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 UltimateAQ-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.
作者
邓阳
蔡骅琳
颜苗
张毕奎
李昕
谭胜蓝
何桂霞
李焕德
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)
出处
《中国医院药学杂志》
CAS
北大核心
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)
关键词
LC-MS/MS
胆汁酸
雷公藤甲素
肝毒性
ICR小鼠
liquid chromatography-tandem spectrometry
bile acids
triptolide
hepatotoxicity
ICR mice
