Potentiometric sensors with plasticized poly(vinyl chloride) membranes based on beta-lactam-tetraalkylammonium ion associates sensitive to penicillin antibiotics are proposed. The physicochemical characteristics (solubility product constants and dissociation constants) of active membrane components and the electrode, transport, and selective properties of the membranes of liquidand solid-contact sensors have been studied. The quantitative characteristics of membrane transport (penetrability, ion flux, and transport rate) have been evaluated. The main charge carriers in the membranes and at the membrane/solution interface have been determined from the membrane transport characteristics. The potentiometric sensors are shown to be applicable to the determination of penicillin antibiotics in biological fluids (blood serum and oral fluid) from patients with urinary tract infection.