中国科学院机构知识库网格
Chinese Academy of Sciences Institutional Repositories Grid
杜仲叶粗提物中高纯度绿原酸的制备工艺研究

文献类型:学位论文

作者苗芹1,2
学位类别硕士
答辩日期2017-05-17
授予单位中国科学院大学
授予地点北京
导师夏传海
关键词绿原酸 高纯度 杜仲叶粗提物 大孔树脂 精制
学位专业生物工程
中文摘要绿原酸(CGA)作为一种重要的天然多酚物质,具有显著的降血压、滋肝润胆、抗菌、预防肿瘤等多种药理活性,是许多中成药的有效成分,近年来在不断受到人们的重视,并被广泛应用于化妆品、食品、医药等行业。因此,研究高纯度绿原酸的制备工艺技术,对我国国民经济和社会发展具有重要意义。
本论文以杜仲叶粗提物为实验原料,通过严谨的实验设计,研究出了一条容易操作,稳定可靠的高纯度绿原酸制备工艺路线,并在此基础上进行了中试放大研究,所得主要研究结果如下:
(1)定量方法的建立
以高效液相色谱仪为检测仪器,建立了定量检测杜仲叶粗提物中绿原酸的方法,并对其进行方法学验证。结果显示,该HPLC法基线稳定,绿原酸与其他成分分离度高,回收率高,因此可以作为杜仲叶提取物中绿原酸的检测和定量分析方法。
(2)大孔树脂初步分离纯化
筛选出ADS-21最佳大孔吸附树脂,并发现该树脂在25 ℃条件时达到最大吸附量,同时满足Langmuir方程和伪二级动力学方程。ADS-21树脂分离纯化粗提物中绿原酸的最佳工艺条件为:1)吸附:吸附液pH=2,温度25 ℃,上样流速2 BV/h,上样浓度3.01 mg/mL,上样体积8 BV;2)解吸:解吸流速2 BV/h,乙醇浓度及用量为40%乙醇3 BV,50%乙醇10 BV。通过大孔树脂吸附解吸一次纯化后,得到绿原酸粗品的纯度由原来的5.01%提高到38.1%,提高了7.62倍,绿原酸回收率为92.8%。
(3)树脂和硅胶除杂精制
采用树脂和硅胶精制绿原酸粗品,通过研究得到X-5树脂除杂最佳工艺条件:pH≈6,当层析柱直径:高(cm:cm)=3.5:35时,料液比为M:V(g:mL)=1:10;硅胶除杂最佳工艺条件:当玻璃层析柱直径:高(cm:cm)=3.5:35时,料液比为M:V(g:mL)=1:100。解吸浓缩液通过硅胶和X-5型树脂吸附除杂后,绿原酸的纯度由38.1%提高到73.2%,产率65.3%。
(4)活性炭脱色精制
以活性炭为脱色材料对绿原酸粗精品进行精制,得到最佳脱色工艺条件为:1)吸附:200目以上活性炭,温度30 ℃,料液比M:V=2:50(g:mL),时间1.5 h;2)解吸:温度50 ℃,乙醇浓度50%,时间5 h,解吸量7 BV,解吸三次。通过活性炭脱色精制后,得到纯度为93.4%接近无色的绿原酸精品,产率为88.7%。
(5)实验室放大研究
实验室放大研究结果表明,放大实验和预实验所得绿原酸精品的纯度和总得率基本一致,均在92%和60%左右。因此,该工艺技术路线总体稳定,实用可靠,适用于从杜仲叶粗提物中工业化制备高纯度绿原酸。
 
英文摘要Chlorogenic acid (CGA), as a kind of natural polyphenols and the effective ingredient of many chinese patent medicines, has received much attention because of its important biological activities, including lowering blood pressure, anti-microbial, antitumor, protecting liver and gallbladder. CGA has been widely applied in pharmaceutical industry, food industry and cosmetics industry. It has great significance to research the preparation of highly pure CGA for our national economy and social development.
In this paper, a reliable, simple and stable technology route of purification and preparation of highly pure CGA from the crude extracts of Eucommia ulmoides (E. ulmodies) leaves was studied scrupulously and rationally. Meanwhile, the scale-up experiment in laboratory was conducted. The main results were shown as follows:
(1) Quantitative detection method of CGA
High performance liquid chromatography (HPLC) was used in this paper. The quantitative detection method of CGA from the crude extracts of E. ulmodies leaves was established and validated with methodologies. The results showed that the HPLC method could be used to quantitative analysis of CGA with its steady baseline, high separation and high average recovery.
(2) Separating with macroporous adsorption resin
ADS-21 was selected as the best resin. And it had the maximum adsorption capacity, satisfying Langmuir equation and Pseudo-two-order kinetics equation at 25 ℃.The optimum technological conditions of ADS-21 were: 1) adsorption: adsorption liquid at pH=2 and 25 ℃ with velocity of flow of 2 BV/h, sample concentration of 3.01 mg/mL and sample volume of 8 BV; 2) desorption: elution velocity at 2 BV/h in the mixture of 3 BV ethanol with 40% concentration and 10 BV ethanol with 50% concentration. The purity of chlorogenic acid would increase 7.62 times from 5.01% to 38.1% and the rate of recovery is 92.8%.
(3) Purifying with resin and silica gel
Further purification and enrichment with resin and silica gel for CGA was conducted. The optimum technological conditions to purify with silica gel was that solid-liquid ratio was M:V(g:mL)=1:100 when the chromatographic was D:H(cm:cm)=3.5:35. And the optimum technological conditions with X-5 resin was that solid-liquid ratio was M:V(g:mL)=1:10 and pH was 6 when the chromatographic was D:H(cm:cm)=3.5:35. The purity of CGA would raise to 73.2% from 38.1% and yield will be 65.3% after purification with resin and silica gel.
(4) Decoloring and purifying with activated carbon
The optimum technological conditions to decolor and purify with activated carbon was: 1) adsorption: 200 mesh powder activated carbon, temperature at 30 ℃, solid-liquid ratio M:V=2:50(g:mL), and duration of 1.5 h; 2) desorption: temperature at 50 ℃, ethanol concentration of 50%, duration of 5 h, desorption quantity of 7 BV and repeated three times. Near colorless CGA product would be got, the purity would raise to 93.4% and yield would be 88.7%.
(5) Scale-up experiment in laboratory
The results of the scale-up experiment in laboratory showed that the purity and total yield is 92% and 60%, consistenting with the results of the preliminary experiment. The extraction technology was stable, reliable and it is applicable to industrial preparation of highly pure chlorogenic acid from crude extracts of E. ulmodies leaves.
 
学科主题化学
源URL[http://ir.yic.ac.cn/handle/133337/21981]  
专题中科院烟台海岸带研究所知识产出_学位论文
作者单位1.中国科学院烟台海岸带研究所
2.中国科学院大学
推荐引用方式
GB/T 7714
苗芹. 杜仲叶粗提物中高纯度绿原酸的制备工艺研究[D]. 北京. 中国科学院大学. 2017.

入库方式: OAI收割

来源:烟台海岸带研究所

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