Related Experiment Video
Updated: May 29, 2026
Simultaneous Quantification of Selected Kynurenines Analyzed by Liquid Chromatography-Mass Spectrometry in Medium Collected from Cancer Cell Cultures
Published on: May 9, 2020
Development and clinical application of a LC-MS/MS method for simultaneous determination of various tyrosine kinase
Lutz Götze1, Axel Hegele, Stephan Klaus Metzelder
1Institute of Laboratory Medicine and Pathobiochemistry, Philipps-University Marburg, Marburg, Germany.
Background:
Increasing numbers of tyrosine kinase inhibitors (TKIs) were studied and approved for therapy of malignancies and other diseases. The aim of this study was to develop and validate a specific, simple and rapid quantification method for various TKIs in human plasma.
Methods:
A simultaneous test for six TKIs (erlotinib, imatinib, lapatinib, nilotinib, sorafenib, sunitinib) was developed using liquid chromatography tandem mass spectrometry in a multiple reaction monitoring mode. After protein precipitation the specimens were applied to the HPLC system and separated using a gradient of acetonitrile containing 1% formic acid with 10 mM ammoniumformiate on an analytic RP-C18 column.
Results:
The calibration range was 10-1000 ng/mL for sunitinib and 50-5000 ng/mL for the other TKIs with coefficients of determination ≥0.99 for all analytes. The intra- and inter day coefficients of variation were ≤15% and the chromatographic run time was 12 min. Plasma specimens were stable for measurement for at least 1 week at 4°C. Clinical applications of the assay are exemplarily discussed.
Conclusions:
This novel high-throughput method is suitable for specific simultaneous determination of different TKIs in routine clinical practice.
Insights
A new method accurately quantifies multiple tyrosine kinase inhibitors (TKIs) in plasma, aiding cancer therapy. This rapid, simultaneous assay is suitable for routine clinical use.
Area of Science:
- Pharmacology
- Analytical Chemistry
- Biochemistry
Background:
- Tyrosine kinase inhibitors (TKIs) are increasingly used for treating malignancies and other diseases.
- Developing efficient methods for monitoring TKI levels in patients is crucial for optimizing therapy.
Purpose of the Study:
- To develop and validate a specific, simple, and rapid quantification method for various TKIs in human plasma.
- To enable simultaneous measurement of multiple TKIs, improving clinical monitoring efficiency.
Main Methods:
- Liquid chromatography tandem mass spectrometry (LC-MS/MS) in multiple reaction monitoring (MRM) mode was employed.
- A simultaneous assay for six TKIs (erlotinib, imatinib, lapatinib, nilotinib, sorafenib, sunitinib) was developed.
- Protein precipitation followed by RP-C18 column separation with a specific mobile phase gradient was utilized.
Main Results:
- The method demonstrated high accuracy with coefficients of determination ≥0.99 for all analytes.
- Intra- and inter-day coefficients of variation were ≤15%, indicating good precision.
- The assay achieved a rapid chromatographic run time of 12 minutes and demonstrated plasma sample stability for at least one week at 4°C.
Conclusions:
- This novel high-throughput method is suitable for the specific and simultaneous determination of different TKIs.
- The assay is well-suited for routine clinical practice, facilitating improved patient management and TKI therapy optimization.
