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采用超快速液相色谱-串联质谱法同时测定大鼠血浆中芪参益气滴丸的8种生物活性成分及其在药代动力学研究中的应用

Simultaneous determination of eight bioactive components of Qishen Yiqi dripping pills in rat plasma using UFLC-MS/MS and its application to a pharmacokinetic study.

作者信息

Shao Yaping, Zhang Wen, Tong Ling, Huang Jingyi, Li Dongxiang, Nie Wei, Zhu Yan, Li Yunfei, Lu Tao

机构信息

School of Pharmacy, China Pharmaceutical University, Nanjing, People's Republic of China.

Tasly Academy, Tianjin Tasly Holding Group Co. Ltd, Tianjin, People's Republic of China.

出版信息

Biomed Chromatogr. 2017 Aug;31(8). doi: 10.1002/bmc.3941. Epub 2017 Apr 9.

Abstract

In this study, a rapid and reliable ultra-fast liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous determination of eight active ingredients, including astragaloside IV, ononin, tanshinol, protocatechualdehyde, protocatechuic acid, salvianolic acid D, rosmarinic acid and ginsenoside Rg , in rat plasma. The plasma samples were pretreated by protein precipitation with acetonitrile. Chromatographic separation was performed on a Waters Acquity UPLC® BEH C column (1.7 μm particles, 2.1 × 100 mm). The mobile phase consisted of 0.1% aqueous formic acid (A)-acetonitrile with 0.1% formic acid (B) at a flow rate of 0.4 mL/min. Quantification was performed on a triple quadruple tandem mass spectrometry with electrospray ionization by multiple reaction monitoring both in the negative and in the positive ion mode. The lower limit of quantification of tanshinol was 2.0 ng/mL and the others were 5.0 ng/mL. The extraction recoveries, matrix effects, intra- and inter-day precision and accuracy of eight tested components were all within acceptable limits. The validated method was successfully applied to the pharmacokinetic study of the eight active constituents after intragastric administration of three doses (1.0, 3.0, 6.0 g/kg body weight) of Qishen Yiqi Dripping Pills to rats.

摘要

本研究建立并验证了一种快速可靠的超快速液相色谱-串联质谱法,用于同时测定大鼠血浆中的八种活性成分,包括黄芪甲苷、芒柄花苷、丹参素、原儿茶醛、原儿茶酸、丹酚酸D、迷迭香酸和人参皂苷Rg 。血浆样品通过乙腈蛋白沉淀法进行预处理。色谱分离在Waters Acquity UPLC® BEH C柱(1.7μm粒径,2.1×100mm)上进行。流动相由0.1%甲酸水溶液(A)-含0.1%甲酸的乙腈(B)组成,流速为0.4mL/min。定量采用三重四极杆串联质谱仪,通过电喷雾电离在负离子和正离子模式下进行多反应监测。丹参素的定量下限为2.0ng/mL,其他成分的定量下限为5.0ng/mL。八种被测成分的提取回收率、基质效应、日内和日间精密度及准确度均在可接受范围内。该验证方法成功应用于大鼠灌胃给予三剂量(1.0、3.0、6.0g/kg体重)的芪参益气滴丸后八种活性成分的药代动力学研究。

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