超高分辨质谱-高速逆流色谱-半制备液相色谱联用快速测定分析红三叶草中环氧合酶-2抑制剂  

Rapid Determination of Cyclooxygenase-2 Inhibitors in Trifolium Pratense L.by High Resolution Mass Spectrometry,High Speed Countercurrent Chromatography and Semi-Preparative Liquid Chromatography

作　　者：侯万超 刘春明[1] 夏建丽 黄彧 吴桐[1] 李赛男[1] HOU Wanchao;LIU Chunming;XIA Jianli;HUANG Yu;WU Tong;LI Sainan(Central Laboratory,Changchun Normal University,Changchun 130032,Jilin,China)

机构地区：[1]长春师范大学中心实验室,吉林长春130032

出　　处：《中华中医药学刊》2020年第7期181-186,共6页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金(31870336,31670358)。

摘　　要：利用超高效液相-质谱联用技术(UPLC-MS)对红三叶草乙醇提取物中化学成分进行分析鉴定,以环氧合酶-2作为生物靶分子,运用超滤质谱技术以"受体-配体"亲和及超滤法作为分析方法筛选具有酶抑制,共鉴定出7个活性化学成分,分别为大豆苷、芒柄花苷、大豆苷元、印度黄檀苷、芒柄花素、德鸢尾素、鹰嘴豆芽素A。利用半制备型高效液相色谱(Semi-preparative HPLC)和高速逆流色谱(High-speed counter-current chromatography,HSCCC)技术对环氧合酶-2抑制剂进行分离和纯化,7种成分纯化至>80%,其中6种成分>92%。结果表明采用超滤质谱-半制备液相色谱-高速逆流色谱联用技术可以快速鉴定、筛选和纯化红三叶草中环氧合酶-2抑制剂。为天然药物的开发提供了理论基础和技术平台,可推广应用于其他酶抑制剂的鉴定和分离。In this study,high performance liquid chromatography coupled with mass spectrometry(UPLC-MS)was used to screen and identify cyclooxygenase-2 inhibitory activity from Trifolium pratense L.extracts.Cyclooxygenase-2 was used as biotargeting molecule,seven major compounds in T.pratense L.were identified as cyclooxygenase-2 inhibitors using ultrafiltration methods,namely Daidzin,Ononin,Daidzein,Sissotrin,Formononetin,Irilone,Biochanin A.The semi-preparative high performance liquid chromatography(semi-preparative HPLC)and high-speed counter-current chromatograph(HSCCC)were applied to separate and isolate the active constituents.These compounds were then isolated by semi-prep HPLC and HSCCC with purity higher than 80%and six ingredients purityhigher than 92%.Our results demonstrated that systematic isolation of bioactive components from T.pratense L.guided by ultrafiltration mass spectrometry-semi-preparative HPLC-HSCCCwas a feasible and efficient technique.It can be extended for the identification and isolation of other enzyme inhibitors.

关 键 词：红三叶草 超滤质谱 半制备型高效液相色谱 高速逆流色谱 环氧合酶-2 

分 类 号：R284.1[医药卫生—中药学]