UPLC-MS/MS法同时测定小鼠血浆中的15种胆汁酸  

Simultaneously determination of 15 bile acids in plasma of mouse by LC-MS/MS

作　　者：冯闫茹玉 杨倩[1] 张萌琳 付春梅[1] 李大鹏 FENG Yanruyu;YANG Qian;ZHANG Menglin;FU Chunmei;LI Dapeng(Sichuan Research Center for Drug Precision Industrial Technology,Sichuan Engineering Laboratory for Plant-Sourced Drug,Key Laboratory of Drug-Targeting and Drug Delivery System of the Education Ministry and Sichuan Province,West China School of Pharmacy,Sichuan University,Chengdu,Sichuan,610041 P.R.China;Shanghai Institute of Materia Medica,Chinese Academy of Sciences,Shanghai,201203 P.R.China;University of Chinese Academy of Sciences,Bejing,100049 P.R.China)

机构地区：[1]四川大学华西药学院,靶向药物与释药系统教育部和四川省重点实验室,四川省植物来源工程实验室,四川省小分子药物精准化工程技术研究中心,四川成都610041 [2]中国科学院上海药物研究所,上海201203 [3]中国科学院大学,北京100049

出　　处：《华西药学杂志》2024年第4期416-421,共6页West China Journal of Pharmaceutical Sciences

基　　金：国家自然科学基金资助项目(批准号:82204513)。

摘　　要：目的采用UPLC-MS/MS同时测定小鼠血浆中15种胆汁酸的含量。方法采用Shim-packGISTC_(18)色谱柱(100mm×2.1mm,2μm),流动相A为水(含0.01%甲酸)、B为乙腈(含0.01%甲酸),梯度洗脱,流速0.3mL·min^(-1),进样量5μL,柱温40℃,内标为胆酸-d4;多反应离子监测,采用负离子模式,电喷雾离子源,血浆样品经预处理后进样。结果0.24~3.2×10^(4)ng·mL^(-1)15种胆汁酸与其峰面积的比值呈良好的线性关系(r^(2)≥0.9900);回收率为83%~110%,日内RSD为0.68%~4.02%,日间RSD为1.53%~8.06%;提取回收率为70.24%~94.25%;样品在-80℃下反复冻融和长期冻存后的稳定性良好。结论所用方法操作简单、结果准确、重复性好,可用于测定小鼠血浆中胆汁酸的含量。OBJECTIVE To simultaneously determine 15 bile acids in mice plasma by UPLC-MS/MS.METHODS The analytical column was Shim-pack GIST C_(18)(100 mm×2.1 mm,2μm).The mobile phase consisted of 0.01%formic acid(A)and acetonitrile with 0.01%formic acid(B)for gradient elution.The injection volume was 5μL with a flow rate of 0.3 mL.min^(-1),the column temperature was maintained at 40 C,and the internal standard was cholic acid-de.Monitoring with multiple reactive ions,using negative ion mode,electrospray ion source.Plasma samples were pre-treated and injected.RESULTSThe 15 bile acids showed good linearity with their peak area ratios in the range of 0.24-3.2×10 ng·mL^(-1)(≥0.9900).The recovery rate was in the range from 83%to 110%;the intra-day RSD was in the range from 0.68%to 4.02%;inter-day RSD was in the range from 1.53%to 8.06%.The recovery rate of extraction was in range of 70.24%-94.25%.The samples remained stable after repeated freeze-thaw and long-term freezing at-80 C.CONCLUSION The method is simple,accurate and reproducible,and can be used for the determination of 15 kinds of bile acids in mouse plasma.

关 键 词：超高效液相色谱-串联质谱 胆汁酸 鹅去氧胆酸 脱氧胆酸 非酒精性脂肪肝炎 血浆 C57BL/6小鼠 靶向代谢组学 

分 类 号：R917[医药卫生—药物分析学]