Online identification of the antioxidant constituents of traditional Chinese medicine formula Chaihu-Shu-Gan-San by LC-LTQ-Orbitrap mass spectrometry and microplate spectrophotometer
Online identification of the antioxidant constituents of traditional Chinese medicine formula Chaihu-Shu-Gan-San by LC-LTQ-Orbitrap mass spectrometry and microplate spectrophotometer
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DOI：
10.1016/j.jpba.2010.05.014
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发表时间：
2010-11-02
影响因子：
3.4
通讯作者：
Zou, Zhong-Mei
中科院分区：
文献类型：
--
作者：
Su, Zhi-Heng;Zou, Guo-An;Zou, Zhong-Mei
柴胡疏肝散（CSGS）是一种含有7种草药的中药配方，临床上已用于治疗胃炎、消化性溃疡、肠易激综合征和抑郁症。但迄今为止，柴胡疏肝散的化学成分尚未得到研究。为了快速鉴定柴胡疏肝散的化学成分，了解与柴胡疏肝散抗氧化活性相关的化学特征，采用液相色谱-电喷雾电离混合线性阱四极杆轨道阱采用LC-LTQ-Orbitrap质谱技术对复杂体系中的化学成分进行在线鉴定，同时通过馏分收集和微孔板读板系统对CSGS的抗氧化谱进行了研究，共鉴定出CSGS中的33种化学成分，其中13种成分在正离子模式和负离子模式下均能检出，20种成分仅在正离子模式下检出，2种成分仅在负离子模式下检出。 96孔板收集HPLC分析和微孔板分光光度计的洗脱液，通过酶标仪分析可以直接考察各组分对DPPH产生的自由基的清除活性。本研究快速筛选了 CSGS 组分对抗氧化活性的贡献，并在线鉴定了相应的活性成分。结果表明，本文建立的 LC-MSn 和 96 孔板测定系统的组合将是一种无需分离和纯化即可将中药化学特征与其生物活性相关联的有用策略。皇冠版权所有 (c) 2010 由 Elsevier B V 出版 保留所有权利。
Chaihu-Shu-Gan-San (CSGS), a traditional Chinese medicine (TCM) formula containing seven herbal medicines, has been used in treatment of gastritis, peptic ulcer, irritable bowel syndrome and depression clinically However, the chemical constituents in CSGS had not been studied so far To quickly identify the chemical constituents of CSGS and to understand the chemical profiles related to antioxidant activity of CSGS, liquid chromatography coupled with electrospray ionization hybrid linear trap quadrupole orbitrap (LC-LTQ-Orbitrap) mass spectrometry has been applied for online identification of chemical constituents in complex system, meanwhile, antioxidant profile of CSGS was investigated by the fraction collecting and microplate reading system As a result, 33 chemical constituents in CSGS were identified Among them, 13 components could be detected both in positive and in negative ion modes, 20 constituents were determined only in positive ion mode and 2 components were only detected in negative ion mode Meanwhile, the potential antioxidant profile of CSGS was also characterized by combination of 96-well plate collection of elutes from HPLC analysis and microplate spectrophotometer, in which the scavenging activities of free radical produced by DPPH of each fraction could be directly investigated by the analysis of microplate reader. This study quickly screened the contribution of CSGS fractions to the antioxidant activity and online identified the corresponding active constituents The results indicated that the combination of LC-MSn and 96-well plate assay system established in this paper would be a useful strategy for correlating the chemical profile of TCMs with their bioactivities without isolation and purification. Crown Copyright (c) 2010 Published by Elsevier B V All rights reserved.