LC-MS/MS法测定人血浆中布洛芬对映体浓度及其药动学研究

Determination of ibuprofen enantiomers in human plasma by LC-MS/MS in pharmacokinetics study

采用LC-MS/MS法测定人血浆中S-(+)-布洛芬(S-IBP)和R-(-)-布洛芬(R-IBP)的浓度,并应用于健康受试者体内药物动力学研究。以萘普生为内标,采用Daicel公司Chiralpak AD-3R(4.6 mm×150 mm,3.0μm)色谱柱,流动相为乙腈-0.01%甲酸水溶液(40∶60),流速为750μL·min^(-1),每个样品的分析时间为23.0 min,样品经电喷雾离子源负离子化后,通过三重四极杆串联质谱仪,在多反应监测模式下测定S-IBP和R-IBP(m/z205.1→161.0)和内标萘普生(m/z 229.1→185.0)的浓度。血浆样品前处理采用甲醇沉淀蛋白。S-IBP和R-IBP的血浆浓度在0.05~30.00μg·mL^(-1)内线性良好,定量下限为0.05μg·m L^(-1)。S-IBP批内、批间精密度(RSD)均在2.2%~4.2%以内,相对偏差(RE)在-12.0%~13.0%以内。R-IBP批内、批间精密度(RSD)均在2.0%~8.2%以内,相对偏差(RE)在-11.5%~10.6%以内。S-IBP和R-IBP血浆样品室温(25℃)放置6 h,反复冻融(-30℃)3次及冰冻(-30℃)保存47天的情况下均稳定。该分析方法简便、特异性高、灵敏度高,可用于受试者空腹口服布洛芬缓释胶囊300 mg后血浆样品中布洛芬对映体S-IBP和R-IBP的药动学研究。

The study aims to establish an LC-MS/MS method for the determination of S-（＋）-ibuprofen（S-IBP） and R-（-）-ibuprofen（R-IBP）, which may be used subsequently to investigate the pharmacokinetics of ibuprofen enantiomers in healthy Chinese volunteers. Naproxen was used as an internal standard. The separation was achieved on a Chiralpak AD-3R column（4.6 mm × 150 mm, 3.0 μm） with a mobile phase consisting of acetonitrile/0.01% formic acid aqueous solution（40∶60） at a flow rate of 750 μL·min^-1 within 23.0 min. Naproxen and the internal standard were measured by a triple-quadrupole mass spectrometer in negative electron electronic spray ion（ESI） mode using multiple reaction monitoring（MRM）. The extracted ions monitored following MRM transitions were m/z 205.1→161.0 for ibuprofen enantiomers and m/z 229.1→185.0 for the internal standard naproxen. Plasma samples were pretreated through methanol precipitation. The calibration curve of S-IBP and R-IBP in human plasma was linear over the concentration rang of（0.05-30.00） μg·mL^-1. The lower limit of quantitation was 0.05 μg·m L^（-1）. The intra-and inter-run precisions of S-IBP at three quality control levels were within 2.2%-4.2%, the relative deviation of the assay was within-12.0%-13.0%. The intra-and inter-run precisions of R-IBP at three quality control levels were within 2.0%-8.2%, the relative deviation of the assay was within-11.5%-10.6%. The plasma samples were stable at room temperature（25 ℃） for 6 h, at-30 ℃ for 47 days and during three freeze-thaw cycles. The method was proved to be convenient, accurate and sensitive, and suitable for the pharmacokinetics study of ibuprofen enantiomers in healthy Chinesevolunteers after a single oral dose of 300 mg ibuprofen extended-release capsule.