大孔树脂纯化滇黄精黄酮工艺优化及纯化前后抗氧化活性比较

Optimization of purification process of flavonoids from P.kingianum Coll.by macroporous resin and comparison of antioxidant activity before and after purification

试验研究滇黄精黄酮的纯化工艺及其抗氧化活性。通过单因素试验结合响应面试验,利用超声辅助溶剂提取法对滇黄精黄酮进行提取工艺优化,通过静态和动态试验,考察树脂种类、粗提液浓度、洗脱剂、洗脱流速对滇黄精黄酮吸附解吸性能的影响,确定最佳纯化工艺条件。采用1, 1-二苯基-2-三硝基苯肼(DPPH)自由基和2, 2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)自由基法,比较纯化前后滇黄精黄酮的抗氧化性。结果显示,滇黄精总黄酮的最佳提取工艺参数为乙醇浓度79%、提取温度61℃、提取时间2 h、液料比20 mL/g,提取率达到0.29%。AB-8大孔树脂纯化滇黄精黄酮效果最好,最佳条件为将40 g/L粗提液上柱,用70%乙醇以2.0 g/L速度洗脱,此条件下滇黄精黄酮纯度提高至5.31%,是粗提物浓度的11.8倍。滇黄精黄酮纯化后对DPPH自由基和ABTS自由基清除率为82.44%、86.22%,远高于纯化前。研究表明,AB-8大孔树脂适合分离纯化滇黄精黄酮,且滇黄精黄酮具有抗氧化活性。

The purification process and antioxidant activity of flavonoids from P.kingianum Coll.were studied.Through a single factor test combined with a response surface test,ultrasonic-assisted solvent extraction was used to optimize the extraction process of flavonoids from P.kingianum Coll..Through static and dynamic experiments,the effects of resin type,crude extract concentration,eluent,and elution flow rate on the adsorption and desorption performance of flavonoids from P.kingianum Coll.were investigated,and the best purification conditions were determined.The antioxidant activity of flavonoids from P.kingianum Coll.before and after purification was compared by DPPH free radical and ABTS free radical methods.The results showed that the optimum extraction parameters were ethanol concentration 79%,extraction temperature of 61℃,extraction time of 2 h,the ratio of liquid to material 20 mL/g,and the extraction rate reached 0.29%.AB-8 macroporous resin had the best effect on the purification of flavonoids from P.kingianum Coll..The best condition was that 40 g/L crude extract was put on the column and 70%ethanol was eluted at 2.0 g/L.Under this condition,the purity of flavonoids from P.kingianum Coll.was increased to 5.31%,which was 11.8 times of the concentration of crude extract.The scavenging rates of DPPH and ABTS radicals after purification were 82.44%and 86.22%,which were much higher than those before purification.The study indicates that AB-8 macroporous resin was suitable for separation and purification of flavonoids from P.kingianum Coll.and the flavonoids have antioxidant activity.