The School of Chemistry and Chemical Engineering, Shandong University, Jinan, Shandong 250100, PR China.
Talanta. 2011 Jan 15;83(3):732-7. doi: 10.1016/j.talanta.2010.10.020. Epub 2010 Oct 23.
Trenbolone (TRE) is a steroid used by veterinarians on livestock to increase appetite and body weight. The use of TRE has been restricted because of its harmful side effect for consumers. To effectively control TRE residue in food and food product, a rapid and convenient immunoassay was developed by preparing an anti-TRE monoclonal antibody. The immunogen and coating antigen were prepared by coupling TRE hapten with carrier proteins via 1-ethyl-3-(dimethylaminopropyl)carbodiimide hydrochloride (EDC) method. The optimized method gave an average IC(50) value of 0.323 ng mL(-1) towards TRE and an average detection limit (LOD) of 0.06 ng mL(-1), which is much lower than the maximum residue levels (2.0 ng g(-1)) accepted by the Joint FAO/WHO Expert Committee on Food Additives (JECFA). The specificity of the antibody was evaluated by measuring cross-reactivity of six structurally related compounds, including 19-nortestosterone (9.7%), testosterone (0.13%), methyltestosterone (<0.01%), methandrostenolone (<0.01%), (+)-dehydroisoandrosterone (<0.001%) and β-estradiol (<0.001%). The recovery rates of the test in detection of TRE-fortified animal tissue, urine and animal feed samples were in the range of 81.3-89.4%, while the intra- and inter-assay coefficients of variation were less than 12.0%.
特伦布酮（TRE）是一种兽医在牲畜上使用的类固醇，以增加食欲和体重。由于其对消费者的有害副作用，TRE 的使用受到限制。为了有效地控制食品和食品中 TRE 残留，通过制备抗 TRE 单克隆抗体开发了一种快速方便的免疫测定法。免疫原和包被抗原通过 1-乙基-3-（二甲基氨基丙基）碳化二亚胺盐酸盐（EDC）法将 TRE 半抗原与载体蛋白偶联制备。优化后的方法对 TRE 的平均 IC（50）值为 0.323ngmL（-1），平均检测限（LOD）为 0.06ngmL（-1），远低于食品添加剂联合专家委员会（JECFA）接受的最大残留限量（2.0ngg（-1））。通过测量六种结构相关化合物的交叉反应性来评估抗体的特异性，包括 19-去甲睾酮（9.7％），睾酮（0.13％），甲基睾酮（<0.01％），美雄酮（<0.01％），（+）-脱氢异雄酮（<0.001％）和β-雌二醇（<0.001％）。在检测 TRE 强化动物组织，尿液和动物饲料样品中的测试回收率在 81.3％至 89.4％之间，而批内和批间变异系数小于 12.0％。
