Study On High Efficient Preparation And Anticancer Activity In Vitro Of Tea Saponin

Tea saponin is a kind of pentacyclic triterpenoid saponins with oleanolic acid structure which exists in the camellia plant?camellia,Camellia oleifera?tea,etc.?seeds It contains hydrophilic sugar and hydrophobic Ligand,and it is a pure natural non-ionic surfactant,and has a variety of biological activity,widely used in agricultural protection,medicine,cosmetic,building materials and other fields.China is the world's largest tea producing country,tea seed is rich in resources,tea saponin content of 18²5%in the de-oiled tea seed cake,but the tea seed is usually produced as a by-product of tea which makes tea saponin Can't be effectively used.In this study,with tea seed cake meal as raw materials,in order to study the effect of tea seed saponin efficient preparation and in vitro anti-cancer activity research,So as to provide scientific basis for the industrial preparation and application of tea saponin in tea seed.The main findings are as follows:?1?Tea saponin extraction:the supercritical CO₂ extraction of tea seed oil after the cake as raw material with ethanol as the extraction solvent,through single factor experiment,orthogonal experiment and the best combination of technology validation experiments,The effects of ratio of material to liquid,volume fraction of ethanol,temperature and time on the extraction rate of tea seed tea saponin were investigated.The results showed that the yield of tea saponin was 16.3%and the process was stable under the optimum conditions of the ratio of material to liquid 1:20,ethanol volume fraction 70%,extraction temperature 70?and extraction for 2 hours.At the same time,based on the optimum ethanol extraction technology,the effects of three methods of hot water extraction,ethanol extraction and ultrasonic assisted ethanol extraction on tea saponin extraction were studied.The results showed that ethanol extraction was better at tea saponin yield,purity and extraction efficiency.?2?Separation and purification of tea saponin:To study the effects of 10 kinds of macroporous adsorption of D101,AB-8,ADS-17,HPD400,HP2MG,SP70,S-8,CAD-40,H-20 and XAD-16N on tea seed cake 90%ethanol reflux extraction after add water to dilute make-up liquid,which with filter vacuum concentration and ethanol recycling after supercritical tea seed oil.Effect of Resin on Static Adsorption and Desorption of Tea Saponin.the optimum parameters of dynamic adsorption and desorption of tea saponin were optimized by single factor experiment,orthogonal experiment and verification experiment.The results showed that D101 resin had good adsorption and desorption effect on tea saponin,The static adsorption capacity and desorption rate of D101 resin were 142.974mg/g and98.02%,respectively.When the yield of tea saponin was mainly investigated,The optimal technological parameters were tea saponin 10mg/mL,sample flow rate 3BV/h,loading volume 6BV,ethanol volume fraction 80%,elution flow rate 3BV/h,eluent volume 5BV,tea saponin yield 74.25%,purity of 84.30%;When the purity of tea saponin was mainly investigated,the optimum technological parameters were as follows:tea saponin 10 mg/m L,sample flow rate 4BV/h,loading volume 7 BV,ethanol volume fraction of 70%,elution flow rate of 3BV/h,the elution volume of 5BV,the tea saponin purity of 97.7%,the yield was72.04%.It is shown that D101 macroporous adsorption resin is a kind of resin which can be used for the separation and purification of tea saponin in tea seed cake.?3?Study on anticancer activity in vitro of tea saponin:The cell viability was determined by MTT assay,the fluorescence intensity of the cells was detected by staining with Rhodamine 123 and the sclerations were measured at different concentrations of tea saponin.The effects of tea saponin anticancer activity in vitro on Hela human cervical cancer cells were studied by using the high purity saponin which extract from tea seed by separation and purification.The results showed that the high purity tea seed saponin extract treated group have?P<0.01?extremely significant anticancer activity compared with the blank control group at the lower concentration of 30?g/ml.In the range of 40⁵0?g/ml,high purity tea seed saponin has a lethal effect on Hela human cervical cancer cells;Indicating that high purity tea saponin have a good effect to prevent cancer cell proliferation,weaken the vitality of cancer cells and even kill.