A Highly Sensitive LC–MS/MS Method Development and Validation of Fedratinib in Human Plasma and Pharmacokinetic Evaluation in Healthy Rabbits

Background: A simple and sensitive quantitation analytical technique by liquid chromatography– tandem mass spectrometry (LC-MS/MS) is essential for fedratinib in biological media with kinetic study in healthy rabbits. Objective: The main objectives of the present research work are the development of the LC-MS/MS method and to validate a procedure for the quantitation of fedratinib and its application to kinetic study in rabbits. Methods: Separation of processed samples was done on zorbax SB C18 column (50mm×4.6 mm) 3.5μm with a movable phase of methanol, acetonitrile and 0.1% formic acid in the ratio of 30:60:10. The movable phase was monitored through column at 0.8 ml/min flow rate. The drug and ibrutinib internal standard (IS) were evaluated by monitoring the transitions of m/z -525.260/57.07 and 441.2/55.01 for fedratinib and IS, respectively in multiple reaction monitoring mode. Results: The linear equation and coefficient of correlation (R²) results were y =0.00348x+0.00245 and 0.9984, respectively. Intra and inter-day precision RSD findings of the developed technique were found in the range of 2.4 - 5.3% for the quality control (QC)-samples (252.56, 1804.0 and 2706 ng/ml). The proposed method was subjected to pharmacokinetic study in healthy rabbits and the kinetic study, fedratinib showed mean AUClast 13190±18.1 hr*ng/ml and Cmax was found to be 3550±4.31 ng/ml in healthy rabbits. Conclusion: The validated method can be applicable for the pharmacokinetic and toxicokinetic studies in the clinical and forensic analysis of fedratinib in different kinds of biological matrices successfully.