HPLC-MS/MS多组分定量测定结合化学计量学研究市售小金丸的质量一致性

Consistency evaluation of commercially Xiaojin Pills using determination of multicomponent contents by HPLC-MS/MS method coupled with chemometrics

目的建立HPLC-MS/MS同时测定小金丸中藁本内酯、原儿茶酸、肌苷、齐墩果酸、苯甲酰新乌头原碱、苯甲酰次乌头原碱、苯甲酰乌头原碱、新乌头碱、次乌头碱、乌头碱10种成分的检测方法,并结合聚类分析、主成分分析(PCA)等化学计量学方法对不同厂家、不同批次小金丸的质量一致性进行评价。方法建立HPLC-MS/MS方法同时测定小金丸中10种活性成分的量。采用Agilent Technologies Zorbax Eclipse XDB-C18色谱柱(150 mm×4.6 mm,5μm);流动相为0.1%甲酸水溶液-乙腈;质谱采用ESI+多反应监测(MRM)模式扫描,并对定量测定结果进行聚类分析和PCA,综合评价市售小金丸质量的差异性。结果方法学验证结果显示,10个化合物在各自质量浓度范围内线性关系良好(R2>0.990 0);定量限为0.01~4.49 ng/m L,平均回收率在94.82%~104.33%。样品测定结果表明A厂家小金丸质量与其他厂家存在较大差异,PCA降维处理后所得荷载图中,苯甲酰新乌头原碱、苯甲酰次乌头原碱、乌头碱、肌苷等组分离原点距离最远,其量的差异是导致一致性差异的主要成分。结论市售小金丸质量一致性较差,各厂家在小金丸生产过程中应加强对制草乌和地龙药材的质量控制。

Objective An HPLC-MS/MS method was established for determination of 10 kinds of components in Xiaojin Pills such as ligustilide, protocatechuic acid, inosine, oleanolic acid, benzoyl neatidine, benzoyl aconitrate, benzoyl aconitine, neaconitine, hypaconitine, aconitine, and combined with cluster analysis, principal component analysis（PCA） and other chemical metrology methods to assess the quality consistency of different batches of Xiaojin Pills from different manufacturers. Methods The HPLC-MS/MS method was established for the simultaneous determination of 10 active ingredients in Xiaojin Pills. The Agilent Technologies Zorbax Eclipse XDB C18 column（150 mm × 4.6 mm, 5 μm） was used; 0.1% formic acid aqueous solution-acetonitrile as mobile phase; The mass spectrum was scanned by ESI＋ multiple reaction monitoring（MRM） mode. And the content analysis was carried out by cluster analysis and principal component analysis, and the difference of the quality of commercially available Xiaojin Pills was evaluated comprehensively. Results The methodological validation results showed that the linear range of 10 compounds was good（R2 0.990 0）. The limit of quantification was 0.01—4.49 ng/m L, and the average recovery was 94.82%—104.33%. The results showed that the quality of Xiaojin Pills in a factory was different from others. Benzoyl aconitine, benzoyl aconitine, aconitine and inosine were the main components of classification. The load chart after PCA reduced dimension processing showed that the content of benzoyl neatidine, benzoylthioate, aconitine and inosine were the most distant from the origin, and the difference in content was the main cause of the difference in quality consistency. Conclusion Considering the poor quality consistency of Xiaojin Pills, the quality control of Aconiti Kusnezoffii Radix and Lonicerae Radix should be strengthened in the production of Xiaojin Pills.