UPLC-MS-MS法测定健康人血浆中阿托伐他汀浓度及药物代谢动力学研究

Determination of atorvastatin in human plasma by UPLC-MS-MS and pharmacokinetics in healthy subjects

目的建立超高效液相色谱-串联质谱法测定健康人血浆中阿托伐他汀浓度，用于药动学研究。方法采用Acquity UPIC^TM BEH C18色谱柱（2．1mm×50mm，1．7μm），柱温：40℃，以乙腈0．05％甲酸（60：40，v／v）为流动相，流速：0．25mL·min^-1；质谱采用电喷雾电离源正源（ESI^＋），选择离子监测（SIR），m／z 440．0（ATV），m／z 129．6（格列齐特）血浆样品经磷酸酸化后，采用叔丁基甲醚提取，氮气挥干，内标法定量，并用于24名健康男性志愿者单剂量口服10mg阿托伐他汀钙片的药动学研究。结果阿托伐他汀浓度在0．385～15．400ng·mL^-1线性关系良好，r^2为0．9974。日内、日间RSD符合方法学要求，单次服用10mg阿托伐他汀钙片后药动学参数AUC0-48、AUC0-∞、Cmax、f1／2分别为（49．6±40．6）ng·h·mL^-1、（54．7±42．0）ng·h·mL^-1、（5．8±3．4）ng·mL^-1、（1．4±0．7）h、（14．8±6．5）h。结论该方法稳定、灵敏度高、专属性强、操作简单，适用于阿托伐他汀血药浓度测定及药动学研究。

Objective To establish an Ultra Performance Liquid Chromatography -tandem MS method for the determination of atorvastatin in human plasma. Methods UPLC separation was performed on Acquity UPLC^TM BEH C18 column （2. 1 mm×50 mm, 1.7 μm）, and the column temperature was 40 ℃. The mobile phase consisted of 60% aceto nitrile, and 40% formic acid （0.05%）. The compound was ionized in the electrospray ionization （ESI^＋ ） ion source of the mass spectrometer and detected in the selected ion recording （SIR） model. Human plasma samples were extracted with chlorform： methyl t butyl ether after phosphoric acidification. The internal standard method was used to quantify atorvastatin. The assay was validated and applied to pharmacokinetic study of atorvastatin in 24 healthy volunteers following a single oral dose of 10 mg atorvastatin calcium tablets. Results The linear range was 0. 385 15. 400 ng mL^-1 with correlation coefficient of 0. 997 4. The RSD of the intra-day and inter-day validation was less than 15%. AUC0-48, AUC0-∞, Cmax,tmax, and t1/2 of atorvastatin were （49.6±40.6） ng · h · mL^-1, （54.7±42.0） ng·h·mL^-1, （5.8±3.4） ng· mL^-1, （1.4±0.7） h, and （14.8±6.5） h, respectively. Conclusion The method is accurate, sensitive, simple and suitable for the determination of atorvastatin in human plasma and pharma- cokinetic study.