Validation of an LC-MS/MS assay for rapid and simultaneous quantification of 21 kinase inhibitors in human plasma and serum for therapeutic drug monitoring

Zaid N Al Shirity; Niels Westra; Kai van Hateren; Thijs H Oude Munnink; Jos G W Kosterink; Paola Mian; Marjolijn N Lub-de Hooge; Daan J Touw; Bahez Gareb

doi:10.1016/j.jchromb.2023.123872

Validation of an LC-MS/MS assay for rapid and simultaneous quantification of 21 kinase inhibitors in human plasma and serum for therapeutic drug monitoring

J Chromatogr B Analyt Technol Biomed Life Sci. 2023 Sep 1:1229:123872. doi: 10.1016/j.jchromb.2023.123872. Epub 2023 Sep 9.

Authors

Zaid N Al Shirity¹, Niels Westra², Kai van Hateren², Thijs H Oude Munnink², Jos G W Kosterink³, Paola Mian², Marjolijn N Lub-de Hooge⁴, Daan J Touw⁵, Bahez Gareb²

Affiliations

¹ Department of Clinical Pharmacy and Pharmacology, University Medical Centre Groningen, University of Groningen, Groningen, the Netherlands. Electronic address: z.al.shirity@umcg.nl.
² Department of Clinical Pharmacy and Pharmacology, University Medical Centre Groningen, University of Groningen, Groningen, the Netherlands.
³ Department of Clinical Pharmacy and Pharmacology, University Medical Centre Groningen, University of Groningen, Groningen, the Netherlands; Department of PharmacoTherapy, -Epidemiology and -Economics, Groningen Research Institute of Pharmacy, University of Groningen, Groningen, the Netherlands.
⁴ Department of Clinical Pharmacy and Pharmacology, University Medical Centre Groningen, University of Groningen, Groningen, the Netherlands; Nuclear Medicine and Molecular Imaging, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.
⁵ Department of Clinical Pharmacy and Pharmacology, University Medical Centre Groningen, University of Groningen, Groningen, the Netherlands; Pharmaceutical Analysis, Groningen Research Institute of Pharmacy, University of Groningen, Groningen, the Netherlands.

PMID: 37716342
DOI: 10.1016/j.jchromb.2023.123872

Abstract

Kinase inhibitors have revolutionized cancer treatment in the past 25 years and currently form the cornerstone of many treatments. Due to the increasing evidence for therapeutic drug monitoring (TDM) of kinase inhibitors, the need is growing for new assays to rapidly evaluate kinase inhibitor plasma concentrations. In this study, we developed an LC-MS/MS assay for the rapid and simultaneous quantification of 21 kinase inhibitors. First, a literature search was conducted to ensure that the linear ranges of the analytes were in line with the reported therapeutic windows and/or TDM reference values. Subsequently, the assay was validated according to FDA and EMA guidelines for linearity, selectivity, carry-over, accuracy, precision, dilution integrity, matrix effect, recovery, and stability. The assay was fast, with a short run-time of 2 min per sample. Sample pre-treatment consisted of protein precipitation with methanol enriched with stable isotope-labeled internal standards (SIL-IS), and the mixture was vortexed and centrifuged before sample injection. Separation was achieved using a C18 column (3 μm,50 × 2.1 mm) with a gradient of two mobile phases (ammonium formate buffer pH 3.5 and acetonitrile). Analyte detection was conducted in positive ionization mode using selected reaction monitoring. The assay was accurate and precise in plasma as well as in serum. Extraction recovery ranged between 95.0% and 106.0%, and the matrix effect was 95.7%-105.2%. The stability of the analytes varied at room temperature and in refrigerated conditions. However, all drugs were found to be stable for 7 days in the autosampler. The clinical applicability of the analytical method (486 analyzed samples between 1 July 2022-1 July 2023) as well as external quality control testing results were evaluated. Taken together, the results demonstrate that the analytical method was validated and applicable for routine analyses in clinical practice.

Keywords: Bioanalysis; Kinase Inhibitors; LC-MS/MS; Therapeutic Drug Monitoring; Tyrosine Kinase Inhibitors; Validation.

MeSH terms

Chromatography, High Pressure Liquid / methods
Chromatography, Liquid / methods
Drug Monitoring* / methods
Humans
Protein Kinase Inhibitors
Reproducibility of Results
Tandem Mass Spectrometry* / methods

Substances

Protein Kinase Inhibitors