title: A simple and sensitive HPLC fluorescence method for determination of tadalafil in mouse plasma.
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 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.
authors.affiliation: Department of Internal Medicine, Division of Nephrology, Clinical Pharmacology and Hypertension, Virginia Commonwealth University, Richmond, VA 23298, USA. cafarthing@vcu.edu
authors.forename: Christine A
authors.initials: CA
authors.lastname: Farthing
authors.forename: Don E
authors.initials: DE
authors.lastname: Farthing
authors.forename: Saisudha
authors.initials: S
authors.lastname: Koka
authors.forename: Terri
authors.initials: T
authors.lastname: Larus
authors.forename: Itaf
authors.initials: I
authors.lastname: Fakhry
authors.forename: Lei
authors.initials: L
authors.lastname: Xi
authors.forename: Rakesh C
authors.initials: RC
authors.lastname: Kukreja
authors.forename: Domenic
authors.initials: D
authors.lastname: Sica
authors.forename: Todd W B
authors.initials: TW
authors.lastname: Gehr
mesh_terms: D000818:Animals; D002243:Carbolines / Q000097:blood* / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004317:Doxorubicin / Q000737:chemistry; D004355:Drug Stability; D016014:Linear Models; D008297:Male; D051379:Mice; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012995:Solubility; D013050:Spectrometry, Fluorescence / Q000379:methods*; D000068581:Tadalafil
chemical_list: D002243:Carbolines; D000068581:Tadalafil; D004317:Doxorubicin
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