%0 Journal Article
%T 酶联免疫吸附法测定水产品中甲基睾酮残留
%A 王强
%J 现代食品科技
%D 2015
%R 10.13982/j.mfst.1673-9078.2015.5.048
%X 建立了测定水产品中甲基睾酮(MT)残留的间接竞争酶联免疫吸附法(icELISA)。采用琥珀酸酐法衍生化制备半抗原MT17，进一步偶联钥孔血蓝蛋白(KLH)制备免疫原MT17-KLH，经动物免疫成功获取特异性识别MT的多克隆抗体（与其它结构类似物交叉反应小于1%）。优化确立了icELISA最佳检测条件：包被原浓度100 μg/L，MT多克隆抗体稀释度1/1.2×105，抗体反应时间40 min，药物稀释液为PBS，水产样品经乙腈提取，N-丙基乙二胺吸附剂(PSA)分散固相萃取(DSPE)净化后用于icELISA测定。本方法的抑制中浓度IC50为3.48 μg/L，检测线性范围为0.77~13.06 μg/L。水产样品加标回收率为81.02~103.19%，相对标准偏差为4.46%~13.14%，测定结果与液相色谱-质谱联用法具有良好的相关性（R=0.9971）。本方法可用于水产品中甲基睾酮残留的快速测定。</br>An indirect, competitive enzyme-linked immunosorbent assay (icELISA) was developed for rapid determination of residual methyltestosterone (MT) in aquatic products. MT was reacted with succinic anhydride to obtain the hapten MT17, which was then coupled to keyhole limpet hemocyanin (KLH) as an immunogen (MT17-KLH). After animal immunization, the polyclonal antibody was successfully obtained and could specifically recognize MT (cross-reactivity with other structurally related chemicals was < 1%). The optimized assay conditions for icELISA were as follows: coating antigen concentration at 100 μg/L; dilution ratio of MT polyclonal antibody at 1:1.2 × 105; 40 min reaction time of antibody; phosphate-buffered saline (PBS) as dilution solution; extraction of aquatic product with acetonitrile, followed by purification by dispersive solid-phase extraction (DSPE) using ethylenediamine-N-propyl (PSA)-bonded sorbent. The half inhibition concentration (IC50) was 3.48 μg/L, while the linear range (IC20~IC80) was 0.77 to 13.06 μg/L. Recoveries from spiked aquatic products were in the range of 81.02% to 103.19%, with relative standard deviation ranging from 4.46% to 13.14%. The results showed good correlation with liquid chromatography-tandem mass spectrometry (R = 0.9971). The method developed in this study is suitable for rapid determination of methyltestosterone residues in aquatic products.
%K 甲基睾酮 酶联免疫吸附法 分散固相萃取 水产品&lt
%K /br&gt
%K methyltestosterone enzyme-linked immunosorbent assay dispersive solid phase extraction aquatic product
%U http://www.xdspkj.cn/ch/reader/view_abstract.aspx?file_no=20150548&flag=1