该文旨在建立一种胶体金免疫层析技术同步测定猪肉和鸡肉组织中呋喃唑酮代谢物、呋喃西林代谢物、氯霉素药物残留的方法。动物肌肉组织中呋喃唑酮代谢物、呋喃西林代谢物、氯霉素经酸化、衍生化及乙酸乙酯提取,并经含吐温-20的Tris-HCl缓冲液稀释后,与胶体金标记的单克隆抗体结合,抑制抗体和硝酸纤维素膜检测线(T线)上的抗原结合,从而导致T线颜色变化。通过比较T线和质控线(C线)颜色深浅,对试样中多兽药残留进行定性判定。试验优化了pH值、抗体添加量、抗原包被量、吐温-20添加量、衍生化试剂浓度和冻干保护剂种类等关键因素,并对检测卡的灵敏度、特异性和稳定性进行检测。结果显示,所建立的免疫层析方法可在35 min内检出猪肉和鸡肉中呋喃西林代谢物、呋喃唑酮代谢物和氯霉素药物,检出限分别为0.5、0.5、0.1 μg/kg,灵敏度≥98%,假阳性率≤2%,假阴性率≤2%,稳定性可达到1年,且与现有标准方法检测结果一致。该方法操作简便、特异性强、灵敏度和准确度高,检测时间短且稳定性好,可用于动物肌肉组织中多兽药残留的高通量快速检测。
The purpose of this paper was to develop a method for the high-throughput determination of furazolidone metabolites, nitrofural metabolites and chloramphenicol drug residues in pork and chicken tissues by colloidal gold immunochromatographic assay (GICA).Furazolidone metabolites, nitrofural metabolites and chloramphenicol drug residues in animal muscle tissues were acidified, derivatized and extracted by ethyl acetate solution and diluted by Tris-HCl buffer with Tween-20, binding to colloidal gold-labeled monoclonal antibodies and inhibiting the binding of antibodies to the antigen on nitrocellulose membrane test line (T-line), which results in a change of the color of T-line.The qualitative determination of poly-veterinary drug residues in the samples was made by comparing the color shades of the T-line and control line (C-line).This test optimized the key factors such as pH value, antibody addition, antigen encapsulation, Tween-20 addition, density of derivatization reagent and type of lyophilized protectant.The results showed that the established immunochromatographic method could detect furazolidone metabolites, nitrofural metabolites, and chloramphenicol in pork and chicken in 35 minutes, and the limit of detection were 0.5, 0.5, 0.1 μg/kg, respectively. The sensitivity ≥98%, the false positive rate ≤2%, the false negative rate ≤2%, the stability can reach 1 year and the method is consistent with the results detected by the existing standard method.The method was simple with strong specificity, high sensitivity, high precision, short detection time and good stability.It could be used for high-throughput rapid detection of multi-veterinary drug residues in animal muscle tissue.
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