Enrichment Of Total Flavonoids By Membrane Separation And Chemical Constituents Research From Rosa Xanthina Lindl.

Objective:Rosa xanthina Lindl in North China are rich in resources,widely distributed,but low in utilization rate.In this paper,the Membrane separation were used to enrich the total flavonoids of Rosa xanthina Lindl.which laid a foundation for the preparation of high content of Rosa xanthina Lindl.HPLC method was established to simultaneously determine rutin,hyperoside and quercetin in the extract of Rosa xanthina Lindl.wcich determining the content of three chemical components;Separating the chemical constituents from the 75%ethanol extract of Rosa xanthina Lindl.with the?-glucosidase inhibitory activity as a standard.It lays the foundation for the further development and utilization of the Rosa xanthina Lindl.Methods:Taking the total flavonoids and membrane flux as the index,the optimum conditions for the enrichment of flavonoids in the Rosa xanthina Lindl.by membrane separation were determined by single factor test method by changing the temperature,pH value and ratio of material to liquid.Through the determination of wavelength and mobile phase in high performance liquid chromatography?HPLC?,high performance liquid chromatography?HPLC?was used to determine the chemical constituents of rutin,quercetin and hyperoside in the extract of Rosa xanthina Lindl.Method,the chromatographic conditions are:column Inertsil-ODS-SP model,mobile phase acetonitrile-0.5%phosphoric acid solution system,flow rate 1.0 ml/min,detection wavelength 360 nm,column temperature 37°C,injection volume 20?l,gradient wash The elution time was 55min.The linearity,precision,stability,repeatability and sample recovery rate were investigated.The 75%ethanol extract of Rosa xanthina Lindl.was isolated by silica gel column chromatography and preparative thin-layer chromatography.The different polar components of Rosa xanthina Lindl.were separated by?-glucosidase inhibitory activity.The structures were identified by ¹H-NMR,¹³C-NMR,DEPT,COSY HSQC and MS.Results:The optimum process conditions for separation of the ultrafiltration membrane are:temperature 45°C,ratio of material to liquid 1:5,pH of the feed liquid 7.0.The content of total flavonoids in the extract of Rosa xanthina Lindl.under the optimal process conditions was 34.0%,and the total flavonoid content was significantly increased,Especially compared with the original alcohol extract 23.46%.Simultaneous determination of Rosa xanthina Lindl.in rutin,quercetin and hyperoside by HPLC method.The precision test results showed the area of rutin,hyperoside and quercetin.The RSD values were 0.78%,0.73%,and 0.83%,respectively.The RSD values of the rutin,hyperoside and quercetin.in the stability experiment were 0.79%,1.53%,and 1.38%,respectively,and the rutin,hyperoside and quercetin.in the calibration were repeated.The RSD values were 0.93%,0.84%,and 1.02%,respectively,which met the experimental requirements.The recoveries of rutin,hyperoside and quercetin were 98.10%,97.80%and 97.50%,RSD values were3.20%,3.00%and 2.80%,rutin,quercetin and hyperoside.The linear relationship is good and R² is greater than 0.9999.The experimental results show that the precision of the instrument is good,the method is reproducible and accurate,and it meets the experimental requirements.The contents of rutin,quercetin and hyperoside in the actual samples of Rosa xanthina Lindl.were 65.15?g/g,1.86?g/g and 13.43?g/g,respectively.The silica gel column chromatography was used to isolate the 75%ethanol extract of Rosa xanthina Lindl.to obtain 10 components of E1-E10.The inhibition of?-glucosidase by E2 and E3 was determined.The Hypoglycemic activity is higher.The chemical composition of E6 component is simpler by TLC.The three components were separated in the next step,and a total of five compounds were obtained.The structure of two of the compounds was characterized,one was determined to be dibutyl phthalate,and the structure of compound 2 was further confirmed.Conclusion:Membrane separation enrichment experiments on the total flavonoids of Rosa xanthina Lindl.showed that ultrafiltration was feasible for the purification of extracts of Rosa xanthina Lindl.After methodological verification,the HPLC method can be used to simultaneously determine the content of rutin,quercetin and hyperoside in the Rosa xanthina Lindl.The method is feasible,accurate and repeatable,and the linear relationship is good.Both compounds were isolated from the Rosa xanthina Lindl.for the first time,and the compound 1 was confirmed to be dibutyl phthalate,and the structure of the compound 2 was further confirmed.Among them,E1,E2,E3 and E9 inhibited?-glucosidase by 78.76,88.35,45.20 and 67.80,respectively,and the hypoglycemic activity was high.It can be speculated that the yellow thorn rose fruit contains chemical components with hypoglycemic activity.