高速逆流色谱法分离制备粉防己中粉防己碱和防己诺林碱研究

Separation and Preparation of Tetrandrine and Fangchinoline from Stephania tetrandra by High-Speed Countercurrent Chromatography

目的粉防己的主要活性成分为粉防己碱和防己诺林碱,为获得一种快速从粉防己中分离制备粉防己碱和防己诺林碱的方法。方法采用不同溶剂体系萃取测试粉防己中粉防己碱和防己诺林碱在上、下相的分配系数,得到最佳溶剂体系,利用高速逆流色谱(High-Speed Countercurrent Chromatography,HSCCC)对两种生物碱进行制备,并测试纯度。结果经优化得到正己烷-乙酸乙酯-甲醇-水-三乙胺(7∶3∶5∶4∶1)的溶剂体系能够满足实验要求,以该体系上相为固定相,下相为流动相,3 mL·min^(-1)流速进行HSCCC分离,能够在70 min内连续分离得到防己诺林碱(F1)和粉防己碱(F2),经HPLC测试纯度分别为97%和95%。结论通过HSCCC快速制备获得高纯度粉防己碱和防己诺林碱,较现有柱色谱和pH梯度逆流色谱等方法节省大量时间。

OBJECTIVE The main active components of Stephania tetrandra are tetrandrine and fangchinoline.This paper aims at obtaining a rapid separation and preparation method of tetrandrine and fangchinoline from Stephania tetrandra.METHODS The distribution coefficients of tetrandrine and fang chinoline in the upper and lower phases of mixture solvents were determined by extraction using different mixture solvents.The two alkaloids were separated and prepared by high-speed countercurrent chromatography(HSCCC)and their purities were also tested.RESULTS The optimized solvent system of n-hexane-ethyl acetate-methanol-water-triethylamine(7∶3∶5∶4∶1)was obtained,which meet the experimental requirement.Afterwards,fangchinoline(F1)and tetrandrine(F2)were obtained by continuously separation within 70 min by HSCCC using upper phase as the stationary phase and the lower phase as the mobile phase at a flow rate of 3 mL·min^(-1).The purities of fangchinoline(F1)and tetrandrine(F2)were determined to be 97% and 95%,respectively.CONCLUSION The present study provided a rapid preparation of tetrandrine and fangchinoline by HSCCC.This is time-saving method in comparison of the existing methods such as column chromatography and pH gradient countercurrent chromatography.