罗汉松种子总黄酮提取工艺优化及抗氧化活性

Optimization of Extraction Process and Antioxidant Activity of Flavonoids from Podocarpus Macrophyllus Seeds

目的采用响应面法优化罗汉松种子总黄酮超声提取工艺,并考查总黄酮的抗氧化活性。方法基于单因素试验,以料液比、超声时间、超声温度为自变量,以总黄酮得率和DPPH自由基的清除率为因变量,采用三因素三水平的响应面分析法,确定最佳提取工艺条件。结果得到最佳工艺参数:在超声频率为40 kHz、超声功率为200 W时,料液比(g/mL)为1∶50、超声温度为70℃、超声时间为2.7 h,其理论预测值总黄酮得率为27.54 mg/g(n=3,相对标准偏差为1.63%)。当添加稀释10倍的总黄酮提取液0.50 mL时,对DPPH自由基的清除率达到87.2%、对Ce(Ⅳ)还原率达到84.2%;当添加稀释10倍的总黄酮提取液0.90 mL时,对OH自由基的清除率为88.1%。结论采用响应面法优化得到的超声波辅助提取工艺稳定、可靠,可用于罗汉松种子总黄酮的提取。罗汉松种子总黄酮可作为优良的天然抗氧化剂资源。

The work aims to optimize the ultrasonic extraction process of flavonoids from Podocarpus macrophyllus seeds by response surface methodology and investigate the antioxidant activity of flavonoids.Based on the single factor test,with the material liquid ratio,ultrasonic time and ultrasonic temperature as independent variables and the yield of flavonoids and DPPH radical scavenging rate as dependent variables,the best extraction process conditions were determined by three-factor and three-level response surface analysis.The optimal process parameters were ultrasonic frequency of 40 kHz,ultrasonic power of 200 W,material liquid ratio(g/mL)of 1:50,ultrasonic temperature of 70℃and ultrasonic time of 2.7 h.The theoretically predicted yield of flavonoids was 27.54 mg/g(n=3,relative standard deviation of 1.63%).When 0.50 mL of flavonoids extract diluted 10 times was added,the DPPH radical scavenging rate was 87.2%and the reduction rate of Ce(Ⅳ)was 84.2%.When 0.90 mL of flavonoids extract diluted 10 times was added,the scavenging rate of OH was 88.1%.The ultrasonic assisted extraction process optimized by response surface methodology is stable and reliable,and can be used to extract flavonoids from Podocarpus macrophyllus seeds.The flavonoids of Podocarpus macrophyllus seeds can be used as excellent natural antioxidant resources.