基于HPLC多指标成分含量测定结合化学计量学的利咽解毒颗粒质量评价

Quality evaluation of Liyan Jiedu granules based on HPLC combined with multi-index content determination and chemometrics

目的建立HPLC法同时测定利咽解毒颗粒中7种成分(R,S)-告依春、直铁线莲宁B、连翘酯苷B、连翘酯苷A、连翘苷、哈巴苷和哈巴俄苷的质量分数,同时采用聚类分析和主成分分析等化学计量学方法对不同生产企业利咽解毒颗粒进行综合评价。方法流动相:乙腈-0.2%磷酸溶液,流速:0.9 mL/min;色谱柱:Agilent Zorbax SB-C18(250 mm×4.6 mm,5μm)柱,柱温:30℃;检测波长分别为230 nm[0~23.0 min检测(R,S)-告依春和直铁线莲宁B]、275 nm(23.0~37.0 min检测连翘酯苷B、连翘酯苷A和连翘苷)和210 nm(37.0~60.0 min检测哈巴苷和哈巴俄苷);采用SPSS 26.0统计软件对7种成分的质量分数进行聚类分析和主成分分析。结果(R,S)-告依春、直铁线莲宁B、连翘酯苷B、连翘酯苷A、连翘苷、哈巴苷、哈巴俄苷分别在1.63~40.75、0.59~14.75、1.01~25.25、6.38~159.50、2.27~56.75、6.86~171.50、1.99~49.75μg/mL范围内线性关系良好(r≥0.9991);平均加样回收率分别为98.52%、97.26%、96.83%、100.0%、98.96%、99.29%、99.03%,RSD分别为1.44%、1.18%、1.01%、0.73%、1.39%、0.88%、1.21%;12批利咽解毒颗粒样品聚为2类,主成分1~3是影响利咽解毒颗粒质量评价的主要因子,化学计量学分析结果表明同一生产企业样品较为稳定,不同企业间样品的整体质量一致性存在一定差异。结论该方法操作便捷、重复性好,可用于利咽解毒颗粒的质量评价。

Objective An HPLC method for simultaneous determination of(R,S)-goitrin,clemastanin B,forsythoside B,forsythoside A,forsythin,harpagide and harpagoside in Liyan Jiedu granules was established.Meanwhile,chemometrics methods such as principal component analysis and cluster analysis were used for comprehensive evaluation of Liyan Jiedu granules from different manufacturers.Methods The determination of(R,S)-goitrin,clemastanin B,forsythoside B,forsythoside A,forsythin,harpagide and harpagoside was executed on an Agilent Zorbax SB-C18 chromatographic column(250 mm×4.6 mm,5μm),and the column temperature was set at 30℃.Acetonitrile-0.2%phosphoric acid solution was used as the mobile phase with gradient elution,and the flow rate was set at 0.9 mL/min.The detection wavelength was set at 230 nm at 0-23.0 min for(R,S)-goitrin and clemastanin B,275 nm at 23.0−37.0 min for forsythoside B,forsythoside A and forsythin,210 nm at 37.0−60.0 min for harpagide and harpagoside.The principal component analysis and cluster analysis were conducted for the content results of seven constituents in Liyan Jiedu granules by SPSS 26.0 statistical software.Results(R,S)-Goitrin,clemastanin B,forsythoside B,forsythoside A,forsythin,harpagide and harpagoside showed good linear relationships within the ranges of 1.63−40.75,0.59−14.75,1.01−25.25,6.38−159.50,2.27−56.75,6.86−171.50,1.99−49.75μg/mL(r≥0.9991),whose average recoveries were 98.52%,97.26%,96.83%,100.02%,98.96%,99.29%,99.03%with the RSDs of 1.44%,1.18%,1.01%,0.73%,1.39%,0.88%,1.21%,respectively.12 batches of Liyan Jiedu granules were clustered into two groups,and the principal components 1−3 were the main factor affecting the quality evaluation of Liyan Jiedu granules.Chemometrics analysis showed that the samples from the same manufacturers were relatively stable,and the overall quality consistency of samples from different manufacturers was different.Conclusion The method is convenient and reproducible,which can be used to evaluate the quality of Liyan Jiedu granules.