The effects of phenothiazine derivatives and calcium on the thermotropic phase transition of bilayers in dipalmitoyl phosphatidylcholine (DPPC) and dipalmitoyl phosphatidic acid (DPPA) liposomes were investigated with differential scanning calorimeter (DSC). Bilayers underwent an abrupt organizational changes at a characteristic temperature when heated. Such temperature-dependent transition was particularly striking and sharp in the bilayers prepared from pure phospholipids. The ability of phenothiazine derivatives to modify the phase transition of phospholipids liposomes was measured by a broadening of the phase transition profile, that transition began to appear at lower temperature than which occurs in untreated liposomes. Calcium ion caused a large upward shift in the transition temperature of DPPC:DPPA (34:66mol%) liposomes. When the liposomes were first incubated with calcium ion followed by phenothiazine derivatives, disappearance of the broad curve centering at 73˚C indicated displacement of calcium ion by phenothiazine derivatives at the anionic site. It is supposed that calcium ion and phenothiazine derivatives might compete with each other on the head group of acidic phospholipid.