Abstract

BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF TADALAFIL IN HUMAN PLASMA USING RP-HPLC

Prashik S. Shimpi* and Dr. Hitendra S. Mahajan

Abstract

Purpose: A simple reverse phase liquid chromatographic analytical method has been developed and validated for estimation of Tadalafil in Human plasma. Methods: The separation was carried out on Eclipse XBD-C8 Plasma 5 μm ( 4.6× 150 mm) as Stationary phase, Mobile Phase: Acetonitrile: Aqueous solution containing 0.012M triethyl amine + 0.020M orthophosphoric acid. (40:60) Flow rate: 0.5 mL/min using uv detector at 225 & 285 nm. Results: The described HPLC method was linear over a concentration range of 100-3200 ng/mL. Sildenafil was used as a internal standard. The Tadalafil showed retention time 9.4 min & sildenafil showed retention time 3.7min respectively. The Accuracy and Precision, recovery, selectivity for Tadalafil was 100-3200 ng/mL respectively. The stability of thedrug spiked human plasma samples during three freeze thaw cycles were stable in plasma for about one month when stored at frozen state. Conclusions: It can be concluded that the developed bioanalytical method is capable of quantifying Tadalafil from spiked human plasma samples. The method meets the requirements of the USFDA Guidelines and can be applied to Bioavailability/ Bioequivalence studies of Tadalafil.

Keywords: Bioanalytical method, development, validation.