摘要
本文有两部分组成。
一、液相色谱-串联质谱法测定血浆中的替加色罗
目的：建立测定人血浆中的替加色罗的快速、灵敏的液相色谱-串联质谱法，并应用于药物动力学研究以及制剂人体生物利用度和生物等效性研究。方法：200μL含替加色罗的血浆样品加入内标苯海拉明，经液一液萃取处理后，以流动相为乙腈：5 mmol／L醋酸铵溶液：甲酸(70：30：0.01，v／v／v)溶解，采用Zorbax XDB-C_(18)柱分离；通过电喷雾离子化四极杆串联质谱，以多反应监测(MRM)方式进行正离子检测，用于监测定量的离子反应分别为m／z 302→m／z 173 (替加色罗)和m／z256→m／z 167(内标苯海拉明)；药动学参数采用非室模型计算。结果：替加色罗测定方法的线性范围分别为0.010～10.0 ng／mL。日内、日间精密度均小于15％，准确度在±15％以内。每个样品测试时间均少于3.0 min。应用上述方法研究比较了健康受试者分别单剂量口服替加色罗6 mg后的药物动力学特点，进行了药物制剂的人体生物利用度和生物等效性研究。结论：本文所建立的定量方法选择性强，灵敏度高，操作简便，血浆用量少，适用于替加色罗制剂的生物等效性评价及临床药动学研究。
二、液相色谱-串联质谱法测定血浆中的异丙嗪
目的：建立测定人血浆中的异丙嗪的快速、灵敏的液相色谱-串联质谱法，并应用于药物动力学研究以及制剂人体生物利用度和生物等效性研究。方法：200μL含异丙嗪的血浆样品加入内标氯雷他定，经乙腈沉淀蛋白处理后，以乙腈-水-甲酸(50：50：0.1，v／v／v)为流动相，采用Zorbax XDB-C_(18)柱分离；通过电喷雾离子化四极杆串联质谱，以多反应监测(MRM)方式进行正离子检测，用于监测定量的离子反应分别为m／z 285→m／z 86(异丙嗪)和m／z 383→m／z 267(内标氯雷他定)；药动学参数采用非室模型计算。结果：异丙嗪测定方法的线性范围分别为0.050～20.0 ng／mL。日内、日间精密度均小于15％，准确度在±15％以内。每个样品测试时间均少于4.0 min。应用上述方法研究比较了健康受试者单剂量口服含异丙嗪12.5mg复方制剂后的药物动力学特点，进行了药物制剂的人体生物利用度和生物等效性研究。
结论：本文所建立的定量方法选择性强，灵敏度高，操作简便，血浆用量少，适用于含异丙嗪复方制剂的生物等效性评价及临床药动学研究。
Two rapid, sensitive and specific LC/MS/MS methods for quantitative analyses of tegaserod and promethazine in plasma were developed and validated.
1．Determination of tegaserod in plasma by LC/MS/MS
Objective: To develop sensitive and specific LC/MS/MS methods for respective determination of tegaserod in human plasma and to study the bioequivalences of different formulations containing tegaserod and clinical investigations of their pharmacokinetics. Method: For determination of the plasma concentration of tegaserod, an aliquot of 200μL plasma was treated by liquid-liquid extraction, then the analytes of interest were analyzed on a Zorbax XDB-C_(18) column with the mobile phase consisted of acetonitrile-5 mmol/L ammonium acetate-formic acid (70: 30: 0．01, v/v/v). A API-4000 tandem mass spectrometer equipped with electrospray ionization source was used as detector and was operated in the positive ion mode. Multiple reaction monitoring (MRM) using the precursor→product ion combination of m/z 302→m/z 173 and m/z 256→m/z 167 was used to respectively quantify tegaserod and diphenhydramine. Results: The linear calibration curves were obtained in the concentration range of 0．010-10．0 ng/mL for tegaserod. The intra-and inter-day relative standard deviation (RSD) across three validation runs over the entire concentration range was less than 15%. Accuracy determined at three QC concentrations was within±15% as terms of relative error (RE). Each plasma sample was chromatographed within 3．0 min. Conclusion: The method were sensitive and convenient, and proved to be suitable for bioequivalence evaluations of different formulations containing tegaserod.
2．Determination of promethazine in plasma by LC/MS/MS
Objective: To develop sensitive and specific LC/MS/MS methods for respective determination of promethazine in human plasma and to study the bioequivalences of different formulations containing promethazine and clinical investigations of their pharmacokinetics. Method: For determination of the plasma concentration of promethazine, an aliquot of 200μL plasma was treated by precipitation. The analytes of interest were separated on a Zorbax XDB-C_(18) column with the mobile phase consisting of acetonitrile-water-formic acid (50: 50: 0．1, v/v/v). A API-4000 tandem mass spectrometer equipped with electrospray ionization source was used as detector and was operated in the positive ion mode. Multiple reaction monitoring (MRM) using the precursor→product ion combination of m/z 285→m/z 86 and m/z 383→m/z 267 was used to respectively quantify promethazine and loratadine. Results: The linear calibration curves were obtained in the concentration range of 0．050-20．0 ng/mL for promethazine. The intra-and inter-day relative standard deviation (RSD) across three validation runs over the entire concentration range was less than 15%. Accuracy determined at three QC concentrations was within±15% as terms of relative error (RE). Each plasma sample was chromatographed within 4．0 min. Conclusion: The method were sensitive and convenient, and proved to be suitable for bioequivalence evaluations of different formulations containing promethazine.
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