用于中药生物指纹图谱分析的固定化DNA固定相亲和色谱法。

Affinity chromatography with immobilized DNA stationary phase for biological fingerprinting analysis of traditional Chinese medicines.

作者信息

Su Xingye, Hu Lianghai, Kong Liang, Lei Xiaoyuan, Zou Hanfa

机构信息

National Chromatographic R.&A. Centre, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.

出版信息

J Chromatogr A. 2007 Jun 22;1154(1-2):132-7. doi: 10.1016/j.chroma.2007.03.050. Epub 2007 Mar 19.

DOI:10.1016/j.chroma.2007.03.050

PMID:17451725

Abstract

A stationary phase for high performance affinity chromatography with immobilization of DNA onto silica gel was prepared and characterized. The effect of the ionic strength, concentration of Mg2+, EDTA and CH3CN in the mobile phase on the retention of alkaloids were investigated. With this stationary phase, biological fingerprinting analysis of traditional Chinese medicines (TCMs) Coptis chinensis Franch and Rheum palmatum L. was performed with both one-dimensional (1-D) and two-dimensional (2-D) chromatography. The 1-D chromatography was performed with isocratic and gradient elution and 2-D chromatography was developed with immobilized DNA column combined with silica monolithic ODS column. It was found that 7 compounds in Coptis chinensis Franch including berberine, palmatine and jatrorrhizine, 14 compounds in Rheum palmatum L. including aloe-emodin, rhein, emodin, chrysophannol-8-O-glucophranoside and physionl-8-O-glucophranoside were active in binding to the immobilized DNA.

摘要

制备并表征了一种用于高效亲和色谱的固定相，该固定相将DNA固定在硅胶上。研究了流动相中离子强度、Mg2+、EDTA和CH3CN的浓度对生物碱保留的影响。使用该固定相，采用一维（1-D）和二维（2-D）色谱法对中药黄连和大黄进行了生物指纹图谱分析。一维色谱采用等度洗脱和梯度洗脱，二维色谱采用固定化DNA柱与硅胶整体ODS柱联用。结果发现，黄连中的7种化合物（包括小檗碱、巴马汀和药根碱）、大黄中的14种化合物（包括芦荟大黄素、大黄酸、大黄素、大黄酚-8-O-葡萄糖苷和大黄素甲醚-8-O-葡萄糖苷）与固定化DNA具有结合活性。