温胆汤颗粒剂质量评价方法的建立

Establishment of Quality Evaluation Method for Wendan Decoction Granules

目的建立评价温胆汤颗粒质量的方法。方法建立15批温胆汤颗粒的高效液相色谱指纹图谱,对其进行相似度分析和共有峰指认与归属;结合聚类分析(HCA)法、主成分分析(PCA)法、正交偏最小二乘判别分析(OPLS-DA)法进行质量评价;建立测定其中鸟苷、尿苷、橙皮苷、6-姜辣素含量的高效液相色谱法。结果15批温胆汤颗粒的指纹图谱相似度均大于0.950;共标记33个共有峰,指认出其中8个色谱峰,分别为16号峰(甘草苷)、19号峰(柚皮苷)、20号峰(橙皮苷)、21号峰(新橙皮苷)、28号峰(甘草酸)、29号峰(川陈皮素)、30号峰(6-姜辣素)、32号峰(橘皮素)。通过HCA将样品聚为5类;通过PCA筛选出7个主成分,累计方差贡献率为93.131%;OPLS-DA与HCA的结果一致,筛选出14个差异组分。高效液相色谱含量测定结果显示,鸟苷、尿苷、橙皮苷、6-姜辣素分别在各自质量浓度范围内与峰面积积分值线性关系良好(R2≥0.9991);精密度、稳定性、重复性试验结果的RSD均小于2.5%(n=6);平均加样回收率分别为99.27%,99.35%,98.51%,99.58%,RSD分别为0.85%,1.52%,1.39%,1.04%(n=6);15批温胆汤颗粒中鸟苷、尿苷、橙皮苷、6-姜辣素的平均含量分别为0.0176%,0.0169%,1.0467%,0.0222%。结论所建立的方法简便快捷、专属性强、重复性好,可反映各化学成分信息,可为温胆汤颗粒的质量控制提供参考。

Objective To establish a method for evaluating the quality of Wendan Decoction Granules.Methods The high-performance liquid chromatography(HPLC)fingerprints for fifteen batches of Wendan Decoction Granules were established,and similarity analysis and identification and attribution of common peaks were performed.Hierarchical cluster analysis(HCA),principal component analysis(PCA),and orthogonal partial least squares-discriminant analysis(OPLS-DA)were used to evaluate the quality of 15 batches of Wendan Decoction Granules.HPLC method used for the content determination of guanosine,uridine,hesperidin,and 6-gingerol was established.Results The fingerprint similarity of fifteen batches of Wendan Decoction Granules was higher than 0.950,with thirty-three common peaks attributed and eight chromatographic peaks identified,namely peak 16(liquiritin),peak 19(naringin),peak 20(hesperidin),peak 21(neohesperidin),peak 28(glycyrrhizic acid),peak 29(tangerine),peak 30(6-gingerol),and peak 32(hesperidin).The samples were divided into five categories by HCA analysis.Seven principal components were screened through PCA,with a cumulative variance contribution rate of 93.131%.The results of OPLS-DA and HCA were consistent,and fourteen differential components were screened.The content determination results by HPLC showed that guanosine,uridine,hesperidin,and 6-gingerol had good linear relationships with peak area integration values within their respective mass concentration ranges(R2≥0.9991).The RSDs of precision,stability,and repeatability test results were all lower than 2.5%(n=6).The average recoveries of guanosine,uridine,hesperidin,and 6-gingerol were 99.27%,99.35%,98.51%,and 99.58%,with RSDs of 0.85%,1.52%,1.39%,and 1.04%(n=6),respectively.The average contents of guanosine,uridine,hesperidin,and 6-gingerol in fifteen batches of Wendan Decoction Granules were 0.0176%,0.0169%,1.0467%,and 0.0222%,respectively.Conclusion The established method is simple,rapiid,highly specific and has good reproducibility,which can reflect information on various chemical components,which can provide a reference for the quality control of Wendan Decoction Granules.