LC-MS/MS法测定人血浆中阿托伐他汀及其代谢物的浓度及在中国健康志愿者中生物等效性研究的应用  

LC-MS/MS determination of atorvastatin and its metabolites in human plasma and bioequivalence in healthy Chinese volunteers

作　　者：肖红梅[1] 刘金凤 郑天东 唐智 陈露露 欧阳冬生[2,3] XIAO Hong-mei;LIU Jin-feng;ZHENG Tian-dong;TANG Zhi;CHEN Lu-lu;OU YANG Dong-sheng(The First Affiliated Hospital of Shaoyang University,Shaoyang Hunan 422000;Department of Clinical Pharmacology,Xiangya Hospital,Central South University,Changsha 410000;Hunan Key Laboratory for Bioanalysis of Complex Matrix Samples,Changsha 410000;School of Pharmacy,Xiangnan Institute,Chenzhou Hunan 423000)

机构地区：[1]邵阳学院附属第一医院,湖南邵阳422000 [2]中南大学湘雅医院临床药理研究所,长沙410000 [3]复杂基质样本生物分析湖南省重点实验室长沙都正生物科技有限公司,长沙410000 [4]湘南学院药学院,湖南郴州423000

出　　处：《中南药学》2023年第11期2842-2847,共6页Central South Pharmacy

基　　金：湖南省复杂基质样本生物分析国际科技创新合作基地(No.2019CB1014);国家自然科学基金项目(No.81803837)。

摘　　要：目的建立可测定人血浆中阿托伐他汀(ATO)及其代谢物浓度的LC-MS/MS分析方法,并将其应用于人体生物等效性研究。方法血浆中目标成分采用叔丁基甲基醚∶乙酸乙酯(1∶1)萃取,以ATO-d5、邻位阿托伐他汀(o-ATO)-d5、对位阿托伐他汀(p-ATO)-d5为内标,流动相A为2 mmol·L^(-1)乙酸铵水溶液-乙腈-甲酸(900∶100∶1),B为乙腈,梯度洗脱。采用电喷雾离子源,正离子模式,MRM监测,ATO和内标的离子选择通道分别为m/z 559.3→440.1和564.2→445.2。对健康受试者口服相同剂量的氨氯地平阿托伐他汀钙片(5 mg/10 mg)受试制剂或参比制剂后的血药浓度进行定量分析。结果ATO和o-ATO的线性范围为0.150~20.000 ng·mL^(-1),p-ATO的线性范围为0.015~2.000 ng·mL^(-1);ATO和o-ATO的定量下限均为0.150 ng·mL^(-1),p-ATO为0.015 ng·mL^(-1)。此外,批内和批间精密度的RSD均小于15%。32名健康受试者口服5 mg/10 mg氨氯地平阿托伐他汀钙片参与了该研究。受试者空腹和餐后状态下服用氨氯地平阿托伐他汀钙片受试制剂和参比制剂的血浆药物最大浓度(Cmax)、时间-浓度曲线下面积(AUC0~t和AUC0~∞)的几何均值比值的90%置信区间均在80%~125%内,两制剂具有生物等效性。结论该方法准确可靠,重复性好,进样过程简单,可用于人血浆中ATO及其代谢物的浓度测定。Objective To establish an LC-MS/MS method to determinate atorvastatin(ATO)and its metabolites in human plasma,and to determine its bioequivalence.Methods Atorvastatin-d5(ATO-d5),ortho-hydroxy atorvastatin-d5(o-ATO),and para-hydroxy atorvastatin-d5(p-ATO)were used as the internal standard.Human plasma samples were extracted by tert-butyl methyl ether:ethyl acetate(1∶1),with the mobile phase A(2 mmol·L^(-1) aqueous ammonium acetate-acetonitrile-formic acid)and B(acetonitrile)with a gradient elution.An electrospray ionization source was also used,and multiple reaction monitoring were operated in the positive mode with a selective channel at m/z 559.3→440.1 and 564.2→445.2 for atorvastatin and its internal standard.Additionally,after the oral administration of the same dose of amlodipine atorvastatin calcium tablets(5 mg/10 mg),the blood concentration of the drug was quantitatively analyzed in healthy volunteers.Results The linear range of ATO and o-ATO was 0.150~20.000 ng·mL^(-1),and that of p-ATO was 0.015~2.000 ng·mL^(-1).The lower limit of quantification of ATO and o-ATO was 0.150 ng·mL^(-1),and that of p-ATO was 0.015 ng·mL^(-1).The linear range of ATO and o-ATO was 0.150~20.000 ng·mL^(-1),and that of p-ATO was 0.015~2.000 ng·mL^(-1).The lower limit of quantification of ATO and o-ATO were 0.150 ng·mL^(-1),and that of p-ATO was 0.015 ng·mL^(-1).Moreover,the relative standard deviation in and between batches was less than 15%.Totally 32 health subjects enrolled the pharmacokinetics study of oral 5 mg/10 mg amlodipine atorvastatin calcium tablets.The 90%confidence intervals of Cmax,AUC0~t and AUC0~∞of amldipine atorvastatin calcium test and reference preparation under fasting and postprandial conditions were wthin 80%~125%.The two preparations were bioequivalent.Conclusion This method is accurate,reliable,simple and reproducible,which can be used to determine the concentration of atorvastatin and its metabolites in human plasma.

关 键 词：液相色谱-质谱联用 阿托伐他汀 邻位阿托伐他汀 对位阿托伐他汀 

分 类 号：R969.1[医药卫生—药理学]