A simple and sensitive HPLC fluorescence method for determination of tadalafil in mouse plasma.
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.
Farthing
Christine A
CA
Department of Internal Medicine, Division of Nephrology, Clinical Pharmacology and Hypertension, Virginia Commonwealth University, Richmond, VA 23298, USA. cafarthing@vcu.edu
Farthing
Don E
DE
Koka
Saisudha
S
Larus
Terri
T
Fakhry
Itaf
I
Xi
Lei
L
Kukreja
Rakesh C
RC
Sica
Domenic
D
Gehr
Todd W B
TW
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
D002243:Carbolines; D000068581:Tadalafil; D004317:Doxorubicin
J Chromatogr B Analyt Technol Biomed Life Sci. 2004 Oct 5;809(2):243-9
15315772
N Engl J Med. 2000 Jun 15;342(24):1802-13
10853004
Eur J Med Res. 2002 Oct 29;7(10):435-46
12435622
J Chromatogr B Analyt Technol Biomed Life Sci. 2005 Aug 5;822(1-2):278-84
15996535
J Chromatogr A. 2005 Feb 25;1066(1-2):89-95
15794558
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Jun 1;852(1-2):403-8
17321225
Cancer. 2003 Jun 1;97(11):2869-79
12767102
Br J Clin Pharmacol. 2006 Mar;61(3):280-8
16487221
Drugs. 2003;63(7):701-30
12656650
Talanta. 2005 Jan 15;65(1):276-80
18969796