液相色谱-串联质谱法测定人血浆中非布司他的浓度

Determination of febuxostat in human plasma by LC-MS/MS

目的建立快速、灵敏的液相色谱-串联质谱法测定人血浆中非布司他的浓度。方法 10名健康受试者单剂量口服试验药物1片(含非布司他80 mg)后,在0~48h内不同时间点分别采集血样,分离血浆,以甲氨蝶呤为内标,血浆样品经乙腈沉淀;采用Agilent Zorbax C_(18)色谱柱(2.1 mm×100 mm,3.5μm),流动相为乙腈:水(含0.1%甲酸)=85:15(V/V),流速为0.25 mL·min^(-1);电喷雾离子源,正离子电离模式,选择性反应监测扫描方式,非布司他和甲氨蝶呤检测的离子对分别为m/z 317.6→m/z261.9和m/z 455.3→m/z 308.7。结果非布司他在5~5 000μg·L^(-1),浓度范围内有良好的线性关系(r^2=0.998 2),定量下限为5μg·L^(-1),准确度及精密度良好。受试者单剂量口服1片非布司他片后,测得非布司他的ρ_(max)、t_(max)、AUC_(0-48h)和t_(1/2)分别为(4 745.6±136.0)μg·L^(-1)、(1.1±0.3)h、(18 561.1±343.6)μg·h·L^(-1)和(13.7±0.7)h。结论本方法快速、灵敏、准确,可用于人血浆中非布司他的浓度检测和人体药动学研究。

AIM To establish a rapid and sensitive LC-MS/MS method for plasma febuxostat measurement in humans. METHODS The pharmacokinetic characteristics of the febuxostat tablet （febuxostat 80 mg） were investigated in 10 healthy male volunteers after single oral dose administration. Serial blood samples were collected in heparinized tubes at given time points during 0 - 48 h after drug administration, and the supernatants were separated with centrifugation. Methotrexate was used as the internal standard and plasma samples were treated with acetonitrile for precipitation. The column was Agilent Zorbax C^s （2.1 mm x 100 mm, 3.5 μm ）, and the mobile phase consisted of acetonitrile-water （ 0.1% formic acid） （85 ： 15, V/V） at a flow rate of 0.25 mL.min^-1. ESI and SRM were used with positive ions scans. The mass transition pairs of m/z 317.6→ m/z 261.9 and m/z 455.3 →m/z 308.7 were used to detect febuxostat and methotrexate, respectively. RESULTS The standard curve of febuxostat was linear over the concentration range of 5 - 5 000 μg.L^-1 （ r^2 = 0.998 2）. The lower limit of quantification was 5 μg.L^-1. This method has acceptable precision and accuracy. The plasma pharmacokinetic parameters of pmax, tmax AUC0-48h and tla of febuxostat were （4745.6 ± 136.0）μg. L^-1, （1.1 ± 0.3） h, （18 561.1 ± 343.6） μg.h.L^-1 and （13.7 ± 0.7） h, respectively. CONCLUSION This method is rapid, sensitive and accurate. It is applicable for measuring plasma febuxostat concentration and other clinical pharmaeokineties study.