天然产物研究与开发 ›› 2014, Vol. 26 ›› Issue (5): 695-698.

• 研究简报 • 上一篇    下一篇

中压液相色谱法快速制备木犀草素-7-O-β-D-葡萄糖醛酸苷单体及鉴别

陈家骄1,2,王凤玉1,2,田旭辉1,邓昌瑞1,黄小萍1,王 东1,2,刘东春1,2*   

  1. 1 沈阳药科大学中药学院;2 沈阳药科大学中韩分子生药学研究室,沈阳110016
  • 出版日期:2014-05-31 发布日期:2014-11-17

Preparation and Identification of Luteolin-7-O-β-D-glucuronide by Preparative Medium Pressure Liquid Chromatography

CHEN Jia-jiao1,2, WANG feng-yu1,2, TIAN Xu-hui1, DENG Chang-rui1, HUANG Xiao-ping1, WANG Dong1,2, LIU Dong-chun1,2*   

  1. 1 School of Traditional Chinese Medicine,Shenyang Pharmaceutical University; 2 Laboratory of China and Korea Molecular Pharmacognosy,Shenyang Pharmaceutical University,Shenyang 110016,China
  • Online:2014-05-31 Published:2014-11-17

摘要: 本文采用二元中压制备液相色谱系统,以装有反相C18(Chromatorex,MB 100-40/75)分离填料的两端具有锥形不锈钢接口的耐压玻璃柱(φ49 mm×H310 mm)作为制备色谱柱,流动相为甲醇(A)/含0.05%冰醋酸水溶液(B),梯度洗脱程序:0~20 min,10%(A);20~230 min,30%(A);230~270 min,60%(A);270~300 min,100%(A),流速:50 mL/min,检测波长254 nm,进样量:200 mL,从抱茎苦荬菜(苦碟子)总黄酮中快速、高效地分离得到木犀草素-7-O-β-D-葡萄糖醛酸苷(L7GU)单体。所得单体经UV、IR、TOF-MS、1H-NMR、13C-NMR方法进行了结构确证。经硅胶和聚酰胺薄层色谱检查,在365 nm紫外灯下均呈单一的黄绿色荧光斑点;HPLC-UV分析表明,在210、254、290、348 nm四个波长下,采用色谱峰面积归一化方法计算产品纯度均在98%以上。所得产品可直接作为对照品用于苦碟子药材及制剂的质量控制。

关键词: 抱茎苦荬菜, 苦碟子, 木犀草素-7-O-&beta, -D-葡萄糖醛酸苷, 分离, 制备液相色谱

Abstract: To establish a rapid and efficient method for isolation and purification of luteolin-7-O-β-D-glucuronide (L7GU) from herb of Ixeris sonchifolia (Bge) Hance,a preparative medium pressure liquid chromatography (MPLC) method was developed and used.The MPLC system consisted two-unit pumps and a glass column (φ49 mm×H310 mm) linked with cone-shaped stainless steel port and packed with C18 particles (Chromatorex,MB 100-40/75).The mobile phase was methanol (A)-water (containing 0.05% acetic acid,B).The gradient elution program was as follows:0-20 min,10%A; 20-23 min,30%A;23-27 min,60%A;27-30 min,100%A.The flow rate was 50 mL/min,and the detection wavelength was set at 254 nm.The structure of isolated L7GU was subsequently confirmed by analyzing its UV,IR,TOFMS, 1H-NMR and 13C-NMR.Silica gel HPTLC,polyamide TLC and HPLC analyses were performed to test and verify the purity of isolated L7GU.One single yellowish green spot under UV at 365 nm was detected in both TLC analyses.The HPLC analyses at detection wavelength of 210 nm,254 nm,290 nm and 348 nm demonstrated the purity of isolated L7GU was more than 98%.The high-purity L7GU prepared by this method can be used as reference substance for the quality control of herb of I.sonchifolia and its proprietary medicine.

Key words: Ixeris sonchifolia, Kudiezi, luteolin-7-O-&beta, -D-glucuronide, isolation, preparative liquid chromatography

中图分类号: 

R284.1 R284.2