摘要
目的构建一种可配合花生蛋白夹心酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)试剂盒实现对于加工食品中花生成分可靠检测的高质量样品前处理方法。方法对9种不同pH的提取缓冲液提取未处理、湿热处理、干热处理花生蛋白效果进行比较分析以确定最优提取缓冲液类型。而后,将吐温-20、二硫苏糖醇(dithiothreitol,DTT)、十二烷基硫酸钠(sodium dodecyl sulfate,SDS)以单独或组合添加的方式加入上一步获得的最优提取缓冲液中,提取3组花生蛋白,并对提取效果与夹心ELISA试剂盒检测花生蛋白回收率进行比较分析以确定最优添加剂配方。结果最终确定添加0.2%SDS和0.1%吐温-20的碳酸盐缓冲液(50 mmol/L,pH 9.6)为最优提取缓冲液。向碳酸盐缓冲液中加入SDS、吐温-20较好地提升了花生蛋白的提取率和回收率,将湿热处理花生蛋白回收率提高了2倍以上。结论本研究实现了对商业加工食品中花生成分的可靠检测,为推动过敏原信息在食品标签的准确标注提供了技术支撑,也为相关研究提供了参考。
Objective To develop a high-quality sample pretreatment method applicable to the peanut proteins sandwich enzyme-linked immunosorbent assay(ELISA)test kit for reliable detection of peanut components in commercially processed food.Methods The extraction effects of proteins from untreated,wet heat-treated,and dry heat-treated peanut extracted by 9 extraction buffers of different pH were compared and analyzed in order to determine the optimal type of extraction buffer.Afterward,Tween-20,dithiothreitol(DTT),and sodium dodecyl sulfate(SDS)as the additives were added separately or in combination into the optimal extraction buffer obtained in the previous step to extract proteins from 3 groups of peanut.The extraction effects and the recoveries of peanut proteins detected by the sandwich ELISA test kit were compared and analyzed to determine the optimal additive formulation.Results Carbonate buffer(50 mmol/L,pH 9.6)with 0.2%SDS and 0.1%Tween-20 was ultimately determined to be the optimal extraction buffer.Adding SDS and Tween-20 into carbonate buffer effectively improved the extraction efficiency and the recovery of the peanut proteins.The recovery of the proteins from wet heat-treated peanut was improved by more than 2 times.Conclusion This study achieves reliable detection of peanut components in commercially processed foods,provides technical support to promote the accurate labeling of allergen information in food labels,and also provides a reference for related research.
作者
季雨
林洪
李振兴
张莉
JI Yu;LIN Hong;LI Zhen-Xing;ZHANG Li(Department of Logistics Support,Ocean University of China,Qingdao 266100,China;College of Food Science and Engineering,Ocean University of China,Qingdao 266000,China;Tongbiao Standard Technical Services Co.,Ltd.,Qingdao Branch,Qingdao 266100,China)
出处
《食品安全质量检测学报》
CAS
2024年第20期246-256,共11页
Journal of Food Safety and Quality
基金
国家自然科学基金项目(32072338)。
关键词
前处理
提取探究
花生
过敏
夹心酶联免疫吸附测定法
加工食品
可靠检测
pretreatment
extracting exploration
peanut
allergy
sandwich enzyme-linked immunosorbent assay
processed food
reliable detection
作者简介
季雨,硕士,主要研究方向为食品安全与质量控制。E-mail:jiyu@ouc.edu.cn;通信作者:李振兴,教授,主要研究方向为食品安全与质量控制。E-mail:lizhenxing@ouc.edu.cn。
