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

目的：采用纳米均相时间分辨荧光免疫技术(amplified luminescent proximity homogeneous assay, Alpha-LISA)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。

Objective A direct competitive amplified luminescent proximity homogeneous assay (Alpha-LISA) 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 perfor-mance 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 screen-ing for CLB contamination in meat or foods in the future.