Burnet Polyphenol Component Analysis And Functional

As a kind of polyphenol mixtures,polyphenols exist in a wide range of plant tissues. Polyphenols have great capacity of trapping and scavenging oxygen free radicals and reduce risk of some diseases(stroke,tumor and cancer,etc.).Through comparing the content of polyphenols,Radix Sanguisorbae ranks the highest among 86 different plant materials.There are abundant plant resources of Radix Sanguisorbae which are widely distributed in many provinces of China.To make full use of Radix Sanguisorbae and its value-added processing,it is of great importance to extract and analyze the polyphenols from Radix Sanguisorbae.In this paper,the following contents of polyphenols from Radix Sanguisorbae were studied:1.In this study,the quantitative method for hydrolysable tannin and condensed tannin were investigated.Vanillin-H₂SO₄ assay was selected to determine the content of condensed tannin of Radix Sanguisorbae.In this paper,(+)-catechin was used as the standard substance,and there is a very good linearity when the concentration of(+)-catechin changing from zero to 0.30mg/mL.The standard curve equation is Y=1.7839Xq-0.0017 and the R² is 0.9989.The Vanillin-H₂SO₄ assay is suitable especially for the new plant resources which do not have the mature product of their standard substance for its sufficient precision and ruggedness.The content of hydrolysable tannin is quantitatively determined by HPLC assay and the standard curve equation of gallic acid and ellagic acid were established too.The RSD of precision and recovery of tis experiments are 0.34%and 0.36%resctively.The content of hydrolysable tannin and condensed tannin were messured as 84.90mg/g and 62.81mg/g respectively,so the total content of polyphenols is 147.71 mg/g in Radix Sanguisorbae.2.The qualitative analysis of oligomeric polyphenols in crude Radix Sanguisorbae extracts can be performed by reversed-phase high-performance liquid chromatograhy coupled with electrospray ionization mass spectrometry(RP-HPLC-ESI-MS).Pseudomolecular ion and fragment information of ESI-MS made it possible to analyze the composition of oligmeric polyphenols directly without standards.By RP-HPLC-ESI-MS,Three monomers,five dimers,three trimers and one tetramer of condensed tannin were tentatively identified in crude Radix Sanguisorbae extract,at the same time,two monomers,two dimers,three trimers,one tetramer and twelve pentamer of hydrolysable tannin were tentatively identified too,but due to the large number of isomers and their similar ESI-MS characters, the isomers are not identified precisely except monomers.In fact,only oligomers with MW below 1000 can be well detected by RP-HPLC-MS,the higher polymers,with more isomers and more diversepolarity,can neither be separated nor be analyzed by RP-HPLC.3.Quadratic regression orthogonal rotary experiments for acetone extraction of polyphenols were used in this paper.And the results were analyzed by DPS.The results indicated that the 3 factors in the experiments-extraction time(X1),acetone concentration(X2),ratio between solvent and material(X3)-had strong influences to the extraction effects.The best extraction conditions are: extraction time(X1) is 30min,acetone concentration(X2) is 70%,ratio between solvent and material(X3) is 28:1.Under these conditions,the average extraction amount is 198.28mg/g,and it's about 4.59%higher than the theoretical calculation value.The equation which showed the relationship between the extraction effect and the 3 factors is: Y=196.10932 + 8.11188X1 + 5.73496X2 + 7.39815Z3 - 4.92490 X1²-5.06702 X2²The results of static adsorption,desorption and the kinetics of static adsorption show that AB-8 macroporous absorbent resin is suitable for Raidix Sanguisorbae polyphenol.The absorbing and eluting rate of AB-8 resin was 89.6%and 92.6%respectively,which possessing higher absorbing and eluting capacities.Through dynamic desorption of polyphenols absorbed on the AB-8 resin,the results showed that absolute ethanol had a strong eluting effect and the polyphenols can be fully eluted by 4BV of absolute ethanol.The symmetry of elution peak was quite well and the recovery rate was 90.4%.After vacuum freeze-drying,the purity of polyphenols can reach 95.3%.4.The antioxidant capability,enzyme inhibition of tyrosinase and protection effects of human skin fibrolast of Radix Sanguisorbae polyphenols were observed in this paper.The reducing power increased with the growing concentration of polyphenols,and the Ferric Reducing/Antioxidant Power(FRAP) value was 30.566mmol FeSO₄/g Polyphenols of Radix Sanguisorbae.Polyphenols from Radix Sanguisorbae showed a high scavenging activity of 1,1- diphenyl-2-picryl-hydrazyl (DPPH·),and the 50%inhibitory concentration(IC₅₀) was 18.86mg/L.For the scavenging activity for hydroxyl radical(·OH) and superoxide anion radical(O₂^-·),the IC₅₀ was 1.258mg/mL, 18.53mg/L,respectivily.At the present of Radix Sanguisorbae polyphenols,activity of the cell multiplication,SOD and GSH-Px of fibrolast increased obviously,and the content of MDA decreased obviously.This activity of Radix Sanguisorbae polyphenols is dependent on it's molecular structure,for the phenolic hudrooxyl group of polyphenols can capture free radicals,interrupt Chain reaction of unsaturated fatty acid and inhibit peroxidation of unsaturated fatty acid,that finally result in the content of MDA decreasing and the activity of fibrolast multiplication has been keeped.Ultraviolet irradiation can excite Langerhans cells to form more melanogenesis and cause to skin pigmentation.Through tyrosinase activity analysis,Polyphenols from Radix Sanguisorbae showed a high inhibition activity of tyrosinase enzyme,and the 50%inhibitory concentration(IC₅₀) was 14.5mg/mL,indicate that Radix Sanguisorbae polyphenols could be used as skin cosmetics active component.