呋喃它酮代谢物时间分辨荧光免疫分析法的建立与应用

发布时间：2018-01-20 15:06

  本文关键词： 呋喃它酮代谢物AMOZ 时间分辨荧光免疫分析 铕标二抗　出处：《分析化学》2016年08期 　论文类型：期刊论文

【摘要】：建立了Eu~(3+)标记的间接竞争时间分辨荧光免疫分析法(Indirect competitive time-resolved fluoroimmunoassay,ic-TRFIA)用于鱼肉样品中呋喃它酮代谢物AMOZ的检测。通过单因素实验考察了包被抗原浓度、抗体稀释倍数、竞争反应时间等参数对方法灵敏度的影响。结果表明,ic-TRFIA的最佳反应条件为:包被抗原浓度为0.25μg/m L,抗体稀释5×10~4倍,最佳竞争时间为50 min。在优化的条件下,方法的检出限(LOD,IC_(10))为0.01 ng/m L,半抑制浓度(IC_(50))为0.26 ng/m L,线性范围(IC_(20)~IC_(80))为0.025~2.83 ng/m L,对鱼样中AMOZ回收率在78.0%~86.0%之间,各样品变异系数均小于15%,与HPLC-MS/MS对比检测结果显示相关性良好。本方法灵敏度高、特异性好,能够满足实际样品的检测需求。
[Abstract]:An indirect competitive time resolved fluorescence immunoassay (Eu~(3) was established for Eu~(3 labeling. Indirect competitive time-resolved fluoroimmunoassay. Ic-TRFIA was used to detect AMOZ of furantadone metabolites in fish meat samples. The concentration of coated antigen and dilution times of antibody were investigated by single factor experiment. The effect of competitive reaction time and other parameters on the sensitivity of the method was studied. The results showed that the optimal reaction conditions of TRFIA were as follows: the concentration of coated antigen was 0.25 渭 g / mL. The best competitive time was 50 mins. Under the optimized conditions, the detection limit of the method was 0. 01 ng/m / L. The half inhibitory concentration (ICS) was 0.26 ng/m / L, and the linear range was 0.025 / 2.83 ng/m / L. The recovery of AMOZ in fish samples was between 78.0% and 86.0%, and the coefficient of variation of each sample was less than 15%. Compared with HPLC-MS/MS, the method has high sensitivity and specificity, and can meet the needs of actual samples.
【作者单位】： 华南农业大学食品学院广东省食品质量安全重点实验室;
【基金】：国家自然科学基金(Nos.31371769,31271865) 广东省科技计划项目(Nos.2014A020219008,2013B040402006,2012A020100002)资助
【分类号】：O657.3
【正文快照】： 1引言呋喃它酮是硝基呋喃类抗生素的一种,曾被广泛应用于饲料添加剂及家禽、家畜、水产等养殖物传染病的预防与治疗[1]。但研究表明,呋喃它酮及其代谢物具有潜在的致癌和诱导机体突变的作用,且其代谢产物能与蛋白质结合,形成比原药化合物更稳定的蛋白结合物而在体内富集。但在，

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