Validated Lc–Ms/Ms Method for Simultaneous Determination of Cabozantinib and Trametinib in Human Plasma with Comprehensive Stability Evaluation and Pharmacokinetic Application

Authors

  • Chikoti Kranti Kumar
  • Jitendra Kumar Gupta
  • Navneet Kumar

Keywords:

LC–MS/MS; Cabozantinib; Trametinib; Stability studies; Quality control; Human plasma; Pharmacokinetics; Therapeutic drug monitoring.

Abstract

 Background: Stability evaluation and quality control implementation are essential components of LC–MS/MS analysis to ensure reliable quantification of drugs in biological matrices during pharmacokinetic investigations. The present study assessed the stability characteristics of Cabozantinib and Trametinib in human plasma under various storage conditions and demonstrated the applicability of an LC–MS/MS assay for pharmacokinetic analysis.

Methods: Stability studies were performed using low-quality control (LQC, 15 ng/mL) and high-quality control (HQC, 800 ng/mL) samples under freeze–thaw, bench-top, auto-sampler, and long-term storage conditions following the recommendations of the US Food and Drug Administration (US FDA), European Medicines Agency (EMA), and International Council for Harmonisation (ICH M10) bioanalytical method validation guidelines. Quality control samples were incorporated into analytical batches to monitor analytical performance, and the LC–MS/MS assay was applied to pharmacokinetic evaluation following oral administration of Cabozantinib and Trametinib.

Results: Both analytes demonstrated excellent stability under all evaluated storage conditions. The measured concentrations remained within ±15% of the nominal values, and the %CV values were below 15%, confirming acceptable stability throughout sample handling and storage. Quality control samples consistently satisfied the predefined acceptance criteria, demonstrating reliable analytical performance during routine analysis. Pharmacokinetic application successfully characterized the plasma concentration–time profiles of both analytes, with Cabozantinib exhibiting a Cmax of 2.96 ± 0.28 µg/mL and Trametinib a Cmax of 32.84 ± 3.15 ng/mL, confirming the suitability of the assay for quantitative pharmacokinetic investigations.


Conclusion: The LC–MS/MS assay demonstrated reliable stability performance, consistent quality control implementation, and successful pharmacokinetic application for the simultaneous determination of Cabozantinib and Trametinib in human plasma. The findings support the use of this analytical approach for routine pharmacokinetic investigations, therapeutic drug monitoring, and future clinical studies involving these targeted anticancer agents.

Downloads

Download data is not yet available.

References

[1] Shah VP, Midha KK, Findlay JWA, et al. Bioanalytical method validation—A revisit with a decade of progress. Pharm Res. 2000;17(12):1551–1557.

[2] Viswanathan CT, Bansal S, Booth B, et al. Quantitative bioanalytical methods validation and implementation. Pharm Res. 2007; 24:1962–1973.

[3] Chambers E, Wagrowski-Diehl DM, Lu Z, Mazzeo JR. Systematic investigation of matrix effects in LC–MS/MS analysis. J Chromatogr B. 2007; 852:22–34.

[4] Jemal M, Xia Y, Whigan DB. Recent advances in LC–MS/MS bioanalysis for pharmaceutical compounds. Bioanalysis. 2021; 13:587–604.

[5] Bottino D, et al. Bioanalytical method validation for quantitative LC–MS/MS assays: Current regulatory perspectives. J Pharm Biomed Anal. 2022; 215:114741.

[6] Wang L, et al. LC–MS/MS determination of kinase inhibitors in human plasma: Applications in pharmacokinetic studies. J Chromatogr B. 2023; 1224:123685.

[7] Ni G, et al. Therapeutic drug monitoring of targeted anticancer agents using LC–MS/MS. Biomed Chromatogr. 2022;36: e5345.

[8] Xu RN, Fan L, Rieser MJ, El-Shourbagy TA. Recent advances in high-throughput quantitative bioanalysis using LC–MS/MS. J Pharm Biomed Anal. 2007; 44:342–355.

[9] Pitt JJ. Principles and applications of LC–MS/MS in clinical laboratories. Clin Biochem Rev. 2009; 30:19–34.

[10] Niessen WMA. Liquid Chromatography–Mass Spectrometry. 3rd ed. CRC Press; 2019.

[11] International Council for Harmonisation (ICH). ICH M10: Bioanalytical Method Validation and Study Sample Analysis. 2022.

[12] U.S. Food and Drug Administration. M10 Bioanalytical Method Validation and Study Sample Analysis Guidance for Industry. 2024.

[13] European Medicines Agency. ICH M10 on Bioanalytical Method Validation. 2022.

[14] Kaza M, Karaźniewicz-Łada M, Kosicka K, et al. Bioanalytical method validation: New FDA guidance versus EMA guideline. J Pharm Biomed Anal. 2019; 165:381–385.

[15] Vazvaei-Smith F, et al. ICH M10 Bioanalytical Method Validation Guideline—One year later. AAPS Pharm SciTech. 2024.

[16] Choueiri TK, Escudier B, Powles T, et al. Cabozantinib versus everolimus in advanced renal-cell carcinoma. N Engl J Med. 2015; 373:1814–1823.

[17] Yakes FM, Chen J, Tan J, et al. Cabozantinib inhibits MET and VEGFR2 signaling. Mol Cancer Ther. 2011; 10:2298–2308.

[18] Nguyen L, Holland J, Miles D, et al. Population pharmacokinetics of cabozantinib. J Clin Pharmacol. 2016; 56:1132–1140.

[19] Lacy S, Hsu B, Miles D, et al. Clinical pharmacokinetics and pharmacodynamics of Cabozantinib. Clin Pharmacokinetic. 2018; 57:1049–1061.

[20] Cabometyx® (Cabozantinib) Prescribing Information. Exelixis Inc.

[21] Choueiri TK, et al. Cabozantinib in advanced renal cell carcinoma: Long-term efficacy and safety. Lancet Oncol.

[22] Apolo AB, et al. Clinical applications of cabozantinib in solid tumors. Clin Cancer Res.

[23] Flaherty KT, Robert C, Hersey P, et al. Improved survival with MEK inhibition in BRAF-mutated melanoma. N Engl J Med. 2012; 367:107–114.

[24] Gilmartin AG, Bleam MR, Groy A, et al. Trametinib: A selective MEK inhibitor with antitumor activity. Clin Cancer Res. 2011; 17:989–1000.

[25] Kim KB, et al. Clinical pharmacology of trametinib. Clin Cancer Res. 2013; 19:560–567.

[26] Falchook GS, Lewis KD, Infante JR, et al. Pharmacokinetics and pharmacodynamics of trametinib. Lancet Oncol. 2012; 13:782–789.

[27] Mekinist® (Trametinib) Prescribing Information. Novartis Pharmaceuticals.

[28] Long GV, et al. Combination therapy involving trametinib in melanoma. N Engl J Med.

[29] Dummer R, et al. Trametinib in metastatic melanoma. Lancet.

[30] Rowland M, Tozer TN. Clinical Pharmacokinetics and Pharmacodynamics. 5th ed.

[31] Gibaldi M, Perrier D. Pharmacokinetics. 2nd ed.

[32] Gabrielsson J, Weiner D. Pharmacokinetic and Pharmacodynamic Data Analysis.

[33] Benet LZ, Hoener BA. Basic pharmacokinetic principles. Clin Pharmacokinetic.

[34] Toutain PL, Bousquet-Mélou A. Plasma concentration-time analysis. J Vet Pharmacol Ther.

[35] Blessy M, Patel RD, Prajapati PN, Agrawal YK. Stability-indicating analytical methods: A review. J Pharm Anal. 2014; 4:159–165.

[36] Bakshi M, Singh S. Development of validated stability-indicating assays. J Pharm Biomed Anal. 2002; 28:1011–1040.

[37] Blessy P, et al. Stability studies in pharmaceutical analysis: Recent trends. Curr Pharm Anal. 2021.

[38] El Orche A, et al. A comprehensive ICH M10 approach for validating LC–MS/MS assays and stability evaluation. Molecules. 2024.

[39] Grebe SK, Singh RJ. LC–MS/MS in clinical laboratories. Clin Biochem. 2011; 44:4–12.

[40] Jemal M. High-throughput LC–MS/MS bioanalysis. Rapid Commun Mass Spectrom.

[41] Jemal M, Ouyang Z. Quantitative LC–MS/MS in drug development. Biomed Chromatogr.

[42] Korfmacher WA. Foundation review of LC–MS/MS in pharmaceutical analysis. Mass Spectrom Rev.

[43] Hopfgartner G, Bourgogne E. LC–MS in drug metabolism studies. Mass Spectrom Rev.

[44] Jemal M. Advances in quantitative bioanalysis using tandem mass spectrometry. J Pharm Biomed Anal.

[45] Roskoski R Jr. Properties of small-molecule kinase inhibitors. Pharmacol Res.

[46] Roskoski R Jr. MEK1/2 inhibitors in cancer therapy. Pharmacol Res.

[47] Roskoski R Jr. VEGFR inhibitors in oncology. Pharmacol Res.

[48] Hanahan D. Hallmarks of cancer: new dimensions. Cancer Discov. 2022.

[49] Bray F, et al. Global cancer statistics. CA Cancer J Clin.

[50] Sung H, et al. Global cancer statistics 2020. CA Cancer J Clin..

Downloads

Published

2023-12-07

How to Cite

1.
Kumar CK, Gupta JK, Kumar N. Validated Lc–Ms/Ms Method for Simultaneous Determination of Cabozantinib and Trametinib in Human Plasma with Comprehensive Stability Evaluation and Pharmacokinetic Application. J Neonatal Surg [Internet]. 2023 Dec. 7 [cited 2026 Jul. 21];12(1):233-47. Available from: https://jneonatalsurg.com/index.php/jns/article/view/10474

Issue

Vol. 12 (2023): Journal of Neonatal Surgery

Section

Original Article

License

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

You are free to:

  • Adapt — remix, transform, and build upon the material for any purpose, even commercially.

Terms:

  • Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.
  • No additional restrictions — You may not apply legal terms or technological measures that legally restrict others from doing anything the license permits.