| Traditional Chinese medicine is a complex mixture. It has many species different process and complex action mechanism. Quality control and evaluation are the difficulties in the modernization of Traditional Chinese Medicine research. Based on the concept of the whole in Chinese Medicine theory, with the development of modern analytical chemistry techniques, The chromatographic fingerprint technology has been widely used in traditional Chinese Medicine and its preparation quality control and evaluation. Especially1H-NMR fingerprints is a more integrated, holistic, comprehensive, systematic techniques. it can detect plant primary and secondary metabolites simultaneously, it is a indispensable method for holistic metabolite analysis.Diversity of origin and species were the difficulty of quality control and evaluation. At present, the way of quality control about Phellodendri Cortex and Alisma orientalis(Sam.) Juzep. almost were derterminate single or multiple active ingredient content by HPLC, it is not enough to reflect herbs multicomponent, multi-target characteristics. So it is badly in need of new quality control methods for studying different areas and different varieties of herbs systematically. Early in our laboratory, based on1H-NMR metabonomics technology has been successfully applied to distinguish Rhizoma Coptidis and "Zangyingchen". This paper draws on our previous work in the laboratory,1H-NMR metabonlomics technology is used to discriminate the geographical oringin of Rhizoma Alismatis and identify Cortex Phellodendri, that aims to strengthen quality control and innovate the model of quality evaluation of TCM.Purposes:In this paper, we choose Alismatis Rhizoma and Phellodendri Cortex as the object of research. Use1H-NMR metabonomic technology to discriminate TCM of different species or geographical origin, reveal the holistic metabolite difference of samples from different geographical origin or different species. This method can provide theory basis for Rhizoma Alismatis and identify Cortex Phellodendri of quality control and evaluation, utilizating species resource and guiding the clinical uses.Methods:1. Estabolish holistic metabolite fingerprint of Phellodendri Cortex and Alismatis Rhizoma by1H-NMR. Combine principal component analysis and partial least squares discriminant analysis to find out the metabolic markers.2. Apply HPLC method to determinate five active component and verify the results of chemometric analysis.Results:1. Four primary metabolites of Alisma orienta/is(Sam.)Juzep extract were identified as23-O-Acetylalisol B, sucrose, β-galactose, and alanine.Though PCA and PLS-DA analyse, score plot shows four different origin samples can be clearly separated. We determine the four metabolites simutanoeusly, the relative content of23-O-Acetylalisol B, sucrose, β-galactose, alanine were0.07%-0.19%.0.67%-6.58%,0.20%-0.56%.0.09%-0.18%, respectively. It is found that the sample collecting from Sichuan was the best quality through quantitive’H-NMR analysis of23-O-Acetylalisol B.2. Eleven primary metabolites of Phellodendri Cortex extract were identified as berberine, palmatine, phellodendrine, chlorogenic acid, caffeic acid, ferulic acid, obacunone, magnoflorine, sucrose, α-glucose, β-glucose, saturated fatty acids and unsaturated fatty acids. PCA and PLS-DA scores plot shows Phel/odendron chinense Schneid. and P.amurense Rupr. can be clearly separated. However, P.chinense Schneid.and P.chinense Schneid.var glabriusculum Schneid. gathered together, can not be effectively distinguished. Through the loading chart, it is found that P. chinense Schneid. has more berberine, phellodendrine, ferulic acid, caffeic acid, chlorogenic acid, P. amurense Rupr. has more magnoflorine, palmatine, obacunone, saturated fatty acids and unsaturated fatty acid and part unidentified sugar. Some metabolites are selected to integral manually and quantitative analysis by1H-NMR, the relative contents of berberine, phellodendrine, caffeic acid, chlorogenic acid, sucrose, α-glucose, β-glucose in Pchinense Schneid.were3.47%.0.44%,0.20%, 2.02%,1.30%,0.32%,0.42%, respectively; the relative contents of berberine, palmatine, phellodendrine,, chlorogenic acid, sucrose, α-glucose, β-glucose in P. amurense Rupr. were0.50%,0.33%,0.15%,0.15%,0.52%,1.58%,0.36%,0.50%, respectively. The result are basically consistent with the1H-NMR metabonomic.3. The determination of berberine, phellodendrine, magnoflorine in P. chinense Schneid. were5.44%,0.45%,0.08%by HPLC analysis, respectively. The dertermination of berberine, phellodendrine, magnoflorine, palmatine, obacunone in P. amurense Rupr. were:0.64%,0.08%,0.31%,0.35%,0.18%by HPLC analysis, respectively. By analysis of variance, phellodendrine, magnoflorine, palmatine, berberine, obacunone in P. amurense Rupr. and P. chinense Schneid. has a significant difference.Conclusions:1. Metabolites of Alisma orientalis(Sam.)Juzep.from four different geographical origin were significantly different.23-acetylalisol B, sucrose, β-galactose, alanine might be as the metabolites maker of discriminating difference origin. The samples from Sichuan origin has best quality. It was consistent with the say that Sichuan are considered the genuine area of Alisma orientalis(Sam.) traditionally. The results of NMR quantitative analysis are basically consistent with the results of chemometrics analysis.2. Metabolites of P.amurense Rupr. and P.chinense Schneid. had significantly difference. Berberine. palmatine, phellodendrine, chlorogenic acid, caffeic acid, ferulic acid, obacunone, magnoflorine, sucrose, a-glucose,(3-glucose, saturated fatty acids and unsaturated fatty acids might be as the metabolites maker for distinguishing two species. The difference between P.chinense Schneid.and P.chinense Schneid.var glabriusculum Schneid. was not significant. P. chinense Schneid. is very rarely, so we suggested that P.chinense Schneid.var glabriusculum Schneid. should be included as a source of P. chinense Schneid. in Chinese pharmacopoeia. |
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