| Rheumatoid arthritis is an autoimmune disease with erosive arthritis as the main clinical manifestation.The basic pathological manifestations are synovitis,pannus formation,and gradual destruction of articular cartilage and bone,eventually leading to joint deformity and loss of function,which can be complicated by lung disease,cardiovascular disease,malignant tumor,and depression.At present,the pathogenesis of RA has not been fully clarified,so there are still no specific treatment measures in clinical practice.Munziq Balgam is a traditional classic prescription of Uyghur medicine.Based on the Uyghur medical body liquid theory,it could mature the abnormal mucus in the human body,treats various diseases caused by abnormal mucus.By restoring the balance of body liquids,organ function and physical state,so as to achieve the purpose of improving clinical symptoms even curing diseases.Munziq Balgam has been developed by Hospital of Xinjiang Traditional Uyghur Medicine into an in-hospital preparation(Z20200034000)that is convenient for clinical adjustment.It has a significant effect on the treatment of Rheumatoid arthritis.However,at present,there are few reports on the chemical composition,basic research and quality evaluation of Munziq Balgam,and the current quality control methods of Munziq Balgam mixture are relatively backward.In order to clarify the basis of active ingredients of Munziq Balgam in the treatment of Rheumatoid arthritis and improve the quality control level,this paper mainly takes Munziq Balgam as the research object.A method for the systematic analysis of chemical constituents and the components absorbed into the blood of Munziq Balgam mixture based on chromatography-mass spectrometry was established,a net work "Functional compounds-Reumatoid arthritis related targets",and a quality control method based on UPLC-MS/MS technology for the determination of curative effect-related multi-index constituents was established.1.Through reviewing a large number of literatures in the early stage,the chemical composition information database of each single herb in Munziq Balgam was established.The volatile components in Munziq Balgam mixture were analyzed by GC-MS combined with computer search of NIST Libraries database,and 5 chemical components were identified;the relative percentage content of each component was calculated by the peak area normalization method.The results show that its main volatile components are 2,4-limonene,Fenchone,Phenylethyl Alcohol,Estragole,and Anethole.The results show that the relative content of aromatic volatile components is relatively high.The water-soluble components in Munziq Balgam mixture were analyzed by UPLC-MS/MS method.First,through the direct injection mass spectrometry system analysis of the reference substance,a data acquisition method suitable for the analysis of phenolic acids,flavonoids and triterpenoid saponins,and a novel data mining method based on high-resolution data characteristic product ions(CPI)combined with neutral loss(NL)filtering were established.From the watersoluble components of Munziq Balgam mixture,150 chemical components were quickly identified and their molecular structures were preliminarily deduced,the structures of 61 compounds were confirmed by comparative analysis with the reference substance.2.A rapid identification and analysis method for the components absorbed into the components absorbed into the rat plasma in Munziq Balgam mixture was established.A total of 43 transitional components in blood were found in the plasma of rats,of which 5 components entered the blood with the prototype structure,including Kaempferol 3-O-glucuronide,Liquiritigenin,Isoliquiritigenin,Glycyrrhetic acid-3-mono-glucuronide,Gycyrrhetic acid.Their structures were verified by comparison with the reference substances.The possible structures of 38 metabolites were deduced based on the molecular element composition given by high-resolution mass spectrometry data,with reference to structural features,cleavage behavior,and correlation analysis between biogenic information and metabolic pathways.Combined with the determination results of relevant functional components,the metabolic relationship between the water-soluble components in Munziq Balgam mixture and the components absorbed into the plasma of rats after oral administration of Munziq Balgam mixture was speculated.3.Through public databases(TCMSP,SwissTargetPrediction,and TargetNet,etc.),the corresponding protein targets of the components absorbed into the plasma of rats after oral administration of Munziq Balgam mixture were predicted.By searching disease databases(OMIM,Genecards,and DrugBank,etc.)to obtain the target information of RA.The intersection of the targets of the components absorbed into the plasma and the RA-related targets are the potential targets for the treatment of RA.Cytoscape was used to construct a network of "Functional compounds-Reumatoid arthritis related targets",which preliminarily clarified the functional components of Munziq Balgam in the treatment of RA,and preliminarily speculated that the functional components of Munziq Balgam play a therapeutic role in the RA signaling pathway.4.According to the research results of the chemical components of Munziq Balgam mixture,combined with the results of the analysis of the components absorbed into the plasma of rats and the results of network pharmacology screening of functional components,a rapid analysis method for simultaneous quantification of its 27 components was established by LC-MS/MS method.After the accuracy,precision,specificity,detection limit,quantification limit,linearity and quantification range,the established LC-MS/MS quantitative analysis method was verified,and the method was stable and reliable.On this basis,the content analysis of 27 functional components in 2 batches of Munziq Balgam mixture produced by Hospital of Xinjiang Traditional Uyghur Medicine was carried out.This method has good adaptability,and can be used as the quality control method of Munziq Balgam mixture after appropriate adjustment in combination with its production process and clinical application. |
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