胶体金免疫层析法快速定量检测猪尿中β2-受体激动剂残留的研究

发布时间：2018-12-13 19:30

【摘要】：传统的胶体金免疫层析方法主要是通过肉眼观察检测线的有无来判断检测结果,对待测物进行定性或者半定量检测,检测结果易受温度、光线等环境因素和检测者的主观因素影响。基于胶体金光学读取仪的胶体金免疫层析快速定量检测方法具有速度快、操作简便、灵敏度高、可定量检测、数据可存储、适合现场快速筛查等优点。本研究旨在建立胶体金免疫层析快速定量检测平台并初步探讨影响其稳定性的主要因素,为推广胶体金免疫层析快速定量检测方法的应用提供理论依据和基础数据。近年来,盐酸氯丙那林作为一种新型的“瘦肉精”使用,该药物在动物体内代谢缓慢且具有蓄积毒性,通过食物链的传递易对人类的健康造成危害。本研究成功建立了基于HG-8胶体金光学读取仪快速定量检测猪尿中氯丙那林残留的方法。通过优化实验参数,该方法能在9分钟内快速分析检测尿样中的氯丙那林,最低检测限为0.15μg/L,在3.0μg/L和20.0μg/L之间有良好的线性关系(R2=0.997),与其它5种β2-受体激动剂没有明显的交叉反应。为了验证本方法的准确性,用本方法与高效液相串联质谱法同时检测20个阴性猪尿加标样品,两种方法的检测结果有良好的相关性。本方法具有便捷灵敏、特异性强的特点,能在现场快速检测尿样中的氯丙那林残留。在动物养殖业中,盐酸克伦特罗、莱克多巴胺和沙丁胺醇等三种β2-受体激动剂(β2-agonist)药物常用于促进动物生长及提高饲养动物的瘦肉率,本研究以克伦特罗、莱克多巴胺、沙丁胺醇等三种胶体金免疫层析快速定量检测试纸条为对象,通过60℃加速保存实验得出影响胶体金定量试纸条稳定性的主要因素是质控线(C线)的稳定性。莱克多巴胺试纸条相比于其它两种试纸条的稳定性较好,而克伦特罗和沙丁胺醇试纸条在加速保存实验中检测线(T线)稳定C线下降明显,导致T/C上升显著。本研究分别在克伦特罗和沙丁胺醇两种试纸条的二抗稀释液中均添加5%(v/v)的WellChampion抗体保护剂,NC膜在60℃条件下干燥24小时,两种试纸条的稳定性均得到了有效提高。同时,本研究建立了胶体金免疫层析快速定量检测平台,检测猪尿中克伦特罗、莱克多巴胺、沙丁胺醇三种β2-受体激动剂的最低检测限分别为0.19μg/L、0.39μg/L和0.83μg/L,回收率在86.6%~109.7%之间,变异系数为4.7%~14.7%;检测猪肉中三种目标物的最低检测限分别为0.43μg/kg、0.52μg/kg和1.01μg/kg,回收率为71.2%~101.6%,变异系数为7.2%~17.9%。对不同地区的135个不同猪尿样本和江西国鸿食品有限公司的710个猪肉样品进行盲检,结果都为阴性,说明目前克伦特罗、莱克多巴胺、沙丁胺醇等3种常用β2-受体激动剂在动物养殖业中得到了有效控制。
[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，

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