【作者】 李平；

【导师】 陈笑艳； 钟大放；

【作者基本信息】 沈阳药科大学 ， 药物分析学， 2007， 硕士

【摘要】 目的建立灵敏、专属、快速的LC／MS／MS法，测定血浆中的纳美芬和阿扑吗啡，并分别用于二者低剂量制剂的临床药动学研究。方法测定人血浆中纳美芬浓度时，200μL血浆样品经沉淀蛋白处理后，以甲醇-5mmol／L醋酸铵（75：25，v／v）为流动相，经Zorbax XDB C₁₈柱分离，采用API 4000型三重四极杆串联质谱仪电喷雾电离源，正离子方式检测，多反应监测（MRM），用于定量分析的离子反应分别为m／z340→m／z268 （纳美芬）和m／z286→m／z 185 （内标，氢吗啡酮）。测定人血浆中阿扑吗啡时，200μL血浆样品经液-液萃取处理后，以乙腈-水-甲酸（40：60：0.1，v／v／v）为流动相，采用Zorbax XDB C₈柱分离，采用TSQ Quantum Ultra型三重四极杆串联质谱仪电喷雾电离源，正离子方式检测，选择反应监测（SRM），用于定量分析的离子反应分别为m／z 268→m／z（237+219）（阿扑吗啡）和m／z 286→m／z 185（内标，氢吗啡酮）。结果测定纳美芬的线性范围为10.0～5000pg／mL，定量下限为10.0 pg／mL。日内、日间精密度（RSD）小于10.1％，准确度（RE）在-3.4％至1.8％范围内。测定阿扑吗啡的线性范围为0.010～20.0ng／mL，定量下限为0.010 ng／mL。日内、日间精密度（RSD）小于7.9％，准确度（RE）在-1.7％至1.0％范围内。结论本文所建立的LC／MS／MS法灵敏、专属、快速，可用于纳美芬和阿扑吗啡的血药浓度测定及二者低剂量制剂的临床药物动力学研究。更多还原

【Abstract】 OBJECTIVE To develop and validate sensitive, rapid and specific liquid chromatographic/tandem mass spectrometric methods for quantitative analyses of nalmefene and apomorphine in human plasma and application to pharmacokinetic studies.METHODS After a simple one-step protein precipitation with methanol, nalmefene and the internal standard hydromorphone were chromatographed on a Zorbax XDB C₁₈ column and detected by MS/MS with a Turbo Ionspray interface on an MDS SCIEX API 4000 tandem mass spectrometer. Multiple reaction monitoring （MRM） adopting the precursor to product ion transitions of m/z 340→m/z 268 and m/z 286→m/z 185 was used to quantify nalmefene and the internal standard, respectively. Apomorphine was first prepared from plasma samples by liquid-liquid extraction, and then separated with the endogenous interferences on a Zorbax XDB C₈ column. The mobile phase consisted of acetonitrile-water-formic acid （40: 60: 0.1, v/v/v） at a flow rate of 0.50 mL/min. A TSQ Quantum Ultra triple quadrupole mass spectrometer, equipped with an electrospray ionization interface, was operated in the positive ion mode. Selective reaction monitoring （SRM） using the precursor to product ion combinations of m/z 268→m/z （237 + 219） and m/z 286→m/z 185 was employed to determine apomorphine and the internal standard hydromorphone, respectively.RESULTS The linear calibration curve was obtained in the concentration range from 10.0 to 5000 pg/mL for nalmefene. The intra- and inter-run precisions determined from QC samples, expressed as relative standard deviation （RSD）, were no more than 6.0% and 10.1%, respectively, while accuracy ranged from -3.4% to 1.8% in terms of relative error （RE）. The standard calibration curve for apomorphine was linear in the concentration range from 0.010 to 20.0 ng/mL in human plasma. Determined from QC samples for apomorphine, the intra- and inter-run precisions were less than 5.6% and 7.9%, respectively, and the accuracy was from -1.7% to 1.0%.CONCLUSION The validated methods were successfully applied to evaluate the clinical pharmacokinetics of low dose of nalmefene and apomorphine.更多还原