Hydrophobic beta-cyclodextrin polymer nanofibers were obtained by the electrospinning of peracetyl-beta-cyclodextrin polymer (Ac beta CDP) from acetone : DMF (3 : 2 v/v) solution. The morphology of Ac beta CDP nanofibers was studied by scanning electron microscopy. The proposed novel hydrophobic beta-CD polymer was synthesized through reaction of water-soluble beta-CD polymer crosslinked with epichlorohydrin (beta-CD-ECH polymer) and acetic anhydride in the presence of pyridine. The peracylation of beta-CD-ECH polymer is verified by FT-IR and solubility data. The final part of this study is focused on incorporating vitamin B-2 (VB2) into the Ac beta CDP nanofiber. In vitro release of VB2 from Ac beta CDP/VB2 nanofiber in model physiological media was investigated; the results exhibited initial burst and then slow drug release. The release of VB2 from Ac beta CDP/VB2 nanofiber is slower than from Ac beta CDP/VB2.