The Research On Extraction, Separation, Purification And Biological Activity Of Dicranopteris Dichotoma Saponin

Medicinal ferns were widely used because of its wide distribution all the time. It’s an abundant resource of medicine in folk. Dicranopteris dichotoma is an acidic soils indicate ferns, widely distributed in China, cheap and easy to get. In this paper, three different extraction methods were used to study Dicranopteris dichotoma saponin. Which were the traditional ethanol water extraction, ultrasonic extraction and microwave extraction, and the extraction process was optimized by response surface, the optimization extraction process is as following:ethanol water extraction:water extraction bath temperature49.51℃, extraction time4.79h, ethanol volume fraction was62.87%; ultrasonic extraction:ultrasonic power204.17W, ultrasonic treatment time was189s, the ethanol concentration was64.32%; microwave extraction:microwave power is616.78W, microwave processing time7.51min, ethanol concentration was40.38%. The comparison result of three different extraction methods of total Dicranopteris dichotoma saponins is ultrasonic extraction (12.49%)> microwave extraction (11.17%)> ethanol bath extract (10.45%).On the basis of the best extraction process, we explored the comparative study of the adsorption and desorption effects of Dicranopteris dichotoma saponin between four different resin, the results show that D-101macroporous is an ideal resin as its big adsorption capacity and desorption rate, more suitable for the separation and purification of Dicranopteris dichotoma saponins. And the optimum adsorption conditions of D-101macroporous is:the flow rate is2BV/h, feed concentration20.50mg/mL, liquid pH value is5; optimal elution condition is:flow rate is2BV/h, eluent3BV,70%ethanol. After purification,the total saponin content was64.90%.On the basis of the best extraction process, we studied the anti-prostate cancer PC-3cells effects of27components which separated by silica gel, the results show that different components have different inhibitory effect on prostate cancer PC-3cells, at a concentration of80mg/ml, the component1and component2have a high inhibition rate of prostate cancer PC-3cells which reached74.01%and91.40%; but some components inhibition rate is low, even appeared negative inhibition rate, indicating that some component play a facilitating role on the growth of prostate cancer PC-3cells. In addition, the first three components of the DPPH radical scavenging effect showed Dicranopteris dichotoma saponins have a good effect on DPPH radical scavenging, as the sample concentration increases, the scavenging rate increases. So we can conclude that Dicranopteris dichotoma extract can be seen as an effective drug for the treatment of prostate cancer, but also an effective antioxidant.By extracting with solvents of different polarity stratified firstly, then further separated by silica gel, reverse phase silica gel, Sephadex LH-20column chromatography and other methods. Through TLC chromatography results,we can see the component A and component B are relatively simple, which are the ethyl acetate extract layer separated by silica gel column, then reverse phase silica gel column. To explore structure of the component A by mass spectroscopy, IR and NMR, we find the substance A is quercetin, while content of quercetin in Dicranopteris dichotoma measured by HPLC was1.87%. We can conclude that there is a high levels of Flavonoid in Dicranopteris dichotoma.