摘要
目的建立超高分辨质谱(UPLC-Q Exactive MS)-半制备型液相色谱(Semi-preparative HPLC)-高速逆流色谱(HSCCC)联用技术对天麻药材中乳酸脱氢酶抑制剂进行快速筛选分离及鉴定的新方法。方法实验以乳酸脱氢酶作为生物靶分子,以超滤技术为研究手段筛选酶抑制剂,采用Semi-preparative HPLC结合HSCCC技术分离纯化筛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。结果从天麻药材中筛选并分离出3个纯度均大于90%的活性成分(天麻素、对羟基苯甲醇和巴利森苷A)。结论利用超高分辨质谱-半制备型液相色谱-高速逆流色谱联用技术可以快速鉴定、筛选、分离天麻药材中活性化合物。该方法对于筛选乳酸脱氢酶酶抑制剂具有快速和灵敏等优势,为肿瘤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)with High Speed counter-current chromatography(HSCCC)hyphenated techniques were used to screen and isolate lactic dehydrogenase inhibitory from Gastrodiae Rhizoma extracts.Methods Lactic dehydrogenase 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 with HSCCC technology,and further structural identification of the monomer active components were carried out with nuclear magnetic resonance spectrum technology.Results Three major compounds in Gastrodiae Rhizoma was identified as lactic dehydrogenase inhibitors using ultrafiltration methods,namely gastrodin,4-hydroxybenzyl alcohol and parishin A.These compounds were then isolated by Semi-preparative HPLC and HSCCC with purity higher than 90%.Conclusion Our results demonstrated that systematic isolation of bioactive components from Gastrodiae Rhizoma guided by UPLC-Q Exactive MS and Semi-preparative HPLC with HSCCC 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 tumour therapeutic drugs.
作者
李彦杰
周旭
庄思远
刘春明
李赛男
LI Yan-jie;ZHOU Xu;ZHUANG Si-yuan;LIU Chun-ming;LI Sai-nan(Central Laboratory,Changchun Normal University,Changchun,Jilin 130032,China)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2021年第10期2336-2340,共5页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金(31870336)
吉林省人才开发基金项目([2018]853)
长春师范大学自然科学基金(长师大自科合字[2020]第011号)。
关键词
天麻
超滤-质谱
高速逆流色谱
半制备型液相色谱
乳酸脱氢酶
Gastrodiae Rhizoma
Ultrafiltration-MS
High Speed Countercurrent Chromatography
Semi-preparative Liquid Chromatography
Lactic dehydrogenase
作者简介
李彦杰(1995-),男(畲族),福建南平人,长春师范大学中心实验室在读硕士研究生,学士学位,主要从事天然药物分析工作;通讯作者:李赛男(1985-),女(汉族),吉林长春人,长春师范大学中心实验室副研究员,博士学位,主要从事天然药物分析工作.
