Bordanaba-Florit Guillermo, Liempd Sebastiaan van, Cabrera Diana, Royo Félix, Falcón-Pérez Juan Manuel
Exosomes Laboratory, Center for Cooperative Research in Biosciences (CIC bioGUNE), Basque Research and Technology Alliance (BRTA), 48160 Derio, Spain.
Metabolomics Platform, Center for Cooperative Research in Biosciences (CIC bioGUNE), Basque Research and Technology Alliance (BRTA), 48160 Derio, Spain.
Metabolites. 2022 Jul 30;12(8):714. doi: 10.3390/metabo12080714.
Steroid hormones play a vital role in the regulation of cellular processes, and dysregulation of these metabolites can provoke or aggravate pathological issues, such as autoimmune diseases and cancer. Regulation of steroid hormones involves different organs and biological compartments. Therefore, it is important to accurately determine their levels in tissues and biofluids to monitor changes after challenge or during disease. In this work, we have developed and optimized the extraction and quantification of 11 key members of the different steroid classes, including androgens, estrogens, progestogens and corticoids. The assay consists of a liquid/liquid extraction step and subsequent quantification by high-resolution liquid chromatography coupled time-of-flight mass spectrometry. The recoveries range between 74.2 to 126.9% and 54.9 to 110.7%, using a cell culture or urine as matrix, respectively. In general, the signal intensity loss due to matrix effect is no more than 30%. The method has been tested in relevant steroidogenic tissues in rat models and it has also been tested in human urine samples. Overall, this assay measures 11 analytes simultaneously in 6 min runtime and it has been applied in adrenal gland, testis, prostate, brain and serum from rats, and urine and extracellular vesicles from humans.
类固醇激素在细胞过程的调节中起着至关重要的作用，这些代谢产物的失调会引发或加剧诸如自身免疫性疾病和癌症等病理问题。类固醇激素的调节涉及不同的器官和生物区室。因此，准确测定它们在组织和生物流体中的水平对于监测激发后或疾病期间的变化非常重要。在这项工作中，我们开发并优化了不同类固醇类别的11种关键成分的提取和定量方法，包括雄激素、雌激素、孕激素和皮质激素。该测定法包括液/液萃取步骤，随后通过高分辨率液相色谱-飞行时间质谱进行定量。分别以细胞培养物或尿液为基质时，回收率范围为74.2%至126.9%和54.9%至110.7%。一般来说，由于基质效应导致的信号强度损失不超过30%。该方法已在大鼠模型的相关类固醇生成组织中进行了测试，也已在人类尿液样本中进行了测试。总体而言，该测定法在6分钟的运行时间内可同时测量11种分析物，并且已应用于大鼠的肾上腺、睾丸、前列腺、大脑和血清，以及人类的尿液和细胞外囊泡。
