The quercetin loaded Eudragit L-100 nanofiber membrane with high ductility and a desired drug release rate was prepared in this work. The morphological characteristics of the Eudragit L-100 nanofibers with different drug loadings amount were observed by scanning electron microscope (SEM). After adding Polyethylene glycol-4000 (PEG-4000), the degree of the fiber breakage decreased and the fiber length increased. Fiber diameter analysis, X-ray diffraction (XRD), thermal analysis (TA) and differential scanning calorimetry (DSC) have demonstrated that the crystallinity of the fiber membrane was significantly reduced after adding PEG-4000. The mechanical property test also showed that the fiber membrane with PEG-4000 had a greater elongation at break. The in vitro release experiments showed that after adding PEG-4000, the drug-loaded fibers showed rapid release at a pH of 7.4. After adopting the strategy of reducing the crystallinity, the ductility of the fiber was enhanced, which could provide a fesibility to enable this nanofiber membrane to be used in sports wound healing treatment. The electrospun Eudragit L-100 nanofiber membranes loaded with quercetin have the potential to be applied in sport wound healing of skin, tissue and joints.