陕西产艾叶中非挥发性成分的初步分析

A preliminary analysis of the non-volatile components in the folium of Artemisae argyi Levl.et Vant.from Shaanxi province

目的用指纹图谱联合多元统计分析陕西产艾叶中的非挥发性成分。方法建立陕西产1年艾叶中非挥发性成分的高效液相色谱(high performance liquid chromatography,HPLC)指纹图谱,用聚类分析进行陕西产艾叶及蕲春产艾叶样本系统分类,用主成分分析(principle component analysis,PCA)联合偏最小二乘法判别分析(partial least squares discriminant analysis,PLS-DA)进行陕西产艾叶及蕲春产艾叶样本指纹图谱主要特征峰比较。结果陕西产艾叶中非挥发性成分指纹图谱样本用甲醇水溶液(50∶50)以料液比1∶60水浴回流提取30 min。HPLC分析参数为:以2 mL·L^(-1)甲酸水溶液(A)-甲醇(B)为流动相梯度洗脱,0~5 min,20%~51%B;5~10 min,51%~60%B;10~15 min,60%B;15~35 min,60%~80%B;35~40 min,80%~95%B;40~50 min,95%B。检测波长为330 nm;流速为0.3 mL·min^(-1);柱温为35℃,进样量为10μL。经验证,该方法精密度、重复性、稳定性均符合2020年版《中华人民共和国药典》(以下简称《中国药典》)相关要求。6个指纹图谱共有峰成分被初步归属;聚类分析发现6批蕲春样本中有5个可与陕北、陕南产艾叶聚为一类,1批可与关中及少部分陕南样本聚为一类;PCA及PLS-DA发现,蕲春产艾叶样本主成分信息与陕西产艾叶大部分重合,无显著分簇特征。结论建立的陕西产艾叶指纹图谱分析方法符合2020年版《中国药典》相关要求;基于指纹图谱共有峰信息进行的聚类分析、PCA及PLS-DA发现陕西产艾叶中非挥发性成分主要特征与蕲春产艾叶无显著性差异。

Objective To analyze the non-volatile components in Artemisia argyi levl.et Vant.from Shaanxi province by high performance liquid chromatography(HPLC)fingerprints combined with multivariate statistical analysis.Methods The HPLC fingerprints of non-volatile components in one-year Artemisae argyi Levl.et Vant.from Shaanxi province was established.Cluster analysis was used to classify the samples of Artemisae argyi Levl.et Vant.from Shaanxi province(Shaanxi)and Qichun area(Qichun).Besides,principal component analysis(PCA)combined with partial least squares discriminant analysis(PLS-DA)was employed to compare the main characteristics of the fingerprints of Artemisia argyi levl.et Vant.from Shaanxi and Qichun.Results The optimal parameters of fingerprints of non-volatile components in Shaanxi Artemisae argyi Levl.et Vant.were as fo-llows:Samples were prepared by 50%methanol with a liquid to material ratio of 1∶60.The mobile phase consisted of 2 mL·L^(-1) formic acid solution(A)and methanol(B).The gradient elution program of mobile phases was as follows:0-5 min,20%-51%B;5-10 min,51%-60%B;10-15 min,60%B;15-35 min,60%-80%B;35-40 min,80%-95%B;40-50 min,[JP1]95%B.The detection wavelength was 330 nm,the flow rate was 0.3 mL·min^(-1),the column temperature was 35℃,and the injection vo-lume was 10μL.The precision,repeatability and stability of the method met with the relevant requirements of the Chinese Pharmacopoeia(2020 edition).The peak components of the 6 fingerprints were preliminarily assigned.The cluster analysis showed that 5 of the 6 batches of Qichun samples could be clustered into the same group with northern and southern Shaanxi samples,and one batch could be clustered with Guanzhong and a small number of southern Shaanxi samples.PCA and PLS-DA showed that most of the principal component information of Qichun samples coincided with Shaanxi Artemisae argyi Levl.et Vant.and there was no statistically significant clustering feature.Conclusion The fingerprint analysis parameters of Artemisae argyi Levl.et Vant.in this paper complied with the Chinese Pharmacopoeia(2020 edition).According to the clustering analysis,PCA and PLS-DA based on the common peak information of the fingerprints,no significant difference was obtained between Artemisia argyi levl.et Vant.samples from Shaanxi and Qichun in terms of non-volatile components.