The interaction of Ca(2+), with dipalmitoyl phosphatidylcholine (DPPC) model membranes was studied in the presence and absence of vitamin D(2) by using Fourier transform infrared spectroscopy. Addition of vitamin D(2) and/or Ca(2+) into pure DPPC liposomes shifts the phase transition to higher temperature, orders and decreases the dynamics of the acyl chains in both phases and does not induce hydrogen bond formation in the interfacial region. Moreover, the dynamics of the head group of the phospholipid decreases in both phases. The addition of vitamin D(2) into DPPC liposomes containing Ca(2+), decreases the effect of Ca(2+) at all the functional groups under investigation. Similarly, the effect of vitamin D(2) also decreases in the presence of Ca(2+). This behavior is dominant at high Ca(2+) concentrations. Our results show how simultaneous presence of vitamin D(2) and Ca(2+) alter the behavior of each other, which is reflected as a decrease in the interactions between the ions and vitamin D(2) within the membrane.