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目的:建立高效液相色谱-二苯基三硝基苯肼(HPLC-DPPH)在线法评价葡萄酒清除自由基的活性,研究其抗氧化活性物质基础。方法:葡萄酒样品经HPLC分离,在柱后与DPPH工作液在三通处混合,于PEEK管中充分反应后,流经检测器记录反应信号,以水溶性维生素E类似物(Trolox)为标准计算活性成分的抗氧化当量(TEAC),比较7种不同地区葡萄酒总抗氧化能力及活性成分差异。结果:通过定性分析得出葡萄酒样品中没食子酸、原儿茶酸、原花青素二聚体B1、龙胆酸、儿茶素、咖啡酸和没食子酸乙酯对DPPH自由基具有清除活性,但原花青素二聚体B2、香草酸、丁香酸和对香豆酸没有清除活性。不同地区葡萄酒总抗氧化能力不同,所含抗氧化活性成分也有较大差异,共有活性成分有7种,其中原花青素二聚体B1与没食子酸活性最强,其次是儿茶素和未知物1,活性较弱的是咖啡酸、原儿茶酸和没食子酸乙酯。结论:葡萄酒中含有多种抗氧化活性物质;该法能够实现对天然产物抗氧化活性的分析,具有在线、无损、高通量筛选、快速分析的特点。
OBJECTIVE: To establish an online method for the determination of free radical scavenging activity in wine by high performance liquid chromatography (HPLC) -piperazine dinitrophenylhydrazine (HPLC-DPPH), and to study its antioxidant basic substance. Methods: The wine samples were separated by HPLC, mixed with DPPH working fluid at the tee position after the column, and reacted fully in the PEEK tube. The reaction signal was recorded through the detector and calculated according to the standard of water soluble vitamin E analogue (Trolox) Antioxidant activity of active ingredient (TEAC), compare the total antioxidant capacity of seven different regions of wine and active ingredient differences. Results: The results showed that gallic acid, protocatechuic acid, proanthocyanidin dimer B1, gentisic acid, catechin, caffeic acid and ethyl gallate had scavenging activity on DPPH radical in wine samples by qualitative analysis. However, Binder B2, vanillic acid, syringic acid and p-coumaric acid. Different regions of the total antioxidant capacity of different wines, the antioxidant activity of the ingredients also have a greater difference, a total of seven kinds of active ingredients, of which proanthocyanidins dimer B1 and gallic acid strongest, followed by catechins and unknown 1, Less active caffeic acid, protocatechuic acid and ethyl gallate. Conclusion: The wine contains a variety of antioxidants; this method can realize the analysis of natural products antioxidant activity, with online, non-destructive, high-throughput screening, rapid analysis.