Rapid validated liquid chromatographic method coupled with Tandem mass spectrometry for quantification of nintedanib in human plasma

Hany W Darwish,Mohamed W Attwa,Adnan A Kadi

doi:10.4314/tjpr.v15i11.23

Hany W Darwish, Mohamed W Attwa + Show 1 more

Open Access

https://doi.org/10.4314/tjpr.v15i11.23

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Abstract

Purpose: To develop and validate a fast, sensitive, and simple liquid chromatographic method coupled with tandem mass spectrometry for the determination of the potent tyrosine kinase inhibitor, ninetedanib (NTB) in plasma, utilizing cyclobenzaprine (CBP) as internal standard (IS). Methods: Separation of the two components (NTB and CBP) was performed on a pentafluorophenyl (PFP) reversed phase column (50 × 2 mm, 3μm) at ambient temperature using isocratic elution with acetonitrile-water (60:40, v/v) containing 0.01 M ammonium formate buffer (pH 4.2) at a flow rate of 0.4 mL/min. NTB and CBP were monitored by a triple quadrupole tandem mass spectrometer with electrospray ionization source in the positive ion mode. The current method was validated following the European Medicines Agency (EMA) guidelines Results: The proposed method allowed rapid and specific quantification of NTB in the calibration range of 2 - 150 ng/mL and determination coefficient of ≥ 0.999. Intra- and inter-day accuracy and precision were < 4 % in all cases. Conclusion: The developed procedure is rapid, specific, reliable, and validated for quantification of NTB in human plasma, and thus can be applied efficiently for the analysis of clinical samples containing NTB. Keywords: Nintedanib assay, Cyclobenzaprine, LC-MS/MS, Validation

Highlights

Nintedanib (NTB, Figure 1(a)), as a powerful tyrosine-kinase inhibitor (TKI), acts by targeting numerous pathways for instance fibroblast growth factor receptor (FGFR), vascular endothelial growth factor receptor (VEGFR)-1n and others
This study was devoted for quantification of NTB in spiked human plasma in presence of cyclobenzaprine (CBP; internal standard (IS), Figure 1(b)) in a very short run time (2 min), which permits the analysis of a lot of clinical samples containing NTB in a reasonable time
A rapid and validated LC-MS/MS method has been developed for the quantification of NTB in human plasma with good linearity

Summary

INTRODUCTION

Nintedanib (NTB, Figure 1(a)), as a powerful tyrosine-kinase inhibitor (TKI), acts by targeting numerous pathways for instance fibroblast growth factor receptor (FGFR), vascular endothelial growth factor receptor (VEGFR)-1n and others. This study was devoted for quantification of NTB in spiked human plasma in presence of cyclobenzaprine (CBP; IS, Figure 1(b)) in a very short run time (2 min), which permits the analysis of a lot of clinical samples containing NTB in a reasonable time. Fifty microliter of CBP working standard solution (1 μgmL-1) was added to different aliquots of NTB working standard solution (1 μgmL-1) and diluted to 1 mL with human plasma to prepare ten NTB concentrations: 2, 4, 6 (low quality control; LQC), 10, 30, 50 (medium quality control; MQC), 80, 100 and 120 (high quality control; HQC), and 150 ng mL-1. NTB concentrations in spiked plasma samples were calculated utilizing the corresponding linear regression equation

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Conflict of Interest