摘要
为提升茶饮料产品品质,对茶饮料浑浊沉淀产生机理和反应条件进行快速精确的分析检测,以表没食子儿茶素没食子酸酯(EGCG)和咖啡碱为试验对象,采用双交联法制备偶联蛋白,通过紫外光谱对偶联蛋白进行分析,计算偶联比,采用酶联免疫吸附试验研究EGCG与咖啡碱相互作用的条件。结果表明:ELISA法测定茶多酚与咖啡碱相互作用,宜选牛血清白蛋白(BSA)及卵清蛋白(OVA)为咖啡碱及EGCG偶联的载体蛋白;咖啡碱-BSA偶联蛋白的最佳包被浓度为2μg/mL,最佳包被条件为4℃过夜;EGCGOVA偶联蛋白最佳工作浓度为1∶1 000;咖啡碱与EGCG反应最佳作用时间为60min。批内重复的变异系数均值为5.26%,批间重复的变异系数均值为6.73%,精密度较高,具有较好的重复性。
In order to improve the quality of tea beverage product,producing mechanism and reaction condition of turbidity and precipitation in it were tested fast and accurately,taking epigallocatechin gallate（EGCG）and coffeic alkali as the test objects,the authors prepared coupling-protein by double crosslinking method,which was analyzed by ultraviolet spectrum,and the coupling rate was calculated either.And the interaction conditions of tea polyphenols and coffeic alkali were studied by ELISA method.Results：for determination of interaction of tea polyphenols and coffeic alkali by ELISA,BSA was the appropriate carrying-protein coupled with coffeic alkali and the appropriate carrying-protein coupled with EGCG was OVA;The best wrapping concentration for coupling-protein coffeic alkali-bsa was 2ug/mL;The best wrapping condition was 4℃and overnight;The best acting concentration for coupling-protein EGCG-OVA was 1：1000;The best reaction time for coffeic alkali with EGCG was 60 minutes.The mean value of coefficient of variations for within-batchs and between batchs respectively are 5.26% and 6.73%,the precision was high and the repeatability was good.
出处
《贵州农业科学》
CAS
北大核心
2014年第4期182-185,共4页
Guizhou Agricultural Sciences
基金
贵州省科学技术基金项目"茶多酚与咖啡碱相互作用的ELISA检测方法的建立"[黔科合J字(2011)2171]
关键词
茶多酚
咖啡碱
相互作用
ELISA检测方法
tea polyphenols
coffeic alkali
interaction
enzyme linked immunusorbent assay
