In situ time-resolved energy dispersive X-ray diffraction (EDXD) was applied to investigate the hydration kinetics of oriented dioleoylphosphocoline (DOPC) and 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) membranes. The long-range order in the investigated lipid multilayers has been found to vary as a function of hydration in a nonmonotic way. In the first stage, water adsorption increased the long-range order along the normal to the lipid bilayer whereas, in the second one, a progressive loss of interbilayer coherence was observed. Possible molecular mechanisms underlying the experimental observations are discussed. (c) 2005 Elsevier B.V. All rights reserved.