色谱

色谱 ›› 2012, Vol. 30 ›› Issue (09): 971-974.DOI: 10.3724/SP.J.1123.2012.05037

• 技术与应用 • 上一篇    

高速逆流色谱法快速分离制备枸杞中莨菪亭

李小多1, 李学刚2, 宋尚华2, 张波2, 刘旭晶2, 叶小利1*   

  1. 1. 西南大学生命科学学院, 重庆 400715; 2. 西南大学药学院, 重庆 400716
  • 收稿日期:2012-05-29 修回日期:2012-07-17 出版日期:2012-09-28 发布日期:2012-09-20
  • 通讯作者: 叶小利,博士,副教授,主要从事药用植物生化工程研究. Tel: (023)68250728, E-mail: yexiaoli@swu.edu.cn.
  • 基金资助:

    国家科技部重大新药创制(2010ZX09401-306-3-10)和“十二五”国家科技支撑计划项目(2011BAI13B02-1).

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

LI Xiaoduo1, LI Xuegang2, SONG Shanghua2, ZHANG Bo2, LIU Xujing2, YE Xiaoli1*   

  1. 1. College of Life Science, Southwest University, Chongqing 400715, China; 2. College of Pharmacy, Southwest University, Chongqing 400716, China
  • Received:2012-05-29 Revised:2012-07-17 Online:2012-09-28 Published:2012-09-20

PDF

460

被引次数

15

摘要： 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。

关键词: 薄层色谱-荧光法, 高速逆流色谱, 枸杞, 莨菪亭

Abstract: 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.

Key words: Lycium barbarum L., scopoletin, thin layer chromatography coupled with fluorometric spectrophotometry method, high-speed counter-current chromatography (HSCCC)

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