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