摘要
目的建立一种筛选葛根(PLR)中酪氨酸酶抑制剂的离线二维液质联用法。方法PLR提取物与酪氨酸酶共同孵育30 min后,经涡流色谱柱(1D)分离,收集大分子流出物(靶酶-配体复合物)。用甲醇沉淀蛋白,将与酪氨酸酶结合的成分解离,解离液经HPLC-MS/MS进行定性(2D)分析,筛选出可与酪氨酸酶结合的组分;最后以筛选出的葛根素为代表对其酪氨酸酶抑制活性进行验证。结果PLR提取物中有3种主要成分与酪氨酸酶有结合作用,经鉴定分别为葛根素、6''-O-木糖葛根素和葛根素芹菜糖苷。结论该方法操作简单,分析速度快,为天然产物中活性成分的快速筛选提供了一种有效的策略。
Objective To establish a 2D LC-MS/MS method for efficient and rapid screening of tyrosinase inhibitors in Puerariae lobatae radix(PLR).Methods PLR extract was incubated with tyrosinase at 37℃ for 30 min.After separated by turbulent flow chromatography column(1D),the enzyme-ligands complexes were obtained.The small molecular components binding to the tyrosinase were dissociated by precipitating protein and then characterized by HPLC-MS/MS(2D).These small molecular components are thought to be potential tyrosinase specific inhibitors.Subsequently,puerarin,one of potential active compounds,was used as a representative to verify its tyrosinase inhibitory activity.Results Using this strategy,puerarin,6''-O-xylosylpuerarin and mirificin were successfully identified as potential tyrosinase specific inhibitors.Conclusion This method is simple to operate and fast to analyze,providing an efficient strategy for rapid screening of bioactives from natural products.
作者
徐敬朴
程新杰
卞广丽
郑丽亚
李德强
XU Jing-pu;CHENG Xin-jie;BIAN Guang-li;ZHENG Li-ya;LI De-qiang(Department of Pharmacy,The Second Hospital of Hebei Medical University,Shijiazhuang 050000,Hebei Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2020年第15期2317-2319,共3页
The Chinese Journal of Clinical Pharmacology
基金
国家自然科学基金资助项目(81603066)
河北省人才培养基金资助项目(A201902012)
河北省中医药管理局课题资助项目(2018124)。