Study On Chemical Composition Of Tea Seed Meal And Theasaponins Purification,Synthesis And Activity Of Derivatives

Tea garden area is large in Qinba mountains, and resource of tea fruit is rich. Production of tea seed meal is increased year by year with expanding the production scale of tea seed oil. Although output of tea seed meal reaches 8,000-10,000 tons per year in Shaanxi province, low utilization is an important question. Depth of processing can effectively extend the industrial chain. The main problems in extraction and separation of tea saponin from tea seed meal are as follows:The raw material contains a large number of impurities, such as polysaccharides, starches, proteins, which leads to viscous material, filtering difficulty, color depth, large energy consumption, high cost, low product purity, less downstream products and narrow application range under high temperature.According to these problems, our study exploited a novel, efficient and economic production process, tandem fixed bed adsorption of continuous foam preliminary separation, based on composition analysis of tea seed meal. Furthermore, Structural modifications of tea saponin had been carried on the preliminary exploration research. The main research results are as follows:1) Composition analysis of tea seed meal. Extracts concentrated by ultrasonic and cold soak with 80% methanol were Step two fractional extraction, including 8 compounds were isolated from ethyl acetate extraction phase.17 compounds were isolated from N-butyl alcohol extracts, including 5 compounds belonging to tea saponin. We constructed a reliable composition analysis method for tea saponin using color UV spectrometry and high performance liquid chromatography (HPLC) and contains theasaponins 39.63%?49.95%.2) Separation process of secondary continuous foam. We analyzed the surface activity characteristics of tea saponin solution, and measured the viscosity, critical micelle concentration CMC and the surface tension yCMC. Under 450 tilt angle condition in foam bed, drainage rate was fastest for improving the concentration ratio. Based on the single factor and response surface method optimized for gas velocity, pore diameter of distribution and solution temperature. In the optimum process conditions of laboratory, the average concentration ratio of 1 level of foam separation process was 5.11 and the average concentration of theasaponins was81.76mg.mL-1. When residual liquid into the secondary foam separation, enrichment ratio was 5.49, the average recovery was 57.77%.The total recovery rate 78.45%, secondary continuous foam separation process of tea saponin concentration of 65.55 mg. ML-1, average enrichment ratio was 4.09. The process achieved the enrichment purpose from the extracts. Purity of product was up to 48.37% after spray drying.3) D101C macroporous resin fixed bed for tea saponin purification process.D101C resin reached adsorption equilibrium within 6 h. Langmuir equation was an appropriate model for adsorption of tea saponin. The calculation results of enthalpy of adsorption ?H and absorption free energy ?G show that adsorption process is spontaneous, the high temperature is not conducive to the adsorption. Fitting the experimental data utilizing mathematic models, showing adsorption process kinetic well relevance to the first-order reaction and tendency affects is liquid film diffusion, particle diffusion.Fixed bed adsorption process of tea saponin in the laboratory was as follows:bed height of 35 cm, high ratio of 15, sample concentration of 14.5 mg.mL-1, material liquid velocity of 2.0 V·h-1, solution temperature of 308 K, pH 6.5, adsorption equilibrium within 6 h. Material liquid was pretreated with 2.0% NaOH solution, and then used 2.0% NaOH solution to wash bed with 3BV. Pigment, polysaccharide and other impurities was eluted by ethanol' water (V/V)=20:80 solvent 4.5BV. Then,80% ethanol as washing solution with 8BV, elution rate reached 94.7%. Purity of product was up to 90.5% after taking off the alcohol and spray drying. Finally, purity of product reached 97.08% with recovery rate of 85.14% after dissolution with 4.5 times the volume of methanol solvent at 75? and crystallization for 12 h at 0?. The purity of the product reached 97.2%, yield of 97.2%.4) Design goal was 50 Kg/day of tea saponin with purity> 95.0%. Scale up design secondary continuous foam separation-fixed bed purification Tandem pilot process based on the laboratory process data. We focused on calculation of amplification process foam separation and fixed bed. Purity of product reached 47.17% in secondary continuous foam separation of pilot process,89.45% in fixed bed adsorption process,97.2% after crystallization. It achieved our expected goals in pilot process.5) Preparation of tea saponin and synthesis process of three saponin derivatives.Tea saponin mixture was obtained by hydrolysis for 6 h in boiling water bath as 2.0 mol/L hydrochloric acid ethanol solution. The mixture was separated by preparation liquid chromatographic. The conditions were as follows: mixture of sample concentration of 65.0 mg·mL-1,0-6 min methanol:water (v:v)=30:70,6?38 min methanol: water (v: v)=40: 60, column temperature under 25?, the preparation of flow rate 2.5mL·min-1. The chromate -graphic peaks were collected by collector. Purity of saponin I was 98.67%, and saponin ? was 98.93%.With tea saponin I as substrate and two trichloroacetic amino sugar as donors, a pan of continuous glycosylation strategy completed simple synthesis of oleanolic acid saponin with disaccharides chain for the first time. Compound 1 showed strong inhibition activity of a-glucosidase and a-amylase. Values of IC50 were 160?M and 180?M. Compounds 1 and 3 showed strong inhibition activity of a-glucosidase and lipase. Values of IC50 were 170?M and 190 ?M.190 ?M and 200?M, respectively. It reveals that ?-L-rhamnopyranosyl and sugar substrate period help to improve the inhibition activity of a-amylase. Specific stereochemistry may inhibit the activity of lipase.