Development of Fast and Simple LC-ESI-MS/MS Technique for the Quantification of Regorafenib; Application to Pharmacokinetics in Healthy Rabbits

Abstract

Background: A simple quantification technique by liquid chromatography–electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) is required for regorafenib in biological matrices with bioavailability studies in healthy rabbits, when compared with reported techniques. Objective: The main aim of the research work is to develop a validated LC-ESI-MS/MS technique for the quantification of regorafenib and application to bioavailability studies in healthy rabbits. Methods: Chromatographic separation was achieved with hypersil-C18 analytical column (50mm×4.6 mm, 4µm) and a mobile phase composition of acetonitrile and 5mM ammonium acetate in the proportion of 70:30. The mobile phase was infused into the column with high pressure to get 0.7 ml/min flow rate. The total retention time of the analyte is promising when compared with the existed methods for regorafenib. Quantitation was processed by monitoring transitions of m/z -483.0/262.0 and 450.0/260.0 for regorafenib and internal standard respectively in multiple reaction monitoring. Results: The linearity equation and correlation coefficient (R2) findings were y =0.9948x+2.6624 and 0.998 respectively. The intra and inter-day precision of the developed technique was found between 1.00 – 8.50% for the QC-samples (2, 4, 240 and 480ng/ml). From bioavailability study, the drug was shown Tmax of 3.688 ± 0.754; average AUC0͢α and AUC0͢t was 6476.81 ± 259.59 and 6213.845 ± 257.892 and Cmax was found to be 676.91 ± 22.045 in healthy rabbits. Conclusion: The developed technique was validated and successfully applied in the pharmacokinetic study of drug (40 mg tablet) administered through oral route in healthy rabbits.

Development and validation of LC-MS/MS method for alpelisib quantification in human plasma: Application to pharmacokinetics in healthy rabbits

“…Water has provided for the rabbits under the study throughout the work and 37.5 mg/kg drug dose has given to healthy rabbits and taken 0.60 ml of blood sample solutions from marginal ear vein of rabbits prior to dosing (zero-time) and at different time intervals between 0 and 24 hours. The resultant solution was exposed to 4,500 rpm in the centrifuge for 15.0 minutes and plasma samples were separated and relocated into labelled polypropylene tubings at −20.0°C (Patel et al, 2011;Ravi et al, 2021). The study was approved by Institutional animal ethical committee with No: 1447/PO/Re/8/11/CPCSEA/18/A.…”