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

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.

Monitoring of imatinib targeted delivery in human leukocytes

“…The correlation of imatinib intracellular concentration in bone marrow cells of CML patients with therapeutic response has also been shown by Zhong et al (2012). Widmer et al (2006) andD'Avolioa et al (2012) observed in PBMĆ s an intra/extracellular concentration ratio of approximately 8, confirming the active uptake of the drug into these cells.…”

Development and validation of an ultra performance liquid chromatography tandem mass method for sildenafil and N-desmethyl sildenafil plasma determination and quantification

“…The extraction procedure based on protein precipitation is simple, rapid and economic; instead of procedures used by other methods that requires an extensive sample preparation: for example, Varshney extracted samples on solid-phase extraction cartridges, which is a long and expensive procedure [23]; other methods use toxic solvents (methy t-butyl ether, diethyl ether) as extraction solutions, that are difficult to use in clinical routine [24,25]. Despite its chemical structure difference to those of the other analytes, the internal standard quinoxaline is cheap and easy to purchase, more than sildenafil-d8 and N-desmethyl sildenafil-d8 [25]; and it was used in many our other chromatographic published methods [26][27][28][29][30][31][32][33][34][35][36]. Furthermore, its matrix effect and recovery are very close to those of sildenafil and its metabolite, making it a good choice as internal standard.…”

Determination of nilotinib in spiked plasma, urine, and capsules by high-performance liquid chromatography with fluorimetric detection

“…Parise et al described an HPLC-mass spectrometry (MS) method for NTB in plasma and serum [7]. Furthermore, simultaneous assay of NTB with other antichronic myeloid leukemia drugs using solid phase extraction and ultra-performance liquid chromatographytandem mass (UPLC-MS/MS) [8], HPLC-UV [9][10], HPLC-MS/MS [11][12][13][14][15], and HPLC-MS [16,17] have been reported.…”