Development and validation of LC-MS/MS method for imatinib and norimatinib monitoring by finger-prick DBS in gastrointestinal stromal tumor patients

Autoři: Valentina Iacuzzi aff001;  Bianca Posocco aff001;  Martina Zanchetta aff001;  Marcella Montico aff004;  Elena Marangon aff001;  Ariana Soledad Poetto aff001;  Mauro Buzzo aff001;  Sara Gagno aff001;  Angela Buonadonna aff006;  Michela Guardascione aff001;  Bruno Casetta aff001;  Giuseppe Toffoli aff001
Působiště autorů: Experimental and Clinical Pharmacology Unit, Centro di Riferimento Oncologico di Aviano (CRO) IRCCS, Aviano, Italy aff001;  Doctoral School in Nanotechnology, University of Trieste, Trieste, Italy aff002;  Department of Chemical and Pharmaceutical Sciences, University of Trieste, Trieste, Italy aff003;  Scientific Directorate, Centro di Riferimento Oncologico di Aviano (CRO) IRCCS, Aviano, Italy aff004;  Doctoral School in Pharmacological Sciences, University of Padua, Padua, Italy aff005;  Medical Oncology Department, Centro di Riferimento Oncologico (CRO) IRCCS, Aviano, Italy aff006;  Polo Tecnologico Pordenone, Pordenone, Italy aff007
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: 10.1371/journal.pone.0225225


The introduction of imatinib, an oral tyrosine kinase inhibitor, as first-line standard therapy in patients with unresectable, metastatic, or recurrent gastro-intestinal stromal tumor (GIST), strongly improved their treatment outcomes. However, therapeutic drug monitoring (TDM) is recommended for this drug due to the large inter-individual variability in plasma concentration when standard dose is administered. A Cmin higher than 760 ng/mL was associated with a longer progression free survival. Thus, a LC-MS/MS method has been developed and fully validated to quantify imatinib and its active metabolite, norimatinib, in finger-prick dried blood spot (DBS). The influence of hematocrit, sample homogeneity, and spot size and the correlation between finger-prick and venous DBS measurements were also assessed. The method showed a good linearity (R2 > 0,996) between 50–7500 ng/mL for imatinib and 10–1500 ng/mL for norimatinib. Analytes were extracted from DBS samples by simply adding to 3 mm-discs 150 μL of acidified methanol containing IMA-D8. The collected extract was then injected on a LC Nexera system in-house configured for the on-line cleanup, coupled with an API-4000 QT. The chromatographic separation was conducted on a Synergi Fusion-RP column (4 μm, 2x50 mm) while the trapping column was a POROS R1/20 (20 μm, 2x30 mm). The total run time was 8.5 min. DBSs stored at room temperature in plastic envelopes containing a silica-gel drying bag were stable up to 16 months.

The proposed method was applied to 67 clinical samples, showing a good correlation between patients’ finger-prick DBS and plasma concentrations, measured by the reference LC-MS/MS method, internally validated. Imatinib and norimatinib concentrations found in finger-prick DBS were adjusted by hematocrit or by an experimental correction factor to estimate the corresponding plasma measurements. At the best of our knowledge, the proposed LC-MS/MS method is the first analytical assay to measure imatinib and norimatinib in DBS samples.

Klíčová slova:

Blood – Blood plasma – Drug administration – Filter paper – Hematocrit – Quality control – Formic acid – Therapeutic drug monitoring


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2019 Číslo 11