Billitti J E, Lasley B L, Wilson B W
Department of Environmental Toxicology, University of California at Davis, Davis, California 95616, USA.
Biol Reprod. 1998 Nov;59(5):1023-8. doi: 10.1095/biolreprod59.5.1023.
This is a report on the development and validation of an ELISA method to determine fecal testosterone levels, and on their evaluation as a biomarker for adverse effects of endocrine-disrupting chemicals on reproductive health using male rodents of the Peromyscus maniculatus and Mus musculus species as an animal model. The ELISA antibody had the highest specificity for testosterone (100%), followed by dihydrotestosterone (57.4%) and androstenediol (0.27%). Radiolabeled testosterone was injected i.p. into three mice. Fecal samples were collected, extracted, and analyzed by liquid scintillation counting. The ELISA was performed to characterize the excretion kinetics and metabolic fate of circulating testosterone. Solubilization of feces with 10% methanol overnight provided an extraction efficiency of 87% for all metabolites; an ethyl ether extraction was more selective for testosterone. The fecal excretion of the testosterone was a biphasic process with a majority of the radioactivity recovered in the first 24 hours. HPLC analysis revealed at least five testosterone metabolites in feces, with most metabolites being less polar than testosterone. This study forms the initial evaluation of what will become a field monitoring tool.
这是一份关于开发和验证一种用于测定粪便睾酮水平的酶联免疫吸附测定（ELISA）方法的报告，以及使用鹿鼠和小家鼠作为动物模型，评估其作为内分泌干扰化学物质对生殖健康不良影响的生物标志物的报告。ELISA抗体对睾酮的特异性最高（100%），其次是二氢睾酮（57.4%）和雄烯二醇（0.27%）。将放射性标记的睾酮腹腔注射到三只小鼠体内。收集粪便样本，进行提取，并通过液体闪烁计数法进行分析。采用ELISA来表征循环睾酮的排泄动力学和代谢命运。用10%甲醇将粪便过夜溶解，对所有代谢物的提取效率为87%；乙醚提取对睾酮的选择性更高。睾酮的粪便排泄是一个双相过程，大部分放射性在最初24小时内回收。高效液相色谱分析显示粪便中至少有五种睾酮代谢物，大多数代谢物的极性比睾酮小。本研究构成了对一种未来将成为现场监测工具的初步评估。
