目的 建立UPLC-MS/MS同时定量检测人血浆中 14 种胆汁酸的浓度,探索环孢素肝毒性与胆汁酸的关系。方法 采用蛋白沉淀法处理血浆样本,使用ACQUITY UPLC HSS T3(2.1 mm×150 mm,1.8 μm)色谱柱,流动相为水(4 mmol·L-1乙酸铵)-95%乙腈,梯度洗脱,流速 0.3 mL·min-1,检测时间15 min。检测系统采用电喷雾离子源,负离子模式。结果 14种胆汁酸的线性范围良好,批内和批间RSD均<15%,准确度均在87%~116%之间,提取回收率在101%~120% 之间。以胆酸(CA)为主的胆汁酸与环孢素的浓度相关性良好,提示环孢素改变患者的肝毒性可能与胆汁酸相关。结论 该方法方便准确、灵敏度高,能够满足人血浆中多种胆汁酸的含量测定,可以用于环孢素引起肝毒性的进一步基础研究。
Abstract
OBJECTIVE To establish a method for simultaneous quantitative determination of 14 bile acids in human plasma by liquid chromatograohy-tandem mass spectrometry (LC-MS/MS) and to explore the correlation between the hepatotoxicity induced by cyclosporine A and bile acids. METHODS Plasma samples were extracted by protein precipitation and bile acids were separated on a Waters ACQUITY UPLC HSS T3(2.1 mm×150 mm,1.8 μm) column with a flow rate of 0.3 mL·min-1. The mobile phase was water (containing 4 mmol·L-1 ammonium acetate) and 95% acetonitrile using gradient elution in 15 min. The detection system used an electrospray ion source in negative ion mode. RESULTS Fourteen bile acids had a good linear relationship, the intra-and inter-assay RSD values were less than 15%, the accuracy was between 87% and 116%, and the extraction recovery rates of the quality was between 101% and 120%. The concentration of cholic acid (CA) was well correlated with cyclosporine A and suggested that bile acids may be associated with the hepatotoxicity induced by cyclosporine A. CONCLUSION This LC-MS/MS method for quantitatively determining various bile acids in human plasma is convenient, accurate and sensitive and can be used for further basic research in hepatotoxicity caused by cyclosporine A.
关键词
胆汁酸 /
高效液相色谱-质谱联用 /
环孢素 /
肝毒性
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Key words
bile acid /
liquid chromatography-electrospray tandem mass spectrometry /
cyclosporine A /
hepatotoxicity
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中图分类号：
R969.3
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参考文献
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脚注
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基金
国家自然科学基金项目资助(81773820,81503140)
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