不同产地玄参饮片高效液相色谱指纹图谱及6种化学成分含量测定

HPLC fingerprints of Radix Scrophul ariae from different areas and content determination of 6 chemical compounds

目的建立玄参饮片的高效液相色谱(HPLC)指纹图谱及6种化学成分同时定量分析的方法。方法采用HPLC测定了16个不同批次的玄参样品,色谱柱为Syncronis C18(250 mm×4.6 mm,5μm),以0.03%磷酸-乙腈为流动相进行梯度洗脱,检测波长210 nm,柱温40℃,流速1 mL·min^(-1),运行时间80 min,建立玄参饮片的指纹图谱并进行含量测定,采用中药指纹图谱相似度评价系统(2012版)对16批样品进行共有峰确认及相似度评价;通过SPSS 21.0统计软件采用主成分分析(PCA)和聚类分析(CA)对HPLC指纹图谱进行模式识别分析。结果以肉桂酸为参照物,建立了玄参饮片的HLPC指纹图谱共有模式,标定了20个共有峰,并指认了其中桃叶珊瑚苷、哈巴苷、哈巴俄苷、安格洛苷C、类叶升麻苷和肉桂酸6个峰,并对其中6种主要成分进行含量测定;对16批样品进行了相似度评价,其相似度为0.881～0.980。结论建立的HPLC指纹图谱结合主成分分析、聚类分析方法,可以客观、有效、全面地用于玄参饮片的质量评价;不同厂家、批号、产地的玄参饮片样品质量不一,应加强中药规范化生产。

Objective To establish the HPLC fingerprints and a simultaneous quantitative analysis method for6 chemical compounds of Radix Scrophul ariae pieces.Methods Totally 16 batches of samples were analyzed on Syncronis C18(250 mm×4.6 mm,5μm)column,with the gradient mobile phase of 0.03%phosphoric acid-acetonitrile,the monitoring wavelength at 210 nm,the column temperature at 40℃,the flow rate of mobile phase at 1 mL·min-1,and detection finished within 80 minutes.The method of key component assaying and fingerprinting was established.Similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine(2012 edition)was used to confirm the common peaks in 16 sample batches.SPSS 21.0 statistical software was used for the principal component analysis and cluster analysis for the HPLC fingerprint.Results The HPLC fingerprint of Radix Scrophul ariae pieces used cinnamic acid as the marker,and all tested samples contained 20 common characteristic peaks,6 of which was verified to be aucubin harpagide,harpagoside,cangoroside,acteoside and cinnamic acid,and the content was determined.In addition,the similarity of 16 sample batches was evaluated,with similarities between 0.881 and 0.980.Conclusion HPLC fingerprint,principal component analysis and cluster analysis methods can objectively,effectively and comprehensively evaluate the quality of Radix Scrophul ariae.Besides,the quality of prepared picess of Radix Scrophul ariae may vary with different producers,different batches and different localities,suggesting that the production procedures should be normalized.