Tea Seed Cake, Tea Saponin Extraction And Purification

The development of the tea saponin may have bright future and remarkable economic benefit because there is abundant tea-seed pancake resource in china. Quantitative determination of tea saponin with purified tea saponin as standard by UV spectrophotometry and reverse-phase high performance liquid chromatography (HPLC) was studied. The optimum technological conditions of cha oil and tea saponins extracting in turn from tea-seed pancake were studied. Two methods have been used in purification of tea saponins. One method was that tea saponin had been decolored with H₂O₂, another was purification with macro porous resins. On the base of the review to the characters of tea saponin and applications in some fields, and on summarized quantitative determination, extraction and purification of tea saponin from tea-seed pancake, the following experiments were carried on:(1) Quantitative analysis of tea saponinA reverse-phase high performance liquid chromatography method (HPLC) for determination of the tea saponin which extracted from tea-seed pancake with Soxhlet extractor was studied. After VP-ODS (150mm×4.6mm, i. d., 5μm) was chosen as analytical column and methyl alcohol at a flow-rate of 0.50 mL/min as mobile phase, the UV signal was detected at 280 nm. The value of abscissa shows the mass concentration, and y-axis, area. The linear equation was Y=3.54×10⁶X+1.15×10⁵. The recovery of teasaponin is 97.93%～102.78%, the relative standard deviation is 0.10%～0.51%。Through method of HPLC determined the content of tea saponin in tea-seed pancake, was 12.706%. The results of the experiments conformed that this method was accurate, rapid, simple and reproducible.(2) Extraction of cha oil and tea saponinThe extraction technology of cha oil and raw tea saponin from tea-seed pancake was studied in this paper, petroleum ether (range of boiling point from 60℃to 90℃) which used in extraction of cha oil as extractant was optimum, and methyl alcohol was the optimum extractant in extraction of tea saponin. The optimum extraction parameters were obtained by a series of simplex factor experiments and the perpendicular design of L₉ (3³) factors, i. e., the optimum extraction paramenters of cha oil were: time at 120 minutes, the liquid-to-solid ratio is 6mL/g , circumfluence velocity of flow was 4minutes/time and the results in the extraction rate of cha oil reached 7.8%; the optimum extraction paramenters of raw tea saponin were: time at 120 minutes, the liquid-to-solid ratio was 6mL/g, circumfluence velocity of flow was 12minutes/time and the results in the extraction rate of tea saponin reaching 18.77% .This method has much more advantages, such as, technique of extraction and separation is simple, the cost is lower, the effective of extraction is higher etc. this method can be used in factory.(3)Purification of tea saponinEffect of different decolorizers on tea saponin decolorization was studied .the optimum condition of hydrolysis found by orthogonal test was: decoloring temperature 60℃, decoloring time 90min, 3% H₂O₂ PH=10With the material of pretreated methyl alcohol extract of tea-seed pancake, the adsorption and desorption effects on tea saponin of macro porous resins were studied, such as AB-8, S-8,X-5,D-4020,NAK-9. The results showed that AB-8 was the suitable resin of purifying tea saponins. The optimum technological conditions as follow :PH=5, flow rate 1BV/h, and 90% ethanol as eluting agent. Resin could be regenerated after every four times uses. The extract was treated by AB-8 resin, then precipitated by acetone, finally, It has been turned into crystal by 90% methyl alcohol .This method was used to purify the tea saponin . the purity of the tea saponins obtained in this way was over 96%, and the yield was over 60%.