基于UPLC-Q Exactive四级杆-轨道阱液质联用法快速建立清热灵颗粒中潜在中药质量标志物(Q-Marker)成分库  

Rapid establishment of Q-Marker database for Qingreling Granules with UPLC coupled with hybrid quadrupole-orbitrap mass spectrometry

作　　者：周秀娟[1] 李燕芳[1] 陈莹[1] 陈晓虎 瞿燕[3] 任晓东[4] 曾锐[1] ZHOU Xiu-juan LI Yan-fang CHEN Ying CHEN Xiao-hu QU Yan REN Xiao-dong ZENG Rui(College of Pharmacy, Southwest University for Nationaities, Chengdu 610041, China Chongqing Food and Drug Control Research Institute, Chongqing 401121, China Pharmacy College, Chengdu University of Traditional Chinese Medicine, Chengdu 610016, China College of Chemistry, Stony Brook University, New York 11794, US)

机构地区：[1]西南民族大学药学院,四川成都610041 [2]重庆市食品药品检验所,重庆401121 [3]成都中医药大学药学院,四川成都610016 [4]纽约州立大学石溪分校化学学院,美国纽约11794

出　　处：《中草药》2017年第1期67-74,共8页Chinese Traditional and Herbal Drugs

基　　金：国家自然基金资助项目(81603309);国家留学基金委资助项目(CSC201500850007);四川省科技支撑计划(2014SZ0131);四川省科技厅基础研究项目(2015JY0009)

摘　　要：目的基于中药质量标志物(Q-Marker)理念,使用UPLC-Q Exactive四级杆-静电场轨道阱高分辨液质联用技术,对清热灵颗粒主要化学成分进行快速识别和鉴定,建立清热灵颗粒的潜在Q-Marker库,为探讨建立中成药清热灵颗粒的系统质量控制方法奠定基础。方法以50%甲醇作为提取液制备供试液,采用ACQUITYUPLC BEHC_(18)色谱柱(100mm×2.1mm,1.7μm)进行分离,以甲醇-0.1%甲酸水溶液为流动相进行梯度洗脱,质谱采用负离子监测模式、全扫描及自动触发二级质谱扫描的功能,对清热灵颗粒中成分进行识别及鉴定。并采用Autodock vina 1.1.2软件,以H5N1禽流感病毒为神经氨酸酶受体,利用AutoDock Tools 1.5.6程序确定靶蛋白的活性口袋,对所选标志物进行分子对接研究。结果通过高分辨UPLC-Q Exactive四级杆-静电场轨道阱高分辨液质联用技术共鉴定了39个化学成分,包括黄酮类化合物25个,苯乙醇苷类6个,有机酸4个,三萜类化合物2个,木脂素类2个。32个化学成分为首次在该制剂中鉴别出,其中15个为专属性成分,建议其中的黄芩苷、汉黄芩苷、连翘酯苷A、甘草苷、连翘酯苷E、连翘酯苷B、异甘草苷、黄芩素、连翘苷、汉黄芩素、甘草素11个成分可作为清热灵颗粒的Q-Marker。结论通过高分辨UPLC-Q Exactive四级杆-静电场轨道阱高分辨液质联用技术,可以快速分析和确定中成药清热灵颗粒的化学成分,建立清热灵颗粒的潜在Q-Marker库。Objective To establish the Q-Marker database for Chinese medicine Qingreling Granules （QG） based on the concept of Q-Marker by the method of ultra high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry （UPLC-quadrupole-orbitrap MS） to recognize and identify the chemical constituents, and study the quality of QG Methods The 50% methanol extract was separated on an Acquity UPLC BEH C18 column （100 mm × 2.1 mm, 1.7μm）, and eluted with a gradient of methanol-water containing 0.1% formic acid. Constituents of QG were identified by HRMS in the negative ion mode using both full scan and two-stage threshold-triggered mass modes. Eleven Q-Marker were docked by Autodock vina 1.1.2 software, using HsN1 avian influenza virus as the Neuraminidase receptor, and the active site of target protein were determined by the Auto Dock 1.5.6 program. Results A total of 39 compounds were identified, including 25 flavonoids, 6 phenylethanoid glycosides, 4 organic acids, 2 triterpenoids, and 2 lignan ingredients. Totally 15 components were specific and 32 chemical compositions were reported for the first time in QG by the method of UPLC-quadrupole-orbitrap MS. Baicalin, wogonoside, forsythoside A, liquiritin, forsythoside E, forsythoside B, isoliquiritin, baicalein, phillyrin, wogonin, and liquirtigenin could be used to establish the Q-Marker database. Conclusion To establish Q-Marker database for Chinese medicine QG and identify the compounds in QG by the method of UPLC-quadrupole-orbitrap MS.

关 键 词：中药质量标志物 UPLC-Q Exactive四级杆-静电场轨道阱高分辨液质联用 清热灵颗粒 成分库 黄芩苷 汉黄芩 苷 连翘酯苷A 甘草苷 连翘酯苷E 连翘酯苷B 异甘草苷 黄芩素 连翘苷 汉黄芩素 甘草素 

分 类 号：R286.02[医药卫生—中药学]