| Quality control of Traditional Chinese Medicine (TCM) is the foundation and the key of realizing standardization, industrialization, modernization and internationalization of TCM. while efficient constituents screening is to supply the prerequisite and foundation for quality control of TCM. However, Study on efficient constituents of TCMs did not gain breakthrough progress, and efficient constituents of most TCMs are still not identified due to the their own complexcities. Understanding that biological essential of the regulative rule of efficient constituents from TCM were affected by environmental factors is lack. Metabolic accumulations (contents) of TCM efficient constituents were remarkably different due to mulriple factors would be affected quality of TCM. This is one of maninly basic scientific issue to discuss the process of metabolic accumulations of TCM efficient constituents, correlation between their efficient constituents and influence facters. Houttuynia cordata Thunb. as TCM of homology of medicine and food, include antioxidant activities, which were showed potential synerigism of phenolics and volatiles, and other multi-activities. The contents of phenolics and volatiles of H. cordata influence quality of H. cordata for factors of geography, climate and growth technology and so on. To discuss the process of metabolic accumulations of phenolics and volatiles of H. cordata and the influences and interactions of them with environmental factors is important significances to uncover efficient constituents screening of H. cordata TCM and steady quality of H. cordata, In this thesis, by multidiscipline intersection interdisciplinary theory from metabolomics, the process of metabolic accumulations of phenolics and volatiles of H. cordat from different regions and culture, correlation between the constituents, and correlation of the constituents with environmental factors of growing soils were investigate using HPLC and GC-MS, etc. The results obtained were following,1. The validated methods developed for simultaneous determination of sixteen phenolics i.e., chlorogenic acid, scopoletin, vitexin, rutin, afzelin, isoquercitrin. narirutin, kaempferitrin, quercitrin, quercetin, kaempferol, chrysosplenol D. vitexicarpin,5-hydroxy-3,3’,4’,7-tetramethoxy flavonoids,5-hydroxy-3,4’,6,7-tetramethoxy flavonoids and kaempferol-3,7,4’-trimethyl ether using HPLC-DAD and simultaneous determination of25volatiles i.e, a-pinene, camphene,/?-pinene,2-methyl-2-pentenal, myrcene,(+)-limonene, eucalyptol,?7-<m?-2-hexenal, y-terpinene, cis-3-hexenyl-acetate,1-hexanol, a-pinene oxide, cis-3-hexen-l-OH, trans-2-hexen-l-OH, decanal, linalool, L-bornyl acetate,β-caryophyllene,2-undecanone, terpiene-4-OH,(-)-bornol,1-decanol, eugenol, isophtol and phytol using GC-MS were successfully applied for ambulatory monitoring of H. cordata samples because of less the sample consumption (only0.05g) which do not influence natural physiological growth of H. cordata. The methods obtained were showed that good linear behaviors over the investigated concentration ranges were observed with the values of r2higher than0.9990for all the analytes, that relative standard deviation of good precision, repeatability and stability were not more than5%, and htat the recoveries measured at three levels varied from98.53to102.25%.2. The accumulation (contents) of the polyphenols, all flavonoids,16phenolics and25volatiles of35H. cordata samples from different regons, and structure properties of microbial communities, enzyme activities and nutritional characteristics of their rhizosphere soils were developed as data-base. The data about phenolics and volatiles of35H. cordata samples from different regons, and PLFAs abundances of microorganism, enzyme activities and nutrition of their growing soils were analyzed. The Analytic results show that the contents of maintly phenolics and volatiles, structure properties of microbial communities of their growing soils, their enzyme activities and nutrition nutritional characteristics were diversity and difference with samples from different regions, and them were interact.3. Primarily to illustrate that, as a whole, growth of H. cordata can promote growth of soil microorganism, and structure properties of soil microbial communities were affected. The results that, as a whole, the PLFAs abundances of all G+, all G", all bacteria, all fungi, all actinomycetes, and all microorganism of growth soil of H. cordata were significantly higher than CK after H. cordata collected different regions were continuously culture two years at same place. Comparison of the PLFAs of soil microorganism collected growth soils of continuously cultural H. cordata, some PLFAs abundances of microorganism would increase (e. g., a-15:0,18:lco9t), and some decrease (e. g.,/-16:0,14:0).4. To identify that metabolic accumulation (contents) of phenolics of H. cordata were affected by environmental facters, and metabolic accumulations (contents) of volatiles of H. cordata were affected by genetic and environmental facters. The results obtained show that contents of phenolics of H. cordata samples were not significantly different, while the constituents and contents of volatiles of H. cordata samples and their trend of change among H. cordata samples were consistent with the H. cordata from different regions when H. cordata, which collected different regions and were significantly different constituents and contents of phenolics and volatiles, were continuously culture at same place.5. The change trends of metabolic accumulation (contents) of phenolics and volatiles at growth process of H. cordata were identified, and are following:(1) the contents of phenolics and volatiles of leavies and rhizomes from H. cordata first increased, and then decreased with time, and their contents were highest at reproductive growth period.(2) The change trends of contents of phenolics of leavies, floweres and rhizomes from H. cordata were following:flower> leaf> rhizomes.(3) the change trends of contents of phenolics from H. cordata flowers were that the contents of volatiles would be increasing from early flowering stage to opening flower stage, then decreasing from opening flower stage to flower-perished period, and the contents of volatiles at opening flower stage were highest.(4) The trends that the contents of volatiles of leavies, floweres and rhizomes from H. cordata flowers change with time were different with different volatiles.6. To discuss that the correlation of the phenolics and volatiles of H. cordata with soil environmental factors (e. g. structure properties of microbial communities, enzyme activities and nutritional characteristics), respectively. It is suggested that H. cordata could promote propagation of soil microorganism and enzyme activities, and activate soil nutrient, which minght be related to metabolites of phenolics and volatiles at growth process of H. cordata. The results of correlation analysis obtained were following:1) One or some constituents of the phenolics and/or volatiles of H. cordata wre significantly correlated with PLFAs abundances of one, or some and or one genre microorganism of growth soils, respectively (P<0.05), and correlation between constituents of the phenolics and/or volatiles of H. cordata and the PLFAs abundances of all bacteria, all fungi, all actinomycetes and all microorganism of growth soil of H. cordata are different with different constituents, respectively. The contents of polyphenols and all flavonoids are significantly positive correlated with the PLFAs abundances of all bacteria, all actinomycetes and all microorganism of H. cordat growth soils (P<0.05), respectively, and significantly negative correlated with the PLFAs abundances of all fungi of H, cordat growth soil (P<0.05), respectively. This suggests that structure properties of microbial communities are affected by the phenolics and/or volatiles of H. cordata.2) Constituents of the phenolics and/or volatiles of H. cordata wre significantly correlated with activities of one or some of catalase, phosphate, urease, polyphenol oxidease, and invertase of H. cordat growth soil (P<0.05), respectively, The contents of polyphenols and all flavonoids are significantly positive correlated with activities of catalase, polyphenol oxidease and phosphate, respectively (P<0.05).3) The contents of polyphenols and all flavonoids are significantly positive correlated with available P, available K and orgamic matter respectively (P<0.05), one or some constituents of the phenolics and/or volatiles of H. cordata wre significantly correlated with one or some of available P, available K, available N and orgamic matter, respectively (P<0.05). For examples, chlorogenic acid wre significantly positive correlated with available P, available K and orgamic matter, respectively (P<0.05), a-pinene and2-undecanone wre significantly positive correlated with available N, respectively (P<0.05), a-pinene, decanal and β-caryophyllene wre significantly positive correlated with orgamic matter, respectively (P<0.05), camphene,2-undecanone,β-pinene, myrcene,(+)-limonene, L-bornyl acetate wre significantly positive correlated with available P, available K, respectively (P<0.05). This suggests that the soil environmental factors can induce and influence metabolic accumulations of the phenolics and volatiles of H. cordata.7. Primarily to discuss that correlation of the phenolics with antioxide activities in H. cordata. The results obtained show that the contents of polyphenols and all flavonoids are significantly positive correlated with scavenging rate of DPPH free radical of H. cordata extracts, respectively (P<0.05). Constituents of the phenolics of H. cordata are positive correlated with scavenging rate of DPPH free radical of H cordata extracts, but not significantly. This suggests that the antioxide activities of H. cordata are due to the contribution of antioxide activities of every the phenolics, and they is organic whole. |
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