The Saponins Stauntonia Purification And Antioxidant Activity

Yemugua, a specialized resource of ZhengAn county in GuiZhou province, belongs to Rosaceae Chaenomelses plant. It implies various organic acids, amino acids, minerals, vitamins, traceelements, and contains some active substances such as trierpenoid saponins, flavoxate , polysaccharides, SOD, and so on.Being a natural plant used as both medicines and foods, it has been applied widely on clinical treat and health protection. At present, the research aimed to Yemugua is concentrated mainly on exploitation of common food , and systematic research on extraction and purification for its active substance remains unreported. The article studied majorly technology conditions of extracting and purifying total saponins, established content determination method of total saponins for Yemugua,posed the best technology parameters suitable to extract and purify for total saponins. In addition, studied initially and tried on its antioxidant capability outside body.The research work and experiment results are as follows:1. Content determination method of total saponins for YemuguaBy selecting oleanolic acid as control article, colorating with 5% vanillin-CH₃COOH and HClO₄, determined absorbability in the point of 545nm with ultraviolet-visible absorption spectroscopy and calculated total saponin content of Yemugua. The results showed that the method is linear in the range of 20～140μg(r=0.9994), absorbability stability within 1h, RSD 0.78%, moreover, the average recovery 102.21%, RSD 2.23%(n=5). The conclusion is that the method is sensitive and reliable, which may be used for the quality control of Yemugua.2. The optimization of extraction conditions for total saponins of YemuguaInfluence factors about extraction ratio of total saponins had been studied by selecting alcohol as solvent and taking method of immersion to withdraw. On the basis of single factor experiment, used orthogonal design to confirm the best extraction parameters. As a result, when alcohol level 75%, solid-liquid proportionl:15,temperature and time respectively 70℃and 60min, the extraction ratio can reach to the highest level.3. The optimization of purification process for total saponins of YemuguaTo use D₁₀₁ macroporous resin (column specification:R15×H200 mm , resin weight :4.000 g) for purifying total saponins of Yemugua, the optimal adsorption conditions were concentration of feed 8.86mg/mL, feeding rate 1 mL/min, PH value 5, the limited feed volume 18mL. The optimal de-adsorption parameters were alcohol concentration 50 %, elution volume both of water and alcohol were 100 mL, elution rate 1.5 mL/min,then the part eluted by 50 %alcohol were gathered. The experimental results showed that total saponins of Yemugua in the solid extract could reach 60.3 %, well above unpurified of 33.5 %, the refine ratio and elution ratio reached respectively 179.2 % and above 80 %.On the whole, the optimal conditions are suitable for separation and purification of total saponins in Yemugua.4. Initial trial on antioxidant capability outside bodyThe experiment studied initially the capability of total saponins in Yemugua to clear hydroxyl free radical (OH) produced by Fenton reaction and to clear superoxide anion (O₂^-·). The results manifested that the capability to clear hydroxyl free radical (·OH) was stronger than to clear superoxide anion (O₂^-·).