BIO-analytical method development and validation of finasteride, tadalafil, and its application to pharmacokinetic studies in RAT plasma by using LC-MS/MS
Abstract
Objective: An easy, quick, precise, active and reproducible LC-MS/MS (Liquid Chromatography Tandem Mass Spectrometry) technique was developed for the bio-analytical method of Finasteride and Tadalafil using Avanafil as internal standard (IS). Methods: This article summarizes the recent progress on bioanalytical liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) methods using waters Symmetry C18 column (150x4.6 mm, 3.5µ) column and mobile phase of 0.1% Perchloric acid and Acetonitrile (ACN) in 60:40. Results: The calibration curve was linear in the range of 12.5-100 ng/ml for Finasteride and 12.5-100 ng/ml Tadalafil. The recovery results of Accuracy and Precision of Finasteride and Tadalafil were 95.10, 96.85, 98.76, 98.81% and 95.77, 97.46, 97.99, 97.01% at different QC (Quality Control) concentration levels. Matrix effect results were within the acceptable limit. An electro-spray ionization source was used to study of Finasteride and Tadalafil at m/z 373.5497→142.0085, m/z 390.4047→128.1138 for Finasteride and Tadalafil, m/z 484.9516→104.5326 for Avanafil were ion pairs of mass analysis. Conclusion: The application denotes all the parameters of system suitability, specificity, linearity and accuracy are in good agreement with USFDA (United States of Food and Drug Administration) guidelines and applied effectively for the investigation of pharmacokinetic studies in rat.
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