Study On Chemical Constituents And Quality Standards Of Radix Polygoni Multiflori Preparata

In comparison with the herbs and natural medicines of other countries,an outstanding characteristic of Chinese materia medica was that Chinese materia medica must be processed before used for clinic.The ancients said,"Processing is as much as training,leading the untrained soldiers cannot overcome the enermy,using the unprocessed drugs is hard to cure the diseases",therefore,the processing is very important to Chinese materia medica.But the basic reason that the processing has great affect to the drug property and efficacy was the change of those internal substances after processed.Study the chemical constituents' changes of before and after processing have important significance to clarify the processing principle of Chinese materia medica and to formulate the quality standard.Heshouwu,the dried root of Polygonum multiflorum Thunb.The crude have the efficacy of detoxication,eliminating carbuncle and loosening bowel to relieve constipation,and used to treat scrofula,sores,carbuncle,rubella,pruritus,constipation and hyperlipidemia.Radix Polygoni Multiflori Preparata,the processed products of Radix Polygoni Multiflori,have the efficacy of reinforcing liver and kidney,tonifying blood and essence,blacken beard and hair, strengthen the tendons and bones,and used to cure hemopenia,etiolate,vertigo,tinnitus, beard and hair graying early,waist and knees lassitude,limb numbness,metrorrhagia and leucorrhea,chronic malaria and body deficiency,hyperlipidemia.This study want to clarify the change of Polygonum multiflorum Thunb.before and after processed and formulate the scientific reasonable qualify evaluation method of Radix Polygoni Multiflori Preparata by extracting and separating the chemical constituents of Radix Polygoni Multiflori Preparata. On this basis,the thin-layer chromatography(TLC) identification method for distinguishing the Radix Polygoni Multiflori and Radix Polygoni Multiflori Preparata was established,the HPLC fingerprint spectrum analysis method of Radix Polygoni Multiflori Preparata have been studied,and the main effective constituents contents in Polygonum multiflorum Thunb. which affected by different processed technology and method have been studied.1 Literature reviewThe research literatures of Polygonum multiflorum Thunb.in the fields of bencaology, preparation,quality control,chemical constituent,pharmacology and clinical application were summed up and analyzed.Based on this the feasible research scheme was formulated,the research investigation index and the researches originating point were determined.2 Experimental research2.1 Study on chemical constituents of Radix Polygoni Multiflori Preparata.Accordingto the modern research,the stilbene glucoside was the main active constitute of Polygonum multiflorum Thunb.,but not the only active constitute,the research shows that its content will decrease after processing.In this experiment,the roots of P.multiflorum were extracted with EtOH,and the EtOH solution obtained evaporated to dryness.The EtOH extract was suspended in water,partitioned successively with,with the resulting EtOAc and n-BuOH solutions evaporated to dryness in vacuo.Ten compounds were isolated by chromatographic fractionation in the EtOAc and n-BuOH extract,and identified by UV,MS,¹H-NMR, ¹³C-NMR.The ten compounds were emodin,physcion,succinie acid,5-hydroxymethyl furfural,4-Hydroxybenzaldehyde,garlic acid,hydroquinone,2,3,5,4'-tetrahydroxy-stilbene-2-O-β-D-glucoside, emodin-8-O-β-D-glucoside,physcion-8-O-β-D-glucoside.The succinic acid and 5-hydroxymethylfurfural were the first found in Radix Polygoni Multiflori Preparata.2.2 Study on quality standard of Radix Polygoni Multiflori Preparata.2.2.1 the thin-layer chromatography(TLC) identification method for distinguishing the Radix Polygoni Multiflori and Radix Polygoni Multiflori Preparata was established.Now the crude and processed product of Polygonum multiflorum Thunb.were identified by the same TLC method.Based on the study of chemical constitutes in Radix Polygoni Multiflori Preparata,this study choose 5-HMF which was the new product after processing as the reference substance,adopt the silica GF254 thin layer plate with the developing solvent of petroleum benzene(60℃-90℃)-EtOAc(1:1),and detected by UV detection at 254 nm,the same color stains were detected on the spectrum of both Radix Polygoni Multiflori Preparata and reference substance,the same color stain was not found in Radix Polygoni Multiflori.2.2.2 Study on the HPLC fingerprint spectrum of Radix Polygoni Multiflori Preparata. Based on the study of 15 Radix Polygoni Multiflori Preparata,the HPLC fingerprint spectrum analysis method of Radix Polygoni Multiflori Preparata was established.2.2.3 Assay of the 2,3,5,4'-tetrahydroxy-stilbene-2-O-β-D-glucoside,emodin-8-O-β-D-glucoside, physcion-8-O-β-D-glucoside in Radix Polygoni Multiflori Preparata.The contents of stilbene glucoside in ten groups Radix Polygoni Multiflori Preparata buying from different places were determined,the results show that the content of stilbene glucoside in was 0.127～4.150%,and there have a great difference between the different place sample,even some not meet the pharmacopoeia of people's republic of chinesestandard that the content of stilbene glucoside in Radix Polygoni Multiflori Preparata can't below 0.70%.2.3 Effect of processing on the main chemical constituents in Polygonum multiflorum Thunb..Aiming at current situation that the disunity of processing technology in different city and the lack of processing technology parameter standard,we study the effect of different processing technology such as black bean sauce steaming,black bean sauce stewing,water stewing,black bean sauce steaming at high pressure,black bean sauce stewing up the lattice and different processing time on the main chemical constituent stilbene glucoside in Polygonum multiflorum Thunb..The results show the contents of stilbene glucoside as: crude＞black bean sauce steaming at high pressure＞black bean sauce stewing＞black bean sauce steaming＞water stewing＞black bean sauce stewing up the lattice.In the processing of black bean sauce stewing,the content of stilbene glucoside in Polygonurn multiflorum Thunb. decreased with processing time.