| ObjectiveThis article takes Radix Polygoni multi flora praeparata(RPMP)as the research object.Based on the research on the chemical composition of RPMP,the traditional classic methods and modern analysis techniques are combined to improve the quality standards of RPMP.To provide a new method and ideas for the establishment of quality grade standards for making RPMP.Methods1.The ethyl acetate fraction of 70%ethanol extract from RPMP was separated by normal phase silica gel,ODS reverse phase silica gel chromatography,LH-20 gel and semi prepared HPLC.The structure of the compound was identified by LC-MS/MS and nuclear magnetic resonance(NMR)spectroscopy,combined with the data reported in the literature.2.On the basis of the study on the chemical components of RPMP,the characteristics,TLC and fingerprint of Radix Polygoni multiflori and Radix Polygoni Multiflori at different processing degrees,the chemical components related to processing time were summarized.3.The 57 batch samples of RPMP were collected from 7 production areas,including Guangdong,Guizhou,Sichuan,Chongqing,Henan,Yunnan and Zhe jiang.Characteristic and microscopic identification,thin layer identification,detection of extracts and content determination.New thin layer identification and fingerprint detection methods related to processing time were added,and a comprehensive evaluation model was adopted to establish a standard for evaluating the quality grade of RPMP.Results1.Nineteen monomer compounds were isolated from RPMP.Succinic acid(Ⅰ),P-hydroxybenzaldehyde(Ⅱ),P-hydroxybenzoic acid(Ⅲ),Protocatechuicaldehyde(Ⅳ),5-hydroxymethyl furoic acid(V),Protocatechuic acid(Ⅵ),Gallic acid(Ⅶ),2-(2-hydroxypropyl)-5-methyl-7-hydroxychromone(Ⅷ),2-methyl-5-carboxymethy1-7-hydroxychromone(Ⅸ),Daidzein(Ⅹ),2,3,5,4’-tetrahydroxy distyrene-2-O-β-D-glucoside(Ⅺ),Emodin methyl ether-8-O-β-D-glucoside(Ⅻ),Emodin-8-O-β-D glucoside(ⅩⅢ),Catechin(ⅩⅣ),Epicatechin(ⅩⅤ),Emodin methyl ether(ⅩⅥ),Emodin(ⅩⅦ),2,5-dimethyl-7-hydroxychromone(ⅩⅧ).Compounds(Ⅲ),(Ⅳ),(Ⅴ),(Ⅵ),and(Ⅷ)were isolated from RPMP for the first time.The appearance or enrichment of compounds(Ⅴ),(Ⅵ),(Ⅶ),(ⅩⅨ)is related to processing RPMP time.2.Supplement the quality standards of RPMP:Based on the quality standards of Polygonum multiflorum based on the current version of the Chinese Pharmacopoeia(Part Ⅰ),the following new test contents have been added:(1)Microscopic characteristics of the powder of RPMP.(2)The character grading characteristics and powder microscopic characteristics of the newly prepared pieces of RPMP.A new TLC identification method of RPMP was added that the powder of RPMP was 1g,70%ethanol was used as the solvent,ultrasonic extraction was used,the GF.54 thin-layer plate,the lower solution of dichloromethane ethyl formate-formic acid(8:5:0.4)was used as the developing agent,and the ultraviolet lamp(254nm)was used for inspection.5-Hydroxymethylfurfural can be used as an index component to evaluate the processing degree of of Polygonum multiflorum Thunb are processed completely.(3)Establishment of Soybean Polygonum multiflorum TLC Identification Method.Soybean Polygonum multiflorum 1g with 70%ethanol as solvent,ultrasonic extraction,GF254 thin-layer plate,lower layer solution of dichloromethane-ethyl formate-formic acid(8:5:0.5)as a developing agent,and sprayed with 1%trichloride Method of heating aluminum solution at 105℃and inspecting under an ultraviolet lamp(365nm).As a thin-layer chromatography identification method of Soybean Polygonum multiflorum,daidzein spots can be used as index components to identify Soybean Polygonum multiflorum and steamed Polygonum multiflorum.(4)The HPLC fingerprint of RPMP was established,belonging to 14 common peaks(the fingerprint of the sample with the missing 5-hydroxymethylfurfural peak belongs to 13 common peaks),9 of which were identified and establishment of gallic acid,5-hydroxymethyl furfural,catechins,epicatechin,stilbene,emodin-8-O-β-D-glc,emodin-8-O-β-D-glc,emodin,emodin methyl ether content determination method.The content range was 0.77-2.56 μg/mg,0.00-3.08 μg/mg,0.43-10.55 μg/mg,0.66～10.04μg/mg,1.19-28.02 μg/mg,0.15-2.70 μg/mg,0.19-1.71 μg/mg,0.40-4.14 μg/mg,0.13-1.88 μg/mg,respectively.3.Based on the established quality standards of RPMP,57 batches of RPMP samples were comprehensively evaluated.Among,the content of 5 batches of stilbene glycosides and 2 batches of free anthraquinone contents were lower than those under the current version of Chinese Pharmacopoeia(PartⅠ).Listed as rejects.The remaining 50 batches can be divided into the following categories:A.5-hydroxymethylfurfural is not detected in the fingerprint and thin-layer chromatography and the identification of the characteristics does not fully meet the description in the Pharmacopoeia.B.A trace amount of 5-hydroxymethyl furfural can be detected by fingerprint,but the spot is not found in thin-layer chromatography,and the identification of the characteristics does not fully comply with the description in the Pharmacopoeia.C.5-Hydroxymethyl furfural can be detected in both fingerprint and thin-layer chromatography,and the identification of the characteristics is consistent with the description in the Pharmacopoeia.Based on the differences in the contents of extracts,stilbene glycosides and free anthraquinone,it can be divided into 2 grades.Conclusion1.Among the 19 compounds isolated from the ethyl acetate extraction site of RPMP,5 compounds(Ⅲ),(Ⅳ),(Ⅴ),(Ⅵ)and(Ⅷ)were first isolated from RPMP,compound(Ⅴ),(Ⅵ),(Ⅶ),(ⅩⅨ)appearance or content enrichment is related to the processing time of RPMP.Combined with the results of preliminary drug efficacy studies,5-hydroxymethylfurfural(ⅩⅨ)can be used as an index component to evaluate the processing degree.Compound(Ⅹ)is daidzein,which is the main component of black bean prepared by Polygonum multiflorum,which can be used to identify Polygonum multiflorum produced by black soybean.2.The quality of RPMP is not only related to the quality of the original medicinal materials,but also related to the processing technology.Based on the research on the chemical composition of RPMP,this study determined the chemical composition related to the processing time and established a detection method.Combining the modern analysis methods of traits and microscopic identification,thin layer identification,content determination,and fingerprints,the quality standard of RPMP decoction pieces,which takes into account the quality evaluation of the original medicinal materials and the evaluation of the processing technology,was established.Through the analysis and comprehensive evaluation of 57 batches of samples Study to determine the evaluation indicators related to the quality grade of RPMP and make the limits of each index,establish evaluation standards for RPMP.The method is simple and reliable,which can provide reference for quality control and grade division of RPMP,can be used to guide the production of RPMP in the market and the circulation in pieces promotes the formation of a high-quality and high-price trading system. |
PDF Full Text Request