PED) and the atomic displacements provide further valuable insight into the molecular vibrations. The most prominent NIR Raman bands at 1,363 cm(-1) and 1,434 cm(-1) are due to C=C stretching (in the quinoline part of mefloquine) and CH(2) wagging vibrations, while the most intense IR peaks at 1,314 cm(-1); 1,147 cm(-1); and 1,109 cm(-1) mainly consist of ring breathings and deltaCH (quinoline); C-F stretchings; and asymmetric ring breathings, C-O stretching as well as CH(2) twisting/rockings located at the piperidine moiety. Since the active agent (mefloquine) is usually present in very low concentrations within the biological samples, UV resonance Raman spectra of physiological solutions of mefloquine were recorded. By employing the detailed non-resonant mode assignment it was also possible to unambiguously identify the resonantly enhanced modes at 1,619 cm(-1), 1,603 cm(-1) and 1,586 cm(-1) in the UV Raman spectra as high symmetric C=C stretching vibrations in the quinoline part of mefloquine. These spectroscopic results are important for the interpretation of upcoming in vitro and in vivo mefloquine target interaction experiments.