【摘要】：中醫(yī)藥是中華民族的寶貴財(cái)富,在醫(yī)療保健等方面有著獨(dú)特的功效。但是中藥的成分十分復(fù)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(yīng)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(yùn)用高效液相色譜(HPLC)對(duì)所得單體進(jìn)行純度分析,利用紫外光譜(UV)、核磁共振光譜(NMR)和質(zhì)譜(MS)對(duì)單體的化學(xué)結(jié)構(gòu)進(jìn)行鑒定,以期為該類藥物的開發(fā)應(yīng)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(shí)驗(yàn)首先用90%乙醇對(duì)丹參藥材進(jìn)行粗提取,得到丹參粗提物,再利用SFC對(duì)粗提物進(jìn)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(duì)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(chǔ)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(jìn)行純度測(cè)定,利用UV、NMR進(jìn)行結(jié)構(gòu)的鑒定。同時(shí)對(duì)該色譜過程的熱力學(xué)規(guī)律進(jìn)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(shí)驗(yàn)首先用70%乙醇對(duì)草豆蔻主要有效成分進(jìn)行提取,再用SFC對(duì)粗提物進(jìn)行分離純化研究,分別考察了色譜柱、改性劑、流速等對(duì)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(chǔ)上得到了制備型SFC色譜分離的條件,由SFC得到的單體,經(jīng)UV和NMR鑒定其結(jié)構(gòu),HPLC檢測(cè)純度。同時(shí)研究了該色譜過程的熱力學(xué)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(shí)驗(yàn)首先對(duì)陳皮藥材進(jìn)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(duì)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(jié)果從陳皮粗提物中分離得到5種黃酮類成分,經(jīng)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(shù),從前胡提取液中分離出高純度的香豆素類活性物質(zhì)。采用C18柱(250 mm×25.4 mm I.D.,10μm),對(duì)洗脫液濃度及洗脫模式、流動(dòng)相流速以及進(jìn)樣量等參數(shù)進(jìn)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(jīng)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(xué)位授予單位】：聊城大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R284.1

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