本文選題：伏馬毒素 + 氯霉素��； 參考：《天津科技大學(xué)》2012年碩士論文

【摘要】：論文采用一種免疫親和凝膠柱對(duì)食品中伏馬毒素B1(FB1)和氯霉素(CAP)殘留的檢測(cè)進(jìn)行了系統(tǒng)研究,用溴化氫活化的瓊脂糖凝膠偶聯(lián)伏馬毒素抗體或氯霉素抗體制備其分別的抗體膠；偶聯(lián)辣根過(guò)氧化氫酶(HRP)抗體制備其HRP膠；將溴化氫活化的瓊脂糖凝膠封閉制備封閉膠。封閉膠與抗體膠混合制備成檢測(cè)層,與HRP膠混合制備成對(duì)照層,裝入檢測(cè)柱,并對(duì)影響實(shí)驗(yàn)的各個(gè)因素(離子強(qiáng)度,pH值等)進(jìn)行了優(yōu)化,建立了一種省時(shí)、靈敏度高、可視化的檢測(cè)方法,可以定性半定量的檢測(cè)食品中的伏馬毒素和氯霉素。 對(duì)于伏馬毒素B1免疫親和凝膠檢測(cè)柱：封閉膠與抗體膠以1：10比例混合可以達(dá)到良好的視覺(jué)顯色結(jié)果,封閉膠與抗體膠1：90混合制備其對(duì)照層,其FB1-HRP以1：8000稀釋用于凝膠檢測(cè)柱的檢測(cè),建立伏馬毒素免疫親和凝膠檢測(cè)柱的最低限量檢測(cè)限(LOD)為40μg/L。對(duì)啤酒、白酒、玉米粉、谷物牛奶中伏馬毒素殘留進(jìn)行了檢測(cè)：啤酒和白酒直接稀釋5倍；玉米粉加入PBS提取離心后稀釋5倍；谷物牛奶離心取上層溶液稀釋10倍。 對(duì)于氯霉素免疫親和凝膠檢測(cè)柱：封閉膠與抗體膠以1：20比例混合是可以達(dá)到良好的視覺(jué)顯色結(jié)果,封閉膠與抗體膠1：100混合制備其對(duì)照層,其CAP-HRP以1：20萬(wàn)稀釋用于凝膠檢測(cè)柱的檢測(cè),建立氯霉素免疫親和凝膠檢測(cè)柱的LOD為1μg/L。對(duì)牛奶、乳粉、魚(yú)塘水、蜂蜜中氯霉素的殘留進(jìn)行了檢測(cè)；牛奶和魚(yú)塘水可以直接進(jìn)行檢測(cè)；乳粉5倍加入PBS溶解；蜂蜜5倍稀釋。 該免疫親和凝膠柱檢測(cè)食品樣品不需要任何有機(jī)溶劑和復(fù)雜的前處理,及大型儀器的輔助,添加3個(gè)不同濃度的伏馬毒素(氯霉素)標(biāo)品,用ELISA方法進(jìn)行了驗(yàn)證,檢測(cè)結(jié)果一致,表明試驗(yàn)所建立的方法具有很好的準(zhǔn)確性,滿足快速檢測(cè)的要求。
[Abstract]:The detection of FB1 and CAP residues in food was systematically studied by an immuno-affinity gel column. The agarose gel coupled with fumaroxin antibody or chloramphenicol antibody was used to prepare its respective antibody glue. Horseradish catalase (HRP) antibody was coupled to prepare the HRP gel, and the agarose gel activated by hydrogen bromide was blocked to prepare the sealant. The sealant and antibody gel were mixed to prepare the detection layer, and the HRP gel was mixed to form the contrast layer. The test column was mounted. The factors affecting the experiment (ionic strength, pH value, etc.) were optimized, and a time-saving and high sensitivity was established. Visual detection method can be used to detect fumaroxin and chloramphenicol in food qualitatively and quantitatively. For Fumatoxin B1 immuno-affinity gel column: the mixture of sealant and antibody glue at 1:10 can achieve good visual color results, and the control layer was prepared by mixing sealant and antibody glue 1:90. The FB1-HRP was diluted with 1: 8000 to detect the gel column. The minimum detection limit (LOD) of the FB1-HRP gel column was 40 渭 g 路L ~ (-1) 路L ~ (-1). The residue of fumarin in beer, liquor, cornmeal and cereal milk was determined: beer and liquor were diluted 5 times directly, corn flour was diluted by 5 times after PBS extraction, and the upper layer solution of cereal milk was diluted by 10 times. For chloramphenicol immunoaffinity gel detection column: the mixture of sealant and antibody glue at 1:20 can achieve good visual color result, and the control layer was prepared by mixing the sealant and antibody glue 1: 100. The CAP-HRP was diluted at 1: 200 000 to detect the gel column. The LOD of the established gel column was 1 渭 g / L. The residues of chloramphenicol in milk, milk powder, fish pond water and honey were detected; milk and fish pond water could be directly detected; milk powder was dissolved by PBS; honey was diluted by 5 times. The immunoaffinity gel column was used to detect food samples without any organic solvent, complex pretreatment, and the aid of large scale instrument. Three different concentrations of chloramphenicol (chloramphenicol) were added to food samples, and the results were verified by Elisa. The results show that the proposed method has good accuracy and meets the requirement of rapid detection.
【學(xué)位授予單位】：天津科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R155.5

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