LC-MS/MS法测定人血清中氟比洛芬含量

LC-MS/MS determination of flurbiprofen in human serum

目的:建立液相色谱-串联质谱法(LC—MS/MS)测定人血清中氟比洛芬的含量。方法:血清经甲醇直接沉淀法去除蛋白质后,以液相色谱分离、电喷雾离子化串联质谱进行检测,氟比洛芬和消炎痛(内标)的 MRM 扫描离子通道 m/z 分别为243.3→198.8,356.3→312.0。二者的保留时间分别为0.9 min,1.6 min。采用 API3000串联质谱仪及 Shimadzu 系列液相色谱仪,Gemini C_(18)柱(50 mm×2.1 mm,5μm),流动相为乙腈-水(18:82,用氨水调 pH 为8.0),流速为0.3 mL·min^(-1),进样体积为3 μL,每个样品的分析时间为3 min。结果:氟比洛芬在0.05～50μg·mL^(-1)范围内线性关系良好(r=0.9995),日内、日间RSD 分别为4.1%～5.5%及4.2%～6.8%,准确度为99.6%～109.5%。结论:本法样品预处理简便快速,检测准确灵敏专一,适用于氟比洛芬药物动力学和生物利用度的研究。

Objective：To establish a liquid chromatography- tandem mass spectrometry（ LC -MS/MS）method for the determination of flurbiprofen in human serum. Method： The serum samples were precipitated with methanol. The drug was determinated by LC -MS/MS using electrospray ionization. Flurbiprofen and its internal standard （IS）indomethacin were detected on multiple reaction monitoring （MRM） by the transitions from the precursor to the product ion （m/z 243.3/198.8 and m/z 356.3/312.0）. The retention time of these two analytes were 0. 9 min and 1.6 min. An API3000 tandem mass spectrometer and an Shimadzu liquid chromatograph were used for all analyses. The analytical column was a Gemini C18 column （50 mm × 2. 1 mm,5 μm）. The mobile phase, acetonitrile - water （ 18： 82） , was used at a flow rate of 0. 3 mL · min^-1. The injection volume was 3 μL and the total run time was 3.0 min. Results ： Calibration curve had good linearity in the range of 0.05 - 50 μg·mL^ -1 （ r = 0. 9995 ）. The intra - and inter- day precision（RSD） were all less than 15% and the accuracy was within 99.6 -101.5%. Conclusion：The established LC - MS/MS method is shown to be sensitive, accurate and simple for the determination of flurbiprofen in human serum. It is suitable for the pharnlacokinetics and bioavailability study of flurbiprofen.