Development and full validation of an LC–MS/MS methodology to quantify capmatinib (INC280) following intragastric administration to rats

Abstract

A highly sensitive, specific and simple LC–MS/MS method for quantification of capmatinib (INC280) in rat plasma was presented. The LC–MS/MS method was validated in terms of specificity and selectivity, linearity, accuracy and precision, matrix effect, extraction recovery, dilution integrity, carryover and stability as per the US Food and Drug Administration's bioanalytical method validation guideline. The validated assay was applied for quantification of capmatinib from a pharmacokinetic study in rats following oral administration at the doses of 1.0, 3.0 and 9.0 mg/kg. The calibration curve ranges from 1 to 2000 ng/ml with desirable linearity and r2 > 0.99. The intra‐ and inter‐batch accuracies were within 99.24–103.59 and 97.76–102.83% with coefficients of variation 5.08–7.36 and 3.18–4.99%, respectively. No significant interference was observed by endogenous peak at the retention time of capmatinib and IS. The assay was free from any matrix effect and showed precise recovery across the calibration curve range, and samples were stable under all experimental conditions. The validated assay was successfully applied to analyze plasma samples of pharmacokinetic study in rat to determine the concentration of capmatinib. In summary, a novel method for analyzing capmatinib in rat plasma has been successfully validated and is now being utilized for quantification of capmatinib from pre‐clinical studies.

HPLC with Fluorescence and Photodiode Array Detection for Quantifying Capmatinib in Biological Samples: Application to In Vivo and In Vitro Studies

“…However, the method has the potential to produce a significantly lower LLOQ by skipping the dilution step we deliberately included (200 μL diluted to 600 μL) to improve the column working life by reducing the amount of injected matrix. Furthermore, the addition of a preconcentration/evaporation step, if needed, fits well with the extraction method used and would further improve the sensitivity to match that of the two reported LC-MS/MS methods which have an LLOQ = 1 ng/mL in rat plasma [ 10 , 11 ]. This sensitivity, however, was not required for the in vivo and in vitro samples tested in the present study and therefore was not investigated.…”

“…The quantification of capmatinib in biological samples in vivo and in vitro is necessary to investigate its pharmacokinetics and potential interactions with other drugs, diet, and supplements. The chromatographic methods for the determination of the capmatinib concentration in biological samples were predominantly performed by LC-MS/MS [ 8 , 10 ] and UPLC-MS/MS [ 11 ]. Zhou et al, developed and validated a fast and reliable UPLC-MS/MS for the determination of capmatinib plasma levels in rats following three oral doses.…”

“…Another study reported the use of LC-MS/MS for the quantification of capmatinib in rat plasma in preclinical studies. The method had a working range of 1–2000 ng/mL [ 10 ]. A recently published study by Ali et al investigated the photophysical and fluorescence characteristics of capmatinib under various experimental conditions of solvents, pH, and temperature, using spectrofluorimetry, with no chromatography [ 12 ].…”

Mechanism of capmatinib degradation in stress conditions including degradation product characterization using ultra‐high‐performance liquid chromatography‐quadrupole‐time of flight mass spectrometry and stability‐indicating analytical method development

“…A suitable analytical method is required for the separation and analysis of such degradants during stability studies of any drug products containing CMT 12–15 . Although in literature bioanalytical methods for quantification of CMT in biological matrices have been reported, there is no stability‐indicating method available to quantify CMT in the presence of its degradation products 15–19 . Furthermore, no data are available on any degradation product of CMT.…”

“…[12][13][14][15] Although in literature bioanalytical methods for quantification of CMT in biological matrices have been reported, there is no stabilityindicating method available to quantify CMT in the presence of its degradation products. [15][16][17][18][19] Furthermore, no data are available on any degradation product of CMT. The present study includes the development of a sensitive, precise and selective RP-HPLC method utilizing Agilent Eclipse C18 column (250 Â 4.6mm, 5 μ) for the estimation of the CMT and its validation per ICH Q2R1 guideline.…”