| 摘要: |
| 【目的】建立β2-受体激动剂类药物多残留间接竞争ELISA检测方法，为动物食源性β2-受体激动剂类药物残留的快速检测提供技术支持。【方法】以沙丁胺醇多克隆抗体为基础，优化间接竞争ELISA的检测条件，建立一种快速检测猪尿中沙丁胺醇、卡布特罗、西布特罗、克伦特罗、溴布特罗、马布特罗、克伦潘特、可尔特罗、吡布特罗、马喷特罗、比托特罗与妥布特罗等12种β2-受体激动剂类药物的间接竞争ELISA检测方法。【结果】建立的β2-受体激动剂类药物间接竞争ELISA检测方法的最佳反应条件为：37 ℃，抗原以1∶10 000倍包被1 h，抗体以1∶40 000稀释作用40 min，酶标抗体以1∶20 000稀释作用40 min。采用建立的间接竞争ELISA最佳反应条件对猪尿中的β2-受体激动剂类药物残留进行检测，结果表明 12种β2-受体激动剂类药物在0.1～20 μg/L范围内线性关系良好，相关系数均大于0.952；检出限为0.15 μg/L，定量限为0.5 μg/L；在0.5与1.0 μg/L的添加浓度下，回收率为68.4%～114.2%，批内、批间RSD分别为0.4%～10.1%和0.4%～12.5%。【结论】所建立的β2-受体激动剂类药物残留ELISA检测方法操作简单、快速，灵敏度高，适用于猪尿中β2-受体激动剂药物的筛选与检测。 |
| 关键词: β2-受体激动剂 间接竞争ELISA 多残留检测 猪尿 |
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| 基金项目:中国动物卫生与流行病学中心创新基金项目（2014IF-0003FF）；国家自然科学基金项目（31772802） |
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| Development of an indirect competitive ELISA for detection of multiple residues of β2-agonist drugs |
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WANG Rui,CAO Xumin,XU Jun,et al
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| Abstract: |
| 【Objective】An indirect competitive ELISA method was established for detection of β2-agonists multiple residues to provide support for the detection of β2-agonists in animal-origin food.【Method】Salbutamol polyclonal antibody was used and a rapid detection method was established for the test of salbutamol,capbuterol,cibuterol,clenbuterol,brombuterol and marbuterol,clenpentero,colterol,pirbuterol,mapenterol,bitolterol and tulobuterol in pig urine.【Result】The optimization conditions for the established indirect competitive ELISA method were temperature 37 ℃，antigen coated with 1∶10 000 times for 1 h,the antibody diluted with 1∶40 000 for 40 min,and the enzyme labeled antibody diluted with 1∶20 000 for 40 min.The results of detecting β2-agonist residues in pig urine showed that the 12 drugs had a good linear relationship within the range of 0.1―20 μg/L,and the correlation coefficients were all greater than 0.952.The detection limit of pig urine sample was 0.15 μg/L and the quantitative limit was 0.5 μg/L.When 0.5 and 1.0 μg/L of standard solutions were added to pig urine,the recovery rates were 68.4%-114.2%,the intra RSDs were 0.4%―10.1% and the inter-RSDs were 0.4%―12.5%.【Conclusion】The established method is simple, rapid and sensitive,and it is suitable for the detection of β2-agonists in pig urine. |
| Key words: β2-agonists indirect competitive ELISA multi-residue determination pig urine |
