Departamento de Toxicología, Centro de Investigación y de Estudios Avanzados Del Instituto Politécnico Nacional (Cinvestav-IPN), Av. IPN No. 2508, Col. San Pedro Zacatenco, Ciudad de México, CP 07360, Mexico.
Instituto Nacional de Salud Pública, Centro de Investigación en Salud Poblacional, Cuernavaca, CP 62100, Mexico.
Anal Biochem. 2020 May 15;597:113670. doi: 10.1016/j.ab.2020.113670. Epub 2020 Feb 29.
Testosterone regulates the male reproductive system and acts directly or indirectly on nearly all systems during fetal, pubertal and adult life. Testosterone homeostasis depends on its synthesis and degradation. The major biotransformation reactions are hydroxylation by different cytochrome P450 (CYP) isoforms. There are no described methods to determine the profile of testosterone-hydroxylated metabolites in human urine. The aim of this study was to develop an analytical method to determine testosterone-hydroxylated metabolites in human urine using UPLC-MS. Seven testosterone-hydroxylated metabolites, androstenedione, and testosterone, were identified by comparison of their t and positive electrospray ionization (ESI) data, with those of analytical standards. The method developed is sensitive, specific, repeatable, and precise. Limits of detection and quantitation for all compounds ranged from 1.360 to 13.054 ng/ml and 4.234-39.679 ng/ml, respectively. The percentages of recovery were between 81.2 and 128.8%. The applicability of the analytical method was confirmed by analysis of urine samples obtained from two groups of healthy men (25-30 and 50-75 years old). All analytes were identified with slightly different metabolites profiles in both groups. In conclusion, the UPLC-MS method developed here was validated for the analysis of testosterone-hydroxylated metabolites in human urine.
睾酮调节男性生殖系统，并在胎儿期、青春期和成年期直接或间接地作用于几乎所有系统。睾酮的动态平衡依赖于其合成和降解。主要的生物转化反应是不同细胞色素 P450（CYP）同工酶的羟化。目前还没有描述的方法来确定人尿中睾酮羟化代谢物的特征。本研究旨在开发一种使用 UPLC-MS 测定人尿中睾酮羟化代谢物的分析方法。通过比较分析物的 t 和正电喷雾电离（ESI）数据与分析标准品的数据，鉴定出 7 种睾酮羟化代谢物，即雄烯二酮和睾酮。所开发的方法具有灵敏度高、特异性强、重复性好、精密度高的特点。所有化合物的检测限和定量限范围分别为 1.360-13.054ng/ml 和 4.234-39.679ng/ml。回收率在 81.2%至 128.8%之间。该分析方法的适用性通过分析两组健康男性（25-30 岁和 50-75 岁）的尿液样本得到了证实。两组均鉴定出具有略有不同代谢物特征的所有分析物。总之，本文建立的 UPLC-MS 方法可用于分析人尿中的睾酮羟化代谢物。
