Simultaneous Quantification of Steroid Hormones Using hrLC-MS in Endocrine Tissues of Male Rats and Human Samples

Abstract

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.

The Correlation of Reactive Changes in the Cortical and Cerebral Substance of the Adrenal Glands of White Rats During the Correction of Testosterone Blockade at the 6th Month of the Experiment

“…Науковці, що вивчають вроджену гіперплазію надниркових залоз та синдром полікістозу яєчників, пов'язують дані патології з впливом метаболітів тестостерону, що утворюються у результаті дефіциту стероїдної 21-гідроксилази, оксидоредуктази P450 та інших ферментів [3,9]. Таким чином, дослідження морфологічних змін надниркових залоз на тлі дефіциту тестостерону є перспективним.…”

Advances in Metabolic Profiling of Biological Samples

“…This Special Issue provides valuable insights into the development of novel sample preparation protocols and analytical methods for rapid metabolite analysis in biofluids and tissues. Specifically, a range of articles is presented [1][2][3], each addressing different aspects of this challenge. Campanella et al [2] proposed a fast and straightforward method for the analysis of saliva by attenuated total reflectance Fourier-transformed infrared spectroscopy (ATR-FTIR) and Raman spectroscopy for large-scale preclinical studies.…”

“…This study proposed a novel method for saliva collection via the deposition of multiple spots onto low-cost polypropylene sheets, revealing reliable and reproducible ATR-FTIR spectra. Bordanaba-Florit et al [1] developed a rapid (6 min runtime per sample) and sensitive method for the quantification of steroid hormone compounds (androgens, estrogens, progestogens, and corticoids) by liquid chromatography-high-resolution mass spectrometry (LC-HRMS). The performance of this methodology was tested in several rat tissues (adrenal glands, testis, prostate, liver, and brain), human urine, and urinary extracellular vesicles.…”