Fluphenazine (FPh) belongs to the phenothiazine family of compounds and exhibits a wide variety of biological effects, including antimutagenic, proapoptotic, antiproliferative and anti-multidrug resistance (MDR) activities. The ability of FPh to interact with lipid membranes can have a significant impact on its biological activities. However, the mechanisms involved in the interaction of FPh with lipid membranes are poorly understood. FTIR–ATR spectroscopy has been used in this study to visualize the interactions between FPh and a model lipid bilayer composed of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). Subsequent interpretation of the temperature-dependent FTIR spectra obtained for FPh:DPPC systems containing different concentrations of FPh was efficiently supported by principal component analysis.