葛根中黄嘌呤氧化酶抑制剂的筛选分离及质谱分析  

Screening and Identification of Xanthine Oxidase Inhibitors from Radix Puerariae Extract Using UPLC-Q Exactive MS

作　　者：夏建丽 侯万超 李赛男[1] 黄彧 吴桐[1] 刘春明[1] XIA Jian-li;HOU Wan-chao;LI Sai-nan;HUANG Yu;WU Tong;LIU Chun-ming(Central Laboratory,Changchun Normal University,Changchun Jilin 130032,China)

机构地区：[1]长春师范大学中心实验室,吉林长春130032

出　　处：《时珍国医国药》2019年第12期2849-2852,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(31870336,31670358);吉林省科技发展计划项目(20180414028GH)。

摘　　要：目的建立液相色谱-高分辨质谱(UPLC-Q Exactive MS)和半制备液相色谱(Semi-preparative HPLC)联用技术对葛根药材中黄嘌呤氧化晦抑制剂进行快速筛选分离及鉴定的新方法。方法实验以黄嘌呤氧化酶作为生物靶分子,以超漆技术为研究手段筛选酶抑制剂,采用Semi-preparative HPLC技术分离纯化篩选所得到的活性成分,利用核磁共振波谱技术对单体活性成分进行进一步结构鉴定。结果从葛根药材中筛选并分离出5个纯度均大于90%的活性成分(葛根素、葛根素-6"-0-木糖苷、大豆苷、6"-0-乙酰基大豆苷和大豆苷元)。结论利用液相色谱-超滤-质谱-半制备高效液相色谱联用技术可以快速鉴定、碎选、分离葛根药材中活性化合物。此方法对于筛选黄嘌吟氧化酶酶抑制剂具有快速和灵敏等优势,为痛风等治疗药物的筛选与开发提供了科学有效的技术支持。Objective In this study,ultra performance liquid chromatography coupled with mass spectrometry-(UPLC-Q Exactive MS)and semi-preparative high performance liquid chromatography(Semi-preparative HPLC)hyphenated techniques were used to screen and isolate xanthine oxidase inhibitory from Radix Puerariae extracts.Methods Xanthine oxidase was used as the biological target molecule in the experiment,enzyme inhibitors were screened with ultrafiltration technology as the research method,and the active components were isolated and purified with semi-preparative HPLC technology,and further structural identification of the monomer active components were canied out with nuclear magnetic resonance spectrum technology.Results Five major compounds in Radix Puerariae was identified as xanthine oxidase inhibitors using ultrafiltration methods,namely puerarin,puerarin-6"-0-xyloside,daidzin,6"-0-acetyldaidzin and daidzein.These compounds were then isolated by Semi-prep HPLC with purity higher than 90%.Conclusion Our results demonstrated that systematic isolation of bioactive components from Radix Puerariae guided by ultrafiltration,UPLC-Q Exactive MS and semi-preparative HPLC hyphenated techniques was a feasible and efficient technique.It can be extended for the identification and isolation of other enzymes inhibitors.This method has the advantages of rapid and sensitive screening for enzyme inhibitors and provides scientific and effective technical support for the screening and development of gout and other therapeutic drugs.

关 键 词：葛根 超滤 液-质联用 半制备高效液相色谱 黄嘌吟氧化酶 

分 类 号：R284.1[医药卫生—中药学]