苦参中黄酮类成分的高效液相指纹图谱及5种成分的含量测定

HPLC fingerprint of flavonoids in Sophora flavescens and determination of five components

采用高效液相色谱法建立苦参中黄酮类成分的指纹图谱,并同时测定3种异戊烯基黄酮(苦参醇I、苦参酮、槐属二氢黄酮G)和2种紫檀素(三叶豆紫檀苷和高丽槐素)的含量。色谱条件采用ULTIMATE XB-C18色谱柱(4.6 mm×250mm,5μm),乙腈-水溶液为流动相进行梯度洗脱,以295 nm为检测波长。共收集了12批苦参和4批同属近缘种山豆根进行测定,运用中国药典委员会"中药色谱指纹图谱相似度评价软件2004AB"进行评价,对黄酮类成分进行主成分分析,并对主要5种成分进行含量测定。建立了苦参黄酮类成分HPLC指纹图谱,标定了13个共有色谱峰,并采用对照品指认了5个主要色谱峰,4批同属近缘种山豆根与苦参黄酮对照图谱明显不同。主成分分析的结果表明,苦参酮、槐属二氢黄酮G等对其质量影响显著。含量测定条件通过方法学验证,平均加样回收率在96.3%～102.3%。该法所建立的苦参黄酮类成分指纹图谱特征性强、方法简便,结合主要成分含量测定可更好地控制其质量,对苦参的鉴定及质量控制具有指导意义和参考价值。

A simple and reliable method of high-performance liquid chromatography with photodiode array detection （HPLC- DAD） was developed to evaluate the quality of a traditional Chinese medicine Sophora flavescens through establishing chromatographic fingerprint and simultaneous determination of five flavonoids, including trifolirhizin, maackiain, kushenol I, kurarinane and sophorafla- vanone G. The optimal conditions of separation and detection were achieved on an ULTIMATE XB-Cls column（4.6 mm ×250mm,5μm） with a gradient of acetonitrile and water, detected at 295 nm. In the chromatographic fingerprint, 13 peaks were selected as the characteristic peaks to assess the similarities of different samples collected from different origins in China according to similarity evalua- tion for chromatographic fingerprint of traditional chinese medicine （2004AB） and principal component analysis（PCA）were used in da- ta analysis. There were significant differences in the fingerprint chromatograms between S. flavescens and S. tonkinensis. Principal com- ponent analysis showed that kurarinone and sophoraflavanone G were the most important component. In quantitative analysis, the five components showed good regression （R 〉 0. 999）with linear ranges, and their recoveries were in the range of 96.3 %-102.3%. This study indicated that the combination of quantitative and chromatographic fingerprint analysis can be readily utilized as a quality control method for S. flavescens and its related traditional Chinese medicinal preparations.