基于体外抗氧化试验的三叶青活性物质提取工艺的研究

Extraction Process of Active Substance from Tetrastigmatis Hemsleyanum Diels Et Gilg. Based on Antioxidant Test in Vitro

为选取最佳的三叶青抗氧化活性物质的提取工艺,分别采用热回流提取法、微波提取法和超声提取法制备三叶青提取物,用DPPH法、铁离子还原能力(FRAP)法、Trolox等效抗氧化能力(TEAC)法和羟自由基清除法综合评价各提取物的体外抗氧化能力,再用福林酚(Folin-Ciocalteu)比色法测定其总酚含量,将不同提取方法所得提取物的总酚含量与抗氧化能力之间做相关性分析,筛选提取方法,用正交试验优化该提取方法的工艺条件。结果表明,热回流提取方法所得三叶青提取物总酚含量最高,其清除羟自由基、ABTS^(+)·、DPPH自由基的能力及还原铁的能力皆优于微波提取法与超声提取法所得提取物;热回流提取法最佳提取工艺是60%(v/v)乙醇、60 ℃提取3次、1.5 h/次。热回流提取法适合于制备三叶青具有抗氧化活性的提取物。

Based on the antioxidant activity in vitro,the best extraction method of active substance from Tetrastigmatis hemsleyanum Diels et Gilg.were screened,and its extraction process was optimized.Heat reflux extraction,microwave extraction and ultrasonic extraction were used to obtain active substance of Tetrastigmatis hemsleyanum Diels et Gilg.The antioxidant activities of the active substance were evaluated by DPPH,Trolox equivalent antioxidant capacity (TEAC),iron ion reduction capacity (FRAP) and hydroxyl radical scavenging methods.Then the content of total phenols was determined by Folin-Ciocalteu colorimetry.In oeder to screen the best extraction method,the correlation between the total phenol content of the extracts and the antioxidant methods was investigated.Orthogonal test was used to optimize the extraction process.The results showed that the total phenol content of the extract obtained by hot reflux extraction method was the highest,and its iron reduction ability,ABTS^(+)· scavenging ability,DPPH free radical scavenging ability and hydroxyl radical scavenging ability were better than those of microwave extraction and ultrasonic extraction.The best extraction process of heat reflux extraction was 60% ethanol,60 ℃ for 3 times,1.5 h/time.The hot reflux extraction method is suitable for the preparation of the extract with antioxidant activity from Tetrastigmatis hemsleyanum Diels et Gilg.