超声波—大孔树脂联用提取富集追风七总黄酮及其清除自由基活性研究

Study on technology of ultrasound-assisted extraction combined macroporous resin purification enrichment for total flavonoids from Geum aleppicum and its radical scavenging activity

采用超声波法提取、大孔树脂富集纯化追风七总黄酮,并测试其清除自由基活性能力。以液料比、乙醇浓度、超声功率、超声温度、超声时间为自变量,总黄酮提取量为因变量,采用单因素试验和响应面设计优化提取工艺;以总黄酮的吸附量和解析率为评价指标,考察纯化追风七总黄酮大孔树脂的吸附性和洗脱参数;采用清除DPPH自由基活性评价追风七中总黄酮的抗氧化能力。结果表明:超声提取追风七总黄酮的最佳工艺为:超声功率300 W、乙醇浓度45%、液料比161(mL/g)、超声时间102min,超声温度73℃,在该条件下总黄酮提取量为17.02mg/g;AB-8树脂富集纯化追风七总黄酮最佳工艺条件为:树脂柱径高比16,上样质量浓度为0.5g/mL,上样量为20mL(2BV),乙醇体积分数40%,洗脱液用量60mL(6BV)。纯化后总黄酮保留率达83.3%,精制倍数达4.5倍,总黄酮含量达61%。清除自由基试验结果表明:总黄酮纯化物的IC50明显小于总黄酮提取物,略大于VC,其DPPH自由基清除IC50值为57.4μg/mL,其IC50大小顺序为VC<总黄酮纯化物<总黄酮提取物。

Taking the total flavonoids fromGeum aleppicum by using ultrasound-assisted extraction and purification techniques,and test the scavenging ability to DPPH of total flavonoids.With extraction quantity of total flavonoids as dependent variable,the independent variables including ratio of liquid to solid,ethanol concentration,ultrasonic power,ultrasonic temperature and ultrasonic time were studied on extraction by single experiment and response surface methodology;the adsorption and desorption of total flavonoids were used to investigate adsorption and elution conditions of macroporous resin;the scavenging ability to DPPH was used to investigate antioxidant activity.The optimal extraction conditions were：ultrasonic power 300 W,ethanol concentration45%,the ratio of liquid to solid161（mL/g）,extraction time 102 min,extraction temperature73 ℃.Under the conditions,the extraction amount of flavonoids was17.02 mg/g.AB-8 macroporous resin purification techniques conditions were：concentration of feed 0.5g/mL,the ratio of diameterheight 1：6,the volume of polyphenols sample 20mL（2BV）,elute ethanol concentration 40%,elute volume 60 mL（6 BV）.After eluted with AB-8resin,the retention rate of total flavonoids was83.3%,the refined multiples was 4.5times,the content of total flavonoids was 61%.The result of experiment on scavenging ability to DPPH showed that IC50 value of purification was obviously less than that of extracts of total flavonoids,and a little higher than that of VC,the scavenging order was VCpurification of total flavonoidsextract of total flavonoids,and the IC50 values of DPPH free radical scavenging was 57.4μg/mL.