Yang Yong, Xu Qiufang, Zhou Lei, Zhong Dafang, Chen Xiaoyan
Center for Drug Metabolism and Pharmacokinetics, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, P. R. China.
J Sep Sci. 2015 Mar;38(6):1026-34. doi: 10.1002/jssc.201401227. Epub 2015 Feb 14.
A high-throughput, specific, and rapid liquid chromatography with tandem mass spectrometry method was established and validated for the simultaneous determination of atorvastatin and its two major metabolites, ortho-hydroxyatorvastatin and para-hydroxyatorvastatin, in human plasma. A simple salting-out-assisted liquid-liquid extraction using acetonitrile and a mass-spectrometry-friendly salt, ammonium acetate, was employed to extract the analytes from human plasma. A recovery of more than 81% for all analytes was achieved in 1 min extraction time. Chromatographic separation was performed on a Kinetex XB C18 column utilizing a gradient elution starting with a 60% of water solution (1% formic acid), followed by increasing percentages of acetonitrile. Analytes were detected on a tandem mass spectrometer equipped with an electrospray ionization source that was operated in the positive mode, using the transitions of m/z 559.3 → m/z 440.2 for atorvastatin, and m/z 575.3 → m/z 440.2 for both ortho- and para-hydroxyatorvastatin. Deuterium-labeled compounds were used as the internal standards. The method was validated over the concentration ranges of 0.0200-15.0 ng/mL for atorvastatin and ortho-hydroxyatorvastatin, and 0.0100-2.00 ng/mL for para-hydroxyatorvastatin with acceptable accuracy and precision. It was then successfully applied in a bioequivalence study of atorvastatin.
建立了一种高通量、特异性强且快速的液相色谱-串联质谱法，用于同时测定人血浆中阿托伐他汀及其两种主要代谢物邻羟基阿托伐他汀和对羟基阿托伐他汀，并进行了方法验证。采用乙腈和质谱友好型盐醋酸铵进行简单的盐析辅助液液萃取，从人血浆中提取分析物。在1分钟的萃取时间内，所有分析物的回收率均超过81%。色谱分离在Kinetex XB C18柱上进行，采用梯度洗脱，起始为60%的水溶液（1%甲酸），随后乙腈比例逐渐增加。在配备电喷雾电离源的串联质谱仪上进行分析物检测，电离源以正模式运行，阿托伐他汀的质荷比跃迁为m/z 559.3 → m/z 440.2，邻羟基阿托伐他汀和对羟基阿托伐他汀的质荷比跃迁均为m/z 575.3 → m/z 440.2。氘代化合物用作内标。该方法在阿托伐他汀和邻羟基阿托伐他汀浓度范围为0.0200 - 15.0 ng/mL、对羟基阿托伐他汀浓度范围为0.0100 - 2.00 ng/mL内进行了验证，准确度和精密度均可接受。该方法随后成功应用于阿托伐他汀的生物等效性研究。
