荧光偏振免疫法测定水产品中的丁香酚  

A fluorescence polarization immunoassay for the detection of eugenol residues in aquatic products

作　　者：王强[1] 王旭峰[1] 张英侠 黄珂[1] WANG Qiang;WANG Xufeng;ZHANG Yingxia;HUANG Ke(South China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Key Laboratory of Aquatic Product Processing,Ministry of Agriculture and Rural Affairs,Guangzhou 510300,China)

机构地区：[1]中国水产科学研究院南海水产研究所,农业农村部水产品加工重点实验室,广州510300

出　　处：《分析试验室》2024年第3期307-313,共7页Chinese Journal of Analysis Laboratory

基　　金：国家重点研发计划“蓝色粮仓科技创新”重点专项(2019YFD0901701);广东省现代农业产业技术体系创新团队建设专项资金(2023KJ151)资助。

摘　　要：将丁香酚(Eul)与丙烯酸甲酯进行衍生化反应,合成了一种新型的丁香酚半抗原4-(4羟基-3-甲氨基苯基)-丁-2-烯酸(Eul-Aca)。采用活性酯法将丁香酚半抗原与乙二胺异硫氰酸荧光素(EDF)偶联,分别制备了同源和异源荧光示踪物。通过比较不同示踪物的抗体稀释度和灵敏度,优化反应时间等检测条件,建立了一种检测水产品中丁香酚的荧光偏振免疫分析法(FPIA)。结果表明:异源示踪物(Eul-AcaEDF)具有更优的检测灵敏度,抗体稀释度为1/200,反应时间仅需5 min。该方法的半抑制浓度(IC_(50))为11.2μg/L,检测线性范围为1.1~111.5μg/L,检出限(LOD)为0.24μg/L。实际样品中丁香酚的加标回收率为80.6%~107.4%,相对标准偏差(RSD)<15%。FPIA检测结果与气相色谱-质谱(GC-MS)法结果具有良好的一致性(r=0.993),适用于水产品中丁香酚的残留检测。Eugenol was derivatized with methyl acrylate to obtain 4-(4-hydroxy-3-methamphenyl)-butyl-2-acrylic acid.The hapten was coupled to fluoresceinthiocarbamyl ethylenediamine(EDF)using the active ester method to produce homologous and heterologous tracers.The influence of tracer structures on assay parameters such as antibody dilution and incubation time were investigated.Furthermore,a fluorescence polarization immunoassay(FPIA)was developed to detect eugenol residues in aquatic products.The results showed that the heterologous tracer(Eul-Aca-EDF)exhibited higher sensitivity with an antibody titer of 1/200.The reaction time of FPIA was only 5 min.The detection limit of eugenol,the IC_(50) value,and the working range were 0.24,11.2 and 1.1-111.5μg/L,respectively.The recoveries of spiked aquatic products were in the range of 80.6%-107.4%,with the relative standard deviations(RSDs)less than 15%.The FPIA also showed a good correlation to the GC-MS method(r=0.993).Therefore,the developed FPIA exhibited potential practicality in the detection of eugenol residues in aquatic products.

关 键 词：荧光偏振免疫分析法 示踪物 丁香酚 水产品 

分 类 号：O657.6[理学—分析化学]