水环境中小分子污染物高灵敏检测方法研究
发布时间：2019-07-01 10:41
【摘要】：水环境质量的好坏与人们健康有重要的关系,随着工农业的快速发展和人们生活水平的不断提高,持久性有机物(POPs)、“三致”物质和内分泌干扰物(EDCs)等残留小分子污染物质污染水体现象日益严重。环境标准中对残留污染物的残留量具有严格的限制要求,常规的检测方法如色谱法、光谱法、化学发光分析等检测方法由于检测精度、操作复杂、检测费用昂贵等难以满足实时在线检测,研究微量有毒有机污染物的现场检测技术对有效预防和控制水环境污染具有重要意义。倏逝波光纤生物传感器(EWA)是一种基于全内反射和荧光免疫分析原理的光纤免疫传感检测系统,采用单多模光纤耦合器和组合锥型结构光纤免疫传感探针的新型设计结构,该系统能够实现快速、准确定量检测水环境中的特征性污染物质。本研究基于EWA及其检测原理,建立了功能型光纤免疫传感探针的修饰方法,制备出高载量、高稳定性、可重复使用的三聚氰胺光纤免疫传感探针和磺胺二甲嘧啶光纤免疫传感探针。在此基础上,成功研制了间接竞争反应模式下水环境中三聚氰胺和磺胺二甲嘧啶的高灵敏检测方法,可以实现快速、简便、有效的定量检测。研究结果表明:(1)本研究制备的三聚氰胺光纤免疫传感探针和磺胺二甲嘧啶光纤免疫传感探针分别对三聚氰胺抗体和磺胺二甲嘧啶抗体具有很好的特异性响应,对其他抗体的非特异性吸附很弱,且光纤免疫传感探针的稳定性强。(2)倏逝波光纤生物传感器对三聚氰胺和磺胺二甲嘧啶的检测标准曲线均符合数学函数Logistic模型,模拟的拟合度R2≥0.99,三聚氰胺的定量检测区间为12.62μg/L~284.18μg/L,检出限为5.14μg/L,IC50值为59.89μg/L;磺胺二甲嘧啶的定量检测区间0.17μg/L~10.73μg/L,检出限为0.05μg/L,IC50值为1.35μg/L。实际样品的加标回收率在86%~116%之间,样品检测的变异系数小于4%,与LC-MS/MS的检测结果相比具有很好的线性相关性。三聚氰胺抗体/磺胺二甲嘧啶抗体对三聚氰胺/磺胺二甲嘧啶结构类似物的交叉反应性小。(3)光纤免疫传感探针的可重复性能测试结果表明,光纤免疫传感探针可重复使用检测300次,检测信号没有明显下降,精密度为1.2%。光纤免疫传感探针的多次重复使用大大降低了检测成本。研究表明,本研究的水环境中小分子污染物的高灵敏检测方法能够满足实际水环境中三聚氰胺和磺胺二甲嘧啶的检测要求,倏逝波光纤生物传感器具有良好的精确度和稳定性,可以实现实时在线、快速、准确检测环境污染物的技术要求,本研究成果利于倏逝波光纤生物传感器在水环境监测领域的普及使用。
[Abstract]:The quality of water environmental quality has an important relationship with people's health. With the rapid development of industry and agriculture and people's living standard, persistent organic matter (POPs), "Three-to-one" and endocrine disruptors (EDCs) have become more and more serious. the method for detecting the residual pollutant in the environmental standard has the strict limitation requirement, and the conventional detection method such as chromatography, spectroscopy, chemiluminescence analysis and the like is difficult to meet the real-time on-line detection due to the detection precision, the complex operation, the high detection cost and the like, The on-site detection of trace toxic organic pollutants is of great significance to the effective prevention and control of water environmental pollution. The evanescent wave optical fiber biosensor (EWA) is an optical fiber immunosensing detection system based on the principle of total internal reflection and fluorescence immunoassay, And the characteristic pollutants in the water environment can be accurately and quantitatively detected. In this study, based on the EWA and its detection principle, the modified method of functional fiber-optic immunosensor probe was established, and a high-load, high-stability, re-usable melamine fiber immunosensor probe and a sulfanilamide fiber-optic immunosensor probe were prepared. On this basis, the high-sensitivity detection method of melamine and sulfanilamide in the water environment of indirect competitive reaction is successfully developed, and the rapid, simple and effective quantitative detection can be realized. The results showed that: (1) The immunosensor probe of the melamine fiber prepared by this study and the immunosensor probe of the sulfanilamide and the sulfanilamide have a good specific response to the antibodies of the melamine and the sulfanilamide, and the non-specific adsorption of the other antibodies is very weak, And the optical fiber immunosensing probe has strong stability. (2) The standard curve of the detection of melamine and sulfanilamide in the evanescent wave fiber biosensor is in accordance with the logistic model of the mathematical function, the simulated fitting degree of R2 is 0.99, the quantitative detection interval of the melamine is 12.62. mu. g/ L to 284.18. m u.g/ L, the detection limit is 5.14. m u.g/ L, and the IC50 value is 59.89. m u.g/ L; The detection limit was 0.05ug/ L, and the IC50 value was 1.35. m u.g/ L. The average recovery of the actual sample was between 86% and 116%. The coefficient of variation of the sample was less than 4%, and it had a good linear correlation with the detection results of LC-MS/ MS. The cross-reactivity of the melamine antibody/ sulfanilamide antibody to the melamine/ sulfanilamide structure analogue is small. (3) The results of the repeatability of the fiber-optic immunosensor probe show that the fiber-optic immunosensor probe can be used repeatedly for 300 times, the detection signal is not significantly reduced, and the precision is 1.2%. The repeated use of the optical fiber immunosensing probe greatly reduces the detection cost. The research shows that the high-sensitivity detection method of small and medium-sized molecular contaminants in the water environment of the present study can meet the detection requirements of melamine and sulfanilamide in the actual water environment, and the evanescent wave fiber biosensor has good accuracy and stability, and can realize real-time on-line and rapid detection, The technical requirements of environmental pollutants are accurately detected, and the research results are beneficial to the popularization and application of the evanescent wave optical fiber biosensor in the field of water environmental monitoring.
【学位授予单位】：内蒙古工业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：X832
本文编号：2508406
[Abstract]:The quality of water environmental quality has an important relationship with people's health. With the rapid development of industry and agriculture and people's living standard, persistent organic matter (POPs), "Three-to-one" and endocrine disruptors (EDCs) have become more and more serious. the method for detecting the residual pollutant in the environmental standard has the strict limitation requirement, and the conventional detection method such as chromatography, spectroscopy, chemiluminescence analysis and the like is difficult to meet the real-time on-line detection due to the detection precision, the complex operation, the high detection cost and the like, The on-site detection of trace toxic organic pollutants is of great significance to the effective prevention and control of water environmental pollution. The evanescent wave optical fiber biosensor (EWA) is an optical fiber immunosensing detection system based on the principle of total internal reflection and fluorescence immunoassay, And the characteristic pollutants in the water environment can be accurately and quantitatively detected. In this study, based on the EWA and its detection principle, the modified method of functional fiber-optic immunosensor probe was established, and a high-load, high-stability, re-usable melamine fiber immunosensor probe and a sulfanilamide fiber-optic immunosensor probe were prepared. On this basis, the high-sensitivity detection method of melamine and sulfanilamide in the water environment of indirect competitive reaction is successfully developed, and the rapid, simple and effective quantitative detection can be realized. The results showed that: (1) The immunosensor probe of the melamine fiber prepared by this study and the immunosensor probe of the sulfanilamide and the sulfanilamide have a good specific response to the antibodies of the melamine and the sulfanilamide, and the non-specific adsorption of the other antibodies is very weak, And the optical fiber immunosensing probe has strong stability. (2) The standard curve of the detection of melamine and sulfanilamide in the evanescent wave fiber biosensor is in accordance with the logistic model of the mathematical function, the simulated fitting degree of R2 is 0.99, the quantitative detection interval of the melamine is 12.62. mu. g/ L to 284.18. m u.g/ L, the detection limit is 5.14. m u.g/ L, and the IC50 value is 59.89. m u.g/ L; The detection limit was 0.05ug/ L, and the IC50 value was 1.35. m u.g/ L. The average recovery of the actual sample was between 86% and 116%. The coefficient of variation of the sample was less than 4%, and it had a good linear correlation with the detection results of LC-MS/ MS. The cross-reactivity of the melamine antibody/ sulfanilamide antibody to the melamine/ sulfanilamide structure analogue is small. (3) The results of the repeatability of the fiber-optic immunosensor probe show that the fiber-optic immunosensor probe can be used repeatedly for 300 times, the detection signal is not significantly reduced, and the precision is 1.2%. The repeated use of the optical fiber immunosensing probe greatly reduces the detection cost. The research shows that the high-sensitivity detection method of small and medium-sized molecular contaminants in the water environment of the present study can meet the detection requirements of melamine and sulfanilamide in the actual water environment, and the evanescent wave fiber biosensor has good accuracy and stability, and can realize real-time on-line and rapid detection, The technical requirements of environmental pollutants are accurately detected, and the research results are beneficial to the popularization and application of the evanescent wave optical fiber biosensor in the field of water environmental monitoring.
【学位授予单位】：内蒙古工业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：X832
【引证文献】
相关会议论文 前1条
1 杜兆林;郑彤;刘丽艳;王鹏;白杨;秦世丽;李一凡;;污水处理厂中抗生素去除情况研究[A];持久性有机污染物论坛2011暨第六届持久性有机污染物全国学术研讨会论文集[C];2011年
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