高速逆流色谱法快速分离制备枸杞中莨菪亭

Preparative isolation and purification of scopoletin from Lycium barbarum L.by high-speed countercurrent chromatography

建立了用高速逆流色谱(HSCCC)从枸杞中快速分离莨菪亭的方法。将枸杞的乙醇提取物经D-101大孔树脂初步纯化后直接进行高速逆流色谱分离,用薄层色谱-荧光法考察了莨菪亭在不同溶剂体系中的分配情况。结果表明,最佳的溶剂体系为氯仿-甲醇-水(10∶7∶3,v/v/v),取上相为固定相,下相为流动相,在主机转速为850 r/min、流速为1.5 mL/min、检测波长为365 nm的条件下,从200 mg样品中一次性分离制备可得到10.2 mg纯度达到98.3%的莨菪亭。制备所得的莨菪亭与对照品的高效液相色谱(HPLC)保留时间一致,且经核磁共振氢谱、碳谱鉴定结构;纯度经HPLC法测定。研究发现,氯仿-甲醇-水(10∶7∶3,v/v/v)体系可连续二次进样而样品的峰形未受明显的影响。实验结果表明用薄层色谱-荧光法可快速选定HSCCC溶剂体系,进而可快速、简便地制备高纯度的莨菪亭。

An effective and rapid method for the separation of scopoletin from Lycium barbarum L.by high-speed counter-current chromatography（HSCCC） was established.The ethyl alcohol extract of the Lycium barbarum L.was initially separated using D-101 macroporous resins and further purified by HSCCC.The thin layer chromatography coupling with fluorometric spectrophotometry（TLC-F） method was used to determine the partitioning coefficient of scopoletin in different solvent systems.The results showed the solvent system of chloroform-methanol-water（10∶7∶3,v/v/v） was the best one for the HSCCC separation.A total of 10.2 mg of scopoletin with high purity（98.3%,analyzed by high performance liquid chromatography（HPLC）） was obtained in one step by the following separation procedures： the upper phase as the stationary phase,the lower phase as the mobile phase,with a flow rate of 1.5 mL/min,with the apparatus rotated at 850 r/min,and detected at 365 nm.The structure of the obtained compound was identified by 1H-nuclear magnetic resonance and 13C-nuclear magnetic resonance.The sample could be injected into HSCCC twice successively and the whole separation was achieved with satisfactory peak resolution.These results suggested that the TLC-F method is useful in measuring the partitioning coefficients of the target compound in HSCCC solvent systems and HSCCC is a fast and convenient method for the separation of scopoletin.