Study On Extraction Process And Lipid-regulation Effects Of Coreopsis Tinctoria

Objectives:1.Study of C.tinctoria aqueous extract and the optimization process on the main chemical components were analyzed.2.Improvement of determination method of total flavonoids content of C.tinctoria.3.Purification process of total flavonoids in extract of C.tinctoria of macroporous resin.4.Establish TLC bioautography method for in vitro screening bioactive substances having glapha glucosidase inhibitory effect.5.To conduct screening experiment of acute toxicity of different extracts of C.tinctoria in order to fulfil clinical application.6.Study on the hyperlipidemia in mice blood lipid regulation effects of C.tinctoria extract with no acute toxicity.Methods:1.To investigate different drying methods of water extracts of C.tinctoria,to further amplify technology research,and to find out different process conditions of the water extract of total flavonoids by UV method.This study is based on Portuguese literature by TLC method and HPLC method to identify to extract the scale-up process in the main components of the qualitative and quantitative study on the preliminary.2.Using naringin as the standard methodological study,and using this method to measure total flavonoids in different extracts of C.tinctoria.3.Using the content of total flavonoids as the index,and using the static adsorption method as well as adsorptionrate and adsorption capacity to screen the best ratio of resin.Moreover,using the rate of recovery as the index to investigate the kind of concentration,sample flow rate,elution,and the process parameters of elution curve agent concentration and determination of total flavone content before and after purification.Then analyzing the main substance of the purified by TLC method,and comparing the purification effect of the main compounds by HPLC method.4.The aqueous extract,alcohol extract and ethyl acetate extract from Coreopsis tinctoria were eluted with methylbenzene-acetic ether-formic acid（9︰7︰3）on the silica gel plate,incubated for 60minutes inα-glucosidase solution which was sprayed on the silica gel plate and were colored with the solution of 2-naphthyl-α-D-glucopyranoside and fast blue B salt（1︰1）as the color developing agent,thusα-glucosidase inhibitors were obtained and identified by resolution in visible light.5.Adopting the acute toxicity test of maximum dosage and calculated the maximum tolerance,converted the maximum tolerance dose with clinical dose（5g dried/60kg weight）to evaluated the safety of different extracts of C.tinctoria.6.Establish the hyperlipidemia model in mice by feeding with high-fat diet for 8 weeks.The mice were then divided into model group,positive drugs simvastatin group,high dose group,middle dose group and low dose group randomly,and the normal mice are still in the normal group.By comparing these two groups,selecting the non-toxic C.tinctoria extract based on acute toxicity test on observation of its effect on model mice four blood lipids TC,TG,HDL-C,LDL-C,AST and ALT content of mice in each group were measured,effect of C.tinctoria extract on mice model of liver function.Results:1.Chlorogenic acid content was the highest in the vacuum drying process,Flavanomarein content was the highest in the vacuum drying process,Flavanokanin content was the highest in the impregnation method without concentration process,Marein content was the highest in the vacuum drying process,Dicaffeoylquinic acid content was the highest in the vacuum drying process,Okanin content was the highest in the vacuum drying process.2.Through the investigation of methodology,using Naringin as the determination of total flavonoids from C.tinctoria in control solution,the detection wavelength was 420nm,and the method has good precision,accuracy.The recovery rate was 99%¹03%.3.Using HPD-100 macroporous separation and purification of total flavonoids content of C.tinctoria extract,to measure the ootimum process for the concentration of sample is20mg·ml^-1,elution flow rate was 3BV·h^-1,eluent concentration was 75%,elution volume was 4BV.4.The white spots on the TLC plate showed the effect ofα-glucosidases inhibition,indicating that the aqueous extract,alcohol extract and ethyl acetate extract from Coreopsis tinctoria contained active ingredients with functions ofα-glucosidase inhibition.5.Using the maximum tolerance to evaluated its safety,dose of the extract of the water extract of impregnation method directly freeze drying,the extract of impregnation concentration after freeze drying,water extract of vacuum drying,the water extract of spray drying,the extract of ethyl acetate and the raffinate extract is equivalent to human dose is 2497 times,2571 times,914 times,4115 times,4582 times,1920 times respectively.6.C.tinctoria raffinate of high,midium and low dose group can reduce the content of TC in cerum of mice（P<0.05）and decrease the content of TG significantly（P<0.01）.The affinate can reduce the content of experimental hyperlipidemia in mice serum AST and the content of ALT significantly（P<0.01）.Conclusion:The content of total flavonoids of different extracts of the laboratory process is between 17%²1%,and after further amplification technology,the total flavonoids content was between 20%³4%.There are differences on the content of C.tinctoria water extracts of different process extract mainly analysis shows that each process of main compounds is different.2.The Method of control samples using naringin can make up for the using of rutin as control article overflow because the content of total flavonoids of ethyl acetate extract cannot be measured.Through the methodology test,the method has good precision and accuracy,the recovery was 99%¹03%.3.Purification of macroporous resin can increase the total flavonoid content of alcohol extracts from 10%to 30%,This process is stable and feasible.4.The established rapid TLC bioautography method for screening the compounds ofα-glucosidases was provided.Marein and okanin have strongα-glucosidases inhibitory effect.5.The maximum of all the extracts of C.tinctoria conversion with people commonly used are more than 100 times,which can explain the clinical medication is security.6.C.tinctoria raffinate can improve hyperlipidemia mice blood lipid levels and liver function.