水環(huán)境中小分子污染物高靈敏檢測(cè)方法研究
發(fā)布時(shí)間：2019-07-01 10:41
【摘要】：水環(huán)境質(zhì)量的好壞與人們健康有重要的關(guān)系,隨著工農(nóng)業(yè)的快速發(fā)展和人們生活水平的不斷提高,持久性有機(jī)物(POPs)、“三致”物質(zhì)和內(nèi)分泌干擾物(EDCs)等殘留小分子污染物質(zhì)污染水體現(xiàn)象日益嚴(yán)重。環(huán)境標(biāo)準(zhǔn)中對(duì)殘留污染物的殘留量具有嚴(yán)格的限制要求,常規(guī)的檢測(cè)方法如色譜法、光譜法、化學(xué)發(fā)光分析等檢測(cè)方法由于檢測(cè)精度、操作復(fù)雜、檢測(cè)費(fèi)用昂貴等難以滿足實(shí)時(shí)在線檢測(cè),研究微量有毒有機(jī)污染物的現(xiàn)場(chǎng)檢測(cè)技術(shù)對(duì)有效預(yù)防和控制水環(huán)境污染具有重要意義。倏逝波光纖生物傳感器(EWA)是一種基于全內(nèi)反射和熒光免疫分析原理的光纖免疫傳感檢測(cè)系統(tǒng),采用單多模光纖耦合器和組合錐型結(jié)構(gòu)光纖免疫傳感探針的新型設(shè)計(jì)結(jié)構(gòu),該系統(tǒng)能夠?qū)崿F(xiàn)快速、準(zhǔn)確定量檢測(cè)水環(huán)境中的特征性污染物質(zhì)。本研究基于EWA及其檢測(cè)原理,建立了功能型光纖免疫傳感探針的修飾方法,制備出高載量、高穩(wěn)定性、可重復(fù)使用的三聚氰胺光纖免疫傳感探針和磺胺二甲嘧啶光纖免疫傳感探針。在此基礎(chǔ)上,成功研制了間接競(jìng)爭(zhēng)反應(yīng)模式下水環(huán)境中三聚氰胺和磺胺二甲嘧啶的高靈敏檢測(cè)方法,可以實(shí)現(xiàn)快速、簡(jiǎn)便、有效的定量檢測(cè)。研究結(jié)果表明:(1)本研究制備的三聚氰胺光纖免疫傳感探針和磺胺二甲嘧啶光纖免疫傳感探針?lè)謩e對(duì)三聚氰胺抗體和磺胺二甲嘧啶抗體具有很好的特異性響應(yīng),對(duì)其他抗體的非特異性吸附很弱,且光纖免疫傳感探針的穩(wěn)定性強(qiáng)。(2)倏逝波光纖生物傳感器對(duì)三聚氰胺和磺胺二甲嘧啶的檢測(cè)標(biāo)準(zhǔn)曲線均符合數(shù)學(xué)函數(shù)Logistic模型,模擬的擬合度R2≥0.99,三聚氰胺的定量檢測(cè)區(qū)間為12.62μg/L~284.18μg/L,檢出限為5.14μg/L,IC50值為59.89μg/L;磺胺二甲嘧啶的定量檢測(cè)區(qū)間0.17μg/L~10.73μg/L,檢出限為0.05μg/L,IC50值為1.35μg/L。實(shí)際樣品的加標(biāo)回收率在86%~116%之間,樣品檢測(cè)的變異系數(shù)小于4%,與LC-MS/MS的檢測(cè)結(jié)果相比具有很好的線性相關(guān)性。三聚氰胺抗體/磺胺二甲嘧啶抗體對(duì)三聚氰胺/磺胺二甲嘧啶結(jié)構(gòu)類似物的交叉反應(yīng)性小。(3)光纖免疫傳感探針的可重復(fù)性能測(cè)試結(jié)果表明,光纖免疫傳感探針可重復(fù)使用檢測(cè)300次,檢測(cè)信號(hào)沒有明顯下降,精密度為1.2%。光纖免疫傳感探針的多次重復(fù)使用大大降低了檢測(cè)成本。研究表明,本研究的水環(huán)境中小分子污染物的高靈敏檢測(cè)方法能夠滿足實(shí)際水環(huán)境中三聚氰胺和磺胺二甲嘧啶的檢測(cè)要求,倏逝波光纖生物傳感器具有良好的精確度和穩(wěn)定性,可以實(shí)現(xiàn)實(shí)時(shí)在線、快速、準(zhǔn)確檢測(cè)環(huán)境污染物的技術(shù)要求,本研究成果利于倏逝波光纖生物傳感器在水環(huán)境監(jiān)測(cè)領(lǐng)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.
【學(xué)位授予單位】：內(nèi)蒙古工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：X832
本文編號(hào)：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.
【學(xué)位授予單位】：內(nèi)蒙古工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：X832
【引證文獻(xiàn)】
相關(guān)會(huì)議論文 前1條
1 杜兆林;鄭彤;劉麗艷;王鵬;白楊;秦世麗;李一凡;;污水處理廠中抗生素去除情況研究[A];持久性有機(jī)污染物論壇2011暨第六屆持久性有機(jī)污染物全國(guó)學(xué)術(shù)研討會(huì)論文集[C];2011年
，本文編號(hào)：2508406
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