Study On Quality Evaluation Of Prunus Mume Sieb.et Zucc. Based On Antioxidant And Xanthine Oxidase Inhibitory Activity

Objective:1.Using total polysaccharides,total flavonoids and total polyphenols as indicators,exploring the differences in total extracts of P.mume from different regions,processing methods and flowering periods.2.Based on GC-MS,HPLC-DAD,and UPLC-Q-TOF-MS/MS,comparing the differences in volatile components and polyphenols of P.mume from different regions,processing methods and flowering periods.3.To study the in vitro activities of P.mume from different regions,processing methods and flowering periods,the antioxidant activity and xanthine oxidase inhibitory activity were used as indicators to analyze the differences of in vitro activities of P.mume from different regions,processing methods and flowering periods.4.Taking P.mume as an example,based on the antioxidant activity in vitro and the inhibitory activity of xanthine oxidase in vitro of 13 polyphenols in P.mume,the quality functions of P.mume were established respectively to evaluate the quality of P.mume.Method:1.The P.mume from different regions and flowering periods were dried to constant weight by different processing methods,and the condensation reflux extraction method was selected to investigate the influence of four factors on the yield of three total extracts of P.mume one by one in combination with the control variates.According to the results of the single factor test,the extraction conditions of total polysaccharides,total flavonoids and total polyphenols were optimized through design response surface test and compared the effects of different processing methods on the total extract of P.mume.2.Based on GC-MS,detecting the volatile components in the alcohol extract of P.mume and qualitatively and quantitatively analyze the polyphenols in P.mume using HPLC-DAD and UPLC-Q-TOF-MS/MS,and analyzing their differences among different regions,processing methods and flowering periods of P.mume.3.The total extracts of P.mume from different regions,processing methods and flowering periods were tested for in vitro antioxidant activity and XOD inhibitory activity using DPPH,total antioxidant,reducing power,and O^2-clearance methods.The differences in antioxidant activity and XOD inhibitory activity of P.mume from different regions,processing methods and flowering periods.4.Introduce the concept of Effective Components Equivalent（ECE）,quantify the role of each component in regulating the biological activity of traditional Chinese medicine,and construct an objective,reliable,simple and feasible method for evaluating the quality of traditional Chinese medicine that links clinical efficacy and safety.Based on the in vitro antioxidant activity and in vitro xanthine oxidase inhibitory activity of 13compounds in P.mume,combined with their content,the effective component equivalents were calculated by multiple regression method,and the quality function of P.mume was established to evaluate the quality of P.mume.Result1.Using Box Benhnken Design response surface methodology,the extraction methods of total polysaccharides,total flavonoids and total polyphenols from P.mume were optimized,and the optimal reflux extraction process for total polysaccharides,total flavonoids and total polyphenols was determined.The results showed that there was a significant growth period variation pattern in the total polysaccharide content of P.mume at different flowering periods,namely fully open>semi open>unopened.The total flavonoid and total polyphenol content of P.mume at different flowering periods also showed a significant growth period variation pattern,namely unopened>semi open>fully open.2.The detection method established based on GC-MS was used to analyze the volatile components in P.mume,and the results showed that there were differences in the volatile components in P.mume at different flowering periods.Qualitative and quantitative analysis of polyphenolic compounds in P.mume was conducted using detection methods based on HPLC-DAD and UPLC-Q-TOF-MS/MS.The results showed that these 13 target compounds were present in P.mume from different regions,processing methods and flowering periods,but there were differences in content.3.The DPPH free radical scavenging and reducing ability of the water extract of P.mume is higher than that of the alcohol extract,but when measuring the superoxide anion scavenging ability and total antioxidant activity,the alcohol extract of P.mume is higher than that of the water extract.And in terms of XOD inhibitory activity,the ethanol extract of P.mume was superior to the water extract,with Yangshucun freeze-dried and unopened ethanol extract having the best activity,with an IC₅₀of 2.397±0.0379 mg/m L.4.Based on the antioxidant activity of 13 polyphenolic compounds,a quality functionforP.mumewasestablished:Q=3.959+280.116X₁+201.888X₂-427.258X₃+110.158X₄+1763.142X₅-191.068X₆-3392.672X₇-27583.701X₈+139866.580X₉-18807.214X₁₀-55613.265X₁₁+11297.690X₁₂+6679395X₁₃.Based on the inhibitory activity of 13 polyphenol compounds xanthine oxidase,thequalityfunctionofP.mumewasestablished:Q=4.169-2914.957X₁-1132.420X₂+6554.196X₃+6175.706X₄+26377.387X₅-9857.363X₆-4668.413X₇-10489.942X₈-29619.234X₉.Conclusion1.The optimal reflux extraction process for total polysaccharides,total flavonoids and total polyphenols in P.mume optimized by response surface methodology;The extraction method optimized by response surface methodology was used,and the results showed that there were differences in total polysaccharides,total flavonoids and total polyphenols among different flowering periods of P.mume,and there was a clear pattern of growth period changes;There is no significant difference in the content of total flavonoids and total polyphenols among different regions and processing methods.2.Based on the analysis methods established by GC-MS,HPLC-DAD and UPLC-Q-TOF-MS/MS,compounds in P.mume from different habitats,processing methods,and flowering periods were analyzed.The volatile components preliminarily identified differed in different sample species and relative percentage content.Different samples all contained 13 polyphenolic compounds,but the content varied.3.Based on the in vitro antioxidant test and XOD inhibitory activity test,the P.mume from different places of origin,different processing methods and different periods showed different antioxidant activity and xanthine oxidase inhibitory activity.4.Based on the antioxidant activity in vitro and the inhibitory activity of xanthine oxidase,the quality function between the content and activity of 13 compounds in P.mume was established.