In order to investigate the magnetic anisotropy of non-aligned Fe nanofibers, electrospinning and heat treatment techniques were successfully used to prepare novel Fe nanofibers. A new method was offered to characterize the magnetic anisotropy only using VSM combining with Mossbauer spectroscopy techniques. It is found that each individual nanofiber was uniform and straight on the whole measurement substrate with little breakage and Mossbauer spectroscopy result shows that all magnetic moments lied in the EA of nanofibers. With simple numerical calculation and fitting with experimental data, the magnetic anisotropy Ha of non-aligned Fe nanofibers was obtained and the result shows that the shape anisotropy dominates the overall magnetic anisotropy in the Fe nanofibers. This method provides a rapid and effective access to analyze and measure the magnetic anisotropy of soft magnetic nanofibers. (C) 2012 Elsevier B.V. All rights reserved.