作　　者： ; ; ;

机构地区： 武汉工业学院食品科学与工程学院

出　　处： 《分析化学》 2009年第4期527-531,共5页

摘　　要： 以自制特异性抗体为核心试剂,建立了以苯甲醛为衍生试剂的呋喃唑酮残留标示物间接竞争ELISA方法。通过方阵滴定法和间接竞争法确定ELISA方法最佳反应条件:抗原最佳包被浓度200μg/L,抗体最佳稀释倍数1∶2.5×105,抗体与药物最佳工作配比40μL∶60μL,最佳竞争时间1h。检出限为0.1μg/L,检测范围0.1～25.6μg/L,线性关系良好。在空白组织中(猪肌肉、猪肝脏、鸡肌肉、鸡肝脏、鱼肉)添加0.4、1.0和5.0μg/kgAOZ,回收率55.8%～96.6%,相对标准偏差均小于20%。测定20份不同组织的空白样品,方法的检出限为0.4～0.5μg/kg。以100mg/kg呋喃唑酮饲喂动物,其组织样品同时经HPLC方法和本方法测定,数据表明本方法具有实际检出能力,测定结果与仪器方法的测定结果相近。通过与德国同类试剂盒比较实验表明,本方法的灵敏度、精密度、准确度接近于进口试剂盒水平,可用于动物可食性组织肝脏和肌肉中AOZ筛选。 Based on the specific polyclonal antibody, an indirect competitive enzyme-linked immunosorbent assay （ie-ELISA） was developed to detect 3-amino-2-oxazolidone（AOZ）, following derivatisation with benzaldehyde. The selection of the optimum ELISA conditions was performed by ic-ELISA and checkbroad. The results of the experiments showed the coating antigen concentration of 200 g/L, the polyclonal antibody dilutions of 1：2.5 × 10^5 and the competition reaction time of 1 h were needed to get to the linear portion of the response curve. Calibration curve was prepared for 3-[ N-phenyl-methylene]-amino-2-oxazolidinone（PAOZ）, a benzaldehyde derivative of AOZ, and good linearity was achieved over the concentration （0. 1 - 25.6 μg/L）. The ic-ELISA method showed lowest detection limits of 0.1 μg/L for PAOZ. Recoveries of AOZ fortified at the levels of 0.4, 1.0 and 5.0 μg/kg ranged from 55.8% to 96.6% in the tissues（swine liver, swine muscle, chicken liver, chicken muscle and fish muscle）. The interassay RSD was 〈 20%. The limits of detection calculated from the analysis of 20 known negative tissues were 0.4 - 0.5 μg/L for AOZ. The same samples obtained from the pigs treated with feed containing 100 mg/kg furazolidone were subjected to this ELISA procedure and HPLC analysis. The results implied a good agreement between AOZ concentrations determined by two methods and proved the reliability of the ELISA for the determination of real samples. The sensitivity, precision and accuracy of this method were coincident with that of Germanic kit. The results showed that this ELISA method was reliable for screening residues of AOZ in animal edible tissues.

关 键 词： 呋喃唑酮 氨基 恶唑烷酮 免疫分析 动物组织 残留

领　　域： [轻工技术与工程] [农业科学] [农业科学] [农业科学]