The Extraction And Purification Of Total Saponins From Shiny-Leaved Yellowhorn Kernel Cake And Its Antioxidant Activity, Antibacterial Property, Anti-hepatoma Activity

Xanthoceras sorbifolia Bunge is a kind of deciduous trees which has multiple uses of edible, medical, ecological, environmental protection, ornamental, timber, and biomass. Currently, the study of Xanthoceras sorbifolia Bunge on its activity sbstabce such as saponin is gradually increasing with the deeper understanding of it. However, there are great differences of category, content and structure of different parts of Xanthoceras sorbifolia Bunge saponin. It is still unclear that the type of saponins monomer structure in Xanthoceras sorbifolia Bunge kernel is consistent with the seed peel or not, and if there exist large differences in biological activity among different monomers. In this paper, the extraction and purification process parameters of total saponin from Xanthoceras sorbifolia Bunge kernel were discussed, and its antioxidant activities, antibacterial characteristics, anti-hepatoma activities also were preliminary studied. This test results will lay a experimental foundation for the further exploration of saponin’s from Xanthoceras sorbifolia Bunge on the anti-hepatoma molecular mechanisms.The results of the study are as follows:1. Single factor and orthogonal experiments were used to analyzed the main influence factors of total saponins extraction from Xanthoceras sorbifolia Bunge’s kernel cake and determined the optimal extraction parameters. The results showed that under the condition of 70% ethanol concentration,50℃ extraction temperature, solid-liquid ratio 1: 20 and the ultrasonic power of 140 W, the yield of total saponins will reach to the highest content of 2.67%.2. The total saponins of Xanthoceras sorbifolia Bunge kernel were separated and purified by using macroporous resin, and the adsorption and desorption properties of several kinds of commonly used resin were discussed. The resin which has good adsorption performance to saponin was choosed, and its effects on the adsorption and desorption properties was studied to determine the optimal condition of purification. The XAD-16 macroporous resin was choosed as the best for its highest ratio of adsorption and desorption on the total saponins purification. The optimum conditions were as follows:the static adsorption time was 12 h, the static desorption time was 4 h, eluent ethanol concentration is 70%, pH=4. The optimum conditions of dynamic adsorption also were as follows:sample flow velocity is 10 mL/min, the proportion of sample volume and column volume is 1.5. The total saponins content of Xanthoceras sorbifolia Bunge was increased from 0.12 mg/mL to 0.52 mg/mL after purification for three times using this resin.3.The antioxidant activity of purified extract of total saponins from Xanthoceras sorbifolia Bunge was studied. Using Vc as a control, the reducing power and the scavenging ability of total saponins on the DPPH free radical, hydroxy radical and superoxide free radical were determined. The results showed that the reducing power and hydroxy radical scavenging effect of total saponins were stronger than Vc, and the abilities of scavenging DPPH free radical and scavenging superoxide were less than Vc.4. After purified, the antibacterial characteristics of total saponins extract from Xanthoceras sorbifolia Bunge were investigated with filter paper osmosis test method. The results showed that there were different degrees of inhibition of Xanthoceras sorbifolia Bunge saponins on the Staphylococcus aureus, Escherichia coli, Salmonella typhi, Bacillus subtilis bacillus, and Pseudomonas bacteria. It had the strongest inhibition on Pseudomonas bacteria, and its MIC was 3.6 mg/mL. Meanwhile it had the weakest inhibition of saponins on Salmonella typhimurium, and its MIC got to 4.8 mg/mL.5. The anti-hepatoma activities of purified extract of total saponins from kernel of Xanthoceras sorbifolia Bunge were preliminary studied. The inhibition of proliferation and apoptosis of total saponins extract on HepG2 cells were evaluated with MTT assay. The ICso of total saponins against HepG2 cells was 9.672±0.3856 mg/L after treatment of 24 h. The apoptosis induction of total saponins extract on HepG2 cells was determined by cell morphology and fluorescence staining method.