Abstract
In the current work, a rapid, specific, sensitive and validated liquid chromatography tandem mass-spectrometric method was developed for the quantification of ponatinib (PNT) in human plasma and rat liver microsomes (RLMs) with its application to metabolic stability. Chromatographic separation of PNT and vandetanib (IS) were accomplished on Agilent eclipse plus C18 analytical column (50 mm × 2.1 mm, 1.8 μm particle size) maintained at 21±2°C. Flow rate was 0.25 mLmin-1 with run time of 4 min. Mobile phase consisted of solvent A (10 mM ammonium formate, pH adjusted to 4.1 with formic acid) and solvent B (acetonitrile). Ions were generated by electrospray (ESI) and multiple reaction monitoring (MRM) was used as basis for quantification. The results revealed a linear calibration curve in the range of 5–400 ngmL-1 (r2 ≥ 0.9998) with lower limit of quantification (LOQ) and lower limit of detection (LOD) of 4.66 and 1.53 ngmL-1 in plasma, 4.19 and 1.38 ngmL-1 in RLMs. The intra- and inter-day precision and accuracy in plasma ranged from1.06 to 2.54% and -1.48 to -0.17, respectively. Whereas in RLMs ranged from 0.97 to 2.31% and -1.65 to -0.3%. The developed procedure was applied for quantification of PNT in human plasma and RLMs for study metabolic stability of PNT. PNT disappeared rapidly in the 1st 10 minutes of RLM incubation and the disappearance plateaued out for the rest of the incubation. In vitro half-life (t1/2) was 6.26 min and intrinsic clearance (CLin) was 15.182± 0.477.
Highlights
Ponatinib (PNT; Fig 1), marketed as (IclusigTM tablets), is a drug taken orally for the management of some types of tumers including chronic Philadelphia chromosome—positive acute lymphoblastic leukemia (Ph+ ALL) and myeloid leukemia (CML) [1].Chimeric breakpoint cluster region-abelson (BCR-ABL) protein is responsible for the activity of ABL tyrosine kinase which is considered the main reason of chronic myeloid leukaemia (CML) [2]
General regulations stated by International Conference on Harmonisation (ICH) [9, 10] and Food and Drug Administration (FDA) guidelines for analytical procedures and methods validation [11] were followed for validation purposes
Calibration curves for PNT in spiked rat liver microsomes (RLMs) and plasma matrices were prepared in the same day of analysis at 11 different concentrations
Summary
Ponatinib (PNT; Fig 1), marketed as (IclusigTM tablets), is a drug taken orally for the management of some types of tumers including chronic Philadelphia chromosome—positive acute lymphoblastic leukemia (Ph+ ALL) and myeloid leukemia (CML) [1].Chimeric breakpoint cluster region-abelson (BCR-ABL) protein (encoded by BCR-ABL oncogene) is responsible for the activity of ABL tyrosine kinase which is considered the main reason of chronic myeloid leukaemia (CML) [2]. Some CML cases have recently reported in PLOS ONE | DOI:10.1371/journal.pone.0164967. Validated LC-MS/MS Method for the Quantification of Ponatinib in Plasma doi:10.1371/journal.pone.0164967.g001 the literature resistant to common TKIs (e.g. dasatinib and imatinib). There was only one reported spectrofluorimetric method to quantify PNT in spiked plasma and urine [4]. In this method, the recovery % of PNT in plasma was around 85%. The proposed procedure was fast, sensitive, specific and reproducible. It is considered the first validated LC-MS/MS for assaying PNT. In this study, validated and reliable LCMS-MS assay for the analysis of PNT in plasma and RLMs is described. To the best of the authors knowledge, the method is the first LC-MS based method of its kind
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