呋喃它酮代谢物时间分辨荧光免疫分析法的建立与应用 |
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引用本文: | 邓丽华,戴尽波,徐振林,杨金易,王弘,肖治理,雷红涛,孙远明,沈玉栋.呋喃它酮代谢物时间分辨荧光免疫分析法的建立与应用[J].分析化学,2016(8):1286-1290. |
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作者姓名: | 邓丽华 戴尽波 徐振林 杨金易 王弘 肖治理 雷红涛 孙远明 沈玉栋 |
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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,广州510642 |
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基金项目: | 国家自然科学基金(.31371769;31271865),广东省科技计划项目(.2014A020219008 |
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摘 要: | 建立了Eu3+标记的间接竞争时间分辨荧光免疫分析法( Indirect competitive time-resolved fluoroimmu-noassay , 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/mL,抗体稀释5×104倍,最佳竞争时间为50 min。在优化的条件下,方法的检出限( LOD, IC10)为0.01 ng/mL,半抑制浓度(IC50)为0.26 ng/mL,线性范围(IC20~IC80)为0.025~2.83 ng/mL,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。
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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 |
Application of Time-resolved Fluroimmunoassay for Determination of Furaltadone Metabolite 3-Amino-5-morpholinomethyl-2-oxazolidinone |
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Abstract: | To detect furaltadone metabolite 3-amino-5-morpholinomethyl-2-oxazolidinone ( AMOZ ) in fish sample, an Eu3+ labeling time-resolved fluoroimmunoassay ( TRFIA ) was developed. The effects of experimental conditions including AMOZA-OVA concentration, dilution of antibody, and reaction time on the sensitivity of TRFIA were explored. The results showed that the optimized assay conditions were as follows:the AMOZA-OVA concentration was 0. 25 μg/mL; the antibody was diluted 5í104 folds, and the competitive reaction time was 50 min. Under optimal conditions, the method showed a detection limit of 0. 01 ng/mL, an IC50 of 0. 26 ng/mL and a linear range (IC20-IC80) of 0. 025-2. 83 ng/mL. The recoveries of AMOZ in fish at three spiked levels ranged from 78 . 0% to 86 . 0%, and the relative standard deviations were less than 15%. Good correlation between the ic-TRFIA and high performance liquid chromatography-tandem mass spectrometry was obtained for spiked food samples. The proposed ic-TRFIA method was suited for the determination of AMOZ residue in food samples. |
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Keywords: | Furaltadone metabolite 3-amino-5-morpholinomethyl-2-oxazolidinone Time-resolved fluoroimmunoassay Europium Ⅲ-labeled antibody |
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