Determination of febuxostat concentration in human plasma by UPLC-MS/MS method and pharmacokinetic application

ZHANG Mei-juan; ZHANG Jian-xiong; LI Jiang-shuo; YIN Hang; LI Xin-gang; DONG Rui-hua

doi:10.3969/j.issn.1672-8157.2023.04.015

Abstract：Objective:To establish a method using ultra high performance liquid chromatography-mass spectrometry(UPLC-MS/MS)for the determination of febuxostat in human plasma,and apply to clinical pharmacokinetic research.Methods:Plasma samples were extracted via protein precipitation with acetonitrile diclofenac sodium was selected as the internal standard(IS).Chromatographic separation was accomplished on a Shimadzu Shim-pack GIST C18 column(2.1 mm×100 mm,2 μm).Febuxostat was separated with aqueous 0.01%formic acid containing 5 mmol·L-1 ammonium formate as mobile phase A,and acetonitrile as mobile phase B.The flow rate was 0.4 mL·min-1,the column temperature was maintained at 40℃.Electrospray ion source in positive and multiple reaction monitoring mode,the quantitative LC-MS/MS was performed with MRM mode using target ions at m/z 317.1→m/z 261.0 for febuxostat and m/z 296.0→m/z 214.0 for IS,respectively.Results:The linear range of febuxostat was 10-8000 ng·mL-1(r>0.999)and showed good linearity.For febuxostat,the intra-and inter-day RSDs were from 4.30%to 8.28%,the relative error was from-2.15%to 6.03%,the relative recovery ranged from 87.24%to 97.77%,the RSDs of matrix effects were lower than 15%.The established method was applied to detect the blood concentration of 16 subjects after received febuxostat.The AUC0-t was(16170.61±2949.47)ng·mL-1·h,AUC0-∞ was(16431.17±2995.22)ng·mL-1·h,Cmax was(4946.25±1439.39)ng·mL-1,Tmax was(2.10±1.51)h,t1/2 was(6.05±1.86)h.Conclusion:The method is simple,rapid and accurate,and has good selectivity,which is suitable for the blood concentration determination and pharmacokinetic study of febuxostat.