In this work, we studied the formation of a complex between the polyene antibiotic amphotericin B (AmB) and dipalmitoylphosphatidylcholine (DPPC) lipid multibilayers by means of X-ray diffraction and FTIR spectroscopy. The complex formation can be detected in multibilayers by characteristic reflection around 2.2 A(-1) . The FTIR spectra indicate that in this case AmB lowers the main phase transition of DPPC. Additionally, with an increase in temperature, a shift of AmB towards the hydrophilic part of the DPPC bilayer is observed. For the first time, such complex formation has been observed by Xray diffraction and discussed. (C) 2014 Elsevier B.V. All rights reserved.