Pilli Nageswara Rao, Inamadugu Jaswanth Kumar, Mullangi Ramesh, Karra Vijaya Kumari, Vaidya Jayathirtha Rao, Rao J V L N Seshagiri
University College of Pharmaceutical Sciences, Jawaharlal Nehru Technological University, Kukatpally, Hyderabad, India.
Biomed Chromatogr. 2011 Apr;25(4):439-49. doi: 10.1002/bmc.1462.
A rapid, simple, sensitive and specific LC-MS/MS method has been developed and validated for the simultaneous estimation of atorvastatin (ATO), amlodipine (AML), ramipril (RAM) and benazepril (BEN) using nevirapine as an internal standard (IS). The API-4000 LC-MS/MS was operated under the multiple-reaction monitoring mode using electrospray ionization. Analytes and IS were extracted from plasma by simple liquid-liquid extraction technique using ethyl acetate. The reconstituted samples were chromatographed on C(18) column by pumping 0.1% formic acid-acetonitrile (15:85, v/v) at a flow rate of 1 mL/min. A detailed validation of the method was performed as per the FDA guidelines and the standard curves were found to be linear in the range of 0.26-210 ng/mL for ATO; 0.05-20.5 ng/mL for AML; 0.25-208 ng/mL for RAM and 0.74-607 ng/mL for BEN with mean correlation coefficient of ≥0.99 for each analyte. The intra-day and inter-day precision and accuracy results were well with in the acceptable limits. A run time of 2.5 min for each sample made it possible to analyze more than 400 human plasma samples per day. The developed assay method was successfully applied to a pharmacokinetic study in human male volunteers.
已开发并验证了一种快速、简单、灵敏且特异的液相色谱-串联质谱（LC-MS/MS）方法，以奈韦拉平作为内标（IS），同时测定阿托伐他汀（ATO）、氨氯地平（AML）、雷米普利（RAM）和贝那普利（BEN）。API-4000 LC-MS/MS在电喷雾电离的多反应监测模式下运行。通过使用乙酸乙酯的简单液-液萃取技术从血浆中提取分析物和内标。将重构后的样品在C(18)柱上进行色谱分离，以1 mL/min的流速泵入0.1%甲酸-乙腈（15:85，v/v）。按照美国食品药品监督管理局（FDA）指南对该方法进行了详细验证，发现标准曲线在以下范围内呈线性：ATO为0.26 - 210 ng/mL；AML为0.05 - 20.5 ng/mL；RAM为0.25 - 208 ng/mL；BEN为0.74 - 607 ng/mL，每种分析物的平均相关系数≥0.99。日内和日间精密度及准确度结果均在可接受范围内。每个样品的运行时间为2.5分钟，使得每天能够分析400多个人类血浆样品。所开发的测定方法已成功应用于人类男性志愿者的药代动力学研究。
