The behavior of 2-(docosylamino)-5-nitropyridine (DCANP) spreading monolayers was investigated as a function of temperature, pH, and subphase composition. The interactions of DCANP monolayers with copper ions dissolved into the subphase at different concentrations were also studied. The investigations were carried out by measuring surface pressure and surface potential-area isotherms. Langmuir-Blodgett (LB) films transferred from pure water and, copper aqueous subphases were also studied by Fourier transform infrared (FTIR), UV-vis, and electron paramagnetic resonance (EPR) spectroscopies, X-ray photoelectron spectroscopy (XPS), ellipsometry, and refractive index measurements. The interactions between copper ions and DCANP molecules were investigated by means of various techniques in the bulk phase, in monolayers, and in LB films. Clear evidence of interactions between DCANP and copper ions was provided only from EPR spectroscopy in the LB films, because of the very small molar ratio of bound/free DCANP in the LB films. The refractive index of Cu-doped LB films was shown to be increased by the presence of metal ions.