© 2022 Elsevier B.V.Rutin has antiviral, anticancer, anti-inflammatory and heart disease protective activities but its poor solubility plays a critical role in limiting its activity. To relieve this problem, we prepared polyvinylpyrrolidone (PVP) electrospun nanofibers as carrier to release rutin for potential applications. Starch-coated copper nanoparticles were also added to the polymeric precursor solution before electrospinning to increase the activity of rutin. The water soluble monodisperse starch-protected zero-valent copper nanoparticles of about 36 nm diameter have been synthesized by chemical reduction route using starch as green capping agent. The resulting starch coated copper nanoparticles were characterized by transmission electron microscopy and energy dispersive X-ray analysis (EDX). It was found that copper nanoparticles were significantly protected by starch coating. PVP solutions incorporated with rutin and starch capped copper nanoparticles were electrospun at different weight percentages. The nanofibers were characterized by using scanning electron microscopy, energy dispersive spectroscopy, and Raman spectroscopy. We confirmed that copper nanoparticles were embedded in PVP nanofibers. The cytotoxic efficacy of the nanofiber involving starch capped copper nanoparticles and rutin against L929 mouse fibroblast cells was compared using a standard MTT assay. The in vitro release studies were performed using rutin-copper loaded nanofibers through dialysis membrane on the Franz diffusion cells. The nanofiber involving starch capped copper nanoparticles and rutin exhibited no cytotoxicity compared to PVP nanofibers.