High-Speed Analysis of Sunitinib and Axitinib in Plasma Using Triple Quadrupole LC/MS/MS (LCMS-8050)

Significance of the Topic

The molecularly targeted drugs sunitinib and axitinib are pivotal in cancer therapy. Accurate measurement of these compounds in plasma is critical for pharmacokinetic studies, therapeutic drug monitoring, and quality control in pharmaceutical research.

Objectives and Study Overview

This study aims to develop a rapid, robust, and sensitive LC/MS/MS method for quantifying sunitinib, its metabolite SU12662, and axitinib in human plasma. By utilizing a high-sensitivity triple quadrupole mass spectrometer (LCMS-8050) and a streamlined sample preparation, the workflow targets high throughput while maintaining analytical performance.

Methodology

• Sample Pretreatment
  • 50 µL human plasma spiked with standards and QC samples.
  • Addition of 100 µL internal standard (imatinib-d8) solution and 100 µL acetonitrile for protein precipitation.
  • Centrifugation at 15 000 rpm, 4 °C for 5 min, followed by dilution of 100 µL supernatant with 100 µL 10 mmol/L ammonium formate.
• Chromatographic Conditions
  • Column: Shim-pack GIS (75×2.1 mm, 3 µm) in reverse-phase mode.
  • Mobile phase: A) 10 mmol/L ammonium formate in water; B) methanol; gradient from 10 % to 80 % B over 3 min.
  • Flow rate: 0.3 mL/min; column temperature: 40 °C; injection volume: 5 µL.
• Mass Spectrometry
  • ESI positive ion mode; probe voltage +4.0 kV; DL 150 °C; block heater 200 °C; interface 300 °C.
  • Nebulizing 3 L/min, drying 5 L/min, heating 15 L/min.
  • MRM transitions: sunitinib 399.40→283.30; axitinib 387.40→356.30; SU12662 371.40→283.30; imatinib-d8 502.50→394.40.

Main Results and Discussion

Calibration curves for all analytes exhibited excellent linearity (r ≥ 0.999) across 3–300 µg/mL (sunitinib, SU12662) and 0.3–30 µg/mL (axitinib). Representative chromatograms revealed separation of isomeric peaks, which were combined for quantitation. Quality control samples showed repeatability below 10 % and accuracy within 100 ± 15 %, meeting bioanalytical validation criteria.

Benefits and Practical Applications

• High-throughput quantitation with a total run time of 5 min per injection.
• Cost-effective sample preparation by eliminating solid phase extraction.
• High sensitivity and selectivity from triple quadrupole MRM analysis.
• Applicable to pharmacokinetic profiling, therapeutic drug monitoring, and preclinical studies.

Future Trends and Potential Applications

The trend toward ultrafast and multiplexed LC/MS/MS workflows will continue, with potential integration of microflow systems and high-resolution mass spectrometry for broader compound coverage. Further miniaturization of sample handling and automation can enhance throughput for clinical and industrial settings.

Conclusion

A straightforward LC/MS/MS method for sunitinib, SU12662, and axitinib in plasma was developed using simple protein precipitation and triple quadrupole detection. The approach delivers rapid, precise, and accurate quantitation, supporting its use in pharmacokinetic and bioanalytical applications.

References

• No specific literature references were cited in the source text.