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Published on: December 7, 2014
Validated LC-MS/MS Method for the Quantitative Determination and Pharmacokinetic Profiling of Fedratinib (TG101348),
Rajesh Kumar Boggavarapu1, Vishwottam Kandikere2, Jithendra Chimakurthy1
1Department of Pharmaceutical Sciences, Vignan's Foundation for Science, Technology and Research, Guntur, Andhra Pradesh, India.
Abstract:
Fedratinib, a selective Janus kinase 2 (JAK2) inhibitor approved by the FDA for the treatment of myeloproliferative neoplasms, particularly myelofibrosis (MF), has clinical relevance. Despite this, no liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been reported for its quantification in biological matrices. This study aimed to develop and validate an LC-MS/MS method for quantifying fedratinib in mice plasma and assessing its pharmacokinetics. The method employed a Sciex 4500 triple quadrupole mass spectrometer with a Shimadzu LC system in positive ionization mode. Chromatographic separation was achieved using a Phenomenex Kinetex C18 column (2.1 × 50 mm, 5.0 μm) with a gradient mobile phase of 5 mM ammonium formate in 0.1% formic acid and acetonitrile at a flow rate of 0.8 mL/min. Telmisartan was used as the internal standard (IS). The method was validated in accordance with USFDA M10 bioanalytical guidelines. Fedratinib and IS were detected at m/z 525.5 → 468.9 and m/z 515.2 → 276.0, respectively. The assay demonstrated excellent sensitivity (LLOQ: 0.5 ng/mL), linearity (0.5 to 1000.0 ng/mL, r2 = 0.99), and accuracy (95.80-104.52%). Stability studies confirmed the analyte's integrity across multiple freeze-thaw cycles. The developed LC-MS/MS method is selective, sensitive, fully validated, and was successfully applied to pharmacokinetic studies.
Insights
A new liquid chromatography-tandem mass spectrometry (LC-MS/MS) method quantifies fedratinib in mouse plasma. This validated assay is crucial for pharmacokinetic studies of fedratinib, a JAK2 inhibitor for myelofibrosis.
Area of Science:
- Pharmacology and Toxicology
- Analytical Chemistry
- Biomedical Science
Background:
- Fedratinib is an FDA-approved Janus kinase 2 (JAK2) inhibitor for myeloproliferative neoplasms, particularly myelofibrosis.
- Accurate quantification of fedratinib in biological matrices is essential for pharmacokinetic studies.
- No validated LC-MS/MS method for fedratinib quantification in biological samples was previously reported.
Purpose of the Study:
- To develop and validate a sensitive and selective LC-MS/MS method for quantifying fedratinib in mouse plasma.
- To assess the pharmacokinetic profile of fedratinib using the developed method.
Main Methods:
- A Sciex 4500 triple quadrupole mass spectrometer coupled with a Shimadzu LC system was used.
- Chromatographic separation was performed on a Phenomenex Kinetex C18 column using a gradient mobile phase.
- Telmisartan was employed as the internal standard (IS), and the method was validated per USFDA M10 guidelines.
Main Results:
- The method exhibited high sensitivity with a lower limit of quantification (LLOQ) of 0.5 ng/mL.
- Excellent linearity (r² = 0.99) over the range of 0.5 to 1000.0 ng/mL and high accuracy (95.80-104.52%) were achieved.
- The method demonstrated selectivity, sensitivity, and stability, proving suitable for pharmacokinetic analysis.
Conclusions:
- A robust and fully validated LC-MS/MS method for fedratinib quantification in mouse plasma was successfully developed.
- This method is suitable for supporting pharmacokinetic studies of fedratinib in preclinical research.
- The validated assay provides a critical tool for understanding fedratinib's behavior in biological systems.
