Analysis Of Differential Components And Anti-Inflammatory And Antioxidant Activities In PMC And PML In Different Harvest Periods

Objective:The quality evaluation of PMC and PML is closely related to the harvesting period and index components.Based on the multivariate statistical analysis method and combined with the activity test,this study aims to obtain the index components of PMC and PML in Guangdong Deqing.Methods:In this study,the materials of PMC and PML in March,June,September and December from Guangdong Deqing were used as research objects and their chemical compositions were identified by UPLC-Q/TOF MS analysis technology.UPLC-Q/TOF MS and ~1H-NMR analysis techniques combined with multivariate statistical analysis methods such as cluster analysis,PCA,PLS-DA to find the differential components in different harvesting periods of PMC and PML and then analyze the dynamic changes of their contents.According to the content and polarity of differential components,the index components of PMC and PML were obtained.The anti-inflammatory activity of PMC and PML in LPS-induced RAW 264.7 mouse macrophages was investigated by NO and cytokine TNF-?.The antioxidant activities of PMC and PML in different harvesting periods were evaluated by the ability of O~2·?·OH and DPPH scavenging.Results:A total of 43 compounds were identified in PMC extracts,such as small molecule sugars,phenolic acids,catechins,stilbenes,anthraquinones and naphthoquinone,flavonoids,terpenoids,Lignins,fatty acids and so on.A total of 51 compounds were identified in PML extracts,including small molecule sugars and phenolic acids,catechins,stilbenes,anthraquinones and naphthoquinone,flavonoids,coumarins,terpenoids,fatty acids and so on.21different components of PMC and 24 different components of PML in different harvesting periods was screened though two techniques of UPLC-Q/TOF MS and ~1H-NMR.Both analytical techniques can be used for quality control of PMC and PML herbs.Catechin,quercitrin,emodin in PMC,polygoacetophenoside,hyperoside,quercitrin,emodin-8-O-?-D-glucoside in PML,these components have high content and high polarity,so they can be used as an index components.The results of anti-inflammatory activity showed that the PMC extracts in different harvesting periods was better in March and December,and the PML extracts was better in December and September.According to the difference in the content of components in different harvesting periods,the related compounds of anti-inflammatory activity in PMC and PML were catechin,(E)-2,3,5,4'-tetrahydroxystilbene-2-O-?-D-glucopyranoside,quercitrin,hyperoside emodin-8-O-?-D-glucoside and emodin.The different harvesting periods of PMC extracts to scavenging activity of O~2·was better in September and June,and the scavenging activities of·OH and DPPH were better in March and September.The scavenging activity in different harvesting periods of PML to O~2·?·OH was superior in June and September,and the scavenging activity of DPPH was superior in December and September.According to the difference of components content in different harvesting period,the components related to antioxidant activity in PMC and PML were estimated to be catechin,hyperoside,quercitrin and emodin.Conclusion:There are significant differences in the chemical composi-tion of PMC and PML in different harvesting periods and they show regular changes.According to the differential component analysis of PMC and PML combined with anti-inflammatory and anti-oxidation test results,PMC can choose catechin,(E)-2,3,5,4'-tetrahydroxystilbene-2-O-?-D-glucopyranosi-de,quercitrin emodin and PML can use polygoacetophenoside,hyperoside,and quercitrin,emodin-8-O-?-D-glucoside as indicators of their quality control.Therefore,plant metabolomics based on UPLC-Q/TOF MS and ~1H-NMR can be used for quality control of PMC and PML in different harvesting periods,and also provide experimental basis for existing quality evaluation methods of Chinese herbal medicines.