| ObjectivesTo scanning the chemical compounds of folium eucommiae.; to examined the seasonal and regional difference of the chemical compositions through the folium eucommiae. from different regions in different seasons; to establish multi-wavelength liquid chromatography fingerprint.Methods and Results1Through the separation and purification of folium eucommiae., nine chemical compositions including geniposide acid, chlorogenic acid methyl ester, chlorogenic acid, caffeic acid,quercetin-xylose-glucoside, rutin and quercetin-3-O-β-D-glucoside, kaempferol-3-O-β-D-glucoside and quercetin were obtained, where the chlorogenic acid methyl ester was extracted from folium eucommiae. for the first time.2Taking chlorogenic acid as reference substance and relative correction factors(RCFs) of geniposidic acid, caffeic acid, rutin and isoquercitrin to chlorogenic acid were determined by HPLC. The contents of five components were determined by external standard method(ESM) and QAMS. Through comparing with the external standard method, the result showed no significant difference between the two methods (P>0.05), which indicated that quantitative analysis of multi-components by single marker method is credible and accurate.3Based on the quantitative analysis of multi-components by single marker method of chlorogenic acid, geniposide acid, rutin, quercetin3-O-β-D-glucoside of folium eucommiae. from different regions in different season, studying the seasonal and regional differences of chlorogenic acid, geniposide acid, rutin, quercetin3-O-β-D-glucoside. The results showed that the contents of chlorogenic acid and geniposide acid increased and then decreased, finally tended to a steady trend; while rutin and quercetin3-O-β-D-glucoside first decreased and then increased. Besides, the contents of the four compositions are quite different among the folium eucommiae. from different regions. As shown in the results, the optimum harvesting time of folium eucommiae. from Zhangjiajie in Hunan province is May and that of Kunming, Yunnan province and Guiyang, Guizhou province is June. Therefore this study has provided theoretical foundation to ensure the quality of folium eucommiae..4A simple and accurate fingerprint method was developed using high-performance liquid chromatography-photodiode array detection (HPLC-DAD) combined with chemometrics methods. The results are as followings:in PC A,27batch folium eucommiae. were respectively gathered in the region A (Zhangjiajie, Hunan,201104-10; Guiyang, Guizhou201106; Tangshan, Hebei201108), the region B (Guiyang, Guizhou,201108-10; Enshi, Hebei,201107; Hechi, Guangxi,201109; Changde, Hunan201109; Shaoyang, Hunan201109), the region C (Kunming, Yunnan201106; Kunming, Yunnan,201108-10; Cili, Hunan201201; Yuxi, Yunnan201110; Changde, Hunan201201). By HCA, when the Euclidean distance is2.27, classifying the27batch of specimens into four categories, where the No.10specimen was separated into a single category and the others were consistent with the figure of PCA. Which indicates that the quality of folium eucommiae. have large relevance with the producing area and selected time. Also, through loadings, selecting geniposide acid, chlorogenic acid methyl ester, chlorogenic acid as key indicators for chemical compounds of folium eucommiae.ConclusionsFolium eucommiae., as a herb with enormous pharmacological activity and rich resource, has a great potential to be developed. However, the quality of folium eucommiae. is easily affected by the product origin and the harvest time. This thesis, through the studies of the composition and multi-wavelength liquid fingerprint of folium eucommiae., has provided an experimental base to establish the quality standard for folium eucommiae., which is a key factor to take full advantage of folium eucommiae. as a valued herb. |
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