同时测定大鼠血浆中阿托伐他汀及其活性和毒性代谢产物的方法建立与应用  

Establishment and application for simultaneous determination method of atorvastatin and its active/toxic metabolites in rat plasma

作　　者：宋玉晨 杨琳[2] 孙明谦[1] 任常英 刘建勋[1] 张颖[1] SONG Yuchen;YANG Lin;SUN Mingqian;REN Changying;LIU Jianxun;ZHANG Ying(Beijing Key Laboratory of Pharmacology of Chinese Materia Medica,Institute of Basic Medical Sciences of Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;National Clinical Research Center for Chinese Medicine Cardiology,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China)

机构地区：[1]中国中医科学院西苑医院基础医学研究所北京市中药药理重点实验室,北京100091 [2]中国中医科学院西苑医院国家中医心血管病临床医学研究中心,北京100091

出　　处：《中国药房》2023年第8期917-922,共6页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81873179);中国中医科学院科技创新工程项目(No.CI2021A04906)。

摘　　要：目的 建立同时测定大鼠血浆中阿托伐他汀(ATV)及其活性代谢产物2-羟基阿托伐他汀酸(2-HAT)、4-羟基阿托伐他汀酸(4-HAT)和毒性代谢产物阿托伐他汀内酯(ATL)的分析方法并应用于药代动力学研究。方法 采用液相色谱-串联质谱(LCMS/MS)法进行分析。采用一步沉淀法对血浆样品进行处理(血浆样品经酸化调节pH值防止构型转化),采用梯度洗脱分析样品,分析时间为5 min;采用电喷雾离子源,以多反应监测模式进行正离子检测,ATV及其代谢产物2-HAT、4-HAT、ATL及内标匹伐他汀的定量离子对的质荷比(m/z)分别为559.3→440.2、575.2→440.3、575.0→440.2、540.9→448.2和422.2→290.0。对分析方法进行全面的方法学考察后测定ATV及其代谢产物2-HAT、4-HAT、ATL的浓度,并采用WinNonlin 6.1的非房室模型计算ATV及其代谢产物的药代动力学参数。结果 方法学考察结果表明,空白血浆的内源性物质不干扰待测成分的测定,标准曲线线性关系良好,ATV、2-HAT、4-HAT和ATL的定量下限分别为0.5、0.5、0.25、0.063 nmol/L。精密度、准确度、回收率、基质效应和稳定性考察均符合生物分析要求。药代动力学分析表明,大鼠灌胃给药后,ATV迅速代谢,主要以ATV和2-HAT的形式暴露于血液循环,内酯型代谢物浓度最低。结论 建立了针对ATV及其活性和毒性代谢产物的准确、快速、灵敏的血药浓度分析方法,运用该方法可全面解析阿托伐他汀钙在大鼠体内的药代动力学特征。OBJECTIVE To establish a method for simultaneous determination of atorvastatin(ATV)and its active metabolites 2-hydroxy atorvastatin acid(2-HAT),4-hydroxy atorvastatin acid(4-HAT)and toxic metabolite atorvastatin lactone(ALT)in rat plasma and apply it for pharmacokinetic study.METHODS LC-MS/MS method was adopted for analysis.The one-step precipitation method was used for processing plasma samples(plasma samples were pretreated by acidification to adjust pH value so as to prevent inversion of configuration),gradient elution was used to analyze the samples,and the analysis time was 5 min.Electrospray positive ionization was adopted,and positive ion scanning was performed in multi-reaction monitoring.The m/z of quantified ion pairs of ATV and its metabolites such as 2-HAT,4-HAT and ATL,and internal standard pitavastatin were 559.3→440.2,575.2→440.3,575.0→440.2,540.9→448.2 and 422.2→290.0,respectively.After conducting a comprehensive methodological investigation of the analytical method,the concentrations of ATV and its metabolites 2-HAT,4-HAT,and ATL were determined,and the pharmacokinetic parameters of ATV and its metabolites were calculated using the noncompartment model of WinNonlin 6.1.RESULTS The results of methodological validation showed that endogenous substances in blank plasma did not interfere with the determination of the components to be tested, and the standard curve had agood linear relationship;the lower limits of quantification for ATV, 2-HAT, 4-HAT and ATL were 0.5, 0.5, 0.25 and 0.063 nmol/L,respectively. The precision, accuracy, recovery, matrix effect and stability investigation were all in line with the requirements ofbiological analysis. Pharmacokinetic analysis showed that after intragastric administration in rats, ATV calcium metabolized rapidly,and was mainly exposed to blood circulation in the form of ATV and 2-HAT, with the lowest concentration of lactone-typemetabolites. CONCLUSIONS The established method is precise, rapid and accurate for plasma concentration analysis of ATV a

关 键 词：阿托伐他汀钙 液相色谱-串联质谱 羟基阿托伐他汀 阿托伐他汀内酯 药代动力学 

分 类 号：R969.1[医药卫生—药理学]