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用于食品样品中多种霉菌毒素检测的免疫亲和96斑点整体阵列的研制。

Development of immune-affinity 96 spots monolith array for multiple mycotoxins detection in food samples.

作者信息

Li Li, Xia Li-Ru, Zhao Yong-Fu, Wang He-Ye

机构信息

Institute of Facilities and Equipment in Agriculture, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.

Institute of Facilities and Equipment in Agriculture, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Sep 1;1029-1030:72-80. doi: 10.1016/j.jchromb.2016.07.012. Epub 2016 Jul 7.

Abstract

In this paper, a novel highly sensitive chemiluminescence immune-affinity 96 spots monolith array was developed to detect deoxynivalenol (DON), zearalenone (ZEN), T-2 toxin (T-2), and fumonisin B1 (FB1) in corn samples. Firstly, the monolith array was prepared through on suit UV-initiated copolymerization using polyethylene glycol diacrylate (PEGDA) as cross-linker, glycidyl methacrylate (GMA) as functional monomer and polyethylene glycol 200 (PEG 200) as the porogen. Subsequently, the four mycotoxins immune-affinity monolith array was prepared by immobilization of DON, ZEN, T-2, and FB1 antibody. The mole ratio of PEGDA/GMA, UV exposure time, and the volume ratio of PEG 200/PEGDA were optimized to improve the performances of the immune-affinity monolith array. For the mycotoxins immune-affinity monolith array based on chemiluminescence detection, the limit of detection was 0.0036ng/mL (DON), 0.0048ng/mL (ZEN), 0.0039ng/mL (T-2), and 0.0017ng/mL (FB1), respectively. The linear response in the range of 0.01-0.1ng/mL (R(2)=0.98). The results showed that the proposed four mycotoxins immune-affinity monolith array was a stable, accurate, and highly sensitive method to determine levels of DON, ZEN, T-2, and FB1 in real samples.

摘要

本文开发了一种新型高灵敏度化学发光免疫亲和96斑点整体阵列,用于检测玉米样品中的脱氧雪腐镰刀菌烯醇(DON)、玉米赤霉烯酮(ZEN)、T-2毒素(T-2)和伏马菌素B1(FB1)。首先,以聚乙二醇二丙烯酸酯(PEGDA)为交联剂、甲基丙烯酸缩水甘油酯(GMA)为功能单体、聚乙二醇200(PEG 200)为致孔剂,通过原位紫外引发共聚制备整体阵列。随后,通过固定DON、ZEN、T-2和FB1抗体,制备了四种霉菌毒素免疫亲和整体阵列。优化了PEGDA/GMA的摩尔比、紫外照射时间以及PEG 200/PEGDA的体积比,以提高免疫亲和整体阵列的性能。对于基于化学发光检测的霉菌毒素免疫亲和整体阵列,检测限分别为0.0036ng/mL(DON)、0.0048ng/mL(ZEN)、0.0039ng/mL(T-2)和0.0017ng/mL(FB1)。在0.01 - 0.1ng/mL范围内呈线性响应(R(2)=0.98)。结果表明,所提出的四种霉菌毒素免疫亲和整体阵列是一种稳定、准确且高灵敏度的方法,可用于测定实际样品中DON、ZEN、T-2和FB1的含量。

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