This study used an electrochemical sensor based on a glassy carbon electrode that was designed with a nanocomposite of silver nanoparticles and p-aminothiophenol functionalized graphene oxide (Ag NPs/PATP/f-GO/GCE) to measure the level of rutin in actual samples taken from flowers of rose species. To create the Ag NPs/PATP/f-GO nanocomposite, the Ag NPs were synthesized using the traditional heating process and chemically combined with acid-treated GO and p-aminothiophenol. Ag NPs/PATP/f-GO nanocomposite's effective production was confirmed by X-ray diffraction (XRD), scanning electron microscope (SEM), and X-ray photoelectron spectroscopy (XPS) investigations. The construction of a sensitive, selective electrochemical system for the identification of rutin was made possible by the synergistic interaction of Ag NPs, PATP, and f-GO nanosheets, as demonstrated by electrochemical studies based on cyclic voltammetry (CV) and square wave voltammetry (SWV) measurements. The suggested sensor demonstrated a linear response of 5–1050 μM. It was determined that the sensitivity and limit of detection (LOD = 3σ/S, where S is the slope of the regression line and σ is the standard deviation of the intercept) were 0.02902 μA/μM and 17 nM, respectively. For the purpose of determining the level of rutin in a genuine sample made from flowers of a rose species (damascene rose) from Nanyang in Henan province, China, the suggested electrochemical rutin sensor's accuracy and capability were evaluated. Results showed that Ag NPs, PATP, f-GO, GCE may be used as dependable rutin sensors to monitor the rutin concentration in samples of rose species flowers with remarkable recovery values (higher than 98.32 %) and relative adequate accuracy (RSD less than 5.41 %).