Bioequivalance and pharmacokinetic study of febuxostat in human plasma by using LC-MS/MS with liquid liquid extraction method

Abstract

A bioequivalence study was proved of generic Febuxostat 80 mg tablets (T) in healthy volunteers.For this purpose, Authors developed a simple, sensitive, selective, rapid, rugged and reproducible liquid chromatography–tandem mass spectrometry method for the quantification of Febuxostat (FB) in human plasma using Febuxostat D7 (FBD7) as an internal standard (IS) was used. Chromatographic separation was performed on Ascentis Express C18 (50x4.6 mm, 3.5 μ) column. Mobile phase composed of 10 mM Ammonium formate: Acetonitrile (20:80 v/v), with 0.8 mL/min flow-rate. Drug and IS were extracted by Liquid- liquid extraction. FB and FBD7 were detected with proton adducts at m/z 317.1→261.1 and 324.2→262.1 in multiple reaction monitoring (MRM) positive mode respectively. The method was validated with the correlation coefficients of (r2) ≥ 0.9850 over a linear concentration range of 1.00-8000.00 ng/mL. This method demonstrated intra and inter-day precision within 2.64 to 3.88 and 2.76 to 8.44% and accuracy within 97.33 to 99.05 and 100.30 to 103.19% for FB. This method is successfully applied in the Bioequivalence study of 9 human volunteers.

Simultaneous determination of febuxostat and its three active metabolites in human plasma by liquid chromatography–tandem mass spectrometry and its application to a pharmacokinetic study in Chinese healthy volunteers

“…HPLC with fluorescence detection (HPLC-FD) [7,8] and LC-MS/MS [9][10][11][12] methods have been previously developed to determine febuxostat in human plasma for pharmacokinetic study of febuxostat. The quantification of metabolites 67M-1, 67M-2 and 67M-4 in human plasma has been accomplished by two groups of researchers using LC-MS/MS methods [6,8].…”

Ultra-High Performance Liquid Chromatographic Determination of Genotoxic Impurities in Febuxostat Drug Substance and Products

“…Some developed methods for the quantification of febuxostat and its related compounds in the bulk and formulated products by UPLC [3,[8][9][10][11][12][13][14]26]. These previously published research articles were failed to explain about the probable genotoxic impurities in febuxostat.…”

“…Other subjects were explained about the determination of febuxostat was performed by liquid chromatograph coupled mass spectrometer (LCMS/MS) in human plasma [3][4][5][6], by LC-ultraviolet (UV) [7], by HPLC [8][9][10], by UPLC [11][12][13], and by UPLC/MS in dog plasma [14]. The related compounds of febuxostat were separated and determined at the level 0.10% w/w level or 1000 µg/ml level by UPLC in literature [13].…”

Febuxostat mitigates genotoxicity induced by ionizing radiation in human normal lymphocytes

“…One hundred microliter solutions of FB at final concentrations of 1, 10, 50, or 100 μM (6, 7) were added to blood samples. The plasma concentration of FB was 10-12 μM after oral ingestion of FB at a dose of 80 mg in healthy humans (8), whereas it was 15-25 μM at an oral dose of 120 mg (9, 10). FB as powder was dissolved in dimethyl sulfoxide (DMSO) as solvent and diluted in RPMI culture medium.…”