A simple and sensitive HPLC fluorescence method for determination of tadalafil in mouse plasma. Academic Article uri icon


  • A simple and sensitive high-performance liquid chromatographic (HPLC) method utilizing fluorescence detection was developed for the determination of the phosphodiesterase type 5 inhibitor tadalafil in mouse plasma. This method utilizes a simple sample preparation (protein precipitation) with high recovery of tadalafil (98%), which eliminates the need for an internal standard. For constituent separation, the method utilized a monolithic C(18) column and a flow rate of 1.0mL/min with a mobile phase gradient consisting of aqueous trifluoroacetic acid (0.1% TFA in deionized water pH 2.2, v/v) and acetonitrile. The method calibration was linear for tadalafil in mouse plasma from 100 to 2000ng/mL (r>0.999) with a detection limit of approximately 40ng/mL. Component fluorescence detection was achieved using an excitation wavelength of 275nm with monitoring of the emission wavelength at 335nm. The intra-day and inter-day precision (relative standard deviation, RSD) values for tadalafil in mouse plasma were less than 14%, and the accuracy (percent error) was within -14% of the nominal concentration. The method was utilized on mouse plasma samples from research evaluating the potential cardioprotective effects of tadalafil on mouse heart tissue exposed to doxorubicin, a chemotherapeutic drug with reported cardiotoxic effects.

published proceedings

  • J Chromatogr B Analyt Technol Biomed Life Sci

altmetric score

  • 7

author list (cited authors)

  • Farthing, C. A., Farthing, D. E., Koka, S., Larus, T., Fakhry, I., Xi, L., ... Gehr, T.

citation count

  • 37

complete list of authors

  • Farthing, Christine A||Farthing, Don E||Koka, Saisudha||Larus, Terri||Fakhry, Itaf||Xi, Lei||Kukreja, Rakesh C||Sica, Domenic||Gehr, Todd WB

publication date

  • October 2010