Extraction Of Triterpene Saponins From Camellia Oleifera Seed Pomace And Their Activities Against ADH

This article embarked on saponin of seed pomace and studied the methods of separation and purification and its inhibition of alcohol dehydrogenase. The main methods and results were as follows:1. The quadratic general spinning design was used to optimize the extraction conditions of tea saponin by ultrasonic-assisted water extraction and ethanol precipitation. The optimal conditions of the extraction of tea saponin were determined to be the ultrasonic extraction time of 38 min, ultrasonic temperature of 50?, pH value of 8.5 and ratio of liquid to the camellia oleifera seed pomace of 8, which contributed to the highest yield of 10.57%. The purity of the tea saponin extract is 84.52%.2. Nuclear magnetic resonance?NMR? method was used to determine the oil content of camellia meal and compared with soxhlet extraction. The research results show that quantitative values of NMR were higher than that of soxhlet extraction. Macroporous adsorption resin AB-8 was selected to purify saponin. After purifying the crude saponin with macroporous adsorption resin AB-8, the finished saponin was separated on silica gel column chromatography and yielded seven fractions?Fr3.1?Fr3.7?.Then they were found to have inhibitory activity of alcohol dehydrogenase and the inhibition ratio was 15.61%?20.36%?4.64%?11.55%?16.17%?6.66%?10.42% separately.3. The in situ intestinal absorption property of Fr3.2 in rats was investigated by means of single-pass intestinal perfusion technique. The results showed that the mechanism of gastrointestinal absorption of Fr3.2 was passive diffusion. The results of HPLC showed that the absorption status of compounds in Fr3.2 was different, the compounds when the retention time is between 40 and 60 min is absorbed more easily, the retention time is less than 40 min compounds have poorer absorption conditions.4. SRI was screened from crude sasanqua saponin by Fe₃SO₄@SiO₂-ADH immobilized enzyme particle. 10 and 27 triterpenoid saponins were found from SRI and Fr3.2 separately by HPLC and LC-TOF/MS. The two sample shared 3 same triterpenoid saponins and their chemical formulas were C₅₄H₈₈O₂₄?C₅₈H₈₉O₂₆?C₅₉H₉₂O₂₆ respectively.5. The alcohol dehydrogenase inhibition characteristics of SRI was studied by Lineweaver-Burk and Dixon plots. The results showed that the IC50 was1.73mg/ml and Ki was 0.445mg/ml. The alcohol dehydrogenase inhibition type of SRI was mixed.