[关键词]
[摘要]
目的 应用指纹图谱和化学计量学分析方法,比较沪地龙药材基原(通俗环毛蚓Pheretima vulgaris、威廉环毛蚓 P. guillelmi或栉盲环毛蚓P. pectinifera)间质量差异,筛选差异性标志物,为建立沪地龙质量标准提供参考。方法 应用高效液相色谱法建立沪地龙水溶性成分指纹图谱,运用“中药色谱指纹图谱相似度评价系统”(2012版)软件比较相似度;应用聚类分析、正交偏最小二乘判别分析和主成分分析法对3个基原间所测得的6种水溶性成分含量进行比较分析,并依据主成分得分对各批次样本进行综合性评价。结果 相似度评价结果,通俗环毛蚓与威廉环毛蚓的相似度为0.987;栉盲环毛蚓与通俗环毛蚓的相似度为0.697,与威廉环毛蚓的相似度为0.717。聚类分析方法不能区分3个基原。正交偏最小二乘判别分析结果显示,通俗环毛蚓和威廉环毛蚓不能区分,但二者能与栉盲环毛蚓区分,差异性成分为尿苷和鸟苷。主成分分析综合评分共有19批样本大于平均分值,其中栉盲环毛蚓11批、通俗环毛蚓6批、威廉环毛蚓2批;产地为上海市13批,江苏省2批,浙江省1批,安徽省2批。结论 沪地龙3个基原间存在差异,其中通俗环毛蚓与威廉环毛蚓在水溶性成分种类和含量方面相似度高,但二者均与栉盲环毛蚓存在差异。
[Key word]
[Abstract]
Objective In order to compare the quality differences between the basal elements of Hudilong herbs (Pheretima vulgaris, Pheretima guillelmi, Pheretima pectinifera), fingerprint and chemometrics analysis methods were applied, and differential markers were screened, so as to provide reference for the establishment of quality standard of Hudilong. Methods The fingerprints of three origins were constructed by High Performance Liquid Chromatography and the similarities among the Hudilong origins were evaluated by using Similarity Evaluation System of TCM Chromatographic Fingerprints (2012). A total of six aqueous extract components were determined, and the results were analyzed by cluster analysis (CA), orthogonal partial least square discriminant analysis (OPLS-DA) and principal component analysis (PCA). The samples in each batch were comprehensively evaluated according to the principal component scores. Results The similarity between the P. vulgaris and P. guillelmi was 0.987, P. vulgaris and P. pectinifera was 0.697, P. pectinifera and P. guillelmi was 0.717. The results of CA showed that three origins could not be well distinguished. OPLS-DA results revealed that the P. vulgaris and P. guillelmi were indistinguishable from each other, and the P. pectinifera was able to distinguish with the two origins. The recurring differentiating components were Uridine and Guanosine. The PCA comprehensive evaluation score of 19 batches of samples were greater than the average score, among which 11 batches of P. pectinifera, six batches of P. vulgaris, and two batches of P. guillelmi. The regions were Shanghai (13 batches), Jiangsu (two batches), Zhejiang (one batch), Anhui (two batches). Conclusion There are differences among three origins of Hudilong. The P. vulgaris and P. guillelmi have high similarity in terms of aqueous extract components kinds and content, but both are distinctly different from the P. pectinifera.
[中图分类号]
R286.2
[基金项目]
国家自然科学基金资助项目(82225047);科技部"中医药现代化"重点研发计划(2022YFC3501703)