UPLC-Q-TOF-MS分析鸡血藤乙酸乙酯提取物中抑制表皮生长因子受体活性成分  

Active constituents of inhibiting EGFR in ehtyl acetate extracts from Jixueteng(Caulis Spatholobi) analyzed by using UPLC-Q-TOF-MS

作　　者：王冬梅[1] 林森森[2] 郑司浩[1] 黄林芳[1] 

机构地区：[1]中国医学科学院药用植物研究所,北京100193 [2]中国药科大学新药筛选中心

出　　处：《北京中医药大学学报》2014年第1期58-62,共5页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(No.81274013);国家自然科学基金重点项目(No.81130069);国家重大科技专项创制新药项目(No.2011ZX09307-002-01)

摘　　要：目的应用均相时间分辨荧光（HTRF）技术，筛选发现具有抑制表皮生长因子受体（EGFR）活性的鸡血藤提取物，同时采用超高效液相色谱．飞行时间质谱联用技术（UPLC—Q—TOF．MS）对其活性部位进行分析和鉴别。方法药材经石油醚渗滤、乙醇提取、乙酸乙酯萃取和水煎煮后，得到4个提取部位。采用HTRF法检测鸡血藤乙酸乙酯提取物对EGFR的抑制作用，计算提取物对EGFR的抑制率。采用ACQUITYUPLCBEH C-18色谱柱，以0．1％甲酸水溶液（A）．乙腈（B）为流动相梯度洗脱，200～400nm扫描，使用ESI离子源，在正离子模式下采集数据。结果首次发现鸡血藤的乙酸乙酯部位显示较强的EGFR抑制活性，IC-18为2．277mg／L，并从其活性部位中分析鉴定出15个化学物，主要成分为黄酮类化合物，其中10个分别是儿茶素．儿茶素，阿福豆素一儿茶素，杨梅素-3＇-O—β—D吡喃木糖苷，芒柄花苷，芒柄花黄素，Afrormosin，表没食子儿茶素，催吐萝芙木醇-9-O-木糖-（1→6）葡萄糖苷，木犀草苷，没食子儿茶素．儿茶素；5个未知成分。儿茶素、表没食子儿茶素、芒柄花黄素为主要活性成分。结论鸡血藤乙酸乙酯提取物所含的黄酮类成分为抑制EGFR活性的主要成分，本研究以期为其在抗癌方面的应用提供理论依据，同时为进一步跟踪分离活性成分奠定基础。Objective To screen the extracts with the inhibitive activity on epidermal growth factor receptor （EGFR） from Jixueteng （Caulis Spatholobi ） by using the technique of homogeneous time- resolved fluorescence （HTRF）, and to analyze and identify these active constituents by applying ultra- performance liquid chromatography/quadrupole time-of-flight mass spectrometry （UPLC/Q-TOF-MS）. Methods The crude material of Jixueteng was percolated with petroleum ether, and then extracted with ethanol and ethyl acetate and boiled in water to obtain 4 extract fractions. The inhibitive effect of ethylacetate extract on ERFR was detected by using HTRF and inhibitive rate was calculated. The UPLC/Q- TOF-MS procedure was performed on the chromatographic column of ACQUITY UPLC BEH Cis, and the mobile phase was 0.1% formic acid solution （A） -acetonitrile （B） scanned in 200 - 400 nm. The data was collected by using ESI ion source in a positive ion mode. Results The ethyl acetate fractions of Jixueteng showed a strong inhibitive activity on EGFR and ICs0 was 2. 277 μg/mL. There were 15 chemicals identified from these active fractions, and their main constituents were flavonoids, among them 10 were as follows ： （epi） catechin- （epi） catechin, myricetin -3＇-O-β-D- xyloside, （epi） afzelechin- （epi） catechin, ononin, formononetin, afrormosin, epigallocatechin, vomifoliol-9-O-β-I）-xylopyranosyl （ 1→6 ） -β-D- glucopyranside, luteolin-7-O-glucoside and （ epi ） gallocatechin-（ epi ） catechin. There were other 5 constituents unknown. The major active constituents are （epi） catechin-（epi） catechin, epigallocatechin and formononetin. Conclusion The flavonoids in ethyl acetate extracts from Jixueteng has a strong inhibitive activity on EGFR. The study hopes to provide theoretic bases for the anticancer application of Jixueteng and further tracking the separation of its active constituents.

关 键 词：鸡血藤 表皮生长因子受体抑制剂 均相时间分辨荧光 有效成分 超高效液相色谱-飞行时 间质谱联用技术 

分 类 号：R284.2[医药卫生—中药学]