双黄连颗粒HPLC指纹图谱及化学模式识别分析

Analysis on HPLC Fingerprint and Chemical Pattern Recognition of Shuanghuanglian Granules

目的建立双黄连颗粒HPLC指纹图谱并进行化学模式识别,为其质量评价提供方法。方法样品经50%甲醇提取后,采用HPLC进行检测,以甲醇-0.2%磷酸水为流动相梯度洗脱,流速0.8mL/min,检测波长278nm,柱温30℃。以对照品及单味药指认共有峰,确定制剂药物的来源归属;对30批样品进行相似度评价、聚类分析及主成分分析。结果双黄连颗粒指纹图谱中确定25个共有峰,通过对照品指认出13个峰。30批样品与对照指纹图谱的相似度为0.827~0.981,聚为两大类。第一主成分贡献率为70.919%,已指认成分有木犀草素、连翘苷、异绿原酸A、黄芩苷、汉黄芩素;第二主成分贡献率为10.835%,已指认成分有隐绿原酸、4,5-O-二咖啡酰奎宁酸、异绿原酸B、新绿原酸、连翘脂素、绿原酸。结论本研究建立的指纹图谱方法稳定、可靠、重复性好,可用于双黄连颗粒的质量控制。

Objective To establish the HPLC fingerprint of Shuanghuanglian Granules and chemical pattern recognition;To provide a method for its quality evaluation.Methods The sample was extracted with 50%methanol and detected by HPLC,methanol-0.2%phosphoric acid as mobile phase with gradient elution;the flow rate was 0.8 mL/min;the detection wavelength was 278 nm;the column temperature was 30℃.According to the comparison of reference substances and single medicinal materials,the common peaks was identified,and the source of the preparation-medicinal materials was determined.Similarity evaluation,cluster analysis and principal component analysis were performed on 30 batches of samples.Results Totally 25 common peaks were identified in the fingerprint of Shuanghuanglian Granules,and 13 peaks were identified by reference substances.The similarity between the 30 batches of samples and the reference fingerprint ranged from 0.827 to 0.981,forming two types.The contribution rate of the first principal components was 70.919%,which identified components were luteolin,forsythin,isochlorogenic acid A,baicalin and wogonin.The contribution rate of the second principal components was 10.835%,which identified components were cryptochlorogenic acid,4,5-O-dicaffeoylquinic acid,isochlorogenic acid B,neochlorogenic acid,forsythiaside and chlorogenic acid.Conclusion The established fingerprint method is stable,reliable and reproducible,which can be used for the quality control of Shuanghuanglian Granules.