Study On The Genuineness Of Aurantii Fructus

Object:Guided by the theory of traditional Chinese medicine,using modern separation and analysis technology,combined with pharmacological efficacy test,the content determination method is established from the effective monomers,and the principal component analysis in pass 3.19 software and the discriminant analysis in SPSS 20.0 software are used to screen out the compatibility ratio between the characteristic medicinal materials of each origin and the effective component groups of each medicinal material,so as to find out the best match experiment In addition,through the pharmacological and pharmacodynamic experiments(small intestine propulsion experiment,spleen deficiency rat model experiment),the best compatibility was initially verified.Finally,through the establishment of neural network model,the best compatibility was further verified,its authenticity was expounded,and the best harvest time was guided.So as to provide a scientific basis for the establishment of objective evaluation quality standards of Aurantii Fructus and clinical use of traditional Chinese medicine.Method:1.The main active components(isonaringin,narigin,hesperidin,neohesperidin,Naringin,Hesperidin,meranzin hydrate,meranzin,marmin,nobiletin,tangeretin and synephrine)in Fructus aurantii were screened by the isolated small intestine smooth muscle test of normal mice.And to establish the dose-response relationship between the active components and the spontaneous contraction of isolated duodenal smooth muscle of normal mice,and to further screen the effective monomers with dose-response relationship and obvious effect.2.To establish a HPLC method for the determination of flavonoid glycosides(isonaringin,narigin,hesperidin and neohesperidin,)and alkaloids(synephrine)in Aurantii Fructus,and to determine the harvest time of Aurantii Fructus with the results as the index,and to use pass 3.19 software and SPSS 20.0 software for the principal component analysis and discriminant analysis,The characteristic medicinal materials from different producing areas were screened out and the content of the decoction was determined.3.By using the isolated duodenal smooth muscle experiment of normal mice,we compared the drugs from different places,and selected the best drugs from different places.Combined with the matching experiment of the effective monomers in the drugs from different places,we preliminarily determined the best matching ratio.4.The experiment of small intestine propulsion in normal mice was used to verify the optimal compatibility.5.In the model experiment of spleen deficiency rats,the best compatibility was verified again through the influence of the best compatibility composition on serum gastrin,plasma acetylcholine,motilin,substance P and vasoactive intestinal peptide.6.Through the construction of a matching model based on neural network,the samples with known content of Fructus aurantii were classified to provide a basis for the rapid search for authentic medicinal materials,so as to elaborate the connotation of authenticity and further guide the harvesting period.Restult:1.Eight active components(isonaringin,hesperidin,neohesperidin,Hesperidin,meranzin hydrate,nobiletin,tangeretin and synephrine)were screened out.Among them,synephrine,neohesperidin and hesperidin have significant dose effect relationship.2.A HPLC method was established for the determination of flavonoid glycosides(isonaringin,narigin,hesperidin and neohesperidin)and alkaloids(synephrine)in Fructus aurantii The results showed that the decocting rate of effective monomers was lower than that of alcohol extract.3.When the concentration is 0.1g/ml and the dosage volume is 10-500 μL in Jiangxi Province,the effect is more obvious than that in other places.The fitting test results of each place show that the trend of composition is consistent with that of Aurantii Fructus in various places.According to the test results,it can be preliminarily considered that the ratio of four flavonoid glycosides in the decoction of Aurantii Fructus in Jiangxi Province is the best,according to the content results It can be calculated that the ratio is: isonaringin: narigin: hesperidin: neohesperidin = 1: 14: 1: 13.4.Compared with the blank group,there was significant difference between the low,middle and high dose groups in the optimal ratio of effective monomers in Fructus aurantii(P < 0.01,P < 0.05).5.Compared with the model group,the best compatibility ratio of effective monomers in Aurantii Fructus has a significant effect on gastrointestinal hormones in the spleen deficiency model rats,which can significantly increase the content of ACh,GAS,SP,MTL and VIP,which is almost the same as the effect of western medicine domperidone.6.he neural network model can be used to classify the samples of Fructus aurantii Immaturus with known content into the best matching ratio and the non optimal matching ratio,and to screen the samples of Fructus aurantii Immaturus quickly.Conclusion:1.In vitro small intestine experiment showed that the active components of Aurantii Fructus were: isonaringin,hesperidin,neohesperidin,Hesperidin,meranzin hydrate,nobiletin,tangeretin and synephrine.However,the active components with significant dose-response relationship were: neohesperidin,Hesperidin and synephrine,.It provides theoretical basis and material basis for the determination of effective monomers in the next step.2.The established HPLC method for the determination of flavonoid glycosides((isonaringin,narigin,hesperidin and neohesperidin,)and alkaloids(synephrine)in Aurantii Fructus is simple,accurate,reliable and reproducible,which can be used as an effective method for the determination of flavonoid glycosides and alkaloids in Aurantii Fructus.It provides a theoretical basis for further investigation of the compatibility ratio of the effective components in Aurantii Fructus and the establishment of the quality standard for objective evaluation of Aurantii Fructus.The best harvest time was determined by content determination,which provided scientific basis for reasonable picking of Aurantii Fructus in the future.The content determination of the selected medicinal materials and their Decoction provides scientific basis and theoretical basis for the next step of monomer fitting experiment.3.The best compatibility ratio of effective monomers in Aurantii Fructus was determined,which indicated that the material basis of Aurantii Fructus was the effect of effective components group according to certain compatibility ratio,which provided scientific basis for further elaborating the connotation of authenticity and clinical medication of traditional Chinese medicine.4.The neural network model is simple,reliable,reproducible and easy to operate.It provides a theoretical reference for the rapid screening of the best proportion of the content of Fructus aurantii,and has a certain practical significance for explaining the connotation of the authenticity of Fructus aurantii and guiding the best harvest time.