Studies On The Extraction, Purification And Hemocytolysis Of Asparagus Saponins

Asparagus saponins are a class of substance with cyclopentanoperhydrophenanthrene nucleus, and have various biological activity. However, Asparagus saponins are belonged to steroid saponins which lead to hematolysis as the injection medicine. This characteristic considerablely confines the application of Asparagus saponins. The method of determining the content of total saponins, the technology of extraction and purification, the hematolysis in vitro of Asparagus saponins and the elementary structure were studied in this paper. The results were as follows:1. To determine the content of total saponins in Asparagus officinalis L. by uv spectral method. Comparing the effect of the chromogenic agent to determine total saponins and establishing the optimal chromogenic method. The samples and sarasaspogenin reacted with three chromogenic agent:vanillin-perchlorie acid-glacial aceticac, sulfuric acid -methanol, perchloric acid. The result shows as follows:the method of using perchloric acid is the optimal, the detective wavelength is 310nm.and the linear range is between 72～216μg. The method is simple, accurate, and it can be used as a quantitative analytical method for Asparagus officinalis L.2. Optimization of the technology for extracting total saponins from Asparagus officinalis L. by ultrasonic wave-assisted was carried out. To investigate the effects of extraction temperature, extraction time and ultrasonic power as well as their pairwise interactions on the yield of total saponins from Asparagus officinalis L., the use of Box-Behnken design combined with response surface methodology was evaluated. The results showed that:the optimization of extraction process ethanol concentration95%, temperature 60℃,extraction time 23min and ultrasonic power 70W, extraction times 2 and the maximum total saponins yield was 23.96mg/g under these conditions.3. Macroporous resin was chosen with static adsorption and desorption experiments, and the optimum purification parameters of saponins from Asparagus officinalis L. were established with dynamic adsorption and desorption experiments. The results showed that: S-8 macroporous resin was found to have a good adsorption and desorption affinity and show an excellent decolor effect. The optimum purification conditions were 1 mg/mL of saponins eluted by pH=1 75% ethanol with a volume of 10 mL/g, the loaded amount was 60 ml/g. The purity of total saponons increased to 50.15% after elution, while the decoloration rate maintained 92.23%.4. Hematolysis of Asparagus saponins was in a dose-dependent, when the concentration of Asparagus saponins <25μg·mL-1,the hematolysis rate<5%. It will not lead to hemolysis as the injecting drug.5. The structure of Asaragus saponins was investigated by Poam reaction, Libermannn Burchard reaction, Ehrish reaction and IR-spectroscopy.It was shown that the nucleus structure of Asaragus saponins was steroidal saponins with furan-type opening ring.