Chemical Modification And Activity Of Paris Polyphylla Var. Chinensis Polysaccharide In Vitro

Plant polysaccharides appear to extensive physiological activities,such as antioxidation,antitumor,antivirus,antibacteria and immunoregulation,et al.Appropriately chemical modification can significantly increase these activities.Paridis Rhizoma is a rare traditional Chinese medicinal material,which has extensive pharmacological effects and significant clinical efficacy.Now,the research of Paridis Rhizoma mainly focuse on the structural analysis,identification and pharmacological effects of its saponins,Paridis Rhizoma polysaccharide was seldom reported,chemical modification of PPCP was not involved.Paridis Rhizoma from Paris polyphylla var.chinensis,distributed in Qinling-Bashan Mountain area,was as test material to obtain the polysaccharide?PPCP?.Detected PPCP by phenol-sulfate method,which was optimized and methodological verificated.Focused on the chemical modification of PPCP by acetylation and sulfation with process optimization,under the optimal conditions,obtained acetylated P.polyphylla var.chinensis polysaccharide?A-PPCP?and sulfated P.polyphylla var.chinensis polysaccharide?S-PPCP?.At the same time,antioxidant and scavenging activities in vrito of PPCP,A-PPCP and S-PPCP were researched.The main contents and results were as following.?1?Cruded by hot water extraction,after deproteinization,alcohol precipitation,dialysised separation and purification,prepared PPCP.The optimized phenol-sulfate method condition for detecting PPCP was built,sulfuric acid dosage was 6.5 mL,phenol dosage was 2.0 mL,reacting temperature was 30?,and reacting duration was30 min,which the dosage of sample was 0.2 mg.Under optimal condition,the average content of PPCP was 91.3%,average recovery rate was 97.8%,and the relative standard deviation was 1.2%.The good faccuracy and stability ensured that this method was feasible for the content analysis of PPCP.?2?PPCP was chemical modified by acetylation and sulfation,and the respective chemical modification process was optimized.The acetylation process condition of PPCP was optimized by Response Surface Methodology,the furnish ratio was 7?1?g?g?,the reaction temperature was 25°C,and the reacting duration was 30 min.Under this process,the average acetyl degree substitution of A-PPCP was 0.342.Optimized the sulfation process condition of PPCP by orthogonal test,the optimal process was as follows,the reagent ratio of sulfuric acid to n-butanol was 3?1?V?V?,reaction temperature was 0°C,and the reacting duration was 1.0 h,the average degree of S-PPCP was 0.402.?3?Two kinds of chemical modification PPCP,which had prepared under the optimized process,were identified by UV and IR.In contrast the characteristic peaks with the ultraviolet and infrared spectrum of PPCP,combined with the detection results of degree of substitution,it was showed that PPCP had been finished chemical modification successfully by acetylation and sulfation.?4?The experiments of scavenging activities on DPPH·,·OH,O₂^-·and NO₂^-in vitro were used to investigate the antioxidant and radicals clearance activities of PPCP,A-PPCP and S-PPCP.Based on mathematical analysis,compared with the IC₅₀0 of PPCP,the IC₅₀ of scavenging activity on DPPH·of A-PPCP and S-PPCP were reduced by34.62%and 36.99%respectively,the IC₅₀ scavenging activity on·OH were decreased by 28.07%and 23.00%respectively,which indicated that both acetylated modification and sulfated modification can effectively increase the scavenging activities on DPPH·,·OH of PPCP in vitro.Compared with PPCP,the IC₅₀ clearning activities on O₂^-·,NO₂^-of A-PPCP were reduced by 9.32%and 21.27%respectively,it was showed that the acetylated PPCP had a certain enhancement effect on the clearance activities of O₂^-·,NO₂^-.On the contrary,compared with PPCP,the IC₅₀ of S-PPCP scavenging activities on O₂^-·,NO₂^-were increased by 13.79%and 8.62%respectively.It was showed that sulfation could reduce the scavenging activities of PPCP on O₂^-·,NO₂^-.Acetylation and sulfation modification of PPCP had firstly carried out,and the respective process condition was optimized.Then the in vitro antioxidant activity and scavenging activity on NO₂^-of PPCP,A-PPCP and S-PPCP were comparative researched.In addition,the phenol-sulfuric acid test condition of PPCP was optimized at the first time.Methodological investigation results showed that the method had high accuracy and good reproducibility.It was concluded that the method can be used for the detection of PPCP.This study is designed to provide a reference for the development and utilization of PPCP.