In the present study a simple, accurate, and sensitive electrochemical sensor was proposed for rapid determination of Papaverine hydrochloride (Papa) as a vasodilators to relaxes veins and arteries. We designed modified electrode which was coated by a multiwall carbon nanotubes (MWCNTs), a thin film of moleculary imprinted polymer and deposition of Au-nanoparticles (Au). This sensing layer was constructed on a treated surface of pencile graphite electrode (PGE) via cyclic voltammetry. AuNP/MIP-sol-gel/f-MWCNT/PGE designed for Papaverine selectivity was electrochemically characterized. The interaction between the imprinted sensor and Papa molecules (as a target) in solution was observed to occur at 0.3 V~ -0.7 V vs (Ag/AgCl) in 5 mM K 3 [Fe(CN) 8 ] as a probe. This sensor demonstrated high sensivity, selectivity, and quick response towards Papa. Electrochemical methods were used to descript this sensor, for instance; Square wave voltammetry (SWV), Electrochemicall impedance voltammetry (EIS), Cyclic voltammetry (CV) and Field emission Scanning electron microscopy (FE-SEM). Furtheremore, Electrochemical sensore and its applications in Bioanalysis were optimized. The calibration curve was plotted in two linear range from 1.0 nM~ 100.0 nM and 100.0 nM ~ 5000.0 nM, with a detection limit of 0.4 nM. The relative standard deviation (RSD %) for six replicate determinations of 1 ?mol Lpapaverine was found to be 3.6% (S/N = 6). Finally, the ability of the electrochemical sensor was successfully applied for determination of target in real samples such as human urine and plasma that were succeeded with satisfactory results. The results indicated that the proposed electrode exhibited efficiently electrocatalytic oxidation for papaverine with relatively high sensitivity and stability.