Quantitation of cortisol and related 3-oxo-4-ene steroids in urine using gas chromatography/mass spectrometry with stable isotope-labeled internal standards
Abstract
A method for the profiling of several important 3-oxo-4-ene urinary steroids is reported. The methodology is combined gas chromatography/mass spectrometry (GC/MS) utilizing stable isotope-labeled internal standards. The following standards were obtained or easily synthesized: [9, 11, 12, 12-2H4]cortisol, [1,2-2H2] and [9, 12, 12-2H2]cortisone, [1,2-2H2]6 beta-hydroxycortisol, and [1,2-2H2]18-hydroxycortisol. We found the following excretions of free steroids for normal adult males and females: cortisol (males mean +/- SD, 35 +/- 13; females mean +/- SD, 23 +/- 13), cortisone (males mean +/- SD, 58 +/- 23; females mean +/- SD, 50 +/- 22), 6 beta-hydroxycortisol (males mean +/- SD, 164 +/- 59; females mean +/- SD, 108 +/- 55), and 18-hydroxycortisol (males mean +/- SD, 148 +/- 55; females mean +/- SD, 71 +/- 30). For 18-hydroxycortisol in particular, the excretions were much higher for males than for females. We found that the larger part of urinary cortisol and cortisone is not free but is released from conjugation by enzymes present in snail digestive juice. Using a pooled urine sample from an equal number of male and female subjects, we found that for cortisol 29% was excreted free, 28% as glucuronide and 43% as other conjugates (probably sulfates). For cortisone 41% was free, 45% beta-glucuronide and 14% as other conjugates. Relatively little (3-8%) of the hydroxylated cortisols were excreted conjugated.
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Two studies report unconjugated excretion of cortisol (30%), cortisone, 20αDHE, 20βDHE, 20βDHF (40-60%); and 20αDHF, 6β-OH-cortisol, 6β-OH-E and 18-OH-cortisol (80-100%).
“Two studies report unconjugated excretion of the following individual steroids: cortisol (30%), cortisone, 20αDHE, 20βDHE, 20βDHF (40–60%); 20αDHF, 6β-OH-cortisol, 6β-OH-E and 18-OH-cortisol (80–100%) [273,274].”
From Human steroid biosynthesis, metabolism and excretion are differentially reflected by serum and urine steroid metabolomes: a comprehensive review (2019) · cites this paper as [273]
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- Authors
- Palermo, M., Gomez-Sanchez, C., Roitman, E., & Shackleton, C. H. L.
- Affiliations
- University of Sassari (IT)
- Corresponding author
- Shackleton, Cedric H.L.
- Venue
- Steroids
- Publisher
- Elsevier BV
- Published
- 1996-10
- Volume
- 61
- Issue
- 10
- Pages
- 583-589
- ISSN
- 0039-128X (ISSN-L)
- OpenAlex
- W2019733623
- Cited by
- 70
- References
- 24
- Citations by year
- 2024: 3 · 2022: 1 · 2019: 2 · 2018: 1 · 2017: 1 · 2015: 1 · 2014: 2 · 2013: 3
- FWCI
- 0.80
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Steroids (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5r01dk034400-10)
- National Institute of Diabetes and Digestive and Kidney Diseases (DK-34400)
- Topics
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
- Physical Sciences › Chemistry › Spectroscopy › Analytical Chemistry and Chromatography
- Life Sciences › Neuroscience › Behavioral Neuroscience › Stress Responses and Cortisol
- Keywords
- urinary cortisol, cortisone, 6β-hydroxycortisol, 18-hydroxycortisol, steroid conjugation, glucuronide conjugates, sulfate conjugates, isotope-labeled internal standards
- MeSH terms
- Adolescent, Adult, Child, Chromatography, Gas, Cortisone, Female, Humans, Hydrocortisone, Male, Mass Spectrometry, Steroids, Reproducibility of Results
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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