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目的:建立人血浆中咪达唑仑、1-羟基咪达唑仑、奥美拉唑和5-羟基奥美拉唑浓度的液相色谱-串联质谱(LC-MS/MS)测定方法,并将其应用于临床试验中咪达唑仑和奥美拉唑及其代谢产物血药浓度的测定。方法:以地西泮为内标,人血浆样品经醋酸乙酯萃取处理后,进行LC-MS/MS分析。采用Agilent C18(4.6 mm×150 mm,5μm)色谱柱,流动相为甲醇-0.015%氨水溶液(甲酸调pH8.20)(80∶20);使用电喷雾离子源(ESI),正离子多反应监测(MRM)模式检测,用于定量分析的咪达唑仑、1-羟基咪达唑仑、奥美拉唑、5-羟基奥美拉唑浓度和内标地西泮的监测离子分别为m/z:326.4→291.2,m/z:342.2→324.0,m/z:346.1→198.1,m/z:362.2→214.0和m/z:285.2→193.1。结果:咪达唑仑、1-羟基咪达唑仑、奥美拉唑和5-羟基奥美拉唑在其线性范围内线性关系良好(r均大于0.99),定量下限分别为0.25,0.15,1.00,0.50 ng·ml-1上。批内精密度(RSD)和批间精密度(RSD)均不大于11.05%,提取回收率稳定。结论:本方法特异性强,灵敏度高,测定结果可靠,适用于临床试验中血浆样本的高通量分析。
Objective: To establish a method for the determination of midazolam, 1-hydroxy midazolam, omeprazole and 5-hydroxy-omeprazole in human plasma by liquid chromatography-tandem mass spectrometry (LC-MS / MS) Its clinical application of midazolam and omeprazole and their metabolites blood concentration determination. Methods: With diazepam as internal standard, human plasma samples were extracted with ethyl acetate and analyzed by LC-MS / MS. The mobile phase consisted of methanol-0.015% aqueous ammonia (formic acid, pH 8.20) (80:20) using an Agilent C18 (4.6 mm × 150 mm, 5 μm) column. Electrospray ionization (ESI) Monitoring (MRM) pattern detection, midazolam, 1-hydroxy midazolam, omeprazole, 5-hydroxy omeprazole concentrations and internal standard diazepam for quantitative analysis were m /z:326.4→291.2,m/z:342.2→324.0,m/z:346.1→198.1,m/z:362.2→214.0 and m / z: 285.2 → 193.1. Results: Midazolam, 1-hydroxy midazolam, omeprazole and 5-hydroxy-omeprazole showed good linearity (r> 0.99) in the linear range. The lower limits of quantitation were 0.25, 0.15, 1.00, 0.50 ng · ml-1. The intra-assay precision (RSD) and inter-batch precision (RSD) were all no greater than 11.05%, and the extraction recovery was stable. Conclusion: The method is specific, sensitive and reliable. It is suitable for high-throughput analysis of plasma samples in clinical trials.