LC-MS/MS-based simultaneous high-sensitivity quantification of clinically relevant high-potency androgens and...

Area of Science:

• Endocrinology
• Analytical Chemistry
• Biochemistry

Background:

• Steroid hormone quantification is vital for diagnosing and monitoring various endocrine disorders.
• Existing assays may lack the sensitivity or breadth to measure the full spectrum of relevant steroids, particularly novel or low-abundance ones.
• 11-oxygenated androgens represent a growing area of research with potential clinical significance.

Purpose of the Study:

• To develop and validate a sensitive and comprehensive assay for quantifying 19 steroids in human plasma or serum.
• To include classical steroids, their metabolites, and a panel of 11-oxygenated androgens.
• To establish a reliable method for clinical applications, especially in conditions with low circulating steroid levels.

Main Methods:

• Development of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay.
• Quantification of 19 steroids including dehydroepiandrosterone (DHEA), androstenedione (A4), testosterone (T), and eight 11-oxygenated androgens.
• Use of 300 μL of human plasma or serum with derivatization for enhanced sensitivity of most steroids.

Main Results:

• The assay accurately quantifies 19 steroids with high sensitivity (lower limit of quantification as low as 3.7 pM).
• It successfully measures classical steroids, their metabolites, and eight 11-oxygenated androgens.
• Validation and cross-validation confirmed the assay's reliability and accuracy.

Conclusions:

• The developed LC-MS/MS assay provides a sensitive and robust method for comprehensive steroid profiling.
• Its high sensitivity and broad analyte coverage are critical for accurate quantification in diverse clinical settings.
• The assay demonstrated clinical applicability in prostate cancer patients, highlighting its utility for monitoring hormone therapy and in conditions with low steroid concentrations.