Separation And Purification Of Tea Saponin From Camellia Cake And Its Performance Research

Tea Saponin is mainly extracted from tea seed meal,which is abundant in China.Tea Saponin,a natural non-ionic surfactant,consists of hydrophilic glycosylation and hydrophobic aglycone,as well as organic acids.It is not only good foaming,excellent emulsifying,but also has the function of killing bacteria,pests and other diseases.Therefore,the tea saponin can be widely used in the daily necessities of life,industrial and agricultural products.At present,there are many methods which the extraction of tea saponin at home and broad.It is popular method that is using the organic solvent to extracte tea saponin from tea cake,the main advantage that is easy to operate and use the existing mature technology for large-scale commercial production.But the tea seed meal contains many ingredients such as fat,protein,sugar,cellulose and so on.The organic solvent can't remove the organic solvent,and the high cost of organic solvent and the pollution of environment.Therefore,this experiment used biological enzyme method to extracte tea saponin,we discussed the influence of lipase,cellulase,protease of tea saponin to extraction effect,and explore the feasibility of this method and optimum extraction conditions.Based on the extraction in aqueous enzymatic method,then carry out further purification of tea saponin.Due to the complexity of the structure of camellia saponin,many components,in various forms,and tea saponin crude and impurities close polarity,it is difficult to remove and separation.This experiment using aqueous enzymatic method as tea saponin crude extracts,and calcium oxide of the crude extract as concentrated and the S-8 macroporous resin purification of tea saponin.In test,we compared the macroporous resin purification of tea saponin with traditional calcium oxide synthesis method extraction of tea saponin and found the two methods can improve the yield and purity of tea saponin.After purification of the crude tea saponin research and obtained certain purity tea saponin.For accurate measurement and analysis of tea saponin content to explore by high performance liquid phase chromatography and establish the high performance liquid phase chromatographic quantitative analysis method of tea saponin.After determining the content of tea saponin,the relatively high purity components by infrared spectroscopy and mass spectrometry.The analysis shows that the separation of tea saponin as camellia saponin B₁,B₂,A₄.Finally,experiments of tea saponin on foaming properties and the decontamination were analyzed.The results show that tea saponin with rich foaming and milder decontamination ability,tea saponin can be used in washing products for daily cleaning.This research mainly obtains the following conclusions:1.A process for extracting tea saponin using enzymatic hydrolysis in this study.The yield of tea saponin was used as an evaluation index accompany with cellulase?lipase and protease act on Camellia Oil Cake.The effects of extraction ratio of solid to liquid 1:20,extraction time 60 min,stroke speed 150r/min on tea saponin yield were investigated by single factor experiment.The extraction parameters of temperature,content of the lipase,cellulase and protease were optimized by response surface methodology.The results showed that the optimal extraction conditions were as follows:extraction temperature 50?,extraction time 1h,content of the lipase,cellulase and protease is 0.31%,0.32%,0.36%.Under these conditions,tea saponin yield was 86.86%±0.92%,recovery percent 87.04%.2.The study purification of the coarse tea saponin by macroporous resin and ascertain the optimal conditions of separation and purification the tea saponin.The important of experiment is that study property of different polarity of macroporous resin D-101?AB-8?S-8?X-5?HPD-100.We choice S-8 macroporous resin that a representative of all macroporous resin and comprehensive analysis capacity of adsorbability and resolving of S-8.Determine the saturated adsorption capacity is1.12mg/ml,the best flow rate on the sample liquid is 3.03ml/min.Tea saponin yield as index,combining the response surface analysis to determine the ethanol elution conditions as eluent,which best concentration is 91.20%,flow rate 7.26ml/min,dosage of ethanol 101.31ml,tea saponin yeld 86.30%.According to the analysis results,we study purification of the content of saponin 47.38%,as a result tea saponin yield81.74%,recovery percent 78.29%,purity 85.36%.3.The study the process purified tea saponin using calcium oxide and S-8macroporous resin.Enzymatic extract water as raw material,tea saponin yield and purity as an indicator,a comprehensive analysis of the different precipitants effects on protein,select the most precipitating agent of calcium oxide through univariate analysis and combination with S-8 macroporous resin concentrated and purified of tea saponin.the optimum amount of its orthogonal analysis for the solution of calcium oxide tea saponin content of 2.5 times volume of tea saponin,the amount of conversion agent ammonium bicarbonate calcium oxide consumption 2 times of tea saponin;S-8 macroporous resin sample volume of 100ml,the volume of the sample extract 150ml,optimal concentration of 1%sodium hydroxide eluent,1.5 times the amount of macroporous resin,the amount of water 1 times the macroporous resin,150ml80%ethanol elution as last eluent agent,measured the yield of tea saponin was76.18%,purity was 87.04%.4.The present study was conducted to extract of tea saponin from Camellia Oil Cake by enzymatic hydrolysis and purified tea saponin using macroporous resin,at last we through LC-MS anlysis it.The yield of tea saponin was used as an evaluation index accompany with cellulase?lipase and protease act on Camellia Oil Cake.The purification of the coarse tea saponin by macroporous resin and ascertain the optimal conditions of separation and purification the tea saponin.Extraction of tea saponin and purification it,different purification were determined by LC-MS.The results showed that the optimal extraction conditions were as follows:extraction temperature50?,extraction time 1h,content of the lipase,cellulase and protease is0.31%,0.32%,0.36%.Under these conditions,tea saponin yield was 86.86%±0.92%,recovery percent 87.04%.According to the analysis results,we study purification of the content of saponin 47.38%,as a result tea saponin yield 81.74%,recovery percent78.29%,purity 85.36%.LC-MS analysis show that the experiment of purity the saponin have three majority ingredient CamelliasaponisB₁,CamelliasaponisB₂ and CamelliasaponisA₄.5.Preliminary studies suggest that tea saponin decontamination ability:tea saponin has excellent foaming,emulsifying ability.Tea saponin has a good effect on the removal of oil,but the color stain ability is poor.Therefore,the tea saponin as a natural non-ionic surface active agent,can be used as a detergent composition for cleaning the tableware.Or as hand washing liquid formula,use.As a kind of natural green sources,has broad application prospects.