Steroid Hormone Analysis Service

What are Steroid Hormones?

Steroid hormones are a class of hormones derived from cholesterol that play vital roles in numerous physiological processes in the body. These hormones are synthesized primarily in the adrenal glands, gonads (testes and ovaries), and placenta during pregnancy. They are characterized by their unique chemical structure, which consists of four interconnected rings of carbon atoms.

There are several types of steroid hormones, each with its specific functions and target tissues. Glucocorticoids, such as cortisol, are involved in regulating metabolism, immune responses, and stress responses. They affect glucose metabolism, protein synthesis, and the immune system, helping the body respond to stress and maintain homeostasis.

Mineralocorticoids, including aldosterone, play a crucial role in regulating electrolyte and water balance in the body. They act on the kidneys, promoting the reabsorption of sodium and the excretion of potassium, thereby controlling blood pressure and fluid balance.

Sex hormones are responsible for the development and maintenance of sexual characteristics and reproductive functions. In males, the primary sex hormone is testosterone, which promotes the development of male secondary sexual characteristics and supports sperm production. In females, estrogen and progesterone regulate the menstrual cycle and play crucial roles in pregnancy and childbirth.

Progesterone is a steroid hormone involved in regulating the menstrual cycle and supporting pregnancy. It prepares the uterus for implantation of a fertilized egg and helps maintain the pregnancy.

Vitamin D, although not traditionally considered a steroid hormone, also falls into this category. It plays a vital role in calcium and phosphorus metabolism and is synthesized in the skin upon exposure to sunlight or obtained through dietary sources. Vitamin D helps regulate bone health, immune function, and cell growth.

Steroid hormones exert their effects by binding to specific receptors in target cells. Once bound, they can enter the cell nucleus and influence gene expression, resulting in the production of specific proteins or the modulation of cellular activities. The regulation of steroid hormone synthesis, release, and metabolism is tightly controlled to maintain hormone balance and ensure proper physiological function.

Steroid Hormone Biosynthesis

The biosynthesis of steroid hormones occurs primarily in specialized endocrine organs, including the adrenal glands, gonads (testes and ovaries), and placenta during pregnancy. The process involves a series of enzymatic reactions that convert cholesterol into specific steroid hormones.

The initial step in steroid hormone biosynthesis is the transport of cholesterol from the cytoplasmic pool (derived from circulating low-density lipoproteins) into the mitochondria or endoplasmic reticulum, where the biosynthetic enzymes are located. Within these organelles, cholesterol undergoes enzymatic modifications to form various intermediates, ultimately leading to the production of specific steroid hormones.

The biosynthetic pathway for steroid hormones involves several key enzymes, including cholesterol side-chain cleavage enzyme (CYP11A1), 3β-hydroxysteroid dehydrogenase (3β-HSD), and aromatase. These enzymes catalyze reactions that introduce specific functional groups and modify the structure of cholesterol, resulting in the formation of different steroid hormones.

The production of specific steroid hormones is regulated by complex mechanisms involving hormonal signals, feedback loops, and the expression and activity of specific biosynthetic enzymes in target tissues. These hormones are released into the bloodstream and transported to target cells, where they bind to specific receptors and exert their biological effects.

Overall, the biosynthesis of steroid hormones is a highly regulated and intricate process that ensures the production of appropriate hormone levels for normal physiological function. Any disruptions in this process can lead to hormonal imbalances and associated health issues.

LC-MS Analysis of Steroid Hormones by Creative Proteomics

Instrumentation Used by Creative Proteomics

• Liquid Chromatography (LC): High-performance liquid chromatography systems such as Agilent 1290 Infinity II and Thermo Fisher Scientific Vanquish Flex provide efficient chromatographic separations.
• Mass Spectrometry (MS): State-of-the-art mass spectrometers like Thermo Fisher Scientific Q Exactive HF-X and SCIEX Triple Quad 6500+ ensure accurate detection and quantification of steroid hormones.

Steroid Hormone Metabolism Analysis Workflow

Sample Collection and Preparation: The first step is to collect appropriate biological samples, such as blood, urine, tissue, or saliva, depending on the study objectives. Samples should be collected using suitable protocols to ensure sample integrity. After collection, samples are processed to extract steroid hormones and their metabolites. This typically involves separation techniques, such as centrifugation or filtration, to obtain the desired sample components.

Metabolite Extraction: Steroid hormone metabolites are extracted from the biological samples using various extraction methods. The choice of extraction technique depends on the target metabolites and the sample matrix. Common extraction methods include liquid-liquid extraction (LLE), solid-phase extraction (SPE), or protein precipitation. These techniques separate the metabolites from interfering substances, enabling subsequent analysis.

Chromatographic Separation: Chromatographic techniques are employed to separate steroid hormone metabolites based on their physicochemical properties. High-performance liquid chromatography (HPLC) or liquid chromatography (LC) coupled with different types of columns, such as reverse-phase or normal-phase, are commonly used. Chromatographic separation enables the resolution of individual metabolites and their quantification.

Detection and Quantification: Once separated, the metabolites are subjected to detection and quantification. Mass spectrometry (MS) is the most commonly used detection technique due to its high sensitivity and specificity. Liquid chromatography-mass spectrometry (LC-MS) is a powerful combination for steroid hormone metabolite analysis. Specific MS techniques, such as tandem mass spectrometry (MS/MS), can be employed for enhanced selectivity and quantification accuracy.

Data Analysis and Interpretation: The obtained data from the detection and quantification step are analyzed and interpreted to determine the metabolic profile of steroid hormones. This involves comparing the detected metabolites with reference standards or databases. Statistical analysis, such as peak integration, calibration curves, and normalization, is performed to quantify the metabolites accurately.

Metabolite Identification: Metabolite identification is a crucial step in steroid hormone metabolism analysis. It involves comparing the mass spectra and retention times of the detected metabolites with reference standards or databases. Advanced techniques, such as high-resolution mass spectrometry (HRMS) and spectral libraries, can aid in the identification of unknown or rare metabolites.

Data Validation and Reporting: Once the analysis is complete, the data are validated to ensure accuracy and reliability. Quality control measures, such as calibration checks, blank samples, and replicate analyses, are performed to verify the analytical performance. Finally, the results are compiled into a comprehensive report, including the identified metabolites, their quantities, and any relevant observations or interpretations.

List of Detectable Steroid Hormone Metabolism Related Metabolites

Sample Requirements of Steroid Hormone Metabolism Analysis

We can analyze a wide range of biological materials including but not limited to cells and solid tissues from humans and animals, such as mice, rats, rabbits. If you need transport your samples to us, please follow the following requirements for different types of sample:

• Blood/plasma: 500ul/sample
• Urine: 1ml/sample
• Tissue: 200mg/sample
• Cells: 1x10⁷/sample
• Feces: 500mg/sample
• Shipment condition: dry ice