Novel free standing and binder free non-enzymatic electrochemical sensors were fabricated using in situ grown copper (Cu) nanoparticles on polyvinylidenefluoride-co-hexafluoropropylene (PVdF-HFP) nanofibers. Morphological images showed that Cu nanoparticles were homogeneously anchored over the PVdF-HFP nanofibers and the elemental composition and structure of the prepared composite nanofiber membranes were identified from elemental analysis and diffraction patterns. The fabricated nanofiber membranes were applied in the quantification of glucose and the non-enzymatic electrooxidation of glucose was facilitated at the surface of Cu nanoparticles that were anchored over the PVdF-HFP nanofibers. The fabricated sensor exhibited the linear range covering from 1 μM to 6.055 mM, high sensitivity of 506.62 μA mM−1 cm−2 and low detection limit of 0.011 μM. Importantly, the PVdF-HFP/Cu membrane exhibited favorable reproducibility, long-term stability, and was relatively insensitive to common interfering species in real time applications. The fabricated electrospun PVdF-HFP/Cu nanofiber membrane offers unique advantages, including simple fabrication, good affinity and selectivity to glucose and quick response, which open up new possibilities for exploring the a variety of electrochemical devices with affordable cost and good stability.