中国中药杂志

2017, v.42(17) 3362-3367

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内皮细胞提取和UHPLC-LTQ-Orbitrap法分析筛选苦碟子注射液中的活性成分
HUVECs extraction and UHPLC LTQ Orbitrap analysis for screening vascular endothelium protective components in Kudiezi injection

刘思燚;张红柱;范菁;尚展鹏;王菲;李玮婕;刘颖;卢建秋;张加余;
LIU Si-yi;ZHANG Hong-zhu;FAN Jing;SHANG Zhan-peng;WANG Fei;LI Wei-jie;LIU Ying;LU Jian-qiu;ZHANG Jia-yu;Beijing University of Chinese Medicine;Center for Drug Evaluation,China Food and Drug Administration;The First Hospital of Fangshan District;Wu Xi App Tec;

摘要(Abstract):

应用细胞生物色谱法及UHPLC-LTQ-Orbitrap质谱联用技术快速筛选鉴定苦碟子注射液中可能的活性成分。选择人脐静脉内皮细胞(huma umbilical vein endothelid cells,HUVEC)作为靶细胞,先使苦碟子注射液中的活性成分与靶细胞特异性结合,经细胞靶点脱敏失活后,应用LC-MS快速鉴定与细胞靶向亲和的化学成分,从而筛选得到苦碟子注射液中可能的活性成分。根据获得的精确相对分子质量数据、保留时间以及结合苦碟子注射液中的化学成分进行鉴定归属。最终,从细胞裂解液中共筛选鉴定出9个化学成分,其中包括倍半萜内酯类成分4个、有机酸类成分3个以及黄酮类成分2个。应用细胞生物色谱法与UHPLC-LTQ-Orbitrap质谱联用技术快速筛选鉴定苦碟子注射液中可能的活性成分,可为苦碟子注射液药效物质基础研究提供一种快捷、有效的方法学借鉴。
An effective method has been employed as a tool for screening active components in Kudiezi injection by using cell chromatography and sensitive UHPLC-HR-MSnmethod. The potential bioactive components in Kudiezi injection could be selectively bound to the HUVECs target cells first. After cell target desensitization and inactivation,the chemical constituents with cell target affinity were identified by LC-MS,so as to screen the possible active components in Kudiezi injection. Based on the accurate mass measurements and the retention time,in total,9 compounds were tentatively identified and characterized,including 4 sesquiterpene lactones,3 phenolic acids and 2 flavonoids. HUVECs biospecific extraction coupled with UHPLC-LTQ-Orbitrap analysis could provide a rapid and efficient method for the identification of potential bioactive components in Kudiezi injection,and provide the reference for further research on its effective materials basis.

关键词(KeyWords): 苦碟子注射液;人脐静脉内皮细胞;细胞生物色谱法;液质联用技术
Kudiezi injection;HUVECs;cell chromatography;UHPLC LTQ orbitrap

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81503244);; 国家重点基础研究发展计划(973)项目(2012CB518406);; 2017年北京市科技新星与领军人才培养专项计划(Z171100001117029)

作者(Author): 刘思燚;张红柱;范菁;尚展鹏;王菲;李玮婕;刘颖;卢建秋;张加余;
LIU Si-yi;ZHANG Hong-zhu;FAN Jing;SHANG Zhan-peng;WANG Fei;LI Wei-jie;LIU Ying;LU Jian-qiu;ZHANG Jia-yu;Beijing University of Chinese Medicine;Center for Drug Evaluation,China Food and Drug Administration;The First Hospital of Fangshan District;Wu Xi App Tec;

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DOI: 10.19540/j.cnki.cjcmm.20170801.001

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