Study On Extraction And Purification Of Tea Saponin From Camellia Oleifera Seed Meal

There is aboundant of the oil-tea camellia seed cake in China which contains tea saponin about 10%~15%.it is a good nonionic surfactant that can make emulsifier,foaming agent,wetting agent,it can be processed into many kinds of high value products.Using camelilla cake as raw material,the process of extraction and purification of tea saponin were investigated in this thesis.The main contents were as follows:(1)Study on determination method of the content of tea saponin.The employed coloring reagents was vanillin-acetic acid and the color determination of saponin content based on aglycone at the wavelength of 554 nm.Optimal color condition for sample solution were obtained as follows:color reagents 1.0mL,temperature60?,reaction time 15 minutes.The color of the samples were stabled within 40 min and the average recovery was 98.28%(RSD=3.86%)with high precision.The method based on spectrophotometry including high accurate,precision and stability,it can be used for quantitative determination of tea saponin.(2)Study on the extraction technology of tea saponin.Using aqueous methanol as an extracting agent.Orthogonal method was used studying the effect of methanol concentration,time,temperature,volume.The optimum extracting conditions were obtained as follows: 3.5 times of amount of 75% methanol,extracted at 60?for 180 minutes.The yield was 11.54% at the conditions listed above.(3)Study on the aqueous two-phase system—precipitation release method of separation and purification of tea saponin process.An aqueous two-phase system based on propanol/ammonium sulfate was used in separation and purification of tea saponin.The optimum separation extraction conditions were obtained as follows:ammonium sulfate concentration 21%,propanol concentration 24%,30? and pH 7 adjustment.Under the optimal conditions,the extration ratio and the content reached 90.32% and 78.12%.Chemical precipitation release method were used for further purifying tea saponin,Ca O was applied,NH4HCO3 was chosed as release reagent.The optimum precipitation release conditions were obtained as follows:the ratio of Ca O to tea saponin was 1.2,The reaction at room temperature for 150 minutes,the ratio of NH4HCO3 to Ca O was 1.5,The reaction at 50? for 120 minutes.The average yield and the content reached 73.21% and 90.41%.This process is simple,reliable and lowcost.It provides a safe and reliable method for industrial production.(4)The tea saponin were treated with a series of test such as color reaction,precipitation,melting point,hydrolysis,spectral,HPLC,MS identification.The results were as follows: the tea saponin were characterised by two features color reaction.precipitation experiment showed a white precipitate appeared,the melting point was about 224~227?,hydrolysis identification showed tea saponin produces the specific aglycone,ultraviolet spectrum identification showed that tea saponin had large absorption peak at 218 nm,infrared spectrum showed the sample and standard characteristic absorption peaks were basically identical;HPLC identification showed the sample chromatograms had no other impurity peak and fitted the standard chromatogram peak time;MS identification showed the sample and standard peak type were great and m/z mainly in 1200~1300.According to the identification,the sample was tea saponin and had a high content.