HPLC-MS method for the simultaneous quantification of the antileukemia drugs imatinib, dasatinib and nilotinib in human peripheral blood mononuclear cell (PBMC).
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.
Laboratory of Clinical Pharmacology and Pharmacogenetics, Department of Infectious Diseases, University of Torino, Amedeo di Savoia Hospital, Turin, Italy. antonio.davolio@unito.it
Antonio
A
D'Avolio
Marco
M
Simiele
Silvia
S
De Francia
Alessandra
A
Ariaudo
Lorena
L
Baietto
Jessica
J
Cusato
Carmen
C
Fava
Giuseppe
G
Saglio
Francesco
F
Di Carlo
Giovanni
G
Di Perri
D000970:Antineoplastic Agents / Q000097:blood*; D001549:Benzamides; D002138:Calibration; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D000069439:Dasatinib; D006801:Humans; D000068877:Imatinib Mesylate; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000188:drug therapy; D007963:Leukocytes, Mononuclear / Q000737:chemistry*; D057230:Limit of Detection; D013058:Mass Spectrometry / Q000379:methods*; D010879:Piperazines / Q000097:blood* / Q000627:therapeutic use; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D011743:Pyrimidines / Q000097:blood* / Q000627:therapeutic use; D012015:Reference Standards; D015203:Reproducibility of Results; D013844:Thiazoles / Q000097:blood* / Q000627:therapeutic use
D000970:Antineoplastic Agents; D001549:Benzamides; D010879:Piperazines; D011743:Pyrimidines; D013844:Thiazoles; D000068877:Imatinib Mesylate; D011505:Protein-Tyrosine Kinases; C498826:nilotinib; D000069439:Dasatinib