Rtudy On Isolation, Purification And The Analyze Method Of Saponin From The Leaves Of Camellia Chrysanth (Hu) Tuyama

Camellia Chrysantha (Hu) Tuyama is a kind of plant which in camellia genus, it is the tea of the chinese specialty and the great ornamental plant of world. The former results suggested, Camellia Chrysantha (Hu) Tuyama has many physiological functions such as depressing cholesterol in blood tumor, elevating organism immunity, restraining tumor and depressing fat, suger and so on. Saponins are the major effective components in the medicine of Camellia Chrysantha (Hu) Tuyama. In this dissertation, we mainly studied the extraction, separating and refining methods of total saponins from Camellia Chrysantha (Hu) Tuyama. Especially the UV spectrophotometry and HPLC analytical methods were developed to measure the total saponins.1. To investigate the technical conditions for the purification of total saponins from Camellia Chrysantha (Hu) Tuyama. The optimum conditions were as follows:Samples were sieved through XAD-16macroporous resin column, washed with water untit appearing of colorless effluent followed by elution with70%ethanol (at avolume of5-foldt hat of resin). After separation, the purity of total saponins in Camellia Chrysantha (Hu) Tuyama increased from about24%to35%.2. In this paper, The total saponins wre refined by organic solvent methods, polyamide resin methods and silica gel column chromatography methods. The results showed that polyamide resin methods was best, the optimum conditions were as follows:Samples were sieved through polyamide resin column, washed with Ethyl acetate untit resin balance followed by elution with70%ethanol (at avolume of5-foldt hat of resin).3. Based on single factor tests, with absorbance value as index, the response surface method (RSM) was used to optimize the determination condition of Total saponins by spectrophotometry. The optimal condition for color reaction was:the reaaction temperature was80℃, adding amount of5%vanillin-glacial acetic acid was0.35mL, adding amount of perchloric acid was1.0mL, the reaction time was25min and wavelength was540nm. Under the condition, the linear range of Ginsenoside Rgl content in samples was ranging from0～114.2μg with r=0.9991. The relative standard deviations of precision and stability were1.36%and2.08%, respectively. The average recovery rates was96.21%.4. A method to determine Ginsenoside Rgl and Ginsenoside Re by high performance liquid chromatography (HPLC) wsa developed. The effects of different chromatographic column, mobile phase composition, injection volumn and flow rate on separation have been studied. The best chromatographic conditions included a Venusil XBP-C18coumn (4.6X250mm,5μm) and a PDA detector. The optimal detection wavelengths were203nm. The flow rate was1.0mL/min. The injection volume was20μL. Under the optimal conditions, calibration curves of two substitutes were establish, the Ginsenoside Rgl correlation coefficients were0.9995and the Ginsenoside Re correlation coefficients were0.9997. The average recovery of Ginsenoside Rgl for96.87%, The average recovery of Ginsenoside Re for93.99%, and RSD of the method was within3.0%. The method is simple, dependable, good recovery rate and repeatability, and it can be used to control the quality of The Camellia Chrysanth (Hu) Tuyama.