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China's tea seed resources are very rich, comprehensive development and utilization of tea seed meal, application prospect obvious economic benefits. This study using spectrophotometry and reversed-phase high performance liquid chromatography (HPLC) to detect tea seed cake, tea saponin, compared two methods of quantitative analysis, and identified by reversed-phase high performance liquid chromatography detection tea The tea saponin content in the seed cake, while using phenol - sulfuric acid method to detect tea seed cake, tea seed polysaccharide. To optimize the extraction of polysaccharides of tea oil, tea saponin, tea seed; using a macroporous resin purification of crude tea saponin, high purity tea saponin; put forward a set of process, is expected to realize from tea seed cake tea oil, tea saponin, tea seed polysaccharide products. The main contents are as follows: (1) the tea saponin quantitative analysis reversed-phase high performance liquid chromatography (HPLC) for determination of tea saponin content: VP-ODS (250 mm × 4.6mm, C18, 5μm) analytical column, methanol - water (v / v = 9/1) as mobile phase, flow rate control in 1.0 mL / min, detection wavelength 280 nm, the concentration of abscissa and peak area for the vertical axis, the working curve for Y = 3.02 × 106X-1.16 × 104, a correlation coefficient of 0.9995. The recovery rate was 97.02% -102.82%. Tea seed cake by RP-HPLC determination of crude tea saponin content of 11.89%. This method has a smaller relative standard deviation, high precision and recovery, easy operation, is a better method of analysis. (2) analysis of the tea seed polysaccharide by phenol - sulfuric acid method Determination of tea seed polysaccharide glucose standard curve, the absorbance is measured at 490 nm, and the linear regression equation: Y = 173.48X-28.262 (Y represents the glucose content / microg X represents absorbance), the regression coefficient r = 0.9965. Experiments tea seed polysaccharide conversion factor f = 4.2783, according to the formula: Y = MCF / M to calculate the purity of tea seed polysaccharide. (3) tea oil, tea saponin extracted based on different physical and chemical properties of tea oil and tea saponin, using different extraction agent and compared to determine the petroleum ether (boiling range 60 ° C to 90 ° C) and methanol solution were respectively extracted tea oil and tea saponin. Soxhlet extraction of tea oil, ultrasonic extraction of crude tea saponin. Petroleum ether extract tea oil yield, low toxicity, and less wear and tear, high recovery rate. The optimization of the experimental conditions for extraction of tea oil, experiments under optimum conditions, ie heating time 2 h, the solid-liquid ratio 7 mL / g, reflux speed 5 min / tea oil extraction rate of 7.5%. Pure methanol as extraction solvent ultrasonic assisted extraction of tea saponin. Better experimental extraction process parameters: ultrasonic frequency 40kHz, extraction solution temperature 40 ° C, extraction time 25 min, liquid to solid ratio of 6 mL / g, ultrasonic power 250W, experiments under this condition, tea saponin was rate of 14.2%. (4) tea saponin purified using macroporous resin AB-8 crude tea saponin adsorption, adjust the the tea saponin solution pH = 5 for the sample, the results show that: the amount of adsorption of 32.1 mg / mL resin adsorption rate of 88% . Elution was carried out with 1% NaOH solution, and the tea saponin is adsorbed impurities and most of the pigment is washed, and then desorbed using 85% -95% methanol, the results show that the sample solution pH = 5: tea saponin, After the adsorption time of 2h, eluting with 1% NaOH solution in addition to impurities and pigments, and then 95% methanol as eluent (eluent and sample solution volume ratio 5:1), the eluate obtained in this condition concentrated to a certain volume, and precipitated by acetone and crystallized from methanol to yield high-purity crystals of tea saponin, measured obtaining a purity of 95.6%. (5) tea seed polysaccharide extracted using ultrasonic tea seed polysaccharide extracted from the Camellia meal. Single factor and orthogonal experiment to get the optimum conditions of ultrasonic assisted extraction of tea seed polysaccharide, extraction time 15 min, the ratio of liquid to solid 16 mL / g, ethanol concentration 50%, extraction temperature 50 ° C C. Experiment, tea seed polysaccharide was 8.5% under the optimum conditions.
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