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A novel spreadsheet method for calculating the free serum concentrations of testosterone, dihydrotestosterone, estradiol, estrone and cortisol: With illustrative examples from male and female populations
Abstract
In humans, testosterone (T), dihydrotestosterone (DHT), estradiol (E2), estrone (E1) and cortisol (C) bind to the serum proteins sex hormone-binding globulin (SHBG), albumin (Alb) and corticosteroid-binding globulin (CBG). Equilibrium dialysis is considered to be the "gold standard" for measuring the free concentrations of these steroids but is technically difficult and not widely available. Based on a mathematical model of the 5-ligand/3-protein binding equilibria, we developed a novel spreadsheet method for calculating the free and bioavailable (free+Alb-bound) concentrations of each steroid in terms of the total steroid and protein concentrations. The model uses 15 association constants K(SHBG-X), K(Alb-X), and K(CBG-X) (X=T, DHT, E2, E1 and C) that have been estimated from a systematic review of published binding studies. The computation of the free and bioavailable concentrations uses an iterative numerical method that can be readily programmed on a spreadsheet. The method is illustrated with six examples corresponding to young men (YM), old men (OM), obese men (Ob M), young women (YM), pregnant women in the 3rd trimester (Preg T3) and oophorectomized women on oral conjugated equine estrogens (CEE). The resulting free hormone concentrations for YM and YW fall within the normal references ranges obtained by equilibrium dialysis for all five hormones. The model also accounts for the competitive binding effects of high estrogen levels on the free T levels in Preg T3. This novel spreadsheet method provides a "user-friendly" approach for estimating the free concentrations of circulating sex hormones and cortisol in men and women.
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In the citing paper
“Among steroid hormones, T has the second highest association constant (Ka), valued at 1–2 × 109 M−1 s−1, whereas DHT has the highest Ka for SHBG (2.69 × 109 M−1 s−1) [16].”
From Role of sex hormone-binding globulin in the free hormone hypothesis and the relevance of free testosterone in androgen physiology (2022) · cites this paper as [16]
In the citing paper
“First, several equations were derived from the general law of mass action. Association constants for albumin and SHBG used in these formulas were obtained from in vitro experiments by linearizing data through Scatchard analysis [16, 208, 212].”
From Role of sex hormone-binding globulin in the free hormone hypothesis and the relevance of free testosterone in androgen physiology (2022) · cites this paper as [16]
In the citing paper
“A wide spectrum of algorithms for free T exists, which can broadly be divided into three categories (Table 6).”
From Role of sex hormone-binding globulin in the free hormone hypothesis and the relevance of free testosterone in androgen physiology (2022) · cites this paper as [16]
In the citing paper
“Circulating T is predominantly bound to sex hormone binding globulin (SHBG) and albumin, with small percentages freely circulating or associated with corticosteroid binding globulin.”
From Challenges in Testosterone Measurement, Data Interpretation, and Methodological Appraisal of Interventional Trials (2016) · cites this paper as [10]
In the citing paper
“Testosterone levels were measured by a radioimmunoassay that has been validated against liquid chromatography tandem mass spectrometry (13) and has a sensitivity of 10 ng/dL.”
From Testosterone induces erythrocytosis via increased erythropoietin and suppressed hepcidin: evidence for a new erythropoietin/hemoglobin set point (2014) · cites this paper as [13]
In the citing paper
“The Testosterone in Older Men with Mobility Limitation trial was a randomized, double-blind, placebo-controlled trial whose design and primary results have been published (13).”
From Testosterone induces erythrocytosis via increased erythropoietin and suppressed hepcidin: evidence for a new erythropoietin/hemoglobin set point (2014) · cites this paper as [13]
In the citing paper
“Eligible participants were randomized to placebo or testosterone gel based on concealed, computer-generated randomization tables, using a block size of 6. Randomization was stratified by age: (a) 65-74 and (b) 75 and older.”
From Testosterone induces erythrocytosis via increased erythropoietin and suppressed hepcidin: evidence for a new erythropoietin/hemoglobin set point (2014) · cites this paper as [13]
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Metadata
- Authors
- Mazer, N. A.
- Affiliations
- Boston University (US)
- Corresponding author
- Mazer, Norman A.
- ORCID
- Mazer, Norman A.
- Venue
- Steroids
- Publisher
- Elsevier BV
- Published
- 2009-06
- Volume
- 74
- Issue
- 6
- Pages
- 512-519
- ISSN
- 0039-128X (ISSN-L)
- OpenAlex
- W2003710746
- Cited by
- 189
- References
- 53
- Citations by year
- 2026: 7 · 2025: 11 · 2024: 8 · 2023: 8 · 2022: 9 · 2021: 11 · 2020: 5 · 2019: 13 · 2018: 13 · 2017: 17 · 2016: 11 · 2015: 11 · 2014: 11 · 2013: 13 · 2012: 11
- FWCI
- 9.84
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Steroids (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5p30ag031679-15)
- National Institute on Aging (U01AG014369, R01AG031206, 1 P30 AG031679, 5UO1AG14369)
- Topics
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Hormonal and reproductive studies
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Endocrinology › Estrogen and related hormone effects
- Keywords
- sex hormone-binding globulin, albumin, corticosteroid-binding globulin, testosterone, dihydrotestosterone, estradiol, estrone, cortisol, competitive binding, pregnancy, oophorectomy, conjugated equine estrogens, binding constant
- MeSH terms
- Adult, Aged, Algorithms, Estradiol, Estrone, Female, Humans, Hydrocortisone, Male, Mathematical Computing, Ovariectomy, Pregnancy, Pregnancy Trimester, Third, Reference Values, Dihydrotestosterone, Testosterone, Reproducibility of Results
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