直接竞争酶联免疫法测定呋喃妥因代谢物

Determination of nitrofurantion metabolite by direct competitive en-zyme-linked immunosorbent assay

中文关键词:  呋喃妥因代谢物  直接竞争酶联免疫检测法  动物组织

英文关键词:nitrofurantoin metabolite  direct competitive enzyme-linked immunosorbent assay  animal tissues

中文摘要:

      目的 建立动物组织中呋喃妥因代谢物1-氨基乙内酰脲的直接竞争酶联免疫(ELISA)检测法, 为检测食品中呋喃妥因代谢物残留提供快速检测的方法。方法 采用活化酯法制备辣根过氧化物酶标记抗原, 并用棋盘法确定酶标抗原和抗体的稀释度, 优化反应条件, 建立检测呋喃妥因代谢物的直接竞争酶联免疫法, 并对其进行方法学评价。结果 该方法的线性方程为Y=-34.723X+158.01(r2=0.9922), 线性检测范围为0.177~11.508 ng/mL, IC50为1.291 ng/mL; 猪肉、鸡肉、鱼肉的最低检测限分别为0.115、0.113、0.109 μg/kg, 加标回收率在82.6%~104.7%之间, 批内变异系数在3.6%~9.3%之间, 批间变异系数在4.3%~12.4%之间。结论 本方法快速准确, 适用于动物组织中呋喃妥因代谢物的检测。

英文摘要:

      Objective To develop a direct competitive enzyme-linked immunosorbent assay (ELISA) for the detection of nitrofurantion metabolite in animal tissues. Methods The horseradish peroxidase labeled antigen was prepared by activated ester method. The dilutions of the enzyme labeled antigen and monoclonal antibody were determined by chequerboard titration, and the reaction conditions were optimized to establish the direct competitive enzyme-linked immunoassay for the detection of nitrofurantion metabolite. Then the assay was evaluated. Results The regression equation was Y=-34.723X+158.01(r2=0.9922) with a linear detection ranging from 0.177 to 11.508 ng/mL. The IC50 was 1.291 ng/mL. The limit of detection (LOD) in pork, chicken and fish samples was 0.115, 0.113 and 0.109 μg/kg, respectively. Recoveries were between 82.6%~104.7% when AHD was spiked to samples. The coefficient of variation of intra-and inter-assay was 3.6%~9.3% and 4.3%~12.4%. Conclusion The ELISA method is fast and accurate, and suitable for the detection of nitrofurantion metabolite residue in animal tissues.