A simple and sensitive HPLC fluorescence method for determination of tadalafil in mouse plasma |
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Authors: | Christine A. Farthing Don E. Farthing Saisudha Koka Terri Larus Itaf Fakhry Lei Xi Rakesh C. Kukreja Domenic Sica Todd W.B. Gehr |
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Affiliation: | 1. Department of Internal Medicine, Division of Nephrology, Clinical Pharmacology and Hypertension, Virginia Commonwealth University, Richmond, VA 23298, USA;2. Division of Cardiology, VCU Medical Center, Virginia Commonwealth University, Richmond, VA 23298, USA |
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Abstract: | 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 C18 column and a flow rate of 1.0 mL/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 2000 ng/mL (r > 0.999) with a detection limit of approximately 40 ng/mL. Component fluorescence detection was achieved using an excitation wavelength of 275 nm with monitoring of the emission wavelength at 335 nm. 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. |
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Keywords: | Tadalafil Doxorubicin HPLC Fluorescence Monolithic column |
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