【作者】 马雪梅；

【导师】 毛子军； 郑春英；

【作者基本信息】 东北林业大学 ， 植物学， 2004， 硕士

【摘要】 杜香是黑龙江省蓄积量丰富的野生植物资源，国内外的研究资料都表明其具有重要的香料价值和药用价值，本文以生长于大兴安岭地区的杜香为研究对象，建立了高效液相色谱法分析杜香有效成分—香豆素和熊果酸的色谱条件，确定了超临界二氧化碳萃取法提取香豆素和精油最佳条件，确定了超声波提取熊果酸的最佳条件，并用硅胶柱分离法对其进行了分离纯化。研究结果如下： 1．高效液相色谱法分析香豆素含量的色谱条件：ODS柱（25cm×4.6mm，5μm）；流动相：乙腈：水（体积比40：60）；检测波长：273nm；进样量：10μL；流速：1.0mL／min；柱温：25℃。高效液相色谱法分析熊果酸含量的色谱条件：流动相：甲醇：水（体积比90：10）；检测波长：210nm；其余条件同上。 2．超临界CO₂萃取法提取香豆素的最佳条件：萃取温度45℃，萃取压力30Mpa，分离温度55℃，其中萃取压力是影响提取率的主要因素。超临界CO₂萃取法的提取率是14.13μg／g，是用乙醇浸提法提取得率（4.26μg／g）的3.32倍。 3．超临界CO₂萃取法提取杜香精油的条件是：萃取温度55℃，萃取压力25Mpa，第一分离釜压力8Mpa，分离釜温度35℃。其中分离釜压力是主要影响因素。采用气相色谱-质谱联用技术（GC-MS），通过标准谱图库检索对萃取物的组分进行分析鉴定，超临界萃取物含有蜡质和脂肪酸等非挥发性杂质。 4．大兴安岭地区杜香不同采集期精油和熊果酸的含量均不同，二者的最高含量均出现在果实成熟的8月份，在生产中其采集期可适当放宽。 5．熊果酸的最佳提取方法是超声提取法，超声提取条件为乙醇浓度95％，超声温度45℃，时间30min，频率40KHz，其中乙醇浓度是主要影响因素。 6．用硅胶柱层析的方法对提取物进行初步分离，洗脱液为氯仿：甲醇（体积比500：1），显色剂为5％的硫酸乙醇溶液，蒸发回收溶剂后得到的浸膏，用95％的乙醇重结晶一次得到纯度为85％的熊果酸粗品。 进一步考察杜香中的有效成分，建立综合利用工艺，提高综合利用率，对于满足当前国际社会对天然药物的需求，充分利用我国丰富的植物资源具有极其重要的意义。更多还原

【Abstract】 Ledum palustre L. is a kind of wild source plant that spreads widely in forestry areas of Heilongjiang province. Its flavor value and medical usefulness value have been paid much attention recently by many researchers. A high performance liquid chromatography (HPLC) method was developed for the determination of ursolic acid and coumarin in Ledum palustre L. Coumarin and essential oil had been extracted by SFE-CO2 methods and the best parameters had been determined. Ursolic acid had been extracted by the ultrasonic method and the best parameters determined. Ursolic acid has been isolated primarily through silica gel column chromatography. The results are as follows:1 .The chromatographic analysis of coumarin was carried out using a Techsphere ODS C18 column(25cmx4.6mm, 5μm); The mobile phase was acetonitrile-water(40:60, v/v); the flow rate was maintained at 1.0mL/min; the wavelength at 273nm; The column temperature was 25℃. The chromatographic analysis of ursolic acid was carried out. The mobile phase was methanol-water(90:10, v/v), the wavelength at 210nm, other conditions were the same as above mentioned.2.The best parameters of extracting coumarin by SFE-CO2 methods were: extraction temperature as 45℃, extraction pressure as 30Mpa, isolation temperature as 55℃, extraction pressure was the main factor of them. The yield was 14.13μg/g by SFE-CO2 methods 3.32 times of ethanol dipping method (4.26μg/g).3.The best parameters of extracting essential oil by SFE-CO2 methods were: extraction temperature as 55℃, extraction pressure as 25Mpa, isolation temperature as 35℃, isolation pressure was the main factor of them. The extraction product had been identified to contain the non-volatilisation component- waxiness and fatty acid by the method of GC-MS, further to analyze through chromatographic figure bank.4.The content of essential oil and ursolic acid was not same in different collection period of the species. Their content was the highest in August when fruit was mature. The collection period can widen during the yield.5.The best method to extract ursolic acid is ultrasonic method, the parameter: ethanol concentration as 95%, the temperature as 45℃, time as 30min, the frequency as40KHz, ethanol concentration is the main factor of them.6. Ursolic acid has been isolated and purified primarily in Ledum palustre L. through silica gel column chromatography with 500:1 MeOH-CHCl3 and recrystallized to obtain 85% ursolic acid.It’s very important to study the effective components further in Ledum palustre L., establish integrate craftwork, enhance the integrate rate of application for satisfying the need of international societies and utilizing plant resource abundantly in our country.更多还原