Title:Development of Fast and Simple LC-ESI-MS/MS Technique for the Quantification of Regorafenib; Application to Pharmacokinetics in Healthy Rabbits
VOLUME: 17 ISSUE: 4
Author(s):Yatha Ravi*, Darna Bhikshapathi, Shankar Cheruku and Bigala Rajkamal
Affiliation:Department of Pharmacy, Mewar University, Chittorgarh, Rajasthan, Department of Pharmaceutics, TRR College of Pharmacy, Hyderabad, Department of Pharmaceutical Analysis, Vijaya College of Pharmacy, Hyderabad, Department of Pharmacy, Mewar University, Chittorgarh, Rajasthan
Keywords:Regorafenib, tyrosine kinase inhibitor, bioavailability studies, LC-MS/MS, matrix effect, metabolites.
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 was 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 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 a 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 AUC0→α and AUC0→t were 6476.81 ± 259.59 and 6213.845 ±
257.892 respectively 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 the drug (40 mg tablet) administered through the oral route in healthy rabbits.
