The invention relates to a preparation method of an iron, cobalt and nitrogen co-doped carbon nanofiber catalyst. The preparation method comprises the following steps: firstly, preparation of an electrostatic spinning precursor solution, respectively adding cobalt salt, zinc salt and iron salt into an organic solvent to obtain a solution A, and adding a high polymer into the organic solvent to obtain a solution B; adding the solution A into the solution B and uniformly mixing; secondly, preparation of polymer nanofibers: preparing the polymer nanofibers by using an electrospinning technology and the electrostatic spinning precursor solution; thirdly, preparation of a catalyst: carrying out heat treatment on the polymer nanofibers to obtain the catalyst. Compared with the prior art, the preparation method of the iron, cobalt and nitrogen co-doped carbon nanofiber catalyst, disclosed by the invention, has the advantages of simple steps, easiness in operation and controllable implementation conditions; the iron, cobalt and nitrogen co-doped carbon nanofiber catalyst can be obtained without a template or activation etching; in addition, the iron, cobalt and nitrogen co-doped carbon nanofiber catalyst has oxygen reduction performance similar to that of commercial platinum carbon, and has the advantages of low cost, high efficiency, no pollution and capability of being used as a cathode catalyst for an alcohol fuel cell.