A UHPLC-MS/MS bioanalytical assay for the determination of BMS-911543, a JAK2 inhibitor, in human plasma.
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.
Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton NJ 08543, United States.
Jane
J
Liu
Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton NJ 08543, United States. Electronic address: long.yuan@bms.com.
Long
L
Yuan
Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton NJ 08543, United States.
Guowen
G
Liu
Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton NJ 08543, United States.
Jim X
JX
Shen
Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton NJ 08543, United States.
Anne-Françoise
AF
Aubry
Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton NJ 08543, United States.
Mark E
ME
Arnold
Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton NJ 08543, United States.
Qin C
QC
Ji
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006575:Heterocyclic Compounds, 3-Ring / Q000097:blood*; D006801:Humans; D053614:Janus Kinase 2 / Q000037:antagonists & inhibitors*; D057230:Limit of Detection; D059625:Liquid-Liquid Extraction / Q000379:methods; D047428:Protein Kinase Inhibitors / Q000097:blood*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
D006575:Heterocyclic Compounds, 3-Ring; C571029: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; D047428:Protein Kinase Inhibitors; C507924:JAK2 protein, human; D053614:Janus Kinase 2
96-Well format; Automation; High-throughput; LC–MS/MS; Liquid–liquid extraction; Quantitation