麻黄中生物碱类成分富集新方法及化学成分分析  

New Enrichment Method and Characterization of Alkaloids in Ephedra sinica

作　　者：史丽颖[1,2] 陈瑶[1] 卢轩[1,2] 冯宝民[1,2] 于大永[1,2] SHI Li-ying;CHEN Yao;LU Xuan;FENG Bao-min;YU Da-yong(School of Life Sciences and Biotechnology,Dalian University,Dalian 116622,China;Institute of Materia Medica,Dalian University,Dalian 116622,China)

机构地区：[1]大连大学生命科学与技术学院,辽宁大连116622 [2]大连大学药物研究所,辽宁大连116622

出　　处：《中国实验方剂学杂志》2018年第21期56-61,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：辽宁省自然科学基金指导计划项目(2016010027-301)

摘　　要：目的：研究麻黄中生物碱类成分富集新方法并对其生物碱类成分进行分析,以期从中发现新的平喘活性成分。方法：为减少非生物碱类成分对定性分析的干扰、提高生物碱类成分分离制备的效率,该研究采用新型固相萃取填料对麻黄提取物中生物碱类成分进行富集,利用高效液相色谱法对富集到的生物碱类成分进行分离制备,采用HPLC-TOF-MS技术对制备的各组分进行分析,应用无标记整合药理学方法筛选各组分对β2受体的激动活性。结果：新型AC18固相萃取填料对麻黄中生物碱类成分的富集具有较高的选择性,采用基于反相/强阴离子交换混合模式的色谱固定相XCharge C18对富集得到的生物碱类成分进行分离,共制备了14个组分,经HPLC-MS分析,共检测出26个生物碱类成分。在筛选的14个组分中有11个组分显示了对β2肾上腺素受体的激动活性。结合这11个组分HPLC-MS分析结果,检识出其中12个生物碱类成分具有β2肾上腺素受体激动活性,其中包括5组生物碱及其异构体：麻黄碱/伪麻黄碱,甲基麻黄碱/甲基伪麻黄碱,去甲基麻黄碱/去甲基伪麻黄碱,（±）-1-苯基-2-亚胺基-1-丙醇,（±）-酪胺甜菜碱。结论：麻黄中活性生物碱同分异构体的分离和鉴定对新的平喘活性成分的发现具有指导意义。Objective： To investigate a new method for the enrichment of alkaloids, analyze the compositions of alkaloids,and look for new anti-asthmatic constituents in Ephedra sinica. Method： In order to reduce the interference by non-alkaloid components on qualitative analysis and to improve the efficiency of separation and preparation of alkaloids,alkaloids in E. sinica were enriched by using new solid phase extraction（ SPE） packing in this study; and the fractions of the enriched alkaloids were prepared by applying high performance liquid chromatography（ HPLC）; HPLC-TOF-MS was used to identify the main components in each fraction; active constituents as β2-adrenergic receptor agonists were characterized by using label-free dynamic mass redistribution assays. Result： The results indicated that AC18 SPE packing had high selectivity for the enrichment of alkaloids in E. sinica. A mixed-mode chromatography method was developed to separate alkaloids based on a mixed-mode reversed phase/strong anion-exchange XCharge C18 column,and 14 fractions were prepared in total.In high performance liquid chromatography-mass spectrometry（ HPLC-MS）,26 alkaloids were identified. Among the 14 fractions screened,11 fractions showed agonistic activity on β2-adrenergic receptor（ β2-AR）. Combined with HPLC-MS results for these 11 fractions,12 alkaloids displayed agonistic activity on β2-AR, including 5 groups of alkaloid and thier isomers： ephedrine/pseudoephedrine, methyl ephedrine/methyl pseudoephedrine,norephedrine/norpseudoephedrine（ ±）-1-phenyl-2-imido-1-propanol and（ ±）-tyramine betaine. Conclusion：The separation and characterization of alkaloid isomers in E. sinica provided guidance for discovering novel antiasthmatic constituents.

关 键 词：麻黄 生物碱 固相萃取 质谱 Β2受体 

分 类 号：R284.1[医药卫生—中药学] R289[医药卫生—中医学]