Thermoplastic polyurethane (PU) polymers with different chemical compositions were synthesized and casted to films, and their water vapor barrier properties at different range of relative humidity (RH) were characterized. The water vapor permeability (WVP) of packaging films is one of their most important properties to identify their suitability for use as packaging materials and is rather a complicated phenomena if the polymer has polar nature. The WVPs of PU films are determined both by permeation measurements which are a steady-state method and water vapor sorption measurements which are a non-steady-state method. Effective permeability (P eff), solubility (S eff), and diffusion (D eff) coefficients of PU films were determined at 23°C within the RH range of 0-97%. It was found that P eff , S eff , and D eff increased with increasing RH gradient due to water vapor and polymer interactions. Microscopic images showed that 1,4-butanediol (BDO) helped to improve porous structure. Castor oil (CO) caused a decrease in the intensity of active absorption sites, namely, the C=O•••H-N hydrogen bonds between chains. Results of two methods were yielded in the same magnitude of order. In most cases, the non-steady-state (sorption) method yields higher WVP values than steady state. At 0➔85% RH, the difference was up to 8-fold. Conditioning and equilibrating of films at 50% RH helped to reach sorption data approximate to permeation data. It was suitable to use sorption measurements to estimate the WVP which is a considerable simplification for polar polymers, e.g., developed PU film.