建立了克伦特罗的直接竞争化学发光酶免疫检测方法,并考察了方法的特异性、灵敏度。最佳反应条件为:包被抗原浓度为25 ng/mL;酶标抗体稀释80 000倍;0.02 mol/L磷酸盐缓冲液(PBS)(pH 7.0),竞争反应时间30 min。方法的IC50为0.09 ng/mL;检出限为0.01 ng/mL;线性范围0.02~7.59 ng/mL;对尿样、猪肉、肝脏以及饲料样品检测的平均回收率分别为94.5%~95.3%、90.7%~94.1%,批内和批间相对标准偏差均小于15%。该方法与HPLC方法的相关性良好,说明其具有较高的灵敏度和特异性,可用于实际样品的检测。

Develop an immunoassay method for determination of clenbuterol.A direct competitive chemiluminescent enzyme immunoassay(dc-CLEIA) was developed for determining clenbuterol.The results showed that the optimized assay conditions for both the highest sensitivity and the best stability were as follows: coating antigen concentration was 25 ng / mL,and dilution fold of antibody enzyme conjugate in 0.02 mol / L pH 7.0 PBS dilution was 80 000.The developed method presented an IC 50 of 0.09 ng / mL,a detection limit of 0.01 ng / mL,a linear range of 0.02 to 7.59 ng / mL.Intra-and inter-batch CV relative standard deviations were below 15%.The analytical recovery rate in urea,pork,liver and fodder sample was 94.5%,95.3%,90.7%,94.1%,respectively.A comparison result between HPLC and the developed assay showed better relativity.The method is sensitive and stable,and it indicates that the dc-CLEIA method can be used to analyze sample.