The present study reports the simultaneous determination of ascorbic acid (AA), dopamine (DA) and uric acid (UA) in 0.1 M phosphate buffer solution (pH = 7.0) using a novel electrode material prepared from oxide ceramic nanofibers by applying a single step chemical vapor deposition method. Electron-transfer kinetics at the electrode/solution interface was studied by standard redox reaction of 5 mM Fe(CN)6 3-/4- in 1 M KCl. Electrochemical and sensing measurements such as cyclic voltammetry and differential pulse voltammetry were performed to detect DA and UA in the presence of AA. The developed electrode was shown to separate the overlapping voltammetric responses of three analytes into the individual voltammetric peaks, totally eliminate the interference from AA, and distinguish DA from UA. Linear relationship was observed between current intensities and concentrations of all three compounds, and the limits of detection (LOD) were reached 0.57 µM, 0.77 µM and 0.84 µM for DA, UA and AA, respectively. The electrode of graphenated nanofibers displayed a very good reproducibility and stability, and was successfully tested for detection of DA, UA and AA in real urine samples.