Determination of drug concentration in body fluids is important issue for clinical studies to arrange treatment of patients. In the present study, we concentrated on the preparation of propranolol-imprinted poly (N-acryloyl-L-phenylalanine) film on a paper surface for spectrophotometric detection of propranolol in human plasma samples. The surface characterization of the imprinted surface was carried out by attenuated total reflectancefourier transform spectroscopy, x-ray photoelectron spectroscopy, scanning electron microscopy and water contact angle measurements. Rebinding isotherms and kinetics were also investigated and maximum adsorption capacity of the imprinted paper surface was found to be 64.3 mg/g with high imprinting factor (4.20). Moreover, the results for selectivity and regeneration ability of the imprinted paper indicated that propranolol selectively interacted with the imprinted paper and had satisfactory reuse without changing its adsorption capacity. Under optimized conditions, the imprinted paper surface had a limit of detection of 0.3 mu g/mL with lower intraday and interday precisions for determination of propranolol. The proposed method was successfully applied to determine propranolol in plasma samples where it showed recoveries ranging from 97.0%-99.5%. The method was also compared with traditional ELISA method and the results showed that the proposed method is sensitive and selective. It is believed that the prepared paper-based molecularly imprinted polymers can be good alternatives to traditional drug assays in clinical practice.