酶联免疫化学发光法快速检测日落黄  

Rapid Detection of Sunset Yellow by Chemiluminescence Enzyme-Linked Immunoassay

作　　者：许龙 许飞悦 徐雨珠 牛靖迪 黄慧 王弯弯 石璐璐 XU Long;XU Fei-yue;XU Yu-zhu;NIU Jing-di;HUANG Hui;WANG Wan-wan;SHI Lu-lu(College of Medicine and Chemistry and Chemical Engineering,Taizhou University,Taizhou 225300,China;Taizhou Food Inspection Institute,Taizhou 225300,China;College of Economics and Management,Taizhou University,Taizhou 225300,China)

机构地区：[1]泰州学院医药与化学化工学院,江苏泰州225300 [2]泰州市食品检验院,江苏泰州225300 [3]泰州学院经济与管理学院,江苏泰州225300

出　　处：《中国调味品》2025年第4期222-227,共6页China Condiment

基　　金：江苏省高等学校自然科学研究面上项目(22KJB360017);江苏省市场监管重点实验室(特医食品风险识别及关键分析技术)开放课题(TSKF-2023005);2023年泰州市“凤城英才”青年科技人才托举项目;泰州学院高层次人才启动基金项目(TZXY2022QDJJ001);泰州学院大学生创新创业训练计划项目(cx250127y)。

摘　　要：日落黄(SY)被广泛用作食用色素,过量和非法使用SY可能会造成潜在的健康风险。因此,建立快速、灵敏、特异的SY检测方法十分有必要。该研究采用活泼酯法合成日落黄(SY)的检测抗原(SY-OVA),以高碘酸盐氧化法制备辣根过氧化物酶(HRP)与抗SY单克隆抗体(mAb)的偶联物(mAb-HRP),并建立直接竞争酶联免疫化学发光法(CLEIA)。该方法在1~50 ng/mL范围内与SY呈现良好的线性关系,最低检测限为0.55 ng/mL,IC_(50)为6.5 ng/mL。在食品和饮料中的回收率为77%~110.2%,变异系数为2.7%~12.6%。实际样品的测定结果显示,样品中SY含量均低于国家限量标准。CLEIA与HPLC的检测结果具有较高的一致性。该研究建立的CLEIA可用于食品和饮料中SY的快速高效检测,为SY的快速检测提供了一种新的途径,具有较好的应用前景。Sunset yellow(SY)is widely used as a edible pigment,and excessive and illegal use of SY may pose potential health risks.Therefore,it is necessary to establish a rapid,sensitive and specific SY detection method.In this study,sunset yellow(SY)detection antigen(SY-OVA)is synthesized using the active ester method,and the conjugate(mAb-HRP)of horseradish peroxidase(HRP)and anti-SY monoclonal antibody(mAb)is prepared by periodate oxidation method.A direct competitive chemiluminescence enzyme-linked immunoassay(CLEIA)is established.This method shows a good linear relationship with SY in the range of 1~50 ng/mL,with the lowest limit of detection(LOD)of 0.55 ng/mL and IC_(50) of 6.5 ng/mL.The recovery rates of SY in food and beverages range from 77%to 110.2%,and the coefficients of variation range from 2.7%to 12.6%.The detection results of actual samples show that the SY content in samples is all lower than the national limit standard.The detection results of CLEIA and HPLC have high consistency.The CLEIA established in this study could be used for the rapid and efficient detection of SY in food and beverages,which has provided a new way for the rapid detection of SY and has good application prospects.

关 键 词：日落黄 酶联免疫化学发光法 高效液相色谱法 饮料 食品 

分 类 号：TS207.3[轻工技术与工程—食品科学]