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An LC-MS/MS Methodological Framework for Steroid Hormone Measurement from Human Serum
Karina Gasbarrino1, Edward Daly2, Stella S Daskalopoulou1
1Department of Medicine, McGill University, Montreal, Quebec, Canada.
Hormone and Metabolic Research = Hormon- Und Stoffwechselforschung = Hormones Et Metabolisme
|March 30, 2022
Summary
New liquid chromatography-tandem mass spectrometry methods accurately measure steroid hormones like testosterone and estradiol in human serum. These advanced techniques offer improved sensitivity and specificity over traditional immunoassays for clinical diagnostics.
Area of Science:
- Endocrinology
- Analytical Chemistry
- Mass Spectrometry
Background:
- Estrogens and androgens are crucial for sexual development and physiological functions.
- Imbalances in these hormones are linked to diseases like osteoporosis, cancer, and cardiovascular/neurologic conditions.
- Current immunoassay methods for steroid hormone analysis lack specificity and sensitivity.
Purpose of the Study:
- To develop highly sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods for quantifying steroid hormones in human serum.
- To address the clinical need for accurate measurement of estrogen and androgen concentrations.
Main Methods:
- Development of three LC-MS/MS methods using a Triple Quadrupole Mass Spectrometer.
- Simultaneous measurement of testosterone, androstenedione, and cortisol.
- Quantification of dehydroepiandrosterone (DHEA) and 17β-estradiol (E2), utilizing derivatizing reagents (Girard's reagent P and dansyl chloride) for enhanced sensitivity.
Main Results:
- Successful quantitative measurement of endogenous steroid hormones in human serum samples.
- Achieved high sensitivity, providing reliable data down to low nanogram/ml and sub-nanogram/ml levels.
- Demonstrated high specificity through unique mass transitions and deuterated internal standards.
Conclusions:
- The developed LC-MS/MS methods are efficient, sensitive, and specific for analyzing steroid hormones in patient sera.
- These methods provide reliable quantification of key hormones, crucial for diagnosing and managing hormone-related pathologies.
- The study highlights the superiority of LC-MS/MS over immunoassays for accurate steroid hormone analysis.
