Departments of Nutrition and Environmental Toxicology, University of California, Davis, California 95616, United States.
Anal Chem. 2013 May 21;85(10):4951-60. doi: 10.1021/ac400016e. Epub 2013 May 13.
In humans, steroids play a broad and vital role in regulation of gene expression, secondary sexual characteristics, maturation, reproduction, cardiovascular health, neurological functions, etc., but imbalance in steroid metabolism is also linked to development and progression of many diseases, such as cancer, neurodegenerative diseases, and cardiovascular diseases. Hence, measurement of steroids in biological samples is essential to monitor human health. Currently, there is radioimmunoassay, gas chromatography-mass spectrometry (GC/MS), and liquid chromatography-mass spectrometry (LC-MS) methods developed for steroid measurements in biological samples. However, these methods require elaborate sample preparation procedures and have concerns(s) related to reproducibility, dynamic range, time, costs, and most importantly the total coverage of steroids. Also currently, there is no method available for comprehensive steroid profiling in a single LC-MS run that includes androgens, corticosteroids, progestogens, estrogens, estrogen metabolites, estrogen conjugates, and estrogen-DNA adducts as well as exogenous steroid derivatives. Here, I present a global steroid metabolic profiling method based on liquid-liquid extraction (LLE) followed by ultra performance liquid chromatography (UPLC)-tandem mass spectrometry (MS/MS) for simultaneous measurement of over 100 indigenous as well as exogenous steroids in about 12 min, without derivatization. The method was successfully applied to determine steroid hormone levels in the breast tissue of healthy women. Overall presence of all major classes of steroids as well as estrogen derivatives was detected in breast tissue.
在人类中，类固醇在基因表达调控、第二性征、成熟、生殖、心血管健康、神经功能等方面发挥着广泛而重要的作用，但类固醇代谢失衡也与许多疾病的发展和进展有关，如癌症、神经退行性疾病和心血管疾病。因此，测量生物样本中的类固醇对于监测人类健康至关重要。目前已经开发出放射免疫测定法、气相色谱-质谱联用（GC/MS）和液相色谱-质谱联用（LC-MS）方法来测量生物样本中的类固醇。然而，这些方法需要精细的样品制备程序，并且存在与重现性、动态范围、时间、成本相关的问题，最重要的是，它们不能全面涵盖类固醇。此外，目前还没有一种方法可以在单次 LC-MS 运行中进行全面的类固醇分析，包括雄激素、皮质类固醇、孕激素、雌激素、雌激素代谢物、雌激素结合物以及雌激素-DNA 加合物和外源性类固醇衍生物。在这里，我提出了一种基于液-液萃取（LLE）的全球类固醇代谢谱分析方法，随后进行超高效液相色谱（UPLC）-串联质谱（MS/MS）分析，可在约 12 分钟内同时测量 100 多种内源性和外源性类固醇，无需衍生化。该方法成功应用于测定健康女性乳腺组织中的类固醇激素水平。在乳腺组织中检测到所有主要类固醇类和雌激素衍生物的存在。
