| Polygonum multiflorum Thunb. is one of the most commonly used traditional Chinese medicines (TCMs). Its dried roots and rattan are well known as "Heshouwu" (Polygoni Multiflori Radix, PMR) and "Shouwuteng"(Polygoni Multiflori Caulis, PMC). The PMR was widely used in clinic for many diseases in raw state or processed form (Polygoni Multiflori Radix Praparata, PMRP). The PMR is used for antioxidation and purgation, whereas the PMRP is used as a tonic and an antiaging agent. PMC had the actions of nourishing blood and calming mind. However, due to differences in harvest time, origin and processing, resulted in the accumulation of the active ingredient in PMR had a great variation and varying quality. The key of manufacturing procedure of PMR is how to ensure its high and uniform quality. This topic through the systematic analysis of the multi-index constituents in PMR, PMRP and PMC, compared the differences between index components in each samples from different origin and commercial herbs, and the index components changes during different harvest time and the processing method. Studied on the differences in chemical constituents on the PMR from different growing stages, regions, processing methods, before and after processed and different parts by UPLC-Triple TOF-MS/MS. Simple and reliable HPLC and fatty acid derivatization GC-MS methods have been developed and validated for the fingerprintings for quality control of PMR. This paper provided basic data and scientific basis to explore the formation process of medicinal materials quality, chose suitable production and appropriate reasonable harvesting and processing.The research mainly included the following contents:1、The literature research of Polygoni Multiflori Radix By consulting the related literature at home and abroad, a literature review of the study was completed.2、Analysis of index components content in PMR, PMRP and PMC The chemical componsitions of PMR in different harvest time, different origin and commercial herbs, different processing method, before and after processed and different parts were analyzed. The principal component analysis (PCA) and Topsis were carried out to have a comprehensive evaluation. UPLC-TQ/MS was developed for simultaneous determination of four kinds components including three stilbenes (stilbene glucoside, polydatin, resveratrol), five anthraquinones (emodin, physcion, emodin-8-β-D-glucopyranoside, rhein, aloe-emodin), five flavonoids (epicatechin, rutin, hyperoside, astragalin,quercetin) and one phenolic acid (gallic acid) in Polygonum multiflorum. The method of MEKC-DAD for simultaneous determination of seven components including stilbene glucoside, emodin, aloe-emodin, rhein, chrysophanol, physcion, catechin. The nucleosides and nucleobases had been simultaneous analyzed and determination by QTRAP LC-MS/MS. The polysaccharide, phospholipids, tannins were detected by UV-VIS; The content of 24 kinds of mineral elements had been detected by ICP-MS. The volatile components were separated and identified by GC-MS. After derivatization by GC-MS analysis of fatty acid components and relative quantity. Results showed that: The dynamic change among the contents of active ingredients in various collecting samples of different harvest time and processing method; There are obvious differences among the contents of active ingredients in various collecting periods samples of different origin and commercial herbs, before and after processed and different parts.3、Studies on the plant metabolomics of Polygoni Multiflori Radix based on UPLC-Triple TOF-MS/MS Studied on the differences in chemical constituents on the PMR from different regions and commercial herbs, growing stages, processing methods, before and after processed and different parts by UPLC-Triple TOF-MS/MS with multivariate statistical analysis. Peak matching, peak alignment and noise filtering were used in analyzing mass spectrometry data. Accurated m/z value analysis of MS data based on software of database search and MS/MS fragment analysis were applied to constituents identification. The results showed that: In different harvest times of PMR, the chemical compositions of traditional harvest time and other harvest times were different. There were 9 differences in the chemical composition showed different variation. The chemical compositions of PMR from different regions had obvious differences. There were 12 differences in the chemical composition showed different variation. It was obvious differences among samples with different processing methods, including PMR of drying and dryed in the sun, There were 15 differences in the chemical composition showed different variation; Chemical compositions of PMR before and after processed were significantly different, There were 14 differences in compositions effected in differentiation except catechin glycosidase, THSG-glucuronide, emdin-O-glucoside sulfat, rendin A in PMR before processed and 5-HMF in PMR after processed; It was obvious differences of chemical constituents in PMR and PMC with the same origin. In addition to quinic acid, gallic acid,3,8-dihydroxy-l-methoxyxanthone, resveratrol-4’-O-β-D-glucopyranosid, rhein, rhaponticin, citreorosein and physcion in PMR, laccaic acid-D-8-O-(6’-O-cinnamyl)-glucopyranoside, pocyanidian-B-7-3-O-gallate, 3,4’,5-trihydroxystlbene-4’-O-β-D-(6"-O-galloyl)-glucopyranoside, cassialoin, epicatechin-(4β-8)-epicatechin-(4β-8)-catechin, aloesone-7-O-β-D-glucopyranoside, physcion-8-O-β-D-gentiobioside in PMC, There were 12 differences in the chemical composition showed different variation.4. To establish the fingerprint chromatogram of Polygoni Multiflori Radix Simple and reliable HPLC and fatty acid derivatization GC-MS methods have been developed and validated for the fingerprintings for quality control of PMR, and the fuzzy cluster analysis and similarity evaluation, the overall description and evaluation of the quality of commercially available of PMR products. The results show that, initially established a HPLC fingerprinting with 19 common peaks as fingerprint information of PMR and fingerprint of PMR fatty acid to 9 common peaks for the characteristic information of the acid, the chromatographic separation is better, to achieve the technical requirements of the fingerprint of traditional Chinese medicine. The method is accurate and reliable, reproducible, and can be used for the evaluation of PMR intrinsic quality and control. |
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