【作者】 陈伟；

【导师】 郁志芳；

【作者基本信息】 南京农业大学 ， 农产品加工与贮藏工程， 2007， 硕士

【摘要】 本文研究了中药杜仲(Eucommia ulmoide oliver)叶片中绿原酸和总黄酮的测定方法及提取分离纯化工艺，研究结果如下：1、改进了杜仲叶片绿原酸和总黄酮测定的方法，采用纸层析一紫外分光光度法测定杜仲叶片中绿原酸含量和纸层析-硝酸铝法测定杜仲叶片中总黄酮含量提高了测定准确性。2、应用正交试验研究微波提取杜仲叶片绿原酸和总黄酮工艺．试验结果显示影响杜仲微波浸提绿原酸的主要因素为料液比，其次是微波功率、提取时间、乙醇浓度；获得的最佳提取工艺条件为乙醇浓度70％、提取时间25min、液料比1∶30、微波功率600W。试验同时表明影响杜仲微波浸提总黄酮的主要因素为乙醇浓度，其次是提取时间、料液比、微波功率；获得的最佳提取工艺条件为乙醇浓度50％、提取时间25min、液料比1∶30、微波功率400W。以获得的方法进行杜仲叶片绿原酸和总黄酮提取，含量分别可达到0.3425％和0.7672％。3、应用正交试验研究超声波提取杜仲叶片绿原酸和总黄酮工艺。试验结果表明影响杜仲超声波浸提绿原酸的主要因素为提取次数，其次是料液比、乙醇浓度、提取时间；最佳提取工艺条件为乙醇浓度30％，提取时间30min，料液比1∶25，提取次数3次。影响杜仲超声波浸提总黄酮的主要因素为提取次数，其次是乙醇浓度、提取时间、料液比。最佳提取工艺条件为乙醇浓度50％，提取时间20min，料液比1∶20，提取次数3次。以获得的方法进行杜仲叶片绿原酸和总黄酮提取，含量分别可达到0.4113％和1.4639％。4、试验研究了杜仲绿原酸、总黄酮的分离纯化工艺。通过对杜仲绿原酸、总黄酮的静态吸附分离，筛选NKAⅡ、S8、HPD600及HPD750吸附树脂；进一步的动态分析获得HPD600为最优纯化材料。优化的杜仲绿原酸、总黄酮的分离纯化工艺为：取pH值2.0原样液15.0ml，上柱吸附，以1.2ml／min流速的去离子水120ml及浓度为10％、30％、50％、70％和90％的乙醇各60ml依次进行梯度洗脱，收集30％和70％乙醇洗脱液为初步纯化的绿原酸、总黄酮。更多还原

【Abstract】 The test methods,extraction, separation and purification techniques of chlorogenic acid and flavonoids in Chinese medicine Eucommia ulmoide oliver were investigated. The results were as following:1. Traditional methods for determinating chlorogenic acid and flavonoids was modified to fit for using Chinese medicine Eucommia ulmoide oliver. Paper-layer chromatography was added to spectrophotomety and paper layer was applied to Al（NO₃）₃ method to make the method more suitable and accurate for measuring chlorogenic acid and flavonoids in Chinese medicine Eucommia ulmoide oliver.2. Orthogonal experiment was designed to investigate the microwave effect on the extraction of chlorogenic acid and flavonoids from Chinese medicine Eucommia ulmoide oliver. The results showed that the most important factor which influenced the extraction of chlorogenic acid was the material-fluid ratio, then microwave power, extraction duration, and finally ethyl alcohol concentration. The best extraction condition was 70% ethyl alcohol, 25min, material-fluid ratio 1:30, and microwave output power 600W. The most important factor which influenced the extraction of flavonoids was ethyl alcohol density, then extraction duration, material-fluid ratio, finally microwave output power. The best extraction condition was 50% ethyl alcohol, 25min extraction, 1:30 material-fluid ratio, and microwave output power 400W. Under above condition the total content of chlorogenic acid could be as high as 0.3425 %, and flavonoids as high as 0.7672%.3. Orthogonal experiment was designed to investigate ultrasonic effect on the extraction of chlorogenic acid and flavonoids from Chinese medicine Eucommia ulmoide oliver. The results showed that the primary factor influencing the extraction of chlorogenic acid was extraction times, next was material-fluid ratio, ethyl alcohol concentration, and lastly extraction duration. The best extraction condition was 30% ethyl alcohol, ultrasonic extract for 30min, 1:25 material-fluid ratio, extracting 3 times. The primary factor influencing the extraction of flavonoids was also extraction times, next was ethyl alcohol concentration, extraction duration, and material-fluid ratio. The best extraction condition was 50% ethyl alcohol, extraction for 25min, material-fluid ratio 1:20, extraction for 3times. Under above condition the total content of chlorogenic acid could be as high as 0.4113%, and flavonoids as high as 1.4639%.4. The separation and purification techniques of chlorogenic acid and flavonoids in Chinese medicine Eucommia ulmoide oliver were tested. NKAII、S8、HPD600 and HPD750 macroporous resin were selected as adsorption carriers for the separation and purification of chlorogenic acid and flavonoids from the extraction of Chinese medicine Eucommia ulmoide oliver, by adopting static absorption and disabsorption performance. Further dynamic absorption and disabsorption performance showed HPD600 was the best. The optimized conditions for the separation and purification of chlorogenic acid and flavonoids from the extraction of Chinese medicine Eucommia ulmoide oliver were: adding 15.0ml original extraction （pH 2.0） to HPD600 colum, then washed with pure water （120ml）, 10%,30%,50%,70% and 90% ethyl alcohol（all were 60ml） sequentialy at the velocity of 1.2ml/min. The 30% and 70% ethyl alcohol were Collected separately as purified chlorogenic acid and flavonoids.更多还原