【作者】 涂云飞；

【导师】 杜先锋；

【作者基本信息】 安徽农业大学 ， 茶学， 2007， 硕士

【摘要】 本论文是在前人研究的基础上进行油茶总皂苷的分离与纯化工艺的研究。论文分四部分进行,如下:第一部分:采用于柱层析制备油茶皂苷对照品。将样品干法上样并用乙酸乙酯-甲醇-正丁醇-水(3:2:5:10)的上相展层;根据Rf值取出相应的条带并用甲醇淋洗收集蒸干,薄层及理化性质检测。结果表明,该方法所制样品由薄层色谱为一点,纯度较高;该方法简便、实用且制备量大,可作为油茶总皂苷对照品;第二部分:利用高效液相色谱及质谱(电喷雾,ESI)对干柱层析法所制备的油茶总皂苷(SQS)进分离及质量分析;结果表明,通过液相色谱及ESI质谱图,干柱层析所得的油茶皂苷为一混合物,可能为含有二至三糖的皂苷,分子量在800 Dalton～1200Dalton范围内,与文献报道相符合;第三部分:通过陶瓷膜微滤去除油茶籽饼粗提液中的杂质,以提高油茶籽籽饼中主要活性成份油茶皂苷的得率与纯度。实验结果表明,以50%乙醇超声提取油茶皂苷,抽滤后,以3%浓度的油茶皂苷进行0.5μm微滤,压力控制在0.1 Mpa～0.15 Mpa,温度控制在40℃左右,油茶总皂苷的转移率约达89.25%,除杂率约14.5%,纯度相对较好;过滤后的透过液清彻,黄亮。作为前处理微滤,应用于工业化,具有一定的可行性;第四部分:通过选用的吸附活性炭纯化油茶皂苷。以总皂苷的得率及纯度为考察指标,确定活性炭吸附富集油茶总皂苷的性能、洗脱参数及再生使用情况。结果表明,水洗之后再以90%乙醇洗脱较好;香草醛—浓硫酸为检测方法检测下,总皂苷解吸得率为62.9%,纯度为87.1%以上,该类活性炭在分离纯化及脱色方面作用较显著。更多还原

【Abstract】 This dissertation studies the separation and purification of SQS based on the previous researching.Four parts divided in this thesis are as follows:PartⅠ:To establish a preparative method for total oil tea saponins as reference substance from the seeds of Camellia Oleiferal Abel.by dry column chromatography.After raw material being extracted with 60%ethanol,the ethanol extracts were evaporated to alcoholess in vacuum.And the solution was extracted by petroleum benzine.Then taking off the upper solution,and adding three times volume of BuOH for extracting,and this time the upper solution was evaporated to dryness in vacuum.The dry powder was fractionated by dry column chromatography on silica gel（100～200 mesh）using the upper solution of EtOAc-MeOH-BuOH-H₂O（3:2:5:10）as the eluent.The collected fraction was taken out by the Rf value,and then put the fraction into glass column to collecting the solution by using MeOH as eluent.After evaporating MeOH with the solution to get the total oil tea-saponins,and detecting the saponins by TLC（Thin-layer chromatography）and physical & chemical properties.The purity of the product quantized by TLC was over 98%. Therefore,the developed method is simple,rapid and the quantity of oil tea saponins also is more than normal preparation methods.All the results showed that the preparation substance of all the oil-tea saponins should be as reference substance.PartⅡ:Used the HPLC（High Performance Liquid Chromatography）and HPLC-MS （Mass Spectrometry）-ESI（Electronic Spray Ionization）to separate and analysis the sasanquasaponin which got from the dry column chromatography.Through the chromatogram of HPLC and the spectrometry of ESI,as a result,the sasanquasaponin was a mixture which might be containing two or three Mono-glycosides.And the molecular weight of the sasanquasaponin was from 800～1200 Dalton,which in accord with the literatures.PartⅢ:Used the ceramic membrane to remove the residues of crude extracts from the cake of C.Oleifera Abel.Seeds for improving the product and purity of the sasanquasaponin.The ultrasonic extraction with the alcohol of 50%was treated by the membrane of 0.5μm pores,the feed concentration was3%（w/v）,the operation pressure was about 0.1 Mpa～0.15 Mpa,and the temperature was about 40℃.Under those conditions,we acquired a better conclusion that the metastasis rate of the total sasanquasaponin was about 89.25%,and the elimination rate was about 14.5%.Also the permeation liquid of the micro-filtration was clarification,so the ceramic membrane was a promising field in the pretreatment of the crude extraction of the sasanquasaponin and a possibility in the Industrialization.PartⅣ:Selected a special Granular activated carbon to purify sasanquasaponin.Using the production & purity as index of examination through all the experiments.And the Vanillin-H₂SO₄ colorimetry was used to analyze the content of total saponins,and the adsorption performance and the elution parameters and the regeneration were investigated. The results showed that the effect of 90%ethanol with a flow rate of 1.0 BV/h was better; and the elution ratio of sasanquasaponin was 92.4%including water elution.And the recovery of the saponins was 62.9%without water elution,the purity of saponins was more than 87.1%.As a result,this Granular activated carbon can be used to refine the sasanquasaponin in the cake of C.Oleifera Abel.Seeds for its simple application process, low price and so on.更多还原