Among of all the pollutants, phenols have the notoriety because of their strong stability, great harmfulness and poor biodegradability [1]. Therefore, determination of phenols by efficient, fast and low cost methods have been developing [2]. In the recent years, the miniaturization of analytical systems has become an interesting research section [3]. In this research, the paper based electrochemical nanobiosensor as simple and fast device was developed by immobilization of the nanoparticles and potato tissue at fiber matrices of the paper disc. For determination of total phenol in aqueous media in this direction, the propylamine functionalized silica nanoparticles was dropped on the paper disc. After being dried at room temperature, the potato tissue in phosphate buffer solution as the source of the polyphenol oxidase was immobilized at fiber matrices of the paper disc via physical adsorption. After drying, this paper disc was placed on the top of the graphite screen printed electrode and was connected to potentiostat/ galvanostat electrochemical device. Electrochemical characterization of this biosensor was studied by cyclic voltammetry and electrochemical impedance spectroscopy methods. After optimization of effective parameters such as pH and potato tissue percentage, phenol measurements were done by differential pulse voltammetry method. This biosensor showed a linear response for phenol in the wide concentration ranges of 0.10-160 µM with a detection limit about 0.07 µM (3) and limit of quantification about 0.23 µM. The selectivity against some interferences and real sample analysis were studied and showed good applicability for this paper based device. This method has some advantages such as simplicity, demanding a little sample for analysis and do not require specialized users.