丹參等中藥材化學成分分離純化方法研究
發(fā)布時間：2019-06-17 09:28
【摘要】：中醫(yī)藥是中華民族的寶貴財富,在醫(yī)療保健等方面有著獨特的功效。但是中藥的成分十分復雜,有效成分的含量比較低而且純化困難,使其在藥理研究和臨床應用上受到了很大的限制,因此建立一種快速分離中藥有效成分的方法尤為重要。本文建立了丹參、草豆蔻藥材有效成分的超臨界流體色譜(SFC)分離純化方法和陳皮、前胡藥材有效成分的制備型高效液相色譜(PHPLC)分離純化方法,運用高效液相色譜(HPLC)對所得單體進行純度分析,利用紫外光譜(UV)、核磁共振光譜(NMR)和質譜(MS)對單體的化學結構進行鑒定,以期為該類藥物的開發(fā)應用提供參考。一、超臨界流體色譜分離純化丹參中5種丹參酮成分實驗首先用90%乙醇對丹參藥材進行粗提取,得到丹參粗提物,再利用SFC對粗提物進行分離純化研究,分別考察了色譜柱、改性劑、流速、溫度、壓力對丹參中5種丹參酮分離過程的影響,確定了比較適宜的色譜條件。然后在分析型SFC的基礎上通過色譜放大公式得到了制備型SFC色譜分離的條件,通過SFC得到的單體,利用HPLC進行純度測定,利用UV、NMR進行結構的鑒定。同時對該色譜過程的熱力學規(guī)律進行了初步研究,說明了在此條件下丹參中丹參酮類成分的色譜分離過程為焓控過程。二、超臨界流體色譜分離純化草豆蔻中4種主要有效成分實驗首先用70%乙醇對草豆蔻主要有效成分進行提取,再用SFC對粗提物進行分離純化研究,分別考察了色譜柱、改性劑、流速等對分離過程的影響,通過比較其有效成分在色譜柱上的保留行為,得到分離草豆蔻有效成分的最優(yōu)條件。然后通過色譜放大公式在分析型SFC色譜條件的基礎上得到了制備型SFC色譜分離的條件,由SFC得到的單體,經UV和NMR鑒定其結構,HPLC檢測純度。同時研究了該色譜過程的熱力學規(guī)律,說明在此條件下草豆蔻4種主要有效成分的色譜分離過程為焓控過程。三、制備高效液相色譜分離純化陳皮中5種黃酮類成分實驗首先對陳皮藥材進行了粗提取,使用85%乙醇提取、乙酸乙酯萃取將陳皮中的黃酮成分分為水溶性和脂溶性兩部分,并對這兩部分分別建立了PHPLC分離純化方法,通過優(yōu)化分離條件,確定了適宜的色譜條件。結果從陳皮粗提物中分離得到5種黃酮類成分,經UV、NMR及MS鑒定分別為橙皮苷、柚皮苷、川陳皮素、3,5,6,7,8,3',4'-七甲氧基黃酮和橘皮素。四、制備高效液相色譜分離純化前胡中3種香豆素成分采用制備型高效液相色譜技術,從前胡提取液中分離出高純度的香豆素類活性物質。采用C18柱(250 mm×25.4 mm I.D.,10μm),對洗脫液濃度及洗脫模式、流動相流速以及進樣量等參數(shù)進行優(yōu)化,確定了適宜的色譜條件。通過該工藝分離可以得到3種成分,經UV、NMR和MS鑒定3種成分分別為白花前胡甲素、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.
【學位授予單位】：聊城大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R284.1
[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.
【學位授予單位】：聊城大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R284.1
【相似文獻】
相關期刊論文 前10條
1 王玉瑾;黃竹芳;安健康;;超臨界流體色譜及其在法醫(yī)毒物分析中的應用(綜述)[J];法醫(yī)學雜志;1993年04期
2 金燈萍;彭國平;陸曉峰;;填充柱超臨界流體色譜在中藥中的分析與制備應用[J];中醫(yī)藥學刊;2006年03期
3 倪坤儀;;超臨界流體色譜進展[J];藥學進展;1989年04期
4 張怡評;洪專;方華;謝全靈;易瑞灶;吳皓;;超臨界流體色譜分離技術應用研究進展[J];中醫(yī)藥導報;2012年07期
5 張清民,顧學裘;新的色譜分離技術—超臨界流體色譜[J];沈陽藥學院學報;1988年03期
6 潘峰云,張亮,杭太俊,安登魁;離子對-超臨界流體色譜在藥物分析中的應用[J];藥學進展;2000年06期
7 鄭衛(wèi);;超臨界流體萃取技術與中藥現(xiàn)代化[J];海峽藥學;2006年03期
8 李振宇;傅青;李奎永;梁圖;金郁;;超臨界流體色譜對吳茱萸中吲哚類生物堿的快速分析[J];色譜;2014年05期
9 楊敏;郭亞東;馬銀海;劉潔;;超臨界流體色譜法測定阿咖酚散中三種組分的含量[J];昆明學院學報;2009年03期
10 黃s鷖，
本文編號：2500889
本文鏈接：http://sikaile.net/shoufeilunwen/mpalunwen/2500889.html
最近更新
教材專著
