锦鹤养心方总黄酮提取工艺优化及抗氧化活性考察

Optimization of Extraction Technology for Total Flavonoids from Jinhe Yangxin Prescription and Investigation of Its Antioxidant Activity

目的：优化锦鹤养心方总黄酮的提取工艺,并对其体外抗氧化活性进行考察。方法：采用回流提取技术,以总黄酮提取量为指标,在单因素试验基础上,利用Box-Behnken试验设计考察乙醇体积分数、液料比、回流时间3个因素对总黄酮提取量的影响,优化总黄酮提取工艺;采用FRAP法、清除1,1-二苯基-2-三硝基苯肼（DPPH）自由基和羟基自由基的方法,对最佳工艺所得总黄酮的抗氧化活性进行评价。结果：最佳提取工艺为加17倍量55%乙醇回流提取2次,每次146 min,总黄酮提取量为74.42 mg·g^-1,与理论预测值74.98 mg·g^-1偏差较小;总黄酮总抗氧化的FRAP值为77.20 mg^-1,清除自由基的能力随着质量浓度的增大而增强,对DPPH和羟基自由基的半数抑制浓度分别为4.96和9.69μg·mL^-1,均优于阳性对照药VC。结论：该提取工艺简便、稳定,适用于锦鹤养心方总黄酮的提取,且锦鹤养心方总黄酮具有较好抗氧化活性,有进一步开发的价值。

Objective： To optimize the extraction process of total flavonoids from Jinhe Yangxin Prescription and investigate its antioxidant activity. Method： Based on the single-factor tests,Box-Behnken design model was applied to investigate the effects of ethanol concentration,time and ratio of liquid to solid on extraction efficiency. Besides,antioxidant activity of total flavonoids in vitro was valuated by FRAP assay,DPPH and hydroxyl free radical scavenging activity assay. Result： Optimum extraction conditions were ethanol concentration 55%,solid-liquid ratio 1∶17,extraction time146 min. Under these conditions,the yield of total flavonoids was 74. 42 mg·g^-1 and the result of trial was agreed with model predictions. The experiments showed that the FRAP value of antioxidative capacity was 77. 20 mg^-1 and herbal pair total flavonoids demonstrated the dose-dependent free radical scavenging and antioxidant activates. Scavenging activity to DPPH free radical and hydroxyl free radical with half inhibitive concentration（IC50） was 4. 96 μg·mL^-1 and 9.69 μg·mL^-1,respectively. And its antioxidative was stronger than vitamin C. Conclusion： Application of this simple and workable technology to extract total flavonoids from Jinhe Yangxin Prescription can improve extraction efficiency. The total flavonoids shows a relatively high antioxidant activity and so it is valuable to be further developed and used.