Development and clinical application of a liquid chromatography-tandem mass spectrometry-based assay to quantify eight tyrosine kinase inhibitors in human plasma

by Selleckchem | Aug 18 2023

Introduction: Tyrosine kinase inhibitors (TKIs) are widely used in tumor treatment. The detection of these medicines by liquid chromatography-tandem mass spectrometry (LC-MS/MS) can avoid the interference of structurally similar compounds.

Objectives: This study aimed to develop and validate a new LC-MS/MS assay for the quantification of eight tyrosine kinase inhibitors in human plasma and to preliminarily evaluate the clinical utility of the therapeutic drug monitoring method.

Methods: Plasma samples were prepared by simple protein precipitation and separated using an ultra-high-performance reversed phase column. Detection was achieved using a triple quadrupole mass spectrometer in the positive ionization mode. The assay was validated against standard guidelines. We reviewed and analyzed the results of 268 plasma samples obtained from patients administered imatinib and other TKIs collected from January 2020 to November 2021 at Zhongshan Hospital. The analytes were separated and quantified within 3.5 min.

Results: The newly developed method demonstrated linearity for the detected drug concentration in the range of 20 to 2000 ng/ml for gefitinib (r2 = 0.991) and crizotinib (r2 = 0.992), 50 to 5000 ng/ml for nilotinib (r2 = 0.991) and imatinib (r2 = 0.995), 1500-150,000 ng/ml for vemurafenib (r2 = 0.998), 1000-100,000 ng/ml for pazopanib (r2 = 0.993), 0.5-100 ng/ml for axitinib (r2 = 0.992) and 5-500 ng/ml for sunitinib (r2 = 0.991) and N-desethyl sunitinib (r2 = 0.998). The lower limit of quantification (LLOQ) was 20 ng/ml for gefitinib and crizotinib, 50 ng/ml for nilotinib and imatinib, 1500 ng/ml for vemurafenib, 1000 ng/ml for pazopanib, 0.5, and 5 ng/ml for sunitinib and N-desethyl sunitinib, respectively. Specificity, precision, accuracy, and stability were tested, and met the requirements of the guidelines. At the same dose, there was no significant difference in plasma drug concentration between the original imatinib medicine and the generic medicine after patent expiration.

Conclusion: We developed a sensitive and reliable method for the quantification of eight TKIs.

Comments:

The developed method in this study utilized liquid chromatography-tandem mass spectrometry (LC-MS/MS) to detect and quantify eight tyrosine kinase inhibitors (TKIs) in human plasma. The use of LC-MS/MS allowed for the accurate measurement of these medicines while minimizing interference from structurally similar compounds.

The researchers prepared plasma samples through simple protein precipitation and separated the analytes using an ultra-high-performance reversed-phase column. Detection was achieved using a triple quadrupole mass spectrometer operating in the positive ionization mode. The method was validated following standard guidelines.

The study analyzed 268 plasma samples obtained from patients who were administered imatinib and other TKIs at Zhongshan Hospital between January 2020 and November 2021. The newly developed LC-MS/MS assay enabled the separation and quantification of the TKIs within a short analysis time of 3.5 minutes.

The results demonstrated linearity for the detected drug concentration within specific ranges for each TKI. The range and correlation coefficient (r2) for each TKI were as follows: 20-2000 ng/ml and r2 = 0.991 for gefitinib and crizotinib, 50-5000 ng/ml and r2 = 0.991 for nilotinib and imatinib, 1500-150,000 ng/ml and r2 = 0.998 for vemurafenib, 1000-100,000 ng/ml and r2 = 0.993 for pazopanib, 0.5-100 ng/ml and r2 = 0.992 for axitinib, and 5-500 ng/ml and r2 = 0.991 for sunitinib and N-desethyl sunitinib. The lower limit of quantification (LLOQ) was determined to be 20 ng/ml for gefitinib and crizotinib, 50 ng/ml for nilotinib and imatinib, 1500 ng/ml for vemurafenib, 1000 ng/ml for pazopanib, and 0.5 ng/ml and 5 ng/ml for sunitinib and N-desethyl sunitinib, respectively.

The method was found to be specific, precise, accurate, and stable, meeting the requirements of the validation guidelines. Additionally, the study compared the plasma drug concentrations between the original imatinib medicine and a generic medicine after patent expiration and found no significant difference at the same dosage.

In conclusion, this study successfully developed a sensitive and reliable LC-MS/MS assay for the quantification of eight TKIs in human plasma. The method's validation and preliminary clinical evaluation demonstrated its potential utility in therapeutic drug monitoring for patients receiving these medications.