比那甫西颗粒的指纹图谱及含量测定方法研究

Study on Fingerprint Chromatograms and Content Determination Method of Binafuxi Granules

目的建立比那甫西颗粒指纹图谱,制定同时测定制剂中没食子酸、秦皮乙素和甘草酸铵含量的方法。方法采用高效液相色谱法(HPLC)建立12批比那甫西颗粒的指纹图谱并进行峰归属,色谱柱为安捷伦TC-C18(4.6 mm×250 mm,5μm),以乙腈-0.05%磷酸溶液为流动相梯度洗脱,检测波长为272 nm。采用中药色谱指纹图谱相似度评价软件(2012版)对12批比那甫西颗粒的指纹图谱进行相似度分析,结合聚类分析、主成分分析对比那甫西颗粒进行质量评价;采用HPLC法测定制剂中没食子酸、秦皮乙素和甘草酸铵的含量。结果12批比那甫西颗粒的HPLC特征图谱,标定共有峰21个,归属5个峰,相似度均>0.9,聚类分析将样品分为3类,主成分分析筛选出3个特异性标志物;3个成分的浓度分别在0.0247~0.6175、0.0375~0.9375、0.0580~1.4500 mg·mL^(-1)范围内与峰面积线性关系良好,平均回收率(n=6)为97.24%~103.02%,RSD为0.64%~1.12%。结论该方法能够全面系统地反映制剂的质量,可用于比那甫西颗粒生产过程的控制和质量控制。

Objective The HPLC fingerprint chromatogram was established,and an HPLC method was established for the content determination of gallic acid,aesculetin,and ammonium glycyrrhizinate in binafuxi granules.Methods HPLC determined the fingerprint chromatogram of 12 batches of binafuxi granules,and the peak attribution was carried out.The Agilent TC-C18 column(250 mm×4.6 mm,5μm)was used with the mobile phase of acetonitrile-0.05%phosphoric acid solution for gradient elution.The detection wavelength was 272 nm.Chinese medicine chromatographic fingerprint similarity evaluation software(2012 edition)was used to analyze the similarity of fingerprint of 12 batches of benafoxi granules,combined with cluster analysis and principal component analysis to compare the quality of benafoxi granules.The contents of gallic acid,aesculetin,and ammonium glycyrrhizinate in the preparation were determined by HPLC.Results Twenty-one common peaks were identified using HPLC characteristics of fingerprints of 12 batches of binafuxi granules,and five were identified.The similarity was higher than 0.9.Cluster analysis divided the samples into three categories,and principal component analysis screened three specific markers.The concentration line ranges of three components were 0.0247—0.6175,0.0375—0.9375,and 0.0580—1.4500 mg·mL^(-1).The average recovery rates were 97.24%—103.02%,with RSD of 0.64%—1.12%(n=6).Conclusion The method can reflect the quality of the preparation comprehensively and systematically,and can be used for the control of the production process and quality of binafuxi granules.