Reutilization of Tacrolimus Extracted from Expired Prograf® Capsules: Physical, Chemical, and Pharmacological Assessment

AAPS PharmSciTech. 2016 Aug;17(4):978-87. doi: 10.1208/s12249-015-0433-7. Epub 2015 Oct 16.


In this study, we investigated whether tacrolimus extracted and purified from the commercial capsules (Prograf® 5 mg) have retained its original quality and activity beyond the capsules expiration date in order to be reused for research purposes after extraction. High-performance liquid chromatography (HPLC) assay method was developed and validated for the quantification of tacrolimus, using cyclosporine A as an internal standard (IS). Moreover, a combination of analytical methods, including nuclear magnetic resonance (NMR), gas chromatography-mass spectrometry (GC-MS), Fourier transform-infrared (FT-IR) spectroscopy, X-ray diffraction (XRD), and differential scanning calorimetry (DSC) were used to assess the quality of extracted/purified tacrolimus. Suppression of murine peripheral-blood mononuclear cells (PBMC) proliferation and the levels of interleukin-2 (IL-2) and interferon gamma (IFN-γ) were also assessed. The data obtained showed no detectable differences in the quality profile between the authentic sample and extracted drug. Also, the results showed that the extracted/purified tacrolimus was able to suppress T cell proliferation, induced by concanavalin A, indicating the retained pharmacological activity. We proved that tacrolimus extracted/purified from expired Prograf® capsuled retains its purity and immunosuppressive activity and can be reused for research and possibly in pharmaceutical manufacturing.

Keywords: extraction; immunosuppression; purity; tacrolimus; validation.

MeSH terms

  • Animals
  • Capsules / chemistry*
  • Capsules / pharmacology*
  • Chromatography, High Pressure Liquid / methods
  • Cyclosporine / chemistry
  • Gas Chromatography-Mass Spectrometry / methods
  • Immunosuppressive Agents / chemistry
  • Immunosuppressive Agents / pharmacology
  • Interferon-gamma / metabolism
  • Interleukin-2 / metabolism
  • Leukocytes, Mononuclear / drug effects
  • Magnetic Resonance Spectroscopy / methods
  • Mice
  • Spectroscopy, Fourier Transform Infrared / methods
  • Tacrolimus / chemistry*
  • Tacrolimus / pharmacology*
  • X-Ray Diffraction / methods


  • Capsules
  • Immunosuppressive Agents
  • Interleukin-2
  • Interferon-gamma
  • Cyclosporine
  • Tacrolimus