盐酸克伦特罗的AlphaLISA检测方法的建立

Quantitative determination of clenbuterol using the Amplified Luminescent Proximity Homogeneous Assay

中文关键词:  盐酸克伦特罗  纳米均相时间分辨荧光免疫技术(AlphaLISA)  竞争反应

英文关键词:clenbuterol  amplified luminescent proximity homogeneous assay (AlphaLISA)  competing reaction

中文摘要:

      目的 采用纳米均相时间分辨荧光免疫技术(amplified luminescent proximity homogeneous assay, AlphaLISA)建立高灵敏检测盐酸克伦特罗(clenbuterol, CLB)残留的方法。方法 在发光微粒上固定CLB, 与游离的CLB共同竞争固定在感光微粒上的CLB单克隆抗体, 优化检测条件并进行方法学考核。结果 该方法的灵敏度为0.04 ng/mL。猪肉样品回收率在83%~109%之间, 牛肉样品回收率在92%~112%之间; 猪肉样品检测的批内变异系数CV<10%、批间变异系数CV<15%。与特布他林、沙丁胺醇的交叉反应率分别为34.5%、25.7%, 与莱克多巴胺、妥布特罗、齐帕特罗无交叉反应。结论 CLB-AlphaLISA检测肉类中克伦特罗残留具有简单、快速、灵敏性高、特异性强、稳定等特点, 具有广阔的应用前景。

英文摘要:

      Objective A direct competitive amplified luminescent proximity homogeneous assay (AlphaLISA) for detecting clenbuterol(CLB) was established. Methods CLB was coated on emitting particles, and competed with sample CLB to anti-CLB which was coated on photosensitive particles. The analytical performance and optinal test conditions of the method were studied. Results The sensitivity of the assay was 0.04 ng/mL. The recoveries of the determination for CLB in pork and beer were respectively 83%~109% and 92%~112%. The CV of intra–and inter-assay were <10% and <15% respectively. The cross-reactivity of the CLB-AlphaLISA with ractopamine, tulobuterol and zilpaterol was negligible, while that with terbutaline and salbutamol was 34.5% and 25.7% respectively. Conclusion The CLB-AlphaLISA had the characteristics of excellent specificity and sensitivity, and good precision and economy. It could be widely used in rapid screening for CLB contamination in meat or foods in the future.