A new method using high performance liquid chromatography coupled with electrospray mass spectrometry is described for the quantification of PBMC concentration of tyrosine kinase inhibitors imatinib, dasatinib and nilotinib. A simple PBMC isolation and extraction procedure were applied on 10-14 mL of blood aliquots. Chromatographic separation of drugs and Internal Standard (quinoxaline) was achieved with a gradient (acetonitrile and water+formic acid 0.05%) on a C18 reverse phase analytical column with 25 min of analytical run, at flow rate of 0.25 mL/min. Mean intra- and inter-day precision for all compounds were 8.76 and 12.20%; mean accuracy was -3.86%; extraction recovery ranged within 79 and 91%. Calibration curves ranged from 50.0 to 0.25 ng. The limit of quantification was set at 0.25 ng for all the analyzed drugs. This novel developed methodology allows a specific, sensitive and reliable simultaneous intracellular determination of the three tyrosine kinase inhibitors imatinib, dasatinib and nilotinib in a single chromatographic run, useful for drugs estimation in PBMC of patients affected by chronic myeloid leukemia.

HPLC-MS method for the simultaneous quantification of the antileukemia drugs imatinib, dasatinib and nilotinib in human peripheral blood mononuclear cell (PBMC).

D'AVOLIO, ANTONIO
First
;
SIMIELE, MARCO;DE FRANCIA, SILVIA;ARIAUDO, ALESSANDRA;BAIETTO, LORENA;CUSATO, JESSICA;FAVA, Carmen;SAGLIO, Giuseppe;DI CARLO, Francesco;DI PERRI, Giovanni
Last
2012

Abstract

A new method using high performance liquid chromatography coupled with electrospray mass spectrometry is described for the quantification of PBMC concentration of tyrosine kinase inhibitors imatinib, dasatinib and nilotinib. A simple PBMC isolation and extraction procedure were applied on 10-14 mL of blood aliquots. Chromatographic separation of drugs and Internal Standard (quinoxaline) was achieved with a gradient (acetonitrile and water+formic acid 0.05%) on a C18 reverse phase analytical column with 25 min of analytical run, at flow rate of 0.25 mL/min. Mean intra- and inter-day precision for all compounds were 8.76 and 12.20%; mean accuracy was -3.86%; extraction recovery ranged within 79 and 91%. Calibration curves ranged from 50.0 to 0.25 ng. The limit of quantification was set at 0.25 ng for all the analyzed drugs. This novel developed methodology allows a specific, sensitive and reliable simultaneous intracellular determination of the three tyrosine kinase inhibitors imatinib, dasatinib and nilotinib in a single chromatographic run, useful for drugs estimation in PBMC of patients affected by chronic myeloid leukemia.
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109
116
http://dx.doi.org/10.1016/j.jpba.2011.10.003
Antineoplastic Agents; blood, Calibration, Cells; Cultured, Chromatography; High Pressure Liquid; methods, Humans, Leukemia; Myelogenous; Chronic; BCR-ABL Positive; drug therapy, Leukocytes; Mononuclear; chemistry, Limit of Detection, Mass Spectrometry; methods, Piperazines; blood/therapeutic use, Protein-Tyrosine Kinases; antagonists /&/ inhibitors, Pyrimidines; blood/therapeutic use, Reference Standards, Reproducibility of Results, Thiazoles; blood/therapeutic use
A. D'Avolio;M. Simiele;S. De Francia;A. Ariaudo;L. Baietto;J. Cusato;C. Fava;G. Saglio;F. Di Carlo;G. Di Perri
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2318/111970
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