Studies On Extraction-Purification And Determination Of Saponins In Ilex Kudingcha C.J.Tseng

Ilex Kudingcha C.J. Tseng is a traditional pharmaceutical vegetation in folk and plants mainly in guangdong, guangxi and hainan province. Research showed that it has distinct pharmaceutical effects, so it has a good prospect of clinical application and a high value to be developed into a new remedy. In order to promote the study biological active components and exploit its pharmaceutical value and provide scientific basis for utilizing Ilex Kudingcha C.J.Tseng resource, The extraction, purification and determination of saponins in Ilex Kudingcha C.J.Tseng were firstly researched, a RP-HPLC method was established for determination the content of oleanolic acid and ursolic acid in Ilex Kudingcha CJTseng simultaneously. At the same time, the technical parameters of sapogenin extraction sapogenin and the solvent system of HPTLC for separation the sapogenins was seeked. The main results are as follows:1. A spectrophotometric method was established for quantitative determination the total content of saponins in Ilex Kudingcha C.J. Tseng. With perchloric acid and vanillin coloration, the result showed that the optimal determination wavelength was 543nm. There was a good linear relationship between the absorbance and the quantity of ginsenoside Re when the concentration of Re(contrast) within the range of 21.4 μg ~ 256.8μg. Regression equation was Y=0.003706X+0.0046 with the correlation coefficient of 0.9998. The average recovery rates were from 97.84% to 100.12%. This method is stable and precise as well as high recovery rates. The method can determinate the total content of saponins in Ilex Kudingcha C.J. Tseng simply and rapidly.2. The extraction process of the total Saponins in Ilex Kudingcha C.J.Tseng was investigated by single factor and orthogonal experiment. The optimum condition for the extraction of the total Saponins in Ilex Kudingcha C.J. Tseng was that alcohol concentration is 80%; temperature is 65°C; extraction time is twice and one hour of each and the ratio of solid to liquid is 1:12.3. The column Chromatography was employed to enhance the content of the totle saponins. In terms of the result of static adsorption and static de-adsorption test, D101 resin was the best one for isolation saponins. The optimal adsorption parameters were feeding rate 2 BV/h, pH value 6, concentration of feed 14.5 mg/Ml. The optimal de-adsorption parameters were alcohol concentration 70%, elution rate 2.5 BV/h, elution volume 3 BV. The purity of final product was 68.92%.4. For the first time, The RP-HPLC method was established for determinating the...