人工栽培关白附中二萜生物碱的HPLC-Q-TOF-MS鉴别  

Identification of diterpenoid alkaloids in the roots of cultured Aconitum coreanum by HPLC-Q-TOF-MS

作　　者：邢贝妮 吴琼[1] 汤庆发[2] 刘静涵[1] 梁敬钰[1] 杨春华[3,4] 

机构地区：[1]中国药科大学天然药物化学教研室,南京210009 [2]南方医科大学中医药学院,广州510515 [3]中国药科大学药物科学研究院,南京210009 [4]美国佐治亚州立大学生物系,亚特兰大30303

出　　处：《中国药科大学学报》2014年第2期192-199,共8页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.81202431);广东省自然科学基金资助项目(No.S2012010009576);中国药科大学天然药物活性组分与药效国家重点实验室资助项目(No.JKGQ 201118)~~

摘　　要：建立了快速鉴定3年生人工栽培的关白附中二萜生物碱类化学成分的分析方法。采用碱化乙醚超声提取关白附药材粉末,提取物以流动相定容制备供试液;样品用Zorbax-SB C18柱分离,以含0.2%甲酸和0.1%三乙胺的缓冲溶液/乙腈为流动相,梯度洗脱,应用ESI-Q-TOF检测,正离子模式下采集数据;根据色谱峰的准确分子量、液相色谱保留时间以及质谱裂解方式比较等方法鉴别化合物。结果表明,借助HPLC-Q-TOF-MS方法从关白附中分离检测出60个成分,根据保留时间和二级质谱裂解规律鉴定了其中26个二萜生物碱,为人工载培关白附的化学成分鉴定和未知化合物发现提供了一种快速方法。与此同时总结了C20-二萜生物碱(Hetisine型、Atisine型)及C19二萜生物碱(Lycoctonine型)在ESI正离子模式下的质谱裂解规律。Our research tentatively summarized the fragment pathways of three different types of diterpenoid alka- loids under electrospray positive mode and established a rapid method for identifying diterpenoid alkaloids in the roots of cultured Aconitum coreanurn （ L^vl. ） Rapaics. The powder of the roots was extracted with diethyl ether. After removing the diethyl ether under reduced pressure, the extract was dissolved in 0. 1% formic acid aqueous solution, separated on HPLC by using an Agilent Zorbax SB-C18 reverse-phase column with a gradient elution of aqueous solution （containing 0. 2% formic acid, 0. 1% triethylamine） and acetonitrile, and then detected by Q-TOF mass spectrometer equipped with electrospray ionization source operated in positive ion mode. Compounds in the roots of Aconitum coreanum were identified by the accurate molecular weight obtained by HRMS, compari- son of the MS fragment pathways and LC retention time with those of reference standards. As a result, 60 compounds were tentatively detected, 26 of which were assigned to known diterpenoid alkaloids. This established method is an accurate and effective way to identify the known constituents and discover the unknown ones from Aconitum coreanum.

关 键 词：关白附 二萜生物碱 HPLC—Q—TOF-MS 

分 类 号：R284[医药卫生—中药学]