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采用微透析采样与电喷雾电离线性四极杆离子阱质谱联用技术鉴定生、制山茱萸在大鼠体内的代谢物。

Identification of metabolites of crude and processed Fructus Corni in rats by microdialysis sampling coupled with electrospray ionization linear quadrupole ion trap mass spectrometry.

机构信息

Research Center of TCM Processing Technology, Zhejiang Chinese Medical University, Hangzhou 310053, PR China.

出版信息

J Pharm Biomed Anal. 2011 Aug 25;56(1):118-25. doi: 10.1016/j.jpba.2011.04.013. Epub 2011 Apr 21.

Abstract

A microdialysis (MD) sampling coupled with electrospray ionization linear quadrupole ion trap mass spectrometry (LTQ-MS(n)) method has been developed for rapid and sensitive analysis of rat microdialysate metabolite profile of Fructus Corni, a well-known Traditional Chinese Medicine (TCM). The purified samples were separated by a reversed-phase HPLC with C₁₈ column under a gradient elution. Parent compounds and metabolites of crude and processed Fructus Corni of Jiu Zheng Pin (JZP, JZP is produced after steaming the crude drug pre-steeped in wine) were detected by the on-line MS(n) detector in negative scan model. The identification of the metabolites and their structural elucidation were performed by comparing the changes in molecular mass and defining sites of biotransformation based on the accurate MS(n) spectral information of diagnostic fragment ions. In this work, we used such strategies for the identification of the parent compounds and metabolites of crude and processed Fructus Corni in rats, and seven parent compounds and three new metabolites of Fructus Corni were found in rats for the first time. This study provides important structural information regarding to the metabolism of crude Fructus Corni and its JZP. Furthermore, this work also demonstrated the possibilities of using microdialysis sampling coupled with LC-MS(n) approach for identification of bioactive compounds from TCM in vivo.

摘要

建立了一种微透析(MD)采样与电喷雾电离线性四极杆离子阱质谱(LTQ-MS(n))联用的方法,用于快速灵敏地分析著名中药(TCM)山茱萸的大鼠微透析液代谢产物谱。在梯度洗脱下,采用 C₁₈反相 HPLC 柱对净化后的样品进行分离。采用在线 MS(n)检测器在负扫描模式下检测原药材和九蒸片(JZP,是将预先用酒浸泡的原药材蒸制而成)炮制的山茱萸的母体化合物和代谢产物。通过比较分子质量的变化和基于准确的 MS(n)谱信息,定义生物转化的部位,对代谢产物进行鉴定和结构解析。在这项工作中,我们使用这些策略来鉴定大鼠中原药材和炮制的山茱萸的母体化合物和代谢产物,首次在大鼠中发现了山茱萸的 7 种母体化合物和 3 种新代谢产物。本研究为山茱萸及其 JZP 的代谢提供了重要的结构信息。此外,这项工作还证明了微透析采样与 LC-MS(n)联用方法用于鉴定体内 TCM 中生物活性化合物的可能性。

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