毛细管电泳—仿生免疫分析检测敌百虫方法研究

发布时间：2018-03-07 20:44

本文选题：敌百虫　切入点：毛细管电泳技术　出处：《山东农业大学》2016年硕士论文　论文类型：学位论文

【摘要】：敌百虫是一种广泛应用于田间农作物的有机磷杀虫剂,具有低毒、高效、水溶性好等优点,被广泛地应用于园艺、农林、畜牧等领域。但是,敌百虫容易经呼吸道及皮肤进入人体,近年来,其潜在地致突变和致癌性引起人们的高度重视。因此,建立一种快速、准确、低成本的敌百虫检测方法迫在眉睫。本课题研究基于毛细管电泳分析技术和仿生免疫分析技术,建立了一种快速检测蔬菜中敌百虫的新方法。研究的主要结果如下:(1)制备合成敌百虫酶标半抗原。敌百虫和琥珀酸酐在无水吡啶中避光反应后,经过萃取、减压蒸馏等步骤得到敌百虫半抗原。采用一步合成法,将辣根过氧化物酶(HRP)与敌百虫半抗原直接连接制备酶标半抗原。(2)敌百虫仿生抗体的制备。敌百虫为模板分子,甲基丙烯酸(MAA)为功能单体,乙二醇二甲基丙烯酸酯(EGDMA)为交联剂,偶氮二异丁腈(AIBN)为引发剂,采用本体聚合法合成仿生抗体,并通过热重分析实验和电镜扫描实验对其进行表征。(3)建立毛细管电泳-仿生免疫分析检测敌百虫新方法。对实验条件进行了系统的优化,结果显示,敌百虫在pH值为7.6的PBS缓冲液中,与酶标敌百虫半抗原竞争反应60 min的条件下,该方法的最低检测限(LOD,IC15)和灵敏度(IC50)分别为和0.13μg/L和0.16 mg/L。本方法对0.65μg/L,0.16 mg/L两个浓度水平的菜豆与黄瓜样品敌百虫添加回收率为78.8%~103%。
[Abstract]:Trichlorfon is a kind of organophosphorus insecticide widely used in field crops. It has the advantages of low toxicity, high efficiency and good water solubility. It is widely used in horticulture, agriculture and forestry, animal husbandry and other fields. Trichlorfon is easy to enter human body through respiratory tract and skin. In recent years, the potential mutagenicity and carcinogenicity of trichlorfon have attracted great attention. Low cost detection of trichlorfon is urgent. This study is based on capillary electrophoresis and bionic immunoassay. A new method for rapid detection of trichlorfon in vegetables was established. The main results are as follows: 1) Synthesis of trichlorfon labeled hapten. Trichlorfon and succinic anhydride were extracted by light avoidance reaction in anhydrous pyridine. The hapten of trichlorfon was obtained by decompression distillation. By means of one-step synthesis, horseradish peroxidase (HRP) was directly linked with trichlorfon hapten to prepare an enzyme-labeled hapten. 2) the bionic antibody of trichlorfon was prepared. Trichlorfon was used as template molecule. Bionic antibody was synthesized by bulk polymerization with MAA as functional monomer, ethylene glycol dimethacrylate (EGDMA) as crosslinking agent and azodiisobutyronitrile (AIBN) as initiator. A new method for the detection of trichlorfon by capillary electrophoresis and bionic immunoassay was established by thermogravimetric analysis and scanning electron microscopy. Trichlorfon reacted with the enzyme labeled trichlorfon hapten in PBS buffer with pH 7.6 for 60 min. The minimum detection limit and sensitivity of this method were 0.13 渭 g / L and 0.16 mg / L, respectively. The recoveries of trichlorfon in two concentrations of 0.65 渭 g / L and 0.16 mg/L were 78.8% and 103%, respectively.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：TS255.7;O657.8

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