LC-MS/MS法同时测定ICR小鼠肝脏中15种胆汁酸及其在雷公藤甲素肝毒性研究中的应用  

Simultaneous determination of 15 bile acids in ICR mice liver by LC-MS/MS and its application in triptolide induced hepatotoxicity research

作　　者：邓阳 蔡骅琳[2] 颜苗[2] 张毕奎[2] 李昕[1] 谭胜蓝 何桂霞[3] 李焕德[2] DENG Yang;CAI Hua-lin;YAN Miao;ZHANG Bi-kui;LI Xin;Tan Sheng-lan;HE Gui-xia;LI Huan-de(Department of Pharmacy, Third Hospital of Changsha ＆ Clinical Application Research Institute of Antibiotics in Changsha, Hunan Changsha 410015, China;Department of Pharmacy, Second Xiangya Hospital of Central South University, Hunan Changsha 410011, China;School of Pharmacy, Hunan University of Chinese Medicine, Hunan Changsha 410208, China)

机构地区：[1]长沙市第三医院药学部长沙市抗菌药物临床应用研究所,湖南长沙410015 [2]中南大学湘雅二医院药学部,湖南长沙410011 [3]湖南中医药大学药学院,湖南长沙410028

出　　处：《中国医院药学杂志》2018年第11期1138-1143,共6页Chinese Journal of Hospital Pharmacy

基　　金：国家自然科学基金(编号：81202985,81573686,81703818)

摘　　要：目的:建立同时测定ICR小鼠肝脏中15种胆汁酸的LC-MS/MS法,将其应用到雷公藤甲素肝毒性研究中。方法:小鼠经腹腔注射1mg·kg^(-1)雷公藤甲素后,进行血清生化、肝病理切片检查。将肝脏用0.1%甲酸-75%乙腈混合液进行匀浆并经样品处理,采用Ultimate~AQ-C_(18)色谱柱,流速0.3mL·min^(-1),进行15种胆汁酸色谱分离。结果:LC-MS/MS法测定的15种胆汁酸在2.8~8 000.0nmol·L^(-1)浓度范围内具有较好的线性关系,加样回收率在85.8%~114.0%之间,日内、日间精密度均在0.7%~10.8%范围内;腹腔注射1mg·kg^(-1)雷公藤甲素后小鼠出现了肝毒性,雷公藤甲素组的胆汁酸浓度有明显上升。结论:该方法准确、稳定性好、灵敏度高,能满足ICR小鼠肝脏中多种胆汁酸的含量测定,为雷公藤甲素肝毒性研究提供了进一步的理论基础。OBJECTIVE To develop a liquid chromatography-tandem mass spectrometry（LC-MS/MS）method for simultaneous determination of 15 bile acids in the liver of ICR mice,and its application to a triptolide induced hepatotoxicity research.METHODS The serum biochemical indices and liver histopathology were detected after intraperitoneal injecting of triptolide at the dose of 1.0 mg·kg^-1.The liver was homogenized with the 0.1% formic acid-75% acetonitrile mixture.Liquid chromatographic separation of 15 bile acids was performed on an UltimateAQ-C18 column,and the flow rate was 0.3 mL·min^-1.RESULTS The linear calibration curves of 15 bile acids were obtained in the ranges of 2.8-8000.0 nmol·L^-1,and the recovery rates of the quality control samples were 85.8%-114.0%.The intra-day and inter-day RSD values were obtained in the ranges of 0.7%-10.8%.The mice developed hepatotoxicity after intraperitoneal injection of triptolide at the dose of 1.0 mg·kg^-1,and the concentrations of bile acids in the TP group increased significantly.CONCLUSION This LC-MS/MS method for quantitatively determining bile acids in the ICR mice liver has high accuracy,sensitivity and stability,and can provide the theoretical basis for the triptolide induced hepatotoxicity research.

关 键 词：LC-MS/MS 胆汁酸 雷公藤甲素 肝毒性 ICR小鼠 

分 类 号：R917[医药卫生—药物分析学]