Levothyroxine (3,5,3',5'-tetraiodothyronine), which is a L-isomer of thyroxine T4 (L-T4), is a synthetic thyroid hormone that is biochemically and physiologically indistinguishable from endogenous T4. It is used as a thyroid hormone replacement drug to treat an underactive thyroid gland. The interaction of L-T4, with zwitterionic dipalmitoyl phosphatidylcholine (DPPC) multilamellar liposomes (MLVs) was studied in the presence (1 mol%, 3 mol%, 6 mol%, 9 mol%, 15 mol%, 24 mol% and 30 mol%) and absence of L-T4 by using two different non-invasive techniques; Fourier Transform Infrared (FTIR) spectroscopy and Differential Scanning Calorimetry (DSC). The results show that L-T4 does perturb the phase transition profile by either decreasing the main transition temperature (T-m) and enthalpy (Delta H) or increasing the width at half height (Delta T-1/2). That means; it changes the physical properties of DPPC bilayers. Addition of L-T4 into pure DPPC liposomes shifts the phase transition to lower temperature, disorder the system in gel phase with opposite effect in liquid-crystalline phase and increases the dynamics of the system in both phases and also causes dehydration of the groups of lipids and the water molecules around.