本文選題：獸藥　切入點：非法添加物　出處：《天津科技大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：本文選擇量子點作為熒光標(biāo)記材料,研制了喹諾酮類獸藥量子點標(biāo)記免疫檢測試紙條和非法添加物三聚氰胺量子點標(biāo)記免疫檢測試紙條,該方法可用于實際樣品的快速檢測。采用活化酯法制備喹諾酮類獸藥包被原(NOR-OVA)。喹諾酮類獸藥量子點標(biāo)記免疫檢測試紙條最佳工作條件為:量子點標(biāo)記抗體偶聯(lián)反應(yīng)的最佳pH值為8.4;量子點(QDs)與活化劑(EDC)與抗體的摩爾比為1:2000:10;包被原的稀釋倍數(shù)為1:40;二抗的稀釋倍數(shù)為1:30;標(biāo)準(zhǔn)品稀釋液為PBST (pH 7.4);工作液添加量為10 μL;硝酸纖維素膜型號為Milipore HF 135s; QDs-Ab偶聯(lián)物添加量為0.5 μL。該試紙條方法檢出限為2 ng/mL,檢測時間為10 min。與5種喹諾酮類獸藥環(huán)丙沙星、恩諾沙星、培氟沙星、氧氟沙星和洛美沙星,有明顯的交叉反應(yīng);與其它的10種喹諾酮類獸藥和5種常見的其它不同種類獸藥沒有明顯的交叉反應(yīng),特異性良好。實際樣品中檢測限為 10 ng/g。采用活化酯法合成三聚氰胺包被原(MEL-BSA)。三聚氰胺量子點標(biāo)記免疫檢測試紙條最佳工作條件為:量子點標(biāo)記抗體偶聯(lián)反應(yīng)的最佳pH值為8.4;量子點(QDs)與活化劑(EDC)與抗體的摩爾比為1:1500:15;包被原的稀釋倍數(shù)為1:80;二抗的稀釋倍數(shù)為1:140;標(biāo)準(zhǔn)品稀釋液為PBS (pH 7.4);工作液添加量為5μL;硝酸纖維素膜型號為Milipore HF 135s,用1%乳粉溶液封閉處理;QDs-Ab偶聯(lián)物添加量為1μL。該試紙條方法檢出限為100 ng/mL,檢測時間為10 min。結(jié)構(gòu)類似物中除了環(huán)丙氨嗪以外,都不存在交叉反應(yīng);與5種常見的其它不同種類獸藥也沒有明顯的交叉反應(yīng),特異性良好。實際樣品中檢測限為400 ng/g。本研究當(dāng)中所建立的喹諾酮類獸藥和三聚氰胺量子點標(biāo)記免疫檢測試紙條樣品前處理簡單、操作簡便、檢測時間短、特異性好,與傳統(tǒng)的膠體金試紙條相比靈敏度更高,能夠適用于現(xiàn)場大量樣品的快速檢測。
[Abstract]:In this paper, Quantum Dots (QDs) were selected as fluorescent labeling materials, and the QDs immunoassay strips for quinolones and the illegal additives, melamine QDs, were developed. This method can be used for rapid detection of real samples. Quinolones coated with NOR-OVAX were prepared by activation ester method. The best working conditions of QDIA strips for quinolones were as follows: QDD-labeled antibody coupling reaction. The optimum pH value is 8.4; the molar ratio of QDs to activator is 1: 2000: 10; the dilution factor of the coating is 1: 40; the dilution multiple of the second antibody is 1: 30; the dilution of the standard sample is PBST / pH 7.4; the working solution is 10 渭 L; the nitrocellulose membrane is coated with nitrocellulose. Model Milipore HF 135s, QDs-Ab conjugate amount 0.5 渭 L. the detection limit of the test strip is 2 ng / mL, the detection time is 10 min. with 5 kinds of quinolones, ciprofloxacin, Enrofloxacin, pefloxacin, ofloxacin and lomefloxacin had significant cross reactions, but not with 10 other quinolones and 5 other common different veterinary drugs. The detection limit in real samples is 10 ng / g. Melamine coated with MEL-BSAA is synthesized by activated ester method. The best working conditions of the test strip for immunological detection of melamine quantum dots are: QD labeled antibody coupling reaction. The optimum pH value is 8.4; the molar ratio of QDs to activator EDC) and antibody is 1: 1500: 15; the dilution factor of the coating is 1: 80; the dilution multiple of the second antibody is 1: 140; the dilution of the standard product is PBS pH 7.4; the working solution is 5 渭 L; the nitrocellulose membrane type is 1: 140; Milipore HF 135s, the addition amount of QDs-Ab conjugate was 1 渭 L. the detection limit of the method was 100ng / mL and the detection time was 10min. There was no cross reaction; there was no significant cross reaction with five other common veterinary drugs. The detection limit in the actual sample is 400 ng / g. The sample of quinolones and melamine quantum dot labeled immunoassay strip established in this study has the advantages of simple pretreatment, simple operation, short detection time and good specificity. Compared with the traditional colloidal gold test strip, it is more sensitive and can be used for rapid detection of a large number of samples in the field.
【學(xué)位授予單位】：天津科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S859.84

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 劉鳳山;;養(yǎng)殖業(yè)獸藥殘留現(xiàn)狀與對策[J];養(yǎng)殖與飼料;2016年09期

2 林黎;張毅;涂小珂;謝麗琪;岳振峰;康海寧;吳衛(wèi)東;羅耀;;液相色譜-串聯(lián)質(zhì)譜法同時測定化妝品中的25種喹諾酮類藥物[J];色譜;2015年03期

3 孟哲;石志紅;呂運開;孫漢文;;超高效液相色譜-高分辨四級桿飛行時間質(zhì)譜法快速篩查乳制品中磺胺類與氟喹諾酮類藥物[J];分析化學(xué);2014年10期

4 鄧思維;鄧劍軍;王婷婷;王羽;康學(xué)軍;;納米纖維固相萃取-高效液相色譜-熒光檢測麻辣燙湯液中喹諾酮類藥物[J];分析化學(xué);2014年08期

5 周梅仙;周業(yè)飛;劉文琪;華春;;高效毛細(xì)管電泳法分離檢測雞蛋中3種喹諾酮類抗菌藥[J];西北農(nóng)業(yè)學(xué)報;2013年03期

6 余從田;;我國食品安全問題產(chǎn)生的原因與對策分析[J];食品工業(yè);2012年06期

7 吳詩聰;;左氧氟沙星注射液致不良反應(yīng)24例分析[J];當(dāng)代醫(yī)學(xué);2012年01期

8 楊玉秀;黃和;謝麗玲;黃國方;曾丹丹;羅林;彭樹峰;;高效液相色譜質(zhì)譜聯(lián)用法測定飼料中的三聚氰胺[J];飼料廣角;2011年22期

9 李昊;唐鏡明;;89例左氧氟沙星注射液不良反應(yīng)病例分析[J];中國衛(wèi)生產(chǎn)業(yè);2011年31期

10 陳潔;;加替沙星注射液的不良反應(yīng)分析[J];基層醫(yī)學(xué)論壇;2011年31期

相關(guān)碩士學(xué)位論文 前3條

1 張蕾;水產(chǎn)品中呋喃唑酮殘留的免疫熒光快速檢測技術(shù)的研究[D];上海師范大學(xué);2014年

2 李春媛;三聚氰胺單克隆抗體制備及其在ELISA檢測中的應(yīng)用[D];吉林大學(xué);2009年

3 成煒;黃曲霉毒素B_1金標(biāo)免疫斑點檢測方法研究[D];華中農(nóng)業(yè)大學(xué);2008年

，

本文編號：1574183