Electrospun carbon nanofibers (ECNFs) have many qualities that make them attractive for applications in various fields of technology. For many applications, it is necessary to modify the surface of CNFs by chemical functionalization. Here, a systematic functionalization of polyacrylonitrile (PAN) based ECNFs by oxidation in concentrated nitric acid was examined.The effects of oxidation on the microstructural, electrical and mechanical characteristics have been studied. The Fourier transform infrared spectroscopy results revealed the time-dependent functionalization of ECNFs. However, according to the scanning electron microscopy there was no considerable change on the surface morphology of CNFs due to the functionalization. While the microstructures as well as electrical and mechanical properties were severely dependent on oxidation duration according to the X-ray diffraction and Raman spectroscopies as well as conductivity and mechanical strength measurements, respectively. Increase in oxidation duration led to more structural disorders and noticeable decreases in electrical conductivity and mechanical strength.