Department of Veterinary Pharmacology and Toxicology, College of Veterinary Medicine, China Agricultural University, Yuan Ming Yuan West Road No.2, Beijing, 100193, China.
Luminescence. 2014 Jun;29(4):393-400. doi: 10.1002/bio.2559. Epub 2013 Aug 12.
A chemiluminescent enzyme immunoassay (CLEIA) was compared to an ultraperformance liquid chromatography tandem mass spectroscopy (UPLC-MS/MS) procedure for the analysis of zeranol and its metabolites in bovine tissue samples. Apparent recoveries from fortified samples by both methods were comparable at 0.5-4.0 µg/kg and a significant correlation was obtained. For CLEIA analysis, hapten mimicking the analyte was first synthesized and conjugated with the carrier protein bovine serum albumin as the immunogen to produce monoclonal antibody. The obtained antibody showed extensive cross-reactivity toward zeranol metabolites (zearalanone). The limit of detection of CLEIA and UPLC-MS/MS was 0.05 µg/kg and 0.5 µg/kg, respectively. Recoveries of both methods for fortified samples were higher than 75.0% with the coefficient of variation less than 15%. These results indicated that the combination of screening with CLEIA and confirmation with UPLC-MS/MS for zeranol and its metabolites would be a reliable method for a large number of bovine samples.
化学发光酶免疫分析（CLEIA）与超高效液相色谱串联质谱（UPLC-MS/MS）法比较，用于分析牛组织样品中的泽那醇及其代谢物。两种方法对加标样品的回收率在 0.5-4.0μg/kg 时相当，并且得到了显著的相关性。对于 CLEIA 分析，首先合成了模拟分析物的半抗原，并与牛血清白蛋白载体蛋白缀合作为免疫原，产生单克隆抗体。所获得的抗体对泽那醇代谢物（玉米赤霉酮）表现出广泛的交叉反应性。CLEIA 和 UPLC-MS/MS 的检测限分别为 0.05μg/kg 和 0.5μg/kg。两种方法对加标样品的回收率均高于 75.0%，变异系数小于 15%。这些结果表明，用 CLEIA 进行筛选，用 UPLC-MS/MS 进行确证，是一种用于大量牛样品的泽那醇及其代谢物的可靠方法。
