金刚烷胺残留化学发光酶免疫法的建立

Establishment of Chemiluminescent Enzyme Immunoassay for Determination of Amantadine Residues

建立金刚烷胺残留的化学发光酶免疫方法。对酶标抗原与磁标抗体的最佳稀释倍数、反应时间、底物发光时间进行优化,建立标准曲线,同时对方法的检测限、准确度和精密度进行评价。结果表明:50%抑制浓度(IC50)为0.481μg/L,线性范围是0.085~2.729μg/L;在动物组织、兽药、饲料中的检测限为0.1μg/kg,样本添加回收率为80.2%~94.5%,批内、批间相对标准偏差均<10%。该方法灵敏度高、操作简便,可广泛用于动物组织、兽药、饲料中金刚烷胺残留量的测定。

The aim of the study is to establish a method for determination of amantadine residues by chemiluminescent enzyme immunoassay( CLEIA).After optimizing the dilution multiple of enzyme - labeled antigen and magnetic labeled antibody, reaction time and substrate luminescence time,a standard curve was established,and finally the detection limit,accuracy and precision of the method were evaluated.The results showed that the 50% inhibitory concentration( IC50 ) was 0.481 μg /L,and the linearity range was 0.085~2.729 μg /L.The detection limit in animal tissues,veterinary drugs and feed were 0.1 μg /kg.The recoveries ranged from 80.2%~94.5%,with the intra-assay and inter-assay coefficients of variation less than 10%.The method is sensitive,simple and it can be widely used for the determination of amantadine residues in animal tissues,veterinary drugs and feed.