Determination of 19 steroid hormones in human serum and urine using liquid chromatography-tandem mass spectrometry
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This paper describes a methodology for simultaneous determination of 19 steroid hormones, viz. estrone, estradiol, estriol, testosterone, 5α-dihydrotestosterone, androstenedione, androstenediol, dehydroepiandrosterone, progesterone, pregnenolone, 17α-OH-progesterone, 17α-OH-pregnenolone, cortisone, cortisol, 11-deoxycortisol, 11-deoxycorticosterone, 11-dehydrocorticosterone, aldosterone, and corticosterone, in 500-µL of urine or serum/plasma. The method was optimized using isotopically labeled internal standards and liquid-liquid extraction followed by detection using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-MS/MS). Dansylation of estrogens significantly improved their sensitivities (~11- to 23-fold) and chromatographic separation. The respective limit of detection (LOD) and limit of quantification (LOQ) of all analytes were 0.04−0.28 and 0.14−0.92 ng/mL in human urine, and 0.11−0.35 and 0.38−1.18 ng/mL in human serum/plasma. Recoveries of all analytes (except for progesterone) fortified at 10, 20, and 200 ng/mL in urine and serum were 80−120%, with standard deviations ranging from 0 to 17.3%. Repeated analysis of similarly fortified urine and serum samples yielded intra-day and inter-day variations of 0−21.7% and 0.16−11.5%, respectively. All analytes except cortisone exhibited weak matrix effects in urine and serum (−13.9−18.2%). The method was further validated through the analysis of the National Institute of Standards and Technology (NIST) plasma Standard Reference Material (SRM1950) with certified concentrations for cortisol, progesterone, and testosterone (coefficient of variation: 3−11%). The developed method was applied in the analysis of urine samples from 20 volunteers, which revealed the occurrence of 16 analytes with detection frequencies (DFs) > 80%. Furthermore, 15 analytes were found in plasma SRM1950, indicating the feasibility of our method in the analysis of steroid hormones in urine and serum/plasma. This method will facilitate analysis of steroid hormones in population-based biomonitoring studies.
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Several sensitive and robust LC-MS/MS methods for multiple steroids have been developed in recent years and many methods meet the requirements for clinical applications.
“Several sensitive and robust LC-MS/MS methods for multiple steroids have been developed in recent years and many methods meet the re quirements for clinical applications [5 -16].”
From Sex- and age-specific reference intervals of 16 steroid metabolites quantified simultaneously by LC-MS/MS in sera from 2458 healthy subjects aged 0 to 77 years (2024) · cites this paper as [11]
The Introduction says LC-MS/MS has been used for steroid analysis in recent years, and the Conclusions describe a validated method for measuring 19 hormones in human serum and urine. The paper does not establish that many such methods meet clinical-application requirements; it describes its own method as suitable for population-based biomonitoring. (LLM, conf=0.86)
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- Authors
- Li, Z.-M., & Kannan, K.
- Affiliations
- Department of Pediatrics, New York University Grossman School of Medicine, New York, NY 10016, USA
- New York University (US)
- Corresponding author
- Kannan, Kurunthachalam
- Venue
- Toxics
- Publisher
- MDPI AG
- Published
- 2022-11-12
- Volume
- 10
- Issue
- 11
- Pages
- 687
- ISSN
- 2305-6304 (ISSN-L)
- PMC
- PMC9699436
- OpenAlex
- W4308969756
- Cited by
- 37
- References
- 28
- Citations by year
- 2026: 10 · 2025: 10 · 2024: 10 · 2023: 6 · 2014: 1
- FWCI
- 2.96
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Toxics (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- National Institute of Environmental Health Sciences (U2CES026542)
- Topics
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical chemistry methods development
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolomics and Mass Spectrometry Studies
- Physical Sciences › Chemistry › Spectroscopy › Analytical Chemistry and Chromatography
- Keywords
- steroid hormones, human serum, human urine, liquid chromatography-tandem mass spectrometry, liquid-liquid extraction, isotope-labeled internal standards, corticosteroids, sex hormones, matrix effects, detection frequency, method validation, sample preparation optimization
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