Study On Ootimization Of Supercritical CO₂ Extraction Process And Activity Evaluation Of "Songge Jiangniaosuan"

Hyperuricemia is a common disease when the blood uric acid content in human body is high,which can cause gouty arthritis,cardiovascular disease,kidney disease and so on.Xanthine oxidase(XO)is a key enzyme in the process of uric acid production.Allopurinol and febuxostat are Hyperuricemia drugs usually used and can inhibit XO activity.,However,long-term use of them can result in a variety of adverse reactions.It is significant to develop safe and effective Chinese medicine to treat Hyperuricemia.In this paper,"Songge Jiangniaosuan Granules"(CFDA-2016L10210)is a Chinese medicine,contains the raw materials of Pueraria lobate root,Pinus massoniana needle,Citrus reticulata peel and thier mixture were established as the research objects,the quantitative methods of total flavonoids and 14 components were successfully established,The Box-Benhnken response surface experiment was designed to optimize the conditions of supercritical CO2 extraction process.And the DPPH activity and XO inhibitory activity of the mixture and 14 components were analyzed according to the established methods.The main research results are shown as follow:(1)Ultra-high performance liquid chromatography(UPLC)successfully quantifies the contents of 14 component compounds and the retention time is determined,including 3'-hydroxy puerarin(5.3 minutes),puerarin(7.4 minutes),3'-methoxy puerarin(8.2 minutes),daidzein(10.2 minutes),ononin(18.0 minutes),daidzein(19.2 minutes),gallic acid(2.0 minutes),3,4-dihydroxybenzoic acid(3.5 minutes),taxifolin(12.6 minutes),kaempferol(22.3 minutes),hesperidin(15.9 minutes),nobiletin(24.7 minutes),3,5,6,7,8,3',4'-heptemthoxyflavone(25.2 minutes),tangeretin(25.7 minutes),with the standard curve correlation coefficient r2>0.999.In addition,quantitative determination and analysis of total flavonoids by UV-vis spectrophotometry is established,with the standard curve correlation coefficient r2>0.999.(2)The optimal conditions for extracting daidzein from Pueraria lobate root with supercritical CO2 are:temperature 51.5?,pressure 31.8 MPa,entrainer ethanol concentration 36.7%,extraction rate 31.3 ?g/g,relative error 1.0%.The best conditions for extracting 3,4-dihydroxybenzoic acid form Pinus massoniana needle with supercritical CO2 are:temperature 57.4?,pressure 29.9 MPa,entrainer ethanol concentration 14.4%,extraction rate 20.6?g/g,relative error 1.0%.The best conditions for extracting 3,5,6,7,8,3',4'-heptamethoxyflavones from Citrus reticulata peel are:temperature 50.4?,pressure 29.7 MPa,entrainer methanol,extraction rate 36.2?g/g,The relative error is 0.6%.(3)The best supercritical extraction conditions for the total flavonoids from the mixture with supercritical CO2 are:temperature 57.3?,pressure 31.3 MPa,entrainer ethanol concentration 12.3%,extraction rate 0.975 mg/g,relative error 1.6%.The experimental results of the mixture extract DPPH activity and XO inhibitory activity showed that the DPPH free radical scavenging rate and total flavonoids correlation coefficients of the extraction at the p=0.01 level was 0.963;The correlation coefficient between the XO inhibition rate of the extraction and daidzein content at the p=0.01 was 0.979.In the test of 14 compounds XO inhibitory activities,the IC50(half maximal inhibitory concentration)of daidzein(1.76 ng/mL)was significantly lower than allopurinol(1.79?g/mL).