| Carex meyeriana Kunth is distributed in northeastern China.Carex meyeriana Kunth has functions of keeping warm and inhibiting bacteria.The flavonoid is one of the main active ingredients in Carex meyeriana Kunth,which has high biological activity and research value.In this thesis,the extraction and purification process of flavonoids of Carex meyeriana Kunth is mainly optimized,and the chemical components and biological activities of flavonoids of Carex meyeriana Kunth are studied.1.Extract the flavonoids of Carex meyeriana Kunth by refluxing with traditional ethanol.The results of the single-factor experiment were determined as follows: ethanol concentration: 50 %,liquid-to-material ratio: 30 g/mL,extraction time: 90 min,extraction temperature: 70 ℃.Based on the single factor experiment,the extraction rate(%)of the total flavonoids of Carex meyeriana Kunth was used as the response value,and the response surface test design was carried out.The best extraction process was 49 % ethanol concentration and 30 mL/g liquid-to-material ratio.The extraction time is 91 min and the extraction temperature is 70 ℃.The optimal extraction rate of total flavonoids in Carex meyeriana Kunth is 6.24 ± 0.04%.2.Through static and dynamic adsorption,select AB from six macroporous resins of AB-8,D101,X-5,S-8,NKA-Ⅱ and polyamide based on the indexes of adsorption rate,desorption rate and recovery rate AB-8 As the best purification resin;using analytic hierarchy process(AHP)combined with response surface method(RSM),the optimal conditions for purification were determined as follows: sample concentration: 3.04 mg/mL,ethanol concentration: 70.33 % ethanol,pH value: 2.93,elution flow rate: 2.95 BV / h.The purity of flavonoids increased from 11.72 ± 0.1% to 56.00 ± 0.68 %.Using the analytic hierarchy process to calculate the weight coefficients of the three purification indicators of recovery rate,purity and decolorization rate calculate the comprehensive score,the purification process conditions are more reliable and scientific.3.Use high-performance liquid chromatography-mass spectrometry(HPLC-MS)to qualitatively analyze the structure of the purified flavonoids.Eight flavonoids and one Phenylpropanoid was identified,namely: catechin,kaempferol-3,7-2-O-glucoside,isoxiafotaside,orientin,luteolin glucuronide,Rutin,luteolin-7-O-β-d glucuronide,luteolin,isochlorogenic acid(dicaffeoylquinic acid).Using retention value comparison method and comparison of peak shape consistency of mixed standards,seven flavonoids and one Phenylpropanoids were identified.The analysis showed that luteolin was in agreement with HPLC-MS.Gallocatechin,epigallocatechin,epicatechin,and chlorogenic acid are the isomers of catechin and isochlorogenic acid in HPLC-MS,respectively;naringin,myricetin,Isorhamnetin does not agree with HPLC-MS.And compared with the reference UV spectroscopy,the results showed that: gallo-catechin,naringin,myricetin,isorhamnetin,chlorogenic acid,five standard flavonoids and an Phenylpropanoids It is consistent with the spectrum of each component in the flavonoids of Carex meyeriana Kunth after purification,and epigallocatechin and epicatechin are inconsistent.4.The antioxidant and bacteriostatic activity comparison of flavonoids of Carex meyeriana Kunth before and after purification.The test results show that the removal rate of flavonoids to DPPH after purification is close to VC,and the removal rate to hydroxyl radicals and reducing power are slightly higher than VC;The purified flavonoids had the strongest inhibitory activity against E.coli.The MIC = 0.875 against E.coli was 3.5 times lower than that of chloramphenicol(MIC 3.12).After purification,the antioxidant capacity and bacteriostatic activity of flavonoids of Carex meyeriana Kunth flavonoids were better than the extracted. |
PDF Full Text Request