發(fā)布時間：2018-12-13 19:30

【摘要】：傳統(tǒng)的膠體金免疫層析方法主要是通過肉眼觀察檢測線的有無來判斷檢測結(jié)果,對待測物進行定性或者半定量檢測,檢測結(jié)果易受溫度、光線等環(huán)境因素和檢測者的主觀因素影響。基于膠體金光學(xué)讀取儀的膠體金免疫層析快速定量檢測方法具有速度快、操作簡便、靈敏度高、可定量檢測、數(shù)據(jù)可存儲、適合現(xiàn)場快速篩查等優(yōu)點。本研究旨在建立膠體金免疫層析快速定量檢測平臺并初步探討影響其穩(wěn)定性的主要因素,為推廣膠體金免疫層析快速定量檢測方法的應(yīng)用提供理論依據(jù)和基礎(chǔ)數(shù)據(jù)。近年來,鹽酸氯丙那林作為一種新型的“瘦肉精”使用,該藥物在動物體內(nèi)代謝緩慢且具有蓄積毒性,通過食物鏈的傳遞易對人類的健康造成危害。本研究成功建立了基于HG-8膠體金光學(xué)讀取儀快速定量檢測豬尿中氯丙那林殘留的方法。通過優(yōu)化實驗參數(shù),該方法能在9分鐘內(nèi)快速分析檢測尿樣中的氯丙那林,最低檢測限為0.15μg/L,在3.0μg/L和20.0μg/L之間有良好的線性關(guān)系(R2=0.997),與其它5種β2-受體激動劑沒有明顯的交叉反應(yīng)。為了驗證本方法的準(zhǔn)確性,用本方法與高效液相串聯(lián)質(zhì)譜法同時檢測20個陰性豬尿加標(biāo)樣品,兩種方法的檢測結(jié)果有良好的相關(guān)性。本方法具有便捷靈敏、特異性強的特點,能在現(xiàn)場快速檢測尿樣中的氯丙那林殘留。在動物養(yǎng)殖業(yè)中,鹽酸克倫特羅、萊克多巴胺和沙丁胺醇等三種β2-受體激動劑(β2-agonist)藥物常用于促進動物生長及提高飼養(yǎng)動物的瘦肉率,本研究以克倫特羅、萊克多巴胺、沙丁胺醇等三種膠體金免疫層析快速定量檢測試紙條為對象,通過60℃加速保存實驗得出影響膠體金定量試紙條穩(wěn)定性的主要因素是質(zhì)控線(C線)的穩(wěn)定性。萊克多巴胺試紙條相比于其它兩種試紙條的穩(wěn)定性較好,而克倫特羅和沙丁胺醇試紙條在加速保存實驗中檢測線(T線)穩(wěn)定C線下降明顯,導(dǎo)致T/C上升顯著。本研究分別在克倫特羅和沙丁胺醇兩種試紙條的二抗稀釋液中均添加5%(v/v)的WellChampion抗體保護劑,NC膜在60℃條件下干燥24小時,兩種試紙條的穩(wěn)定性均得到了有效提高。同時,本研究建立了膠體金免疫層析快速定量檢測平臺,檢測豬尿中克倫特羅、萊克多巴胺、沙丁胺醇三種β2-受體激動劑的最低檢測限分別為0.19μg/L、0.39μg/L和0.83μg/L,回收率在86.6%~109.7%之間,變異系數(shù)為4.7%~14.7%;檢測豬肉中三種目標(biāo)物的最低檢測限分別為0.43μg/kg、0.52μg/kg和1.01μg/kg,回收率為71.2%~101.6%,變異系數(shù)為7.2%~17.9%。對不同地區(qū)的135個不同豬尿樣本和江西國鴻食品有限公司的710個豬肉樣品進行盲檢,結(jié)果都為陰性,說明目前克倫特羅、萊克多巴胺、沙丁胺醇等3種常用β2-受體激動劑在動物養(yǎng)殖業(yè)中得到了有效控制。
[Abstract]:The traditional colloidal gold immunochromatographic method is mainly to judge the test results by observing the existence or not of the detection line with the naked eye, and to carry out qualitative or semi-quantitative tests on the test objects. The test results are susceptible to temperature. Environmental factors such as light and subjective factors of the examiner. The rapid quantitative detection method of colloidal gold immunochromatography based on colloidal gold optical reader has the advantages of high speed, simple operation, high sensitivity, quantitative detection, data storage, and suitable for field rapid screening. The purpose of this study was to establish a rapid quantitative detection platform for colloidal gold immunochromatography and to explore the main factors affecting its stability, and to provide theoretical basis and basic data for popularizing the rapid quantitative detection method of colloidal gold immunochromatography. In recent years, clopramine hydrochloride has been used as a new type of "lean meatloin". The drug is metabolized slowly and has accumulative toxicity in animals. It is harmful to human health through the transmission of food chain. A rapid and quantitative method for the determination of chlorpromazine residues in porcine urine was established based on HG-8 colloidal gold optical reader. By optimizing the experimental parameters, the method can be used for rapid analysis and detection of clopramine in urine samples within 9 minutes. The minimum detection limit is 0.15 渭 g / L, and the linear relationship between 3.0 渭 g / L and 20.0 渭 g / L is good (R2P 0.997). There was no obvious cross reaction with other 5 尾 2 receptor agonists. In order to verify the accuracy of this method, 20 negative pig urine samples were simultaneously detected by this method and high performance liquid chromatography tandem mass spectrometry. The results of the two methods have good correlation. The method is convenient, sensitive and specific, and can be used for rapid detection of chlorpromazine residues in urine samples. In animal husbandry, clenbuterol hydrochloride, ractopamine and salbutamol are three kinds of 尾 _ 2-receptor agonists (尾 _ 2-agonist), which are often used to promote the growth of animals and increase the lean meat rate. Ractopamine, salbutamol and other three kinds of colloidal gold immunochromatography were used for rapid quantitative detection of strip. The stability of quality control line (C line) was found to be the main factor affecting the stability of colloidal gold quantitative strip by accelerated preservation at 60 鈩，

本文編號：2377090