LC-MS/MS测定小鼠肝脏中10种胆汁酸浓度的方法与应用  

Determination method of ten kinds of bile acids and its application in mice liver by LC-MS / MS

作　　者：沈淑娇[1] 张志荣[1] 曾金[1] 陈文文[1] 过林[1] 贺敏[1] 吴雨[1] 蒋健[1] 裘福荣[1] 

机构地区：[1]上海中医药大学附属曙光医院临床药代动力学实验室,201203

出　　处：《肝脏》2015年第4期285-290,294,共7页Chinese Hepatology

基　　金：国家自然科学基金项目(81173118);国家科技部新药重大创制基金资助项目(2012ZX09303009);上海中医临床重点实验室基金资助项目(c10dz2220200)

摘　　要：目的建立LC-MS/MS法同时测定小鼠肝脏中10种游离型与结合型胆汁酸的浓度。方法肝组织样品匀浆经乙腈沉淀蛋白后进行LC-MS/MS分析,空白基质经活性炭(100 mg/m L)吸附获得,色谱柱为Zorbax Eclipse XDB-C18柱(5μm,150 mm×4.60 mm),流动相为乙腈-含0.05%甲酸和4 mmol/L乙酸铵溶液,梯度洗脱,流速1 m L/min,进样10μL,采用电喷雾离子源(ESI)、负离子的多级反应模式(MRM)监测。结果在定量范围内,各胆汁酸的线性关系良好,检测时间较短(14 min),精密度、准确度、提取回收率等均符合要求。结论建立的方法灵敏度高、专属性好,可用于肝组织样品中胆汁酸浓度的测定。Objective To develop a high-performance liquid chromatography tandem mass spectrometry （LC-MS/ MS） method for simultaneous determination of 10 kinds of free and conjugated bile acids in mice liver. Methods Liver samples （homogenized in deionized water） were precipitated as protein by acetonitrile （100mg/ml） to be analyzed by LC- MS/MS. Blank matrix was prepared with 100mg/ml activated charcoal. Analytes were separated on a Zorbax Eclipse XDB- C18 column （5~m, 150ram ~ 4. 60mm） with gradient elution of acetonitrile and 4mmol/L ammonium acetate in aqueous solution （contained 0.05~ formic acid）, of which flow rate was Iml/min and injection volume was 10/11. Detection was carried out by multiple reaction monitoring （MRM） in negative ion mode. Results The method had a good linear relationship in quantitative range, with a short run time as 14min, of which precision, accuracy, extraction recovery and other parameters met requirement. Conclusion The proposed method, proved to be sensitive and selective, is suitable for concentration determination of bile acids in mouse liver.

关 键 词：胆汁酸 LC—MS/MS 小鼠 肝 浓度测定 

分 类 号：R575[医药卫生—消化系统]