Study On Extraction, Separation And Antioxidative Function Of Oleanolic Acid In Chaenomeles Speciosa S. Nakai

Chaenomeles speciosa s. Nakai is a kind of fruit with a medicinal function, it's rich in nutrients and abundant of active substances, and it's a kind of fruit with high food value and nutritional health protection effects, but fresh Chaenomeles speciosa s. Nakai is seldom eaten because its strong acerb taste, there are mainly papaya wine, fruit vinegar, preserved fruit , candied fruit and other traditional foods at present. People pay more and more attention on physical health as people's living standards improved, and more and more attention are payed on the development and research of functional food.The main purpose of this study is treat Chaenomeles speciosa s. Nakai as raw material to get a simple, green, easy-to- industrialization extraction process of the oleanolic acid by analyzing its nutrition and identifying its effective active ingredient of oleanolic acid by extraction and purification, and treat oleanolic acid as a comparison to carry out anti-cancer, anti-oxidation and other biological research on its extract. This study is to provide a theoretical and experimental foundation for the processing of Chaenomeles speciosa s. Nakai and further application of oleanolic acid.1. Chaenomeles speciosa s. Nakai nutrients were determined, which contains 86.937% of water, 2.879% of total sugar, 1.085% of reducing sugar, 3.956% of organic acids, 0.918% of pectin, 2.228% of tannin, crude fiber of 2.125%, 1.041% of protein, 0.206 of oleanolic acid, 0.113% of total flavonoids, and 0.124% of ursolic acid.2. Extracted oleanolic acid from Chaenomeles speciosa s. Nakai by ultrasonic assisted extraction and ethanol as solvent. Established orthogonal test according to the indexes of oleanolic acid in Chaenomeles speciosa s. Nakai's extract, the extraction studies showed: the order of the four factors'influence on the extraction is A> C> B> D, which is ethanol concentration effected most, extraction time was the second, followed by extraction temperature, solid to liquid ratio on the experimental results for the smallest. The best combination of technology under ultrasonic assisted extraction method was A₃B₁C₁D₁, which was 95% of ethanol concentration, 40℃of extraction temperature, 10min of extraction time, 1:20 of solid to liquid ratio. 3. Separation and purification of l oleanolic acid through macroporous resin. Purification experiment on the effect of oleanolic acid of Chaenomeles speciosa s. Nakai to get the best static condition of elution through choosing macroporous resin models, static adsorption and desorption rate, dynamic adsorption, elution concentration. Selected D4020 macroporous resins, with 95% ethanol as eluent eluted samples three times, the purity of what got was 84%, and RSD was 0.78%.4. Chaenomeles speciosa s. Nakai results showed that oleanolic acid has significant antioxidant activity(1) Scavenging free radical. When the concentration of Chaenomeles speciosa s. Nakai oleanolic acid was 1.0mg/mL, the scavenging on hydroxyl radical was largest, the clearance rate was 65.9%, followed by the 0.8mg/mL of concentration group, the minimum was the 0.2mg/mL of concentration group, which was 28.63%. The results showed that the Chaenomeles speciosa s. Nakai oleanolic acid on hydroxyl radical and the concentration had a clear dose-effect relationship from the Fenton reaction. It also showed had a strong scavenging on hydroxyl radical. Chaenomeles speciosa s. Nakai oleanolic acid inhibited O2·- which was from pyrogallol autoxidation, and the inhibition rate increased when the concentration did, which indicated that the inhibition had a visible relationship with dose-concentration, inhibitory effect on O2·- was strongest when oleanolic acid in Chaenomeles speciosa s. Nakai was 1.0mg/mL, which reached 63.83%, The minimum inhibition rate was 0.2mg/mL ,which was only 20.75%; The strongest inhibition on DPPH radical was 60.86% when Chaenomeles speciosa s. Nakai oleanolic acid was 1.0mg/mL, The minimumwas 0.2mg/mLwhich is only 17.31%, the results indicated that Chaenomeles speciosa s. Nakai oleanolic acid had a visible inhibition on DPPH radical, and the inhibition rate increased when the concentration did, showed that the inhibition had an evident dose-effect relationship with concentration.(2) Anti-aging effects on aging mice. This dissertation studied on the anti-aging effects and immune function of oleanolic acid Chaenomeles speciosa s. Nakai. Using D-galactose making aging model of mice, then the mice were given different dosages of oleanolic acid of Chaenomeles speciosa s. Nakai and treated ginsenosides as a positive comparison to observe the effect on the phenomenon of aging in mice and immune organs. The results showed that comparing with the aging model mice, Chaenomeles speciosa s. Nakai oleanolic acid can increase SOD activity which was the serum of aging mice and reduce MDA content significantly, the thymus index and spleen index increased, and the effect became more significant when adding the drug dose, it indicatied that Chaenomeles speciosa s. Nakai oleanolic acid can enhance the antioxidant capacity and immunity, which was the anti-aging effedts.(3) Studied effect of the oleanolic acid of Chaenomeles speciosa s. Nakai on tumor cell growth by MTT, the results showed that: Chaenomeles speciosa s. Nakai oleanolic acid can inhibit MCF-7 human breast cancer cell proliferating effectively; with the increase of oleanolic acid concentration, the growth inhibition enhanced by a dose-effect relationship.