丹参等中药材化学成分分离纯化方法研究

发布时间：2019-06-17 09:28

【摘要】：中医药是中华民族的宝贵财富,在医疗保健等方面有着独特的功效。但是中药的成分十分复杂,有效成分的含量比较低而且纯化困难,使其在药理研究和临床应用上受到了很大的限制,因此建立一种快速分离中药有效成分的方法尤为重要。本文建立了丹参、草豆蔻药材有效成分的超临界流体色谱(SFC)分离纯化方法和陈皮、前胡药材有效成分的制备型高效液相色谱(PHPLC)分离纯化方法,运用高效液相色谱(HPLC)对所得单体进行纯度分析,利用紫外光谱(UV)、核磁共振光谱(NMR)和质谱(MS)对单体的化学结构进行鉴定,以期为该类药物的开发应用提供参考。一、超临界流体色谱分离纯化丹参中5种丹参酮成分实验首先用90%乙醇对丹参药材进行粗提取,得到丹参粗提物,再利用SFC对粗提物进行分离纯化研究,分别考察了色谱柱、改性剂、流速、温度、压力对丹参中5种丹参酮分离过程的影响,确定了比较适宜的色谱条件。然后在分析型SFC的基础上通过色谱放大公式得到了制备型SFC色谱分离的条件,通过SFC得到的单体,利用HPLC进行纯度测定,利用UV、NMR进行结构的鉴定。同时对该色谱过程的热力学规律进行了初步研究,说明了在此条件下丹参中丹参酮类成分的色谱分离过程为焓控过程。二、超临界流体色谱分离纯化草豆蔻中4种主要有效成分实验首先用70%乙醇对草豆蔻主要有效成分进行提取,再用SFC对粗提物进行分离纯化研究,分别考察了色谱柱、改性剂、流速等对分离过程的影响,通过比较其有效成分在色谱柱上的保留行为,得到分离草豆蔻有效成分的最优条件。然后通过色谱放大公式在分析型SFC色谱条件的基础上得到了制备型SFC色谱分离的条件,由SFC得到的单体,经UV和NMR鉴定其结构,HPLC检测纯度。同时研究了该色谱过程的热力学规律,说明在此条件下草豆蔻4种主要有效成分的色谱分离过程为焓控过程。三、制备高效液相色谱分离纯化陈皮中5种黄酮类成分实验首先对陈皮药材进行了粗提取,使用85%乙醇提取、乙酸乙酯萃取将陈皮中的黄酮成分分为水溶性和脂溶性两部分,并对这两部分分别建立了PHPLC分离纯化方法,通过优化分离条件,确定了适宜的色谱条件。结果从陈皮粗提物中分离得到5种黄酮类成分,经UV、NMR及MS鉴定分别为橙皮苷、柚皮苷、川陈皮素、3,5,6,7,8,3',4'-七甲氧基黄酮和橘皮素。四、制备高效液相色谱分离纯化前胡中3种香豆素成分采用制备型高效液相色谱技术,从前胡提取液中分离出高纯度的香豆素类活性物质。采用C18柱(250 mm×25.4 mm I.D.,10μm),对洗脱液浓度及洗脱模式、流动相流速以及进样量等参数进行优化,确定了适宜的色谱条件。通过该工艺分离可以得到3种成分,经UV、NMR和MS鉴定3种成分分别为白花前胡甲素、白花前胡丁素和白花前胡素E。
[Abstract]:Traditional Chinese medicine (TCM) is a valuable asset of the Chinese nation and has a unique effect in medical and health care. However, the composition of traditional Chinese medicine is very complex, the content of effective components is relatively low and purification is difficult, so it is very limited in pharmacological research and clinical application, so it is particularly important to establish a method for rapid separation of effective components of traditional Chinese medicine. In this paper, a method for separation and purification of active components of Salvia miltiorrhiza and cardamom by (SFC) and (PHPLC) was established. The purity of the monomer was analyzed by high performance liquid chromatography (HPLC), and the chemical structure of the monomer was identified by UV (UV), nuclear magnetic resonance spectroscopy (NMR) and mass spectrometry (MS). In order to provide reference for the development and application of this kind of drugs. First, the crude extract of Salvia miltiorrhiza was obtained by 90% ethanol, and then the crude extract of Salvia miltiorrhiza was studied by SFC. The effects of chromatographic column, modifier, flow rate, temperature and pressure on the separation process of five tanshinones in Salvia miltiorrhiza were investigated, and the suitable chromatographic conditions were determined. Then the chromatographic separation conditions of the prepared SFC were obtained by chromatographic amplification formula on the basis of analytical SFC. The purity of the monomer obtained by SFC was determined by HPLC and the structure was identified by UV,NMR. At the same time, the thermodynamic law of the chromatographic process was studied, and it was shown that the chromatographic separation process of Salvianone from Salvia miltiorrhiza was enthalpy control process. 2. Separation and purification of four main active components from Cardamom by Supercritical fluid Chromatography the main active components of nutmeg were extracted with 70% ethanol, and then the crude extract was separated and purified by SFC. The effects of chromatographic column, modifier and flow rate on the separation process were investigated respectively. by comparing the retention behavior of the active components on the chromatographic column, the optimal conditions for the separation of the effective components of nutmeg were obtained. Then the separation conditions of prepared SFC chromatography were obtained by chromatographic amplification formula on the basis of analytical SFC chromatographic conditions. The monomer obtained from SFC was identified by UV and NMR, and the purity was detected by HPLC. At the same time, the thermodynamic law of the chromatographic process was studied, and it was shown that the chromatographic separation process of the four main effective components of nutmeg was enthalpy control process. Third, five flavonoids from Chen Pei were separated and purified by high performance liquid chromatography (HPLC). Firstly, the flavonoids in Chen Pei were extracted with 85% ethanol and ethyl acetate extractive. the flavonoids in Chen Pei were divided into two parts: water solubility and fat solubility. PHPLC separation and purification methods were established for these two parts, and the suitable chromatographic conditions were determined by optimizing the separation conditions. Results five flavonoids were isolated from the crude extract of Chen Pei. Identified by UV,NMR and MS, they were hesperidin, naringin, Chuan Chen et, 3,5, 6, 7, 3, 4 / 7 methoxyflavones and tangerine, respectively. they were identified as hesperidin, naringin, 3, 5, 6, 7, 3, 4, 7, 7, 3, 4, 7, 7, 3, 4 and 4, respectively. 4. Three coumarins from Radix Bupleurum were separated and purified by high performance liquid chromatography (HPLC). The high purity coumarins were separated from the extract of Radix Bupleurum by preparation high performance liquid chromatography (HPLC). A C18 column (250 mm 脳 25.4 mm I.D., 10 渭 m) was used to optimize the eluent concentration, eluting mode, mobile phase flow rate and injection volume, and the suitable chromatographic conditions were determined. Three components were isolated by this process. The three components were identified by UV,NMR and MS as white anthesis, proanthocyanidin and E.
【学位授予单位】：聊城大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R284.1

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