大孔吸附树脂对虎杖中有效成分的分离纯化研究
发布时间：2018-07-17 16:04
【摘要】：虎杖(Polygonum cuspidatum Sieb.et Zucc.)是我国的传统中药材之一,2000年版的《中国药典》中被收录于第一部。虎杖中的生物活性成分有芪类化合物和黄酮类化合物等,现代药理学研究证明,芪类成分中白藜芦醇(Resveratrol)的生理活性主要表现在抗氧化、抗自由基、抗衰老、抗癌、抗心血管疾病等方面;黄酮类化合物(Flavonoids)具有镇痛、抗病毒、降血脂、降血糖、抗氧化等多种医用功效。目前,从虎杖中分离纯化其中有效成分的研究越来越受到人们的关注。本论文研究了虎杖中白藜芦醇和黄酮类化合物的分离纯化,主要取得以下结果:一、采用酶解法提取虎杖中的白藜芦醇,通过单因素和正交设计实验优化得到白藜芦醇最优提取工艺条件如下:提取温度为45℃,提取时间为5.0 h,酶用量0.50%(m/m),提取液p H为6.5,白藜芦醇提取率为75.66%,含量为3.80%,较原料药中的含量(0.4%)提高了约10倍。二、考察了32种MAR及其混合床(MARs)对虎杖粗提物中白藜芦醇的纯化效果,结果表明,MARs LZ-94+LZ-78+LZ-76对白藜芦醇的纯化效果最好,优化得出最佳吸附条件为:吸附液浓度为0.25mg/m L、吸附温度45℃、吸附液体积60.0m L、吸附液p H=3.0、吸附时间9.0 h;最佳解吸条件为:解吸温度55℃、解吸液乙醇体积分数70%,解吸时间2.5h,解吸液p H=4.0。得出MARs对白藜芦醇的吸附率97.96%,解吸率80.55%,最终白藜芦醇回收率达到78.60%,纯度达到19.32%,是白藜芦醇粗提物的5.08倍。MARs对白藜芦醇的吸附动力学研究表明MARs对白藜芦醇的的吸附过程符合准二级动力学模型,大孔区和中孔区的吸附过程均符合Intra-particle-diffusion模型,预示着吸附过程由粒内扩散过程控制,微孔区中的吸附过程由Homogeneous particle-diffusion模型控制;Freundlich模型更适合描述MARs对白藜芦醇的热力学吸附行为,说明MARs对虎杖中白藜芦醇的吸附行为为多分子层物理吸附。三、由于在前部分对虎杖中的白藜芦醇分离纯化后,纯度较低,所以本部分采用虎杖粗提物预处理法辅助MARs对白藜芦醇进行分离纯化。首先研究了萃取方法和工艺,通过比较不同有机溶剂及混合溶剂对白藜芦醇的萃取效果,得出以有机溶剂E/D=3/1为萃取剂时对白藜芦醇萃取效果较好,萃取水相与有机相最佳体积比Vw/Vo=2/3、萃取时间间隔为60min、萃取5次最为适宜。该条件下,白藜芦醇萃取率为88.81%,纯度为66.50%。该萃取方法能够在保证白藜芦醇苷被充分水解成白藜芦醇的前提下,保证白藜芦醇最少量的分解损失,且由于E和D均是国际化学界公认的绿色溶剂,环境友好。之后,将萃取后所得样品分别采用硅胶柱层析技术和MARs技术进行分离纯化,比较了两种分离纯化方法,发现经硅胶柱层析法分离纯化后白藜芦醇的回收率为61.60%,纯度为73.70%,经MARs技术纯化白藜芦醇后白藜芦醇的回收率为81.10%,纯度为87.12%。因此在纯化相同质量白藜芦醇的前提下,MARs纯化技术优于硅胶柱层析法。四、采用超声辅助法提取虎杖中黄酮类化合物,通过单因素实验和中心组合设计以及响应面法优化黄酮类化合物的提取工艺,得出最优提取条件如下:提取液乙醇体积分数为60%,提取温度45℃、超声提取时间34 min、超声功率80W、固液比1/20(g/m L)、最佳提取次数为5次,该条件下虎杖中黄酮类化合物的提取率达到94.50%,其初始提取物的纯度为31.00%。五、考察32种MAR及MARs对虎杖中黄酮类化合物的分离纯化效果,结果得出单一MAR LZ-72对虎杖中黄酮类化合物的分离纯化效果最佳,最佳吸附条件为:吸附原液浓度为2.50mg/m L,吸附温度30℃,吸附液为10%的乙醇溶液、吸附液体积60.0 m L、吸附时间20 h;最佳解吸条件为:解吸温度55℃、解吸液体积为110 m L,解吸液乙醇体积分数为90%,解吸附时间为20 h,最终产物纯度达到82.50%,回收率达到89.70%。LZ-72对黄酮类化合物的吸附动力学研究表明该吸附过程符合准二级动力学模型,大孔区和中孔区的吸附过程均符合Intra-particle-diffusion模型,预示着吸附过程由粒内扩散过程控制,微孔区中的吸附过程由Homogeneous particle-diffusion模型控制;Freundlich模型更适合描述LZ-72对黄酮类化合物的热力学吸附行为,说明LZ-72对虎杖中黄酮类化合物的吸附行为是多分子层物理吸附。
[Abstract]:Polygonum cuspidatum Sieb.et Zucc. (Polygonum cuspidatum) is one of the traditional Chinese medicinal materials in China. The 2000 edition of China pharmacopoeia is included in the first part. The bioactive components in the Polygonum cuspidatum are composed of Astragalus compounds and flavonoids. Modern pharmacological studies have shown that the physiological activity of resveratrol (Resveratrol) in the Astragalus is mainly manifested in the physiological activities of the Radix Polygoni cuspidatum. Antioxidative, anti free radicals, anti aging, anticancer, anti cardiovascular diseases, flavonoids (Flavonoids) have many medical effects, such as analgesic, antiviral, antilipidemic, hypoglycemic, antioxidation and other medical effects. At present, the research on the separation and purification of effective components from Polygonum cuspidatum has attracted more and more attention. This paper studies resveratrol from the Polygonum cuspidatum The main results are as follows: 1. The optimum extraction conditions of resveratrol from Polygonum cuspidatum were obtained by enzymatic hydrolysis. The optimum extraction conditions of resveratrol were optimized by single factor and orthogonal design. The extraction temperature was 45, the extraction time was 5 h, the amount of enzyme was 0.50% (m/m), the extract P H was 6.5, resveratrol. The extraction rate was 75.66%, the content was 3.80%, which was about 10 times higher than the content of the raw material (0.4%). Two, the purification effect of resveratrol from the crude extracts of the Polygonum cuspidatum was investigated by 32 kinds of MAR and its mixed bed (MARs). The results showed that the best purification effect of MARs LZ-94+LZ-78+LZ-76 to resveratrol was best, and the optimum adsorption condition was 0.. The concentration of adsorption solution was 0.. 25mg/m L, adsorption temperature 45 C, adsorption liquid volume 60.0m L, adsorption liquid p H=3.0, adsorption time 9 h; the optimum desorption conditions are: desorption temperature 55, ethanol volume fraction 70%, desorption time 2.5h, desorption solution P H=4.0. obtained MARs to resveratrol adsorption rate 97.96%, the desorption rate of 80.55%, the final recovery of resveratrol reached 78.60%, the purity of up to 78.60%, purity up to 78.60%, purity up to 78.60%, purity up to 78.60%, purity up to the purity of up to the purity of up to 78.60%, the purity of the purity of up to 78.60%, the purity of the purity of up to 78.60%, the purity of the purity of up to 78.60%, the purity of the purity reached 78.60%. By 19.32%, the study on the adsorption kinetics of Resveratrol by 5.08 times.MARs to resveratrol shows that the adsorption of resveratrol conforms to the quasi two kinetic model. The adsorption process of the macroporous and mesoporous regions conforms to the Intra-particle-diffusion model, indicating that the adsorption process is controlled by the intragranular diffusion process and in the microporous region. The adsorption process is controlled by the Homogeneous particle-diffusion model, and the Freundlich model is more suitable for describing the thermodynamic adsorption behavior of MARs on resveratrol. It shows that the adsorption behavior of resveratrol in the Polygonum cuspidatum by MARs is a multi molecular physical adsorption. Three, the purity is low after the separation and purification of resveratrol in the former part of the Polygonum cuspidatum. The extraction of Resveratrol by MARs was used in this part. First, the extraction method and process were studied. By comparing the extraction effects of different organic solvents and mixed solvents on resveratrol, it was found that the extraction effect of resveratrol was better when the organic solvent E/D=3/1 was used as the extractant, and the extraction of water phase and organic matter was obtained. The optimum volume ratio is Vw/Vo=2/3, the interval of extraction time is 60min, and the extraction 5 times is the most suitable. Under this condition, the extraction rate of resveratrol is 88.81%. The purity of the extraction method is 66.50%., which can guarantee the minimum decomposition loss of resveratrol under the premise that the resveratrol is fully hydrolyzed into resveratrol, and the E and D are both internationalized. The green solvents recognized by the academic community were environmentally friendly. After that, the samples were separated and purified by silica gel column chromatography and MARs technology, and two methods were compared. The recovery rate of resveratrol was 61.60% after the silica gel column chromatography was purified and the purity was 73.70%. After the purification of Resveratrol by MARs technology, the purified resveratrol was purified by silica gel column chromatography. The recovery rate of resveratrol is 81.10%, the purity is 87.12%., so the purification technology of MARs is better than silica gel column chromatography on the premise of purifying the same quality resveratrol. Four, the flavonoids in Polygonum cuspidatum are extracted by ultrasonic assisted method, and the extraction of flavonoids is optimized by single factor experiment and central combination design and response surface method. The optimum extraction conditions are as follows: the ethanol volume fraction of the extract is 60%, the extraction temperature is 45, the ultrasonic extraction time is 34 min, the ultrasonic power is 80W, the solid-liquid ratio 1/20 (g/m L), the optimum extraction times is 5 times. The extraction rate of flavonoids in the Polygonum cuspidatum is 94.50%, the purity of the initial extract is 31.00%. five, and 32 kinds of MAR and MA are investigated. Rs for the separation and purification of Flavonoids from the Polygonum cuspidatum, the result is that single MAR LZ-72 is the best for the separation and purification of flavonoids in the Polygonum cuspidatum. The best adsorption conditions are: the concentration of the original solution is 2.50mg/m L, the adsorption temperature is 30, and the adsorption liquid is 10% of the ethanol solution, the volume of the adsorbent is 60 m L, the adsorption time is 20 h, and the best desorption strip is the best. The results are as follows: the desorption temperature is 55 C, the solution volume is 110 m L, the volume fraction of ethanol is 90%, the desorption time is 20 h, the purity of the final product reaches 82.50%, the recovery rate reaches 89.70%.LZ-72 to the adsorption kinetics of flavonoids, which shows that the adsorption process is in accordance with the quasi two kinetics model, the adsorption process of the large pore and the mesoporous areas. According to the Intra-particle-diffusion model, the adsorption process is controlled by the intragranular diffusion process. The adsorption process in the microporous area is controlled by the Homogeneous particle-diffusion model, and the Freundlich model is more suitable for describing the thermodynamic adsorption behavior of LZ-72 to flavonoids, and the adsorption of flavonoids in the Polygonum cuspidatum by LZ-72 It is a physical adsorption of a multi molecular layer.
【学位授予单位】：兰州理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O647.3;R284.2
[Abstract]:Polygonum cuspidatum Sieb.et Zucc. (Polygonum cuspidatum) is one of the traditional Chinese medicinal materials in China. The 2000 edition of China pharmacopoeia is included in the first part. The bioactive components in the Polygonum cuspidatum are composed of Astragalus compounds and flavonoids. Modern pharmacological studies have shown that the physiological activity of resveratrol (Resveratrol) in the Astragalus is mainly manifested in the physiological activities of the Radix Polygoni cuspidatum. Antioxidative, anti free radicals, anti aging, anticancer, anti cardiovascular diseases, flavonoids (Flavonoids) have many medical effects, such as analgesic, antiviral, antilipidemic, hypoglycemic, antioxidation and other medical effects. At present, the research on the separation and purification of effective components from Polygonum cuspidatum has attracted more and more attention. This paper studies resveratrol from the Polygonum cuspidatum The main results are as follows: 1. The optimum extraction conditions of resveratrol from Polygonum cuspidatum were obtained by enzymatic hydrolysis. The optimum extraction conditions of resveratrol were optimized by single factor and orthogonal design. The extraction temperature was 45, the extraction time was 5 h, the amount of enzyme was 0.50% (m/m), the extract P H was 6.5, resveratrol. The extraction rate was 75.66%, the content was 3.80%, which was about 10 times higher than the content of the raw material (0.4%). Two, the purification effect of resveratrol from the crude extracts of the Polygonum cuspidatum was investigated by 32 kinds of MAR and its mixed bed (MARs). The results showed that the best purification effect of MARs LZ-94+LZ-78+LZ-76 to resveratrol was best, and the optimum adsorption condition was 0.. The concentration of adsorption solution was 0.. 25mg/m L, adsorption temperature 45 C, adsorption liquid volume 60.0m L, adsorption liquid p H=3.0, adsorption time 9 h; the optimum desorption conditions are: desorption temperature 55, ethanol volume fraction 70%, desorption time 2.5h, desorption solution P H=4.0. obtained MARs to resveratrol adsorption rate 97.96%, the desorption rate of 80.55%, the final recovery of resveratrol reached 78.60%, the purity of up to 78.60%, purity up to 78.60%, purity up to 78.60%, purity up to 78.60%, purity up to the purity of up to the purity of up to 78.60%, the purity of the purity of up to 78.60%, the purity of the purity of up to 78.60%, the purity of the purity of up to 78.60%, the purity of the purity reached 78.60%. By 19.32%, the study on the adsorption kinetics of Resveratrol by 5.08 times.MARs to resveratrol shows that the adsorption of resveratrol conforms to the quasi two kinetic model. The adsorption process of the macroporous and mesoporous regions conforms to the Intra-particle-diffusion model, indicating that the adsorption process is controlled by the intragranular diffusion process and in the microporous region. The adsorption process is controlled by the Homogeneous particle-diffusion model, and the Freundlich model is more suitable for describing the thermodynamic adsorption behavior of MARs on resveratrol. It shows that the adsorption behavior of resveratrol in the Polygonum cuspidatum by MARs is a multi molecular physical adsorption. Three, the purity is low after the separation and purification of resveratrol in the former part of the Polygonum cuspidatum. The extraction of Resveratrol by MARs was used in this part. First, the extraction method and process were studied. By comparing the extraction effects of different organic solvents and mixed solvents on resveratrol, it was found that the extraction effect of resveratrol was better when the organic solvent E/D=3/1 was used as the extractant, and the extraction of water phase and organic matter was obtained. The optimum volume ratio is Vw/Vo=2/3, the interval of extraction time is 60min, and the extraction 5 times is the most suitable. Under this condition, the extraction rate of resveratrol is 88.81%. The purity of the extraction method is 66.50%., which can guarantee the minimum decomposition loss of resveratrol under the premise that the resveratrol is fully hydrolyzed into resveratrol, and the E and D are both internationalized. The green solvents recognized by the academic community were environmentally friendly. After that, the samples were separated and purified by silica gel column chromatography and MARs technology, and two methods were compared. The recovery rate of resveratrol was 61.60% after the silica gel column chromatography was purified and the purity was 73.70%. After the purification of Resveratrol by MARs technology, the purified resveratrol was purified by silica gel column chromatography. The recovery rate of resveratrol is 81.10%, the purity is 87.12%., so the purification technology of MARs is better than silica gel column chromatography on the premise of purifying the same quality resveratrol. Four, the flavonoids in Polygonum cuspidatum are extracted by ultrasonic assisted method, and the extraction of flavonoids is optimized by single factor experiment and central combination design and response surface method. The optimum extraction conditions are as follows: the ethanol volume fraction of the extract is 60%, the extraction temperature is 45, the ultrasonic extraction time is 34 min, the ultrasonic power is 80W, the solid-liquid ratio 1/20 (g/m L), the optimum extraction times is 5 times. The extraction rate of flavonoids in the Polygonum cuspidatum is 94.50%, the purity of the initial extract is 31.00%. five, and 32 kinds of MAR and MA are investigated. Rs for the separation and purification of Flavonoids from the Polygonum cuspidatum, the result is that single MAR LZ-72 is the best for the separation and purification of flavonoids in the Polygonum cuspidatum. The best adsorption conditions are: the concentration of the original solution is 2.50mg/m L, the adsorption temperature is 30, and the adsorption liquid is 10% of the ethanol solution, the volume of the adsorbent is 60 m L, the adsorption time is 20 h, and the best desorption strip is the best. The results are as follows: the desorption temperature is 55 C, the solution volume is 110 m L, the volume fraction of ethanol is 90%, the desorption time is 20 h, the purity of the final product reaches 82.50%, the recovery rate reaches 89.70%.LZ-72 to the adsorption kinetics of flavonoids, which shows that the adsorption process is in accordance with the quasi two kinetics model, the adsorption process of the large pore and the mesoporous areas. According to the Intra-particle-diffusion model, the adsorption process is controlled by the intragranular diffusion process. The adsorption process in the microporous area is controlled by the Homogeneous particle-diffusion model, and the Freundlich model is more suitable for describing the thermodynamic adsorption behavior of LZ-72 to flavonoids, and the adsorption of flavonoids in the Polygonum cuspidatum by LZ-72 It is a physical adsorption of a multi molecular layer.
【学位授予单位】：兰州理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O647.3;R284.2
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