High-Speed Sensitive Analysis of Sunitinib and Related Metabolites Through UPLC- MS/MS Featuring CORTECS™ Premier C18 Columns With Maxpeak™ High Performance Surfaces (HPS) Technology

Significance of the Topic

Tyrosine kinase inhibitors (TKIs) such as Sunitinib play a crucial role in cancer therapy by targeting cell growth pathways. Monitoring Sunitinib and its active metabolite N-desethyl Sunitinib is essential to optimize dosing, minimize toxicity, and improve patient outcomes.

Objectives and Study Overview

This work aimed to develop a rapid, sensitive, and reproducible UPLC-MS/MS assay for quantifying Sunitinib and its main active metabolite in human plasma. The method leverages CORTECS™ Premier C18 columns with MaxPeak™ High Performance Surfaces technology and detection on the Waters™ Xevo™ TQ Absolute system. The assay covers clinically relevant concentration ranges and achieves analysis in under two minutes.

Methodology and Instrumentation

• Sample Preparation
  • Stock solutions of Sunitinib, N-desethyl Sunitinib, and deuterated internal standards prepared in DMSO.
  • Calibration and quality controls spiked into plasma, followed by protein precipitation using acetonitrile (3× plasma volume).
• Chromatography
  • System: Waters ACQUITY UPLC I-Class PLUS with CORTECS Premier C18 column (1.6 μm, 2.1×50 mm) featuring MaxPeak HPS technology.
  • Mobile phases: aqueous formic acid and methanolic formic acid gradient.
  • Total runtime: <2 minutes per injection.
• Mass Spectrometry
  • Detector: Waters Xevo TQ Absolute in positive electrospray ionization mode.
  • Multiple reaction monitoring (MRM) optimized for each analyte and internal standard.

Main Results and Discussion

• Reproducibility
  • Retention time and peak area %RSD <5% over 10 consecutive injections, demonstrating robust chromatography.
• Sensitivity Enhancement
  • MaxPeak HPS configuration yielded a fivefold increase in peak height and area compared to a traditional stainless-steel system.
• Linearity
  • Calibrated range: 0.1–100 ng/mL for both Sunitinib and N-desethyl Sunitinib in plasma.
  • Correlation coefficients: R² >0.997 (Sunitinib) and >0.994 (metabolite).
• Lower Limit of Quantitation (LLOQ)
  • LLOQ at 0.01 ng/mL plasma (12.5 pg/mL on column) with accuracy within ±15% and precision ≤20% RSD, meeting FDA bioanalytical guidelines.

Benefits and Practical Applications of the Method

• High throughput: sub-two-minute runs support large sample volumes in clinical laboratories.
• Enhanced sensitivity and dynamic range enable reliable therapeutic drug monitoring (TDM).
• Minimized metal-analyte interactions via MaxPeak HPS technology improve signal quality.

Future Trends and Potential Applications

Expansion of high-surface UPLC column technologies to other TKIs and small molecule drugs.
Integration with automated sample preparation platforms for streamlined workflows.
Broader clinical adoption facilitating personalized dosing strategies in oncology.

Conclusion

The described UPLC-MS/MS method combines CORTECS Premier C18 columns with MaxPeak HPS technology and Xevo TQ Absolute detection to deliver a rapid, sensitive, and reproducible assay for Sunitinib and its active metabolite. This workflow is well suited for therapeutic drug monitoring in clinical settings, offering significant improvements over traditional systems.

Reference

• Kim A, Balis FM, Widemann BC. Sorafenib and Sunitinib. The Oncologist. 2009;14(8):800–805.
• Faivre S, Niccoli P, Castellano D, Valle JW, Hammel P, Raoul JL, et al. Sunitinib in Pancreatic Neuroendocrine Tumors: Updated PFS and OS From a Phase III Randomized Study. Ann Oncol. 2017;28(2):339–343.
• Takasaki S, Kikuchi M, Kawasaki Y, Ito A, Arai Y, Yamaguchi H, et al. Severe Toxicity Induced by Accumulation of Active Sunitinib Metabolite in a Japanese Patient With Renal Cell Carcinoma: A Case Report. J Med Case Rep. 2017;11(1).
• Shah D, Smith K, Yang J, Hancock P. Analysis of Fourteen Organic Acids in Various Beverages Using ACQUITY UPLC H-Class PLUS and ACQUITY QDa Mass Detector. Waters Application Note. 2021.
• Waters Corporation. CORTECS Columns Applications Notebook. 2014.
• Layton C, Rainville P. Advantages of Using MaxPeak HPS Technology for the Analysis of Targeted Cancer Growth Inhibitor Therapies. Waters Application Note. 2022.
• Retmana IA, Beijnen JH, Sparidans RW. Chromatographic Bioanalytical Assays for Targeted Covalent Kinase Inhibitors and Their Metabolites. J Chromatogr B. 2021;1162:122466.
• Jolibois J, Schmitt A, Royer B. A Simple and Fast LC-MS/MS Method for the Routine Measurement of Cabozantinib, Olaparib, Palbociclib, Pazopanib, Sorafenib, Sunitinib and Its Main Active Metabolite in Human Plasma. J Chromatogr B. 2019;1132:121844.
• Westerdijk K, Krens SD, Graaf WTA, Mulder SF, Herpen CML, Smilde T, et al. The Relationship Between Sunitinib Exposure and Both Efficacy and Toxicity in Real-World Patients With Renal Cell Carcinoma and Gastrointestinal Stromal Tumour. Br J Clin Pharmacol. 2020;87(2):326–335.
• Zhang M, Liu X, Chen Z, Jiang S, Wang L, Tao M, et al. Method Development and Validation for Simultaneous Determination of Six Tyrosine Kinase Inhibitors and Two Active Metabolites in Human Plasma/Serum Using UPLC–MS/MS for Therapeutic Drug Monitoring. J Pharm Biomed Anal. 2022;211:114562.
• de Bruijn P, Sleijfer S, Lam MH, Mathijssen RHJ, Wiemer EAC, Loos WJ. Bioanalytical Method for the Quantification of Sunitinib and Its N-Desethyl Metabolite SU12662 in Human Plasma by UPLC–MS/MS. J Pharm Biomed Anal. 2010;51(4):934–941.
• U.S. Food and Drug Administration. Bioanalytical Method Validation Guidance for Industry. 2018.