A UHPLC-MS/MS bioanalytical assay for the determination of BMS-911543, a JAK2 inhibitor, in human plasma.

Abstract

Herein we report a rapid, accurate and robust UHPLC-MS/MS assay for the quantitation of BMS-911453, a Janus kinase 2 inhibitor under clinical development for the treatment of myeloproliferative disorders, in human plasma. A systematic method development approach was used to optimize the mass spectrometry, chromatography, and sample extraction conditions, and to minimize potential bioanalytical risks. The validated method utilizes stable-isotope labeled (13)C4-BMS-911543 as the internal standard. Liquid-liquid extraction was used for sample preparation. Chromatographic separation was achieved within 2min on a Zorbax Extend-C18 column with an isocratic elution. BMS-911543 and its internal standard were detected by positive ion electrospray tandem mass spectrometry. The assay range was from 1 to 500ng/mL, and the standard curve was fitted with 1/x(2) weighted linear regression. The intra-assay precision was within 5.0% CV and the inter-assay precision was within 2.6% CV. The inter-assay mean accuracy, expressed as percents of theoretical, was between 99.8% and 102.3%. The assay has high recovery (∼80%) and minimal matrix effect (0.95-1.00). BMS-911543 was stable in human plasma for at least 24h at room temperature, 90 days at -20°C, and following three freeze-thaw cycles. The validated method was successfully applied to sample analysis in clinical studies.

• Authors: Jane Liu, Long Yuan, Guowen Liu, Jim X Shen, Anne-Françoise Aubry, Mark E Arnold, Qin C Ji
• Journal: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (J Chromatogr B Analyt Technol Biomed Life Sci) Vol. 991 Pg. 85-91 (Jun 01 2015) ISSN: 1873-376X [Electronic] Netherlands
• PMID: 25930207 (Publication Type: Journal Article, Validation Study)
• Copyright: Copyright © 2015 Elsevier B.V. All rights reserved.

Chemical References

• Heterocyclic Compounds, 3-Ring
• N,N-dicyclopropyl-4-((1,5-dimethyl-1H-pyrazol-3-yl)amino)-6-ethyl-1-methyl-1,6-dihydroimidazo(4,5-d)pyrrolo(2,3b)pyridine-7-carboxamide
• Protein Kinase Inhibitors
• JAK2 protein, human
• Janus Kinase 2

Topics

• Chromatography, High Pressure Liquid (methods)
• Heterocyclic Compounds, 3-Ring (blood)
• Humans
• Janus Kinase 2 (antagonists & inhibitors)
• Limit of Detection
• Liquid-Liquid Extraction (methods)
• Protein Kinase Inhibitors (blood)
• Reproducibility of Results
• Tandem Mass Spectrometry (methods)