化学发光酶免疫分析测定鱼肉中呋喃它酮代谢物方法研究  

Determination of Furaltadone Metabolite in Fish by Chemiluminescence Enzyme Immunoassay

作　　者：戴尽波[1] 徐振林[1] 刘凤银[1] 杨金易[1] 孙远明[1] 王弘[1] 雷红涛[1] 沈玉栋[1] 

机构地区：[1]华南农业大学食品学院,广东省食品质量安全重点实验室,广州510642

出　　处：《分析化学》2015年第6期871-875,共5页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(Nos.31371769,No.31271865);广东省科技计划项目(Nos.2012A020100002,No.2013B040402006);广州市科技计划项目(No.2013J4100053)资助~~

摘　　要：建立了呋喃它酮代谢物5-吗啉甲基-3-氨基-2-唑烷基酮（AMOZ）间接竞争化学发光酶免疫分析（ic CLEIA）检测方法,通过单因素实验优化了包被原浓度、抗体稀释倍数、反应缓冲体系及浓度、竞争反应时间等参数,结果表明ic CLEIA最佳反应条件为：包被抗原浓度为10 ng/m L,抗体稀释60000倍,最佳竞争时间为50 min,体系缓冲液0.01 mol/L PBS（p H 7.4）。在优化的条件下,本方法的线性检测范围为0.026~3.52μg/L,IC50为0.29μg/L,检出限（LOD,IC10）为0.012μg/L。对鱼肉样品的平均添加回收率在101.4%~115.5%之间。建立的ic CLEIA方法可用于实际样品中AMOZ残留检测。Indirect competitive chemiluminescence enzyme immunoassay （icCLEIA） detection for furaltadone metabolite 5-morpholinomethyl-3-amino-2-oxazolidone （AMOZ） was established. The effects of the coating antigen concentration, antibody dilution, immunoreaction time, reaction buffer system on the performance of the assay were studied and optimized. The result showed that the optimized assay conditions were as follows： the coating antigen concentration was 10 ng/mL; the polyclonal antibody was diluted 60000 times, immunoreaction time was 50 min and the buffer solution was 0.01 mol/L phosphate buffer solution （ pH 7.4）. Under the optimal conditions, the linear detection range of the developed ieCLEIA was 0. 026-3.52 μg/L, the IC50 was 0.29 μg/L and the limit of detection （ IC10 ） was 0. 012 μg/L. The average recoveries of AMOZ of spiked fish ranged from 101.4% to 115.5%. The icCLEIA is applicable for the detection of trace AMOZ in fish samples.

关 键 词：呋喃它酮 5-吗啉甲基-3-氨基-2-唑烷基酮 间接竞争化学发光酶免疫分析 多克隆抗体 鱼肉 

分 类 号：O657.3[理学—分析化学]