改良的ABTS^+法及其在优化抗氧化活性物质提取中的应用

Application of modified ABTS^+ assay method to monitor antioxidant extraction from fruit

目的 对测定抗氧化活性的ABTS+ 法进行改良 ,以提高分析速度、降低分析成本 ,并将该方法用于水果中抗氧化活性物质提取条件的优化。方法 在酶标板上以Trolox为标准对照 ,测定抗氧化物质对预先制备的自由基ABTS+ 的清除能力。用不同浓度的乙醇和重亚硫酸钠为提取液 ,在不同的温度和溶剂 水果比条件下 ,提取水果blackcurrant中的抗氧化活性物质 ,寻找从单位重量水果中提取抗氧化活性水平较高的条件。结果 该方法的测量误差小于 0 2 ,测得的 14种标准样品的抗氧化活性与文献报道基本一致。用乙醇从水果blackcurrant中提取抗氧化活性物质的条件为 :在 17℃上下、每克水果用 2 0毫升、6 7%左右浓度的乙醇进行提取。而用重亚硫酸钠从水果blackcurrant中提取抗氧化活性物质的条件为 :在 30℃上下、每克水果用2 0毫升、30 0mg L左右浓度的重亚硫酸钠进行提取。结论 改良的ABTS+ 法测定物质的抗氧化活性 ,方法简便可行 ,可用于优化抗氧化活性物质的提取 。

Objective Modify the ABTS + method to measure the antioxidant activity of compound. Apply this method to optimize antioxidant extraction from fruit.Methods In microplate, Trolox served as reference. Antioxidant activity was measured according to their ability to scavenge ABTS + in the reaction mixture. Under different temperatures, solvent solid ratios, solvent concentrations, antioxidants were extracted.Results The standard error for measuring Trolox antioxidant activity was under 0.2. The antioxidant activities of fourteen standard samples were consistent with that reported in literatures. Using ethanol as solvent, at about 17℃, for 1 gram of blackcurrant 20ml of 67% ethanol was appropriate. While using sodium metabisulphite as solvent, at about 30℃, for 1 gram of blackcurrant 20ml of 300mg/L metabisulphite would be appropriate.Conclusion Modified ABTS + method for measuring antioxidant activity was simple and feasible. This method could be applied to monitor antioxidant extraction from fruit and high throughput screening for antioxidant.