高效液相色谱-二苯基三硝基苯肼在线检测葡萄酒中抗氧化活性成分  

On-line HPLC-DPPH Assay for Analysis of Antioxidant Compounds in Grape Wine

作　　者：邢航[1] 张媛婷[1] 鲁雪峰[1] 侯晓虹[2] 毕开顺[1] 李清[1] 

机构地区：[1]沈阳药科大学药学院,沈阳110016 [2]沈阳药科大学制药工程学院,沈阳110016

出　　处：《中国实验方剂学杂志》2013年第21期127-131,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：复方酸枣仁汤及红酒提取物开发(2009226010);2012年辽宁省高等学校中药质量控制关键技术创新团队项目(LT2012018)

摘　　要：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： A reversed phase high performance liquid chromatography （RP-HPLC） separation coupled with 2, 2-diphenyl-l-picryhydrazyl （DPPH） assay was used for the screening of multiple antioxidant compounds and the antioxidant basis of grape wine. Method： The red grape wine was passed through HPLC to enter into the PEEK coil contained the DPPH working solution, and signals were recorded at 517 nm. The Trolox equivalent antioxidant capacities （TEAC） were determined to compare the total antioxidant capacity and the antioxidant activity of the active compounds among the seven grape wines. Result： By qualitative analysis, gallic acid, protocatechuic, proeyanin B1, gentisic acid, catechin, caffeic acid and ethyl gallate were active ingredients, but proeyanin B2, vanillic acid, syringic acid, p-coumaric acid did not have the screening activity. The total antioxidant capacity and activity compositions were different among the seven grape wines, and the activity of procyanin B2, gallic acid, catechin and the unknown were better than eaffeie acid, protocateehuic and ethyl gallate. Conclusion： Grape wines contain some amount of antioxidant compounds, and the on-line HPLC-DPPH assay, which is on-line, no-loss, high-flux and fast analysis, can be a powerful technique for screening analysis of antioxidant compounds in grape wine.

关 键 词：葡萄酒 高效液相色谱一二苯基三硝基苯肼(HPLC—DPPH) 抗氧化活性 在线筛选 

分 类 号：R284.1[医药卫生—中药学]