| Chinese herbal compounds refers to the traditional Chinese medicine mixture formed by compatibility theory under the guidance of the basic theory of Chinese medicine.It has been used in clinic widely.In the course of the struggle between the working people and the disease for thousands of years,the traditional famous Chinese medicine prescription has been realized through practice,and is constantly recognized by the clinical screening,which is the crystallization of the wisdom of the Chinese nation.Many classic Chinese medicine prescription names have been transformed by modern technology and have been included in the Chinese Pharmacopoeia.But the curative effect,quality control and drug safety of the compounds are always the hot spot of attention at home and abroad.The reason is that the composition of the compound related to quality control is not necessarily the component of drug effect.The single or multi chemical index component of the western medicine model is not enough to explain the quality of the compound prescription of the traditional Chinese medicine and the significance of the quality control.It is difficult to reflect the true curative effect of the compound.It is also unclear about the effective substances and their action mechanism of compound efficacy,which is the difficulty of quality control.It has always been the primary scientific problem of Chinese medicine to find the research methods suitable for the evaluation of the complex system of traditional Chinese medicine,and to clarify the basis and the essence of the effect of the effect of traditional Chinese medicine.The preliminary work of the project group is to study of the dismantling prescription for single research,and select 5 quality control components that can be absorbed into the blood by prototyping in Astragalus membranaaceus(Fisch.)Bge.var.mongholicus(Bge.)Hsiao.4 effective components are screened out in Angelica sinensis(Oliv.)Diels.One of the known components of the Senkyunolide I can be absorbed into the blood by the prototype,and the combination of Astragalus membranaaceus(Fisch.)Bge.var.mongholicus(Bge.)Hsiao and Angelica sinensis(Oliv.)Diels has not been studied.In addition,it is not clear that the effect of the compound on the quality control components of the compound after compatibility is the superposition of the quality control components selected by two single Chinese medicinal materials,or the increase or decrease of other components after compatibility.It is worth further study.The preliminary research technology and results provide technical and data support for the follow-up study of the team.In this context,our working is focus on the screening of the quality control components of Danggui Buxue Decoction.The aim and method of the study were to study the decoction of Danggui Buxue Decoction on the basis of the previous study.The composition and change of the compound prescription were systematically studied,and the composition of the compound was selected and the mechanism of its differentiation was expounded.Then the quality control components of Danggui Buxue Decoction were selected,providing a reference method for screening the quality control components of Chinese herbal medicine and its compound.The traditional Chinese medicine decoction was used as the carrier to decoct the decoction of Danggui Buxue Decoction with the guidance of sequential metabolism.First,the physical environment stability of Danggui Buxue Decoction was investigated.Secondly,the metabolism of Danggui Buxue Decoction was studied by the body closed enteric ring method and the abdominal aorta by the HPLC method.The composition of blood samples was compared and analyzed by modern chromatography with high performance liquid chromatography and ultra high performance liquid chromatography-mass.The change process of the pharmacodynamic components of the Danggui Bu Xue Decoction was studied,and the quality control components of the decoction were selected.Research results1 Study on the chemical composition of Danggui Buxue Decoction(1)Establishment of characteristic chromatogram by high performance liquidchromatography(HPLC)method of Danggui Buxue Decoction a HPLC method was used to establish the characteristics of Dangggui Buxue decoction.The chromatographic peaks of the 14 compounds were determined and 5 known compounds were identified bystandard.These five compounds were Calycosin-7-O-β-D-glucoside,Calycosin,ononin,Formononetin,and Senkyunolide I.(2)Multi component research of Danggui Buxue Decoction(DBT)based on the LC-MS/MS Based on the known chromatographic conditions,the mixed solution of 5 kinds of standards and the multicomponent of Danggui Buxue Decoction were analyzed,and the characteristics of the decoction were established,and 31 compounds were obtained,including flavonoids,phthalosides and saponins.The identified components of Danggui Buxue Decoction by reference standard products were:,Calycosin-7-O-β-D-glucoside,Calycosin,ononin,Formononetin,and Astragaloside IV.The remaining 26 peaks are determined by combination of literature and accurate molecular weight and cracking rule.2 Study on tihe stability of gastrointestinal tract based on high performance liquid chromatography(HPLC)After incubation with artificial enzyme containing gastric juice and artificial enzymatic enteric liquid,the relationship between the relative peak area ratio and the composition with time is analyzed,and the drug components are not changed basically in the gastric juice 2h and in the intestinal fluid 4h under the general condition.The 14 components of Danggui Buxue Decoction were not changed almostly in the artificial gastrointestinal tract,which ensured the stability of the decoction of Danggui Buxue in the environment of the digestive tract,which was beneficial to the study of the next multicomponent sequential metabolism.3 Study on the sequential multicomponent absorption and metabolism of Danggui Buxue Decoction(1)Study on the changes of intestinal bacteria,intestinal wall enzyme,intestinal asepsis and liver metabolism by HPLC.The intestinal bacteria,intestinal wall enzymes,intestinal bacteria and liver metabolism were analyzed by HPLC,and 7 components were found to be absorbed into the blood circulation through prototyping,and 5 of them were known components.The 2 new component is intestinal metabolites,and the 2 new component is liver metabolites.Based on the theory of sequential metabolism and combined with high performance liquid chromatography(HPLC),the metabolic continuity of the Danggui Buxue Decoction was preliminarily investigated,and 7 prototype components were selected as the quality control component of Danggui Buxue Decoction.(2)The analysis of the multicomponent sequential metabolic changes of Danggui Buxue Decoction based on LC-MS/MS The components analysis of Danggui Buxue Decoction through LC-MS/MS,Calycosin-7-O-beta-D-glucoside,Ferulic acid,Ononin,Calycosin,6"-O-malonate-glycetein-7-O-beta-D-glucoside,Senkyunolide I,Formononetin,Astragaloside IV,isomer of(6aR,11aR)-3-hydroxy-9,10-dimethoxy-pterocarpan,14 components,such as Astraisoflavan-7-O-beta-D-glucopyranoside,Z-ligustilide,E-ligustilide,Senkyunolide A,and(6aR,11 aR)-3-hydroxy-9,10-dimethoxy-pterocarpan were selected which can be absorbed into the blood by prototyping.The other components in the intestinal bacteria,liver metabolism and intestinal wall enzyme in different degrees of metabolic transformation,the main types of the transformation of Danggui Buxue Decoction mainly include reduction,glucuronization,demethylation,deglyacylation,hydroxylation and so on.Conclusion Through the theory of sequential metabolism,we selected the Calycosin-7-O-β-D-glucoside.Ferulic acid.Ononin、Calycosin、6"-O-malonate-glycetein-7-O-β-D-glucoside、Senkyunolide I、Formononetin、Astragaloside Ⅳ、isomer of(6aR,11aR)-3-hydroxy-9,10-dimethoxy-pterocarpan、Astraisoflavan-7-O-β-D-glucopyranoside、Z-ligustilide、E-ligustilide、Senkyunolide A、(6aR,11 aR)-3-hydroxy-9,10-dimethoxy-pterocarpan as qualitu control components.It is also found that some of the metabolites of the prototype components can be found in the liver intestine and absorbed into the body circulation in the form of metabolites,which can be traced to the metabolites of the prototype components by other techniques.On this basis,the selected effective components provide data support for further quantitative analysis of Danggui blood enriching decoction.innovation point(1)First,we used sequential metabolic theory to study the effective components of Danggui Buxue Decoction and select quality control components.(2)The dynamic change process of the composition of Danggui Buxue Decoction was studied by the method of collecting blood from the body closed entero abdominal aorta,and the mechanism of the change in the liver intestine was expounded. |
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