乙肝宁颗粒高效液相色谱指纹图谱研究

Study on HPLC Chromatographic Fingerprint of Yiganning Granula

目的建立乙肝宁颗粒高效液相色谱(HPLC)指纹图谱,为乙肝宁颗粒整体质量评价提供依据。方法以10批乙肝宁颗粒为样品,采用"中药色谱指纹图谱相似度评价系统"(2004年A版)软件建立HPLC指纹图谱并进行相似度评价。色谱柱为Agilent ZORBAX Eclipse XDB-C18(4.6 mm×250 mm,5μm),流动相:乙腈-0.1%磷酸溶液(梯度洗脱),流速:1.0 mL·min^-1,检测波长分别为230 nm(0~72 min)、290 nm(72~80 min);230 nm(80~90 min),柱温:35℃,进样量:20μL。并采用SIMCA-P+软件进行主成分分析(PCA)及正交偏最小二乘判别分析(OPLS-DA)。结果建立了乙肝宁颗粒HPLC指纹图谱,共标定11个共有峰,并指认了绿原酸、芍药苷和槲皮素3个共有峰。10批乙肝宁颗粒图谱相似度为0.900~0.996。PCA及OPLS-DA分析表明10批样品可聚为3类。结论所建立的方法简便、快速、重复性好,为乙肝宁颗粒整体质量评价提供参考。

Objective To establish a HPLC fingerprint of Yiganning granula,and to provide reference for quality control of Yiganning granula.Methods Using 10 batches of Yiganning granula as samples,the HPLC fingerprint was established and the similarity was evaluated by the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System"(2004 A edition) software.The column used in the research was Agilent ZORBAX Eclipse XDBC18(4.6 mm×250 mm,5 μm),the mobile phase was acetonitrile-0.1 % phosphoric acid solution(gradient elution),the flow rate was 1.0 mL·min^-1,the detection wavelength was 230 nm(0~72 min),290 nm(72~80 min) and 230 nm(80~90 min).The column temperature was 35℃,and the injection volume was 20 μL.In this study,principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) were performed on Yiganning granula using SIMCA-P+ software.Results The HPLC fingerprint of Yiganning granula was established,and 11 common peaks were calibrated.Three common peaks,including chlorogenic acid,paeoniflorin,and quercetin were identified.The similarities of the 10 batches of Yiganning granula were 0.900~0.996.The results of PCA and OPLS-DA analysis showed that 10 batches of samples could be divided into three groups.Conclusion The method is simple and repetitive,and provides a reference for the overall quality evaluation of Yiganning granula.