Experimental Studies On Quality Control And Anti-inflammatory Profile-effect Of Emilia Prenanthoides DC.

Emilia prenanthoides DC.(EP)is a traditionally used medicinal plant found in most southeast and south regions of Guizhou province in china which has the efficacy of clearing away heat and toxic matter to treat bruises,red and white dysentery and pyocutaneous disease.The traditional Chinese Medicine derived from the deried of E.prenanthoides which belonging to asteraceae family,and has been listed in “Chinese materia medica”,“the flora of China”,“the flora of guizhou”,“the flora of Guangzhou” “the flora of Fujian” and will be included in the 2013 edition of the “guizhou province traditional Chinese medicine and national medicine quality standard”.Determination of Chemical Constituents and Determination of Multiple Index Components in Identification of chemical composition.Part? Study on the Identification and Determination Chemical Constituents of Emilia prenanthoides DC..According to the study on chemical component of Emilia sonchifolia(L.)DC..The related literatures were reviewed and combined with the research results of the chemical composition of E.sonchifolia(L.)DC.In this study,the chemical composition of the EP medicinal materials was identified by HPLC,through comparsion the retention time and the UV absorption spectrum between the standard compound.Combined with the total ion flow diagram from the UPLC--ESI-MS/MS,comparing the relative molecular mass and ion debris of the reference substance with the sample to prove the above conclusion.7 compounds were identified from EP for the first time.The compounds are as follows: Chlorogenic acid,Vicenin-2,Hyperoside,quercitrin,Isoquercitrin,rutin and protocatechuic acid.The contents of the 7 compounds in EP were determined by HPLC simultaneous.Results show that the method is accurate,reliable,simple and EP from different regions in guizhou province have the same chemical composition,but the determination of component content in 7 has a certain difference.Part? Study on the Fingerprint Chromatogram of Emilia prenanthoides DC.HPLC analysis method was used to study the Chromatographuc Fingerprint of EP,and the and similarity analysis(SA),hierarchical clustering analysis(HCA)and principal compinent analysis(PCA)were used to evaluated the common chromatographic peaks area by the SPSS.The reference chromatogram was generated from the fingerprints of the 12 batchses of EP samples and 14 common fingerprint peaks were found contains Chlorogenic acid,Vicenin-2,Hyperoside,quercitrin,rutin and Isoquercitrin.The different origins of EP's fingerprints all have higher similarity,every characteristic peaks have higher resolution.The results of HCA show that the EP from similar origin was classified into one class,and the quality of the EP might be related to the growth environment,harvest season and storage conditions.The results of PCA show that the first 4 principal components contained 91.297% of the information for the original 12 indexes,moreover A2,A3(Clorogenic acid),A4,A9(rutin),A13 and A14(Qercitrin)were the main ingredients affecting the quality of EP.Part ? Study of the spectrum-effect RelationshipThe EP has obvious anti-inflammatory effects though the modles of Xylene-induced ear edema mice model and Carrageenan-induced paw edema rats model with three doses of high,medium and low doses.Carrageenan-induced paw edema mice model was used on the comparative research of anti-inflammatory effects for the 8 batches of EP with large differences.The model was evaluated by analyzing paw swelling rate,concentrations of superoxide dismutase(SOD),malondialdehyde(MDA),tumor necrosis factor alpha(TNF-?),prostaglandin E2(PGE2).The anti-inflammatory research of EP from different origions showed that Carrageenan-induced paw edema mice model control group had significantly difference(p<0.01)compared with the control group and positive control group all of the five phamacodynamics indexes,also showed that oral administration EP extracts from different origions has different effect on anti-inflammatory.According to the results of EP fingerprints and pharmacodynamics activities,Partial Least Aquars Regression(PLSA),Gray Correlation Analysis(GRA)and Artificial Neural Network algorithm(ANN)were used to find the primary active components in finggerprint representing the anti-inflammatory activities.As the results of Grey relational analysis showed the average grey correlation degree between the 5 pharmacodynamics indexes with the 13 values of peak areas,we can obtain the order as follow: A3 > A1 > A14 > A4 > A13 > A11 > A9 > A10 > A6 > A8 > A12 > A2> A5> A7;PLAR results were showed in these 14 common peaks areas,A3(Clorogenic acid),A4?A9(rutin),A13 and A14(quercitrin)were proformances higher correlation with these five indicators.ANN acquired the similar results as the PLAR.According to the result of GRA,PLSR,and ANN,the contribution to anti-inflammatory mainly the A3(Clorogenic acid),A4?A9(rutin),A10(Hyperoside)A13 and A14(quercitrin).That is to say the main compounds contribute the property of anti-inflammatory may be the 6 compounds.Has proved that we use the content of quercitrin as a quality control program is scientific.Part ? studies on quality control of Emilia prenanthoides DC.According to the research methods of Pharmaceutical standard in 2015 version of “Chinese Pharmacopeia” general rule.EP was identified by TLC,The content of moisture,total ash,acid insoluble ash,pesticide residues and heavy metals,water and alcohol extract from EP was detected.With quercetin and chlorogenic acid as reference substance to establish a TLC identification method of EP,the spots were clear,well separated.A convenient HPLC method with good specificity,linearity,repeatability,precision and recovery can be applied to control the quality for quercetin content as the major ingredients of EP.The draft quality standard of EP is established and the methods are simple and effective.In addition,Good linearity,high precision,appropriate repeatability and stability method was established to determine the total flavonoids in EP,the content of total flavonoids in EP ranged from 23.16 ~ 56.52 mg·g-1.Under the optimal extraction through response surface methodology,the extration ratio was 77.69%.