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.
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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)
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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
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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)
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