A robust and rapid UPLC–MS/MS method has been developed, optimized and validated for determination of amlodipine (AML), indapamide (IND) and perindopril (PRN) in human plasma. A positive electrospray ionization mode was used in a Xevo TQD LC–MS/MS instrument. A single sample preparation step using extraction technique was applied to extract the three analytes from plasma samples. There was no need to extract indapamide from blood samples in a further step. Extraction of the three drugs and internal standards was done using a solvent mixture composed of methyl tertiary butyl ether, dichloromethane and ethyl acetate. The prepared samples were analyzed using an Acquity UPLC HSS C₁₈ (100 × 2.1 mm, 1.7 μm) column. Ammonium acetate and methanol, pumped at a flow rate of 0.3 ml/min, were used as a mobile phase. Method validation was done as per the US Food and Drug Administration guidelines. Linearity was achieved in the range of 0.2–15 ng/ml for AML, 0.5–50 ng/ml for IND and 0.5–120 ng/ml for PRN. Accuracy and precision were estimated and found to be within the acceptable ranges. The rapid chromatography permits analysis of many samples per batch, making the method suitable for clinical and pharmacokinetic investigations. The developed and validated method was applied to estimate AML, IND, and PRN in a fasting bioequivalence study in healthy human volunteers.

中文翻译：

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新型LC-MS / MS方法测定人血浆中的氨氯地平，吲达帕酰胺和培哚普利：在生物等效性研究中的应用

已经开发，优化和验证了一种强大而快速的UPLC-MS / MS方法，用于测定人血浆中的氨氯地平（AML），吲达帕胺（IND）和培哚普利（PRN）。Xevo TQD LC-MS / MS仪器使用正电喷雾电离模式。应用了使用提取技术的单个样品制备步骤，以从血浆样品中提取三种分析物。无需在进一步的步骤中从血液样本中提取吲达帕胺。使用由甲基叔丁基醚，二氯甲烷和乙酸乙酯组成的溶剂混合物萃取三种药物和内标。使用Acquity UPLC HSS C ₁₈分析准备的样品（100×2.1 mm，1.7μm）色谱柱 以0.3 ml / min的流速泵送的乙酸铵和甲醇用作流动相。方法验证已按照美国食品药品监督管理局的指导进行。AML的线性范围为0.2-15 ng / ml，IND的线性范围为0.5-50 ng / ml，PRN的线性范围为0.5-120 ng / ml。估计准确性和精密度，并发现在可接受的范围内。快速色谱允许每批分析许多样品，使该方法适用于临床和药代动力学研究。在健康人志愿者的禁食生物等效性研究中，已开发和验证的方法用于估计AML，IND和PRN。