两种新型有机污染物的环境生物分析方法研究

发布时间：2018-05-29 20:26

本文选题：新型有机污染物 + 电化学免疫传感器　；参考：《江苏大学》2017年硕士论文

【摘要】：新型有机污染物是人工合成或天然存在的物质,它们尚未在环境中被有效检出,但具有潜在的进入环境并引起已知或可疑的生态毒理效应和健康效应的可能性。包括抗生素,个人护理产品,表面活性剂,人造甜味剂,农药和各种工业添加剂等。这些有机污染物广泛存在于环境中,但缺乏完整的生态毒理学数据。针对该类污染物一般采用色谱分析方法,该方法准确性好、灵敏度高。但样品前处理复杂、所用仪器较为昂贵、且不能实现快速分析。与之相反,生物分析方法仪器简单,操作简便,无需复杂的样品处理,且样品用量少并能实现快速分析。本论文选择邻苯二甲酸二丁酯(Dibutyl phthalate,DBP)与四溴双酚A双(2-羟乙基)醚(Tetrabromobisphenol-A bis(2-hydroxyetyl)ether,TBBPA-DHEE)这两种新型有机污染物,分别构建了灵敏、简单、可靠的分析方法,并成功应用于实际环境样品的分析。具体内容如下:(1)针对DBP,采用间接竞争方式,建立了一种交流阻抗电化学免疫传感器。本研究分别以MWCNTs@GONRs纳米复合材料及DBP抗原、抗体修饰GCE,并以NADH促进AuNP的催化沉淀,使电极阻抗层层增大,从而达到信号放大的效果,成功构建了一种电化学免疫传感器并应用于实际水体环境中DBP的检测。研究结果表明,在最优实验条件下,当DBP浓度在5~500 ng mL-1浓度范围内,ECL信号强度与DBP浓度的对数呈良好的线性关系,3倍噪声时其检测限(LOD)为7.0 ng mL-1。与传统ELISA方法(LOD,70.0 ng mL-1)相比具有更高的灵敏度。该电化学免疫传感器耐受性良好,被成功应用于实际环境中DBP的分析。(2)目前缺乏对四溴双酚A(Tetrabromobisphenol-A,TBBPA)衍生物TBBPA-DHEE的快速检测方法,多为复杂的仪器检测,而此种化合物被认为是目前毒性最强的衍生物。因此,建立一种简单、灵敏、可靠的分析方法,对其在环境中的污染特征进行调查,继而评估其生态风险非常必要。基于此,本研究合成了一种新的半抗原,制备了可同时识别TBBPA-DHEE和TBBPA MHEE(TBBPA副产物)的多克隆抗体,并以此建立了高灵敏的酶联免疫分析方法(ELISA)。该方法的检测限可达0.7018 ng m L-1,有较好的准确度(回收率,83-133%)。以所建方法,对环境水体中的此种污染物进行了分析,并首次发现了环境水体中TBBPA-DHEE的存在。(3)针对TBBPA-DHEE,使用上述多克隆抗体,建立了一种高灵敏化学发光免疫分析方法(Chemical luminescence immunoassay assay,CLEIA)。该方法的检测限为0.8010 ng mL-1,IC50值为8.9985 ng mL-1。以所建方法,对环境水体中的此种污染物进行了分析。并与建立的ELISA进行了对比,结果基本相符。本研究为后续针对TBBPA-DHEE进行化学发光研究提供了参考数据。
[Abstract]:New organic pollutants are synthetic or natural substances that have not yet been effectively detected in the environment, but have the potential to enter the environment and cause known or suspicious ecotoxicological and health effects. Including antibiotics, personal care products, surfactants, artificial sweeteners, pesticides and various industrial additives. These organic pollutants are widespread in the environment, but lack complete ecotoxicological data. The chromatographic analysis method is generally used for this kind of pollutants. This method has good accuracy and high sensitivity. But the sample pretreatment is complicated, the instrument used is more expensive, and can not achieve rapid analysis. On the contrary, the biological analysis method is simple, easy to operate, without complex sample processing, and can be quickly analyzed with less sample consumption. In this paper, two new organic pollutants, dibutyl phthalate DBP (dibutyl phthalate DBP) and Tetrabromobisphenol-A (2-hydroxyetyldimethylethyl) ether (Tetrabromobisphenol-A bisol), TBBPA-DHEE), were selected. A sensitive, simple and reliable analytical method was constructed and successfully applied to the analysis of environmental samples. The main contents are as follows: (1) an AC impedance electrochemical immunosensor is established for DBP by indirect competition. In this study, MWCNTs@GONRs nanocomposites and DBP antigens and antibodies were used to modify GCEs, and NADH was used to promote the catalytic precipitation of AuNP, which increased the electrode impedance layer by layer, thus achieving the effect of signal amplification. An electrochemical immunosensor was successfully constructed and applied to the detection of DBP in real water environment. The results showed that under the optimal experimental conditions, the detection limit was 7.0 ng mL ~ (-1) when the signal intensity of DBP was 3 times as much as the logarithm of DBP concentration in the range of 5 ~ 500ng mL-1. Compared with the conventional ELISA method, it has a higher sensitivity than that of the traditional ELISA method (70.0 ng ml -1). The electrochemical immunosensor has good tolerance and has been successfully applied to the analysis of DBP in practical environment. At present, it lacks a rapid method for the detection of tetrabromobisphenol A (Tetrabromo bisphenol A) derivative TBBPA-DHEE, most of which are complex instrument detection. This compound is considered to be the most toxic derivative at present. Therefore, it is necessary to establish a simple, sensitive and reliable analytical method to investigate its pollution characteristics in the environment and then evaluate its ecological risk. Based on this, a new hapten was synthesized and polyclonal antibodies which could recognize both TBBPA-DHEE and TBBPA MHEE(TBBPA by-products were prepared, and a highly sensitive enzyme-linked immunosorbent assay (Elisa) was established. The detection limit of this method is up to 0.7018 ng mL ~ (-1), and the method has good accuracy (recovery rate is 83-133). Based on the established method, this pollutant in environmental water was analyzed, and the existence of TBBPA-DHEE in environmental water was found for the first time. Aiming at TBBPA-DHEE, a highly sensitive chemiluminescence immunoassay method was developed for chemical luminescence immunoassay assayay CLEIAA (CLEIAA) based on the above polyclonal antibodies. The detection limit of this method was 0.8010 ng mL -1 and IC50 was 8.9985 ng ml -1. Based on the established method, this pollutant in environmental water was analyzed. The results are in good agreement with the established ELISA. This study provides reference data for chemiluminescence studies on TBBPA-DHEE.
【学位授予单位】：江苏大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：X830

【参考文献】