Jordan Center for Pharmaceutical Research, P.O. Box 950435, Amman 11105, Jordan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Feb 15;917-918:36-47. doi: 10.1016/j.jchromb.2013.01.001. Epub 2013 Jan 9.
A simple liquid chromatography/ion trap mass spectrometry method for the quantification of amlodipine and atorvastatin with its metabolites, ortho and para hydroxy atorvastatin, simultaneously in human plasma was developed. Analytes with internal standard were extracted by protein direct precipitation with acetonitrile. Adequate chromatographic separation was achieved using Phenomenex Synergi 4u polar-RP 80A (150mm×4.6mm, 4μm) column in the isocratic elution mode and the eluent was water/methanol (14:86%, v/v) adjusted by trichloroacetic acid to pH 3.2 which was delivered isocratically at constant flow rate of 0.50mL/min. Standard solutions for the analytes were prepared using amlodipine besylate, atorvastatin calcium, ortho-hydroxy atorvastatin dihydrate monosodium salt, para-hydroxy atorvastatin disodium salt, and pravastatin sodium as an internal standard. The method validation intends to investigate specificity, sensitivity, linearity, precision, accuracy, recovery, matrix effect and stability according to USFDA guideline. Standard calibration levels were prepared by pooled human plasma to attain final dynamic range of 0.2-20.0ng/mL for amlodipine, 1.5-150ng/mL for atorvastatin, 1.0-100.0ng/mL for ortho-hydroxy atorvastatin and 0.2-20.0ng/mL for para-hydroxy atorvastatin. Clinical bioequivalence study was successfully investigated by the application of this validated bioanalytical method in order to evaluate bioequivalence of two commercial products 10mg amlodipine/80mg atorvastatin in a single dose. In this study, 29 healthy volunteers were participated in randomized, two periods, double blend, open label cross over design. Pharmacokinetic parameters of C(max), AUC(0-t) and AUC(0-∞) were calculated to compare a test product with CADUET(®) reference product.
建立了一种简单的液相色谱/离子阱质谱法，用于同时定量人血浆中的氨氯地平和阿托伐他汀及其代谢物邻位和对位羟基阿托伐他汀。用乙腈直接沉淀蛋白提取分析物和内标。在等度洗脱模式下，使用 Phenomenex Synergi 4u 极性-RP 80A（150mm×4.6mm，4μm）柱实现了充分的色谱分离，洗脱液为水/甲醇（14:86%，v/v），用三氯乙酸调至 pH 3.2，以 0.50mL/min 的恒定流速等度输送。分析物的标准溶液使用氨氯地平苯磺酸盐、阿托伐他汀钙、邻位羟基阿托伐他汀二水合物单钠盐、对位羟基阿托伐他汀二钠盐和普伐他汀钠作为内标制备。方法验证旨在根据美国 FDA 指南研究特异性、灵敏度、线性、精密度、准确度、回收率、基质效应和稳定性。通过混合人血浆制备标准校准水平，以达到氨氯地平的最终动态范围为 0.2-20.0ng/mL，阿托伐他汀为 1.5-150ng/mL，邻位羟基阿托伐他汀为 1.0-100.0ng/mL，对位羟基阿托伐他汀为 0.2-20.0ng/mL。通过应用该验证后的生物分析方法成功进行了临床生物等效性研究，以评估两种商业产品 10mg 氨氯地平/80mg 阿托伐他汀单剂量的生物等效性。在这项研究中，29 名健康志愿者参加了随机、两周期、双交叉、开放标签的交叉设计。计算 C(max)、AUC(0-t)和 AUC(0-∞)的药代动力学参数，以比较试验产品与 CADUET®参考产品。
