【作者】 李帅；

【导师】 吴鸣建；

【作者基本信息】 郑州大学 ， 应用化学， 2012， 硕士

【摘要】 木蝴蝶,别名千张纸、玉蝴蝶、云破纸等,是一种常见中药材,是紫葳科木蝴蝶属的成熟种子。具有润肺、止咳、舒肝、利咽、和胃、生肌等功效。木蝴蝶的有效成分主要为黄酮类化合物,以千层纸苷、木蝴蝶苷A、黄芩苷、木蝴蝶苷B等为主。目前,有关木蝴蝶的研究主要集中在有效成分的含量测定、分离鉴定及其药理作用方面,其中有效成分含量测定主要是利用HPLC或RP-HPLC对某一成分进行测定,仅有本研究室的胡殿丽采用黄芩苷为标准品,测定了木蝴蝶醇提物的总黄酮含量,而未见以木蝴蝶苷A为标准品测定木蝴蝶总黄酮的含量和采用大孔吸附树脂法分离纯化木蝴蝶醇提物的报道。随着人们对植物药物的研究开发,木蝴蝶醇提取物越来越多的应用于医药、食品等领域,对其质量也提出了更高的要求。所以本文以此为基础,选择木蝴蝶种子为原料,得到了木蝴蝶苷A,并进行了结构鉴定和纯度测定,并对大孔吸附树脂分离纯化木蝴蝶总黄酮的工艺进行系统研究。主要研究内容如下：1.木蝴蝶种子经干燥、粉碎,用70%的乙醇提取,提取液经浓缩、醇沉、离心、上硅胶色谱柱,采用氯仿：甲醇系统梯度洗脱,分离出了木蝴蝶苷A。经13CNMR和’HNMR进行结构鉴定,通过熔点测定、紫外吸收、RP-HPLC法进行含量测定,最终确定其纯度为98%。2.以自制的木蝴蝶苷A为标准品,采用紫外分光光度法,测得木蝴蝶苷A的标准工作曲线为C (μg/mL)=16.7491 A-0.2975, R2=0.9998,线性工作范围为1.70-17.0μg/mL。通过方法学考察得知该方法重复性、稳定性良好,精密度实验相对标准偏差较小,加标回收率较高,可以用来测定木蝴蝶总黄酮的含量。3.采用静态吸附与动态吸附相结合的方法比较5种不同类型的大孔吸附树脂的吸附解吸性能,综合考虑发现：HPD100树脂的吸附与解吸效果优于其它树脂。因此,选择HPD100树脂分离纯化木蝴蝶总黄酮。4.考察了HPD100树脂吸附木蝴蝶总黄酮的工艺条件,结果发现：当上样浓度为5.303mg/mL, pH值为2-3,流速为3.0mL/min时,HPD100树脂对木蝴蝶总黄酮的吸附率较大；水洗液用量为3BV时,水溶性杂质可以基本除尽；以90%乙醇为解吸剂,用量为8BV时,木蝴蝶总黄酮解吸效果较好。在此条件下,经一次分离纯化后木蝴蝶总黄酮纯度达到88.3%,且工艺稳定,重现性好。5.对HPD100树脂吸附木蝴蝶总黄酮的动力学与热力学进行考察,结果发现：在150min时,HPD100树脂吸附木蝴蝶总黄酮的过程基本达到吸附平衡；HPD100树脂吸附木蝴蝶总黄酮的动力学符合拟2级动力学模型；Freundlich等温吸附方程更适合描述HPD100树脂吸附木蝴蝶总黄酮的过程,且吸附过程的焓变△H均为负值,绝对值小于4OKJ/moL,吸附过程是一个放热的物理吸附过程。吉布斯自由能变AG小于0,吸附过程能够自发进行。熵变AS小于0,吸附过程是一个熵减小的过程。更多还原

【Abstract】 Semen Orxyli, which is the mature seed of Oroxylum indicum (L.) Vent, has been used a traditional Chinese medicine for its abilities of moistening lung, relieving cough, smoothing liver, eliminating phlegm, promoting new growth of tissues etc. A number of academic studies show that the main chemical components of Oroxylum indicum are Oroxin A, Oroxin B,Tetuin, Baicalin and so on. So far, the research of Oroxylum indicum has mainly focused on the determination, separation and pharmacological actions of the active components.The contents of the active components were determined by using HPLC or RP-HPLC method to test a single component. Only Dian-li Hu has determined the contents of total flavonoids with Baicalin as a chemical reference substance. But the determined of total flavonoids with Oroxin A as a standard and purification of total flavonoids from Oroxylum indicum by macroporous adsorption resins have not been reported. With the development of plant-based drugs, The ethanol extracts from Oroxylum indicum become used more and more in pharmaceutical industry, food industry and other fields, and their qualities are required of higher standard. Based on this, this paper selects Oroxylum indicum as the raw material, obtains Oroxin A and then carries out the structural identification and purity determination, makes the systematical research on the technical conditions of separation and purification total flavonoids from Oroxylum indicum. The main content includes:1. The mature seeds of Oroxylum indicum were first dried,crushed and extracted by 70% ethanol, the extracts after being condensed, alcohol precipitation and centrifugation were separated by silica gel column chromatography. The eventual result is that the Oroxin A was found. Then its structure was identified by 13CNMR and 1HNMR, and the purity of Oroxin A was determined by melting point, UV absorption and RP-HPLC. The result showed that the purity of Oroxin A was 98%.2. The standard working curve is C (μg/mL)=16.7491 A-0.2975, R2= 0.9998,which was obtained by ultraviolet spectrophotometry with home-made Oroxin A as a standard. At the same time, the studying of methodology showed the method was inaccurate, reliable and could be repeated. It could be used to determine total flavonoids of Oroxylum indicum.3. Static adsorption and dynamic adsorption method were combined to study the five different types of macroporous adsorption and desorption properties The results showed that HPD100 resin have good adsorption capacity than other resins. As a result, HPD100 resin was selected to separate and purify the total flavonoids of Oroxylum indicum.4. The purification of the process conditions total flavonoids of Oroxylum indicum attached by HPD100 resin found that when the concentration of the sample is 5.303mg/mL, pH value is 2-3, velocity of flow is 3.0 mL/min,the adsorption rate of the total flavonoids of Oroxylum indicum is relatively big,when the amount of water used is 3 BV,the water-soluble impurities can be basically divisible, using the 90% ethanol as the desorption agent,the amount used is 8BV, the desorption effect of the flavonoids is relatively good. Under such condition, the purity of the total flavonoids of Oroxylum indicum can reach to 88.3% after division and purification with stable process and good reproducibility.5. The kinetic and thermodynamic experimental results of studying HPD100 resin’s adsorption of Oroxylum total flavonoids show that the process basically reaches equilibrium at 150 min, and its kinetics follows the pseudo-second-order kinetic model. Besides, Freundlich isotherm adsorption equation is more suitable for the whole process, and the enthalpy change AH in adsorption process is negative and its modulus is less than 40KJ/mol. The adsorption process is an exothermic physical process. The Gibbs free energy AG is less than 0, so the adsorption process can occur spontaneously. the entropy change AS are less than 0, which indicates that the adsorption reduces the degrees of the entire system’s freedom.更多还原