基于HPLC指纹图谱和多成分含量测定的辽藁本质量评价研究

Quality evaluation of Ligusticum jeholense based on HPLC fingerprint and multi-component analysis

目的建立辽藁本的HPLC指纹图谱及多成分含量测定方法。方法采用Thermo Acclaim TM 120 C18色谱柱(250 mm×4.6 mm,5μm);以乙腈(A)-0.1%甲酸水溶液(B)为流动相,梯度洗脱;体积流量1.0 mL/min;检测波长280 nm;柱温为30℃,同时测定15批辽藁本指纹图谱及相关成分含量,结合聚类分析、主成分分析及偏最小二乘法判别分析(partial least squares discriminant analysis,PLS-DA)对辽藁本进行全面分析。结果15批辽藁本指纹图谱相似度大于0.958,共标定了12个共有峰,指认出绿原酸、阿魏酸、洋川芎内酯I、洋川芎内酯H、洋川芎内酯A、藁本内酯、欧当归内酯A 7个成分,对前6个成分进行定量分析,其质量分数分别为0.108%~0.524%、0.168%~0.268%、0.016%~0.029%、0.004%~0.007%、0.056%~0.033%、0.590%~1.293%。聚类分析将15批药材分为3类,主成分分析筛选出4个主成分,其累积方差贡献率为87.94%,表明这4个主成分可包含原有数据的大多数信息,PLS-DA法标记出药材中的5个差异性成分。结论建立的指纹图谱及多成分含量测定方法适用于辽藁本的质量评价,为辽藁本质量控制及相关经典名方开发提供参考。

Objective To establish the HPLC fingerprint and muti-component analysis of raw Ligusticum jeholense.Methods Thermo Acclaim TM 120 C18 column were performed on HPLC analysis.the mobile phase was acetonitrile(A)-0.1%formic acid(B)for gradient elution.The flow rate was 1.0 mL/min,the wavelength was 280 nm and the column temperature was 30℃.The content and fingerprint of relative ingredients of 15 batches were measured simultaneously,combined with cluster analysis,principal component analysis and partial least squares discriminant analysis(PLS-DA),which were applied in comprehensive analysis.Results The similarity of 15 batches of L.jeholense was greater than 0.985,and 12 common peaks were calibrated,Chlorogenic acid,ferulic acid,senkyunolide I,senkyunolide H,senkyunolide A,ligustilide,levistilide A were identified.The first six components were quantitatively analyzed,and their mass fractions were 0.108%—0.524%,0.168%—0.268%,0.016%—0.029%,0.004%—0.007%,0.056%—0.033%and 0.590%—1.293%,respectively.Cluster analysis classified the 15 batches of medicinal materials into three categories,principal component analysis picked out four main ingredients,and the cumulative variance contribution rate was 87.94%,indicating that the main ingredients contained the most information of the raw statistics.PLS-DA screened out five difference components.Conclusion The established fingerprint and multi-component quantitative analysis can be applied to the quality assessment of L.jeholense,which also can provide reference for the quality control and the development of related prescriptions of L.jeholense.