UPLC-Q-TOF-MS分析姜黄乙酸乙酯提取物抑制表皮生长因子受体活性的成分  

Analysis of chemical constituents with anti-epidermal growth factor receptor activity from Curcuma longa L by 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,南京210009

出　　处：《中国新药杂志》2014年第2期149-153,共5页Chinese Journal of New Drugs

基　　金：国家"重大新药创制"科技重大专项(2011ZX09307-002-01);国家自然科学基金(81274013;81130069);人事部留学人员择优资助项目(2009-2011)

摘　　要：目的：应用均相时间分辨荧光（homogeneous time-resolved fluorescence,HTRF）技术,筛选发现具有抑制表皮生长因子受体（epidermal growth factor receptor,EGFR）活性的姜黄提取物,同时采用超高效液相色谱与串联四级杆飞行时间质谱仪联用技术（ultra-performance liquid chromatography/quadrupole time-offlight mass spectrometry,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法检测姜黄乙酸乙酯提取物对ERFR的抑制作用,计算提取物对EFFR的抑制率。采用ACQUITY UPLC BEH C18色谱柱,以0.1%甲酸水溶液（A）-乙腈（B）为流动相梯度洗脱,200~400 nm扫描,使用ESI离子源,在正离子模式下采集数据。结果：姜黄的乙酸乙酯部位显示较强的EGFR抑制活性,并从其活性部位中分析鉴定出9个化学物,主要成分为姜黄素类化合物,其中6个分别是香豆酸、姜黄酮、双去甲氧基姜黄素、去甲氧基姜黄素、姜黄素、1-（4-羟基-3,5-二甲氧基苯基）-7-（4-羟基-3-甲氧基苯基）-1,6-庚二烯-3,5-二酮,3个未知成分。结论：研究发现姜黄乙酸乙酯提取部位具有较强的EGFR抑制活性,IC50为3.621μg·mL-1,根据Q-TOF-MS测定的相对分子质量及正离子信息,鉴定了姜黄乙酸乙酯提取物主要为姜黄素类成分,其中以姜黄素、去甲氧基姜黄素、双去甲氧基姜黄素为主要活性成分,说明姜黄素类化合物为抑制EGFR活性的主要成分,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 active ingredients from Curcuma longa L extracts with epidermal growth factor receptor（EGFR） inhibitory activity by homogeneous time-resolved fluorescence technology（HTRF）,and to determine the constituents from Curcuma longa L by ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry（UPLC/Q-TOF-MS）.Methods：The corresponding four extraction parts were obtained after herbs percolation with petroleum ether,ethanol extract,ethyl acetate extraction and boiling water.HTRF method was used to detect the inhibiting effects of ethyl acetate extraction on EGFR,and the inhibition rate was calculated.The chromatographic separation was performed on a C 18 column（50 mm × 2.1 mm,1.7 μm） witha gradient elution of 0.1% formic acid water（A）-acetonitrile（B）.The mass spectrometer equipped with electros pay ionization source was used as defector and data was collected under the positive ion modes.Results：The ethyl acetate part of Curcuma longa L showed a strong inhibiting activity.Nine kinds of constituents were analyzed and identified,including the main active component curcumin,and coumaric acid,turmerone,bisdemethoxycurcum,demethoxycurcumin,curcumin as well as 3 unknown constituents.Conclusion：Parts of Curcuma longa L extracted with ethyl acetate has significant EGFR-inhibiting activity with a IC 50 value of 3.621 μg·mL-1.The information of positive ion and relative molecular mass from Q-TOF-MS determination indicates that Curcuma longa L contains active curcumin ingredients inhibiting EGFR.These findings will provide a theoretical basis for their anti-cancer application and a foundation for further separation of active ingredients.

关 键 词：表皮生长因子受体抑制剂 均相时间分辨荧光 姜黄 有效成分 UPLC Q-TOF-MS 

分 类 号：R979.1[医药卫生—药品]