Dual-bioactivity-based liquid chromatography-coupled quadrupole time-of-flight mass spectrometry for NF-κB inhibitors and β2AR agonists identification in Chinese Medicinal Preparation Qingfei Xiaoyan Wan
Dual-bioactivity-based liquid chromatography-coupled quadrupole time-of-flight mass spectrometry for NF-κB inhibitors and β2AR agonists identification in Chinese Medicinal Preparation Qingfei Xiaoyan Wan
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DOI：
10.1007/s00216-012-6332-9
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发表时间：
2012-11-01
影响因子：
4.3
通讯作者：
Bai, Gang
中科院分区：
文献类型：
--
作者：
Cheng, Binfeng;Hou, Yuanyuan;Bai, Gang
传统中药制剂是治疗复杂疾病的有效多靶点策略，但其活性成分不明确且难以鉴定。本研究建立了一种基于超高效液相色谱/四极杆飞行时间质谱联用双活性（NF-κ B和β 2-肾上腺素能受体）荧光素酶报告基因分析系统的快速检测中药制剂中多种活性成分的方法。采用该方法对临床治疗哮喘的中药制剂清肺消炎丸进行了分析。使用NF-κ B和β（2）-肾上腺素能受体活性荧光素酶报告基因测定系统筛选潜在的抗炎和痉挛成分，并根据飞行时间质谱数据同时鉴定。对一种β 2肾上腺素能受体激动剂（麻黄碱）和四种结构类型的NF-κ B抑制剂（牛蒡子苷元衍生物、胆酸衍生物、绿原酸和芥子酸）进行了表征。络石苷被认为是一种新的NF-κ B抑制剂。进一步的细胞因子和趋化因子检测证实了潜在的NF-κ B抑制剂的抗炎作用。超高效液相色谱/四极杆飞行时间质谱联用技术与双活性人体细胞功能评价系统相结合，为快速筛选治疗复杂疾病的中药制剂中的活性成分提供了一种简单有效的方法。
Traditional Chinese medicine (TCM) preparations have been used as an effective multitarget strategy for the treatment of complex diseases; however, their bioactive constituents are undefined and difficult to identify. In this study, a simple and dual-target method based on ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry combined with dual-bioactive (NF-kappa B and beta(2)-adrenergic receptor) luciferase reporter assay systems was developed for the rapid determination of various bioactive compounds of TCM preparations. Qingfei Xiaoyan Wan, a TCM preparation used for the clinical therapy of asthma, was analyzed with this method. Potential anti-inflammatory and spasmolytic constituents were screened using NF-kappa B and beta(2)-adrenergic receptor activity luciferase reporter assay systems and simultaneously identified according to the time-of-flight mass spectrometry data. One beta(2)-adrenergic receptor agonist (ephedrine) and four structural types of NF-kappa B inhibitors (arctigenin derivatives, cholic acid derivatives, chlorogenic acid, and sinapic acid) were characterized. Tracheloside was considered a new NF-kappa B inhibitor. Further cytokine and chemokine detection confirmed the anti-inflammatory effects of the potential NF-kappa B inhibitors. The integration of ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry and dual-bioactive human cell functional evaluation systems proved to be a simple and effective strategy for the rapid screening of various bioactive compounds in TCM preparations used to treat complex diseases.