邻苯二甲酸二甲酯免疫层析快速检测试纸的研制
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朱帆(1991-),男,硕士研究生,研究方向:食品质量安全免疫分析技术 通讯作者:沈玉栋(1977-),男,博士,教授,研究方向:食品质量安全快速检测,生物分析与传感

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广东省科技计划(2013B040402006,2014A020219008);国家自然科学基金项目(31371769);广州市科技计划(2011Y2-00007,2013J4100053);广东省优秀博士学位论文作者项目


Development of a Colloidal Gold Immunochromatographic Strip for the Rapid Determination of Dimethyl Phthalate
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    摘要:

    邻苯二甲酸酯类塑化剂是一类具有生殖毒性的环境激素类物质,通过违规添加或迁移而造成食品污染问题颇为严峻,开发适合现场的塑化剂快速监测方法非常重要。本研究以邻苯二甲酸二甲酯(DMP)为对象,建立了DMP的胶体金标记免疫层析检测试纸方法。采用柠檬酸三钠还原氯金酸制备了胶体金,将其标记DMP多克隆抗体,制备了金标抗体;采用棋盘法和单因素试验确定最佳包被浓度和二抗浓度分别为3000 μg/mL和1600 μg/mL,抗体浓度为稀释1.4倍,基于此建立了DMP免疫层析快速检测方法,其检测限为0.4 μg/mL,5 min内肉眼可观察到检测结果,与其它结构功能类似物无明显交叉反应,特异性良好,白酒样品添加检测结果与GC-MS法相符。本法快速简单,稳定可靠,适用于实际样品中DMP的现场快速检测。

    Abstract:

    Phthalate ester plasticizer, a type of environmental hormone that is toxic to reproductive systems, has become a serious health threat because of its illegal transport and addition into foods intended for human consumption. Thus, development of an in situ testing method for the rapid detection of the plasticizer is essential. Herein, a colloidal gold-labeled immunochromatographic test paper method was developed for the rapid detection of dimethyl phthalate (DMP). To prepare the gold-labeled antibodies, anti-DMP polyclonal antibodies were labeled with colloidal gold prepared by the reduction of HAuCl4 using trisodium citrate. The chessboard method and the single-factor test were used to determine the optimal coating concentration and secondary antibody concentration, which were revealed to be 3000 μg/mL and 1600 μg/mL, respectively. The antibody was diluted 1.4-fold. The limit of detection of the developed assay was 0.4 μg/mL, and the visual detection time was within 5 min. Cross-reaction experiments showed no interference from substances with similar structures, indicating the high specificity of the method. The results from strip testing of baijiu liquor were consistent with those from GC-MS. This proposed immunochromatographic assay is fast, simple, and reliable, providing a valid method for the rapid in situ detection of DMP in foods.

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朱帆,邓丽华,朱彬,杨金易,徐振林,孙远明,王弘,沈玉栋.邻苯二甲酸二甲酯免疫层析快速检测试纸的研制[J].现代食品科技,2017,33(3):279-284.

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  • 收稿日期:2016-02-23
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  • 在线发布日期: 2017-04-01
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