New chloramphenicol-based poly(ester-ether)urethane bioconjugates intended for biomedical applications are presented. Structure-property relationship in the tested biomaterials is established by cross-inspection of the ATR-FTIR investigation, DSC analysis, thermogravimetric analysis, surface morphology characterization, water sorption and/or desorption properties, and antimicrobial efficiency of the obtained polyurethane biomembranes. The estimated values of activation energy for thermal degradation under nitrogen fall between those found for poly(ether urethane)s and poly(ester urethane)s. SEM microphotographs show that the resulted morphologies of polyurethanes are different due to the diverse polyether segments which determine the supramolecular architecture of the self-assembled films. The estimated moisture diffusion coefficients, sorption capacity are dependent on polyurethane network microstructure. The monolayer sorption and average pore size values were estimated by applying BET model. The values of BET area and monolayer capacity are correlated with antibacterial activities. The synthesized biocidal polyurethanes bearing covalently attached chloramphenicol are effective at inactivation of tested bacteria: Sarcina lutea and Escherichia coli.