生附子中C_(19)型二萜生物碱的高速逆流色谱分离及其结构鉴定

Isolation and structure identification of C_(19) diterpenoid alkaloids from Aconitum carmichaeli by high-speed counter-current chromatography

应用高速逆流色谱法分离制备了生附子中的3个C19型二萜生物碱类化合物。以正己烷-乙酸乙酯-甲醇-水(3∶5∶4∶5,v/v/v/v)为两相溶剂系统,上相为固定相,下相为流动相,在主机转速850 r/min、流动相流速2.0mL/min、检测波长235 nm条件下进行分离制备;一次性从90 mg附子总碱粗提物中分离制备得到15.3 mg北草乌碱,35.1 mg中乌头碱和22.7 mg次乌头碱,经高效液相色谱分析,测得它们的纯度分别为97.9%、96.2%和99.2%。并应用波谱(电喷雾离子质谱、核磁共振氢谱和核磁共振13 C谱)解析法确定了它们的结构。利用该方法可以对生附子中的二萜类生物碱成分进行快速的分离和纯化。

Three C19 diterpenoid alkaloids were isolated and purified from the lateral roots of Aconitum carmichaeli Debx.（Fuzi in Chinese） by high-speed counter-current chromatography（HSCCC）.A mixture of n-hexane-ethyl acetate-methanol-water（3∶5∶4∶5,v/v/v/v） was used as the two phase solvent system.The lower phase was used as the mobile phase and was operated at a flow rate of 2.0 mL/min,while the apparatus was rotated at 850 r/min,and the detection wavelength was at 235 nm.Under these conditions,15.3 mg of beiwutine,35.1 mg of mesaconitine and 22.7 mg of hypaconitine were obtained from 90 mg of crude extract in one-step separation with the purities of 97.9%,96.2% and 99.2%,respectively,determined by high performance liquid chromatography.The structures of these three compounds were identified by electrospray ionization mass spectrometry（ESI-MS）,1H-nuclear magnetic resonance（1H-NMR） and 13C-NMR.The results indicate that HSCCC is a powerful technique for the purification of diterpenoid alkaloids from the lateral roots of Aconitum carmichaeli Debx.