A simple and sensitive HPLC fluorescence method for determination of tadalafil in mouse plasma

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 878; no. 28; pp. 2891 - 2895
• Main Authors: Farthing, Christine A., Farthing, Don E., Koka, Saisudha, Larus, Terri, Fakhry, Itaf, Xi, Lei, Kukreja, Rakesh C., Sica, Domenic, Gehr, Todd W.B.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15-10-2010; Elsevier
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Carbolines - blood; Carbolines - chemistry; Chromatography; Chromatography, High Pressure Liquid - methods; Doxorubicin; Doxorubicin - chemistry; Drug Stability; Drugs; Flow rate; Fluorescence; Fundamental and applied biological sciences. Psychology; General pharmacology; HPLC; Inhibitors; Linear Models; Male; Medical sciences; Mice; Monolithic column; Pharmacology. Drug treatments; Reproducibility of Results; Sensitivity and Specificity; Solubility; Spectrometry, Fluorescence - methods; Standard deviation; Tadalafil; Wavelengths; Fluorescence; Tadalafil; Doxorubicin; HPLC; Monolithic column; Antineoplastic agent; Biological fluid; Vasodilator agent; 3',5'-Cyclic-GMP phosphodiesterase; HPLC chromatography; Esterases; Phosphoric diester hydrolases; Determination; Blood plasma; Isomerases; Quantitative analysis; DNA topoisomerase (ATP-hydrolysing); Enzyme; Rodentia; Enzyme inhibitor; Topoisomerase II inhibitor; Phosphodiesterase inhibitor; Phosphodiesterase 5 inhibitor; Vertebrata; Antibiotic; Carboline derivatives; Mammalia; Mouse; Animal; Hydrolases; Anthracyclins
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2010.07.022

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 1.0
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.