LC?MS/MS Bioanalytical Procedure for Quantification of Imatinib Mesylate in Rat Plasma: Development and Application to Pharmacokinetic Study


Author(s): Milind Dharmraj Kamble*, Minal Chaudhari, Ganesh Tapadiya, Bharat Dhokchawle and Chhaya Sonar

Introduction: In this research, a fit-for-purpose LC-MS/MS method for quantification of Imatinib Mesylate in rat plasma was developed and utilized for the pharmacokinetic study. Imatinib extraction was done and isolated from plasma samples using protein precipitation method. Imatinib quantification was done using Liquid Chromatography (LC) tandem Mass Spectrometry (MS) with Electro Spray Ionization (ESI) and Multiple Reaction Monitoring (MRM) in positive ionization mode.

Objective: A simple and fast fit for purpose LC-MS/ MS method was developed and utilized for the quantification of Imatinib mesylate and was applied to a pharmacokinetic study.

Methods: All the sample preparation was acquired and accepted through protein precipitation. High performance chromatographic partition was achieved on a PURITAS PNCN, (100 × 4.6 mm, 5 μm) CHROMACHEMIE analytical column by using an isocratic elution. Pump A (40%): 0.1% Formic acid in 5 mM Ammonium Formate Solution Pump B (60%): 0.1% Formic acid in acetonitrile at a flow rate of 0.8 mL/ min. for 5 mintues.

Results: The retention time of Imatinib mesylate and its internal standard, Verapamil was 3.21 ± 0.8 min and 3.64 ± 0.5 min, respectively. The total run time was 5.0 minutes. The elution detection was obtained with +ve electrospray ionization multiple reaction monitoring of the ion transitions at m/z 494.40 → 394.20 for Imatinib and second mass transitions were monitored: Imatinib at m/z 494.40 → 217.20 while internal standard Verapamil was selected for m/z 455.30 → 165.10. The method was developed and validated over the concentration range of 50- 5000 ng/mL for Imatinib mesylate, with correlation coefficient greater than 0.9991. The extraction recovery was more than 105.37% and the matrix effect was not significant. The intra and inter-day precisions were below 4.65% and accuracies ranged from 91.7 to 102.0%. The quantification limit for Imatinib was 5.05 ng/mL. Imatinib mesylate was demonstrated to be stable in rat plasma under the tested conditions.

Conclusion: The developed LC-MS/MS fit-for purpose procedure for the quantification of Imatinib Mesylate in rat plasma can be used for pharmacokinetic studies in preclinical applications.