Extraction And Purification Process Of Tea Saponin And Its Effects On Alcohol Dehydrogenase Activity

Tea saponin is saponin mixture which is Oleanolic acid type, and the content of tea saponin in Camellia cake is about 10%-18%. In order to get the tea saponin that can dispel the effects of alcohol by activating alcohol dehydrogenase (ADH), the extraction and purification technics tea saponin were studied. Deionized water extraction was used to extract tea saponin from Camellia cake and vanillin-sulfuric acid method to detect the content of tea saponin. Two-step purification was used to get highly pure tea saponin by doing a further purification to tea saponin crude extract. Compare with acetone purification, the tea saponin that was got by two-step purification has a better effect on activating ADH. The results of the studies were listed as follows:1. The first step is using water extraction to extract tea saponin from Camellia cake. And by using single factor experiments and orthogonal experiments, the best extraction process was got, which was water extraction temperature 80℃, extraction time 2 h, solid-liquid ratio 1:12, and extracted three times. And in this condition, the yield of tea saponin is 13.88%.2. By using product purity as index, the two-step purification was established. Tea saponin was obtained in high purity (purity> 87%), and at the same time, crude tea polysaccharides was obtained too. High-purity tea saponin was got by this green process, and it do have an active effect on alcohol dehydrogenase. And the tea cake resource would have a good utilization. The specific steps include:1) Ethanol precipitation purification process:by using the purification of tea saponin as index, single factor test and orthogonal test were used to get the best extraction process:95% ethanol, precipitation reaction time 1.5h, the ratio of concentrated liquid to ethanol 1:4. In this condition, the purity of tea saponin is 58.98%. And the crude tea polysaccharides was obtained in this step.2) Calcium oxide precipitation purification process:by using the purification of tea saponin as index, the selection of precipitation, the dosage of precipitation, precipitating time, the amount of NH4HCO3, transition time, the times of precipitation and transition on the degree of purity of tea saponin were studied. And a good condition of CaO precipitation was found, in this condition, the yield of tea saponin is 10.28%, the purity of tea saponin is 87.58%.3. And the infrared spectra of tea saponin was studied.The tea saponin that obtained by the acetone purification and the two-step purification were studied on the activiation on effective of ADH. The result showed that the tea saponin that obtained by two-step purification had a better effect. And the higher purity tea saponin, the more significant of activating effect to ADH. And the more amount of tea saponin, the more significant of activating effect to ADH. And the tea saponin that obtained by acetone purification had no significant effect on activating ADH.