Zhang Dai, Zhou Lifang, Lei Yajing, Zhou Zhan, Zhou Jie, Chen Shuqing
Institute of Drug Metabolism and Pharmaceutical Analysis, College of Pharmaceutical Sciences, Zhejiang University, 866 Yuhangtang Road, Zijingang Campus, Hangzhou, Zhejiang, 310058, China.
Hangzhou EPIE Bio-detection Technology Limited, Hangzhou, 310051, China.
Environ Sci Pollut Res Int. 2017 Mar;24(8):7042-7050. doi: 10.1007/s11356-017-8405-y. Epub 2017 Jan 14.
Diethylstilbestrol (DES) is used as a kind of animal feed additive and affects people's health through the food chain. The purpose of this study is to detect the residue level of DES in 576 human urine samples directly. DES-BSA was used to immunize Balb/c mice. The monoclonal antibody was produced by hybridoma that was screened through cell fusion techniques. Finally, we developed the indirect competitive ELISA method to analyze 576 human urine samples from Zhejiang Province, China. The IC of this method was 3.33 ng/mL. The LOD and LOQ were 0.16 and 0.54 ng/mL. Linear range of the standard curve was from LOD to 12.50 ng/mL. There was no cross-reactivity with two kinds of estrogens and two structural analogs with DES. Five hundred seventy-six urine samples were analyzed by the indirect competitive ELISA method, and the detection rate was 98.78%. The mean concentration and geometric mean were 4.70 and 3.50 ng/mL. The indirect competitive ELISA method based on monoclonal antibody was sensitive and reliable for the detection of DES in human urine samples. The results warned us to pay more attention to human health and food safety.
己烯雌酚(DES)被用作一种动物饲料添加剂,并通过食物链影响人体健康。本研究的目的是直接检测576份人类尿液样本中DES的残留水平。用DES - BSA免疫Balb/c小鼠。通过细胞融合技术筛选出杂交瘤细胞,进而制备单克隆抗体。最后,我们建立了间接竞争ELISA法来分析来自中国浙江省的576份人类尿液样本。该方法的半数抑制浓度(IC)为3.33 ng/mL。检测限(LOD)和定量限(LOQ)分别为0.16和0.54 ng/mL。标准曲线的线性范围为从检测限到12.50 ng/mL。该方法与两种雌激素以及两种与DES结构类似物均无交叉反应。采用间接竞争ELISA法对576份尿液样本进行分析,检出率为98.78%。平均浓度和几何平均浓度分别为4.70和3.50 ng/mL。基于单克隆抗体的间接竞争ELISA法用于检测人类尿液样本中的DES灵敏且可靠。研究结果警示我们要更加关注人类健康和食品安全。