Age-Stratified Reference Ranges for Directly Measured Serum Free Testosterone in Community-Dwelling and Healthy Men

Direct measurement of serum free testosterone (FT) may help evaluate hypogonadism in men. However, up to present, availability of reference ranges for measured FT (mFT) is limited. To establish age-stratified reference ranges for mFT in healthy and community-dwelling adult men. Serum samples of 1202...

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Published in:The journal of clinical endocrinology and metabolism
Main Authors: Walravens, Joeri, Snaterse, Gido, Narinx, Nick, Reyns, Tim, Van Uytfanghe, Katleen, Vanderschueren, Dirk, Wu, Frederick, Kaufman, Jean-Marc, Antonio, Leen, Fiers, Tom, Lapauw, Bruno
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Language:English
Published: United States 10.09.2025
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ISSN:1945-7197, 1945-7197
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Abstract Direct measurement of serum free testosterone (FT) may help evaluate hypogonadism in men. However, up to present, availability of reference ranges for measured FT (mFT) is limited. To establish age-stratified reference ranges for mFT in healthy and community-dwelling adult men. Serum samples of 1202 Caucasian men were collected from large population cohort studies. Free testosterone was measured using equilibrium dialysis followed by liquid chromatography tandem-mass spectrometry (LC-MS/MS). Sex-hormone binding globulin was measured using immunoassay and total testosterone was measured using LC-MS/MS. Reference ranges per age decade and for men aged 18 to 39 years with BMI < 30 kg/m² were established following Clinical & Laboratory Standards Institute guidelines. Reference ranges were established as the 2.5th and 97.5th percentiles. The reference range for mFT in healthy, non-obese men aged 18 to 39 years was 184-749 pmol/L. mFT levels showed a decrease with aging of -4.5 pmol/L per year. Having a BMI of 30 kg/m² and 35 kg/m² resulted in a decrease in mFT of 14.4% and 22.2%, respectively, compared to a reference BMI of 22 kg/m². This study provides age-stratified reference ranges for mFT in adult men, established in the largest cohort to date, using published methodology. Our results also show that mFT levels are influenced by age and BMI. Future work should focus on whether age-stratified or reference ranges for young men should be employed, and how mFT can best be implemented in clinical practice including the improved application of calculated FT.
AbstractList Direct measurement of serum free testosterone (FT) may help evaluate hypogonadism in men. However, up to present, availability of reference ranges for measured FT (mFT) is limited. To establish age-stratified reference ranges for mFT in healthy and community-dwelling adult men. Serum samples of 1202 Caucasian men were collected from large population cohort studies. Free testosterone was measured using equilibrium dialysis followed by liquid chromatography tandem-mass spectrometry (LC-MS/MS). Sex-hormone binding globulin was measured using immunoassay and total testosterone was measured using LC-MS/MS. Reference ranges per age decade and for men aged 18 to 39 years with BMI < 30 kg/m² were established following Clinical & Laboratory Standards Institute guidelines. Reference ranges were established as the 2.5th and 97.5th percentiles. The reference range for mFT in healthy, non-obese men aged 18 to 39 years was 184-749 pmol/L. mFT levels showed a decrease with aging of -4.5 pmol/L per year. Having a BMI of 30 kg/m² and 35 kg/m² resulted in a decrease in mFT of 14.4% and 22.2%, respectively, compared to a reference BMI of 22 kg/m². This study provides age-stratified reference ranges for mFT in adult men, established in the largest cohort to date, using published methodology. Our results also show that mFT levels are influenced by age and BMI. Future work should focus on whether age-stratified or reference ranges for young men should be employed, and how mFT can best be implemented in clinical practice including the improved application of calculated FT.
Direct measurement of serum free testosterone (FT) may help evaluate hypogonadism in men. However, up to present, availability of reference ranges for measured FT (mFT) is limited.CONTEXTDirect measurement of serum free testosterone (FT) may help evaluate hypogonadism in men. However, up to present, availability of reference ranges for measured FT (mFT) is limited.To establish age-stratified reference ranges for mFT in healthy and community-dwelling adult men.OBJECTIVETo establish age-stratified reference ranges for mFT in healthy and community-dwelling adult men.Serum samples of 1202 Caucasian men were collected from large population cohort studies. Free testosterone was measured using equilibrium dialysis followed by liquid chromatography tandem-mass spectrometry (LC-MS/MS). Sex-hormone binding globulin was measured using immunoassay and total testosterone was measured using LC-MS/MS. Reference ranges per age decade and for men aged 18 to 39 years with BMI < 30 kg/m² were established following Clinical & Laboratory Standards Institute guidelines. Reference ranges were established as the 2.5th and 97.5th percentiles.METHODSSerum samples of 1202 Caucasian men were collected from large population cohort studies. Free testosterone was measured using equilibrium dialysis followed by liquid chromatography tandem-mass spectrometry (LC-MS/MS). Sex-hormone binding globulin was measured using immunoassay and total testosterone was measured using LC-MS/MS. Reference ranges per age decade and for men aged 18 to 39 years with BMI < 30 kg/m² were established following Clinical & Laboratory Standards Institute guidelines. Reference ranges were established as the 2.5th and 97.5th percentiles.The reference range for mFT in healthy, non-obese men aged 18 to 39 years was 184-749 pmol/L. mFT levels showed a decrease with aging of -4.5 pmol/L per year. Having a BMI of 30 kg/m² and 35 kg/m² resulted in a decrease in mFT of 14.4% and 22.2%, respectively, compared to a reference BMI of 22 kg/m².RESULTSThe reference range for mFT in healthy, non-obese men aged 18 to 39 years was 184-749 pmol/L. mFT levels showed a decrease with aging of -4.5 pmol/L per year. Having a BMI of 30 kg/m² and 35 kg/m² resulted in a decrease in mFT of 14.4% and 22.2%, respectively, compared to a reference BMI of 22 kg/m².This study provides age-stratified reference ranges for mFT in adult men, established in the largest cohort to date, using published methodology. Our results also show that mFT levels are influenced by age and BMI. Future work should focus on whether age-stratified or reference ranges for young men should be employed, and how mFT can best be implemented in clinical practice including the improved application of calculated FT.CONCLUSIONThis study provides age-stratified reference ranges for mFT in adult men, established in the largest cohort to date, using published methodology. Our results also show that mFT levels are influenced by age and BMI. Future work should focus on whether age-stratified or reference ranges for young men should be employed, and how mFT can best be implemented in clinical practice including the improved application of calculated FT.
Author Snaterse, Gido
Narinx, Nick
Reyns, Tim
Vanderschueren, Dirk
Kaufman, Jean-Marc
Lapauw, Bruno
Antonio, Leen
Fiers, Tom
Walravens, Joeri
Van Uytfanghe, Katleen
Wu, Frederick
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  givenname: Gido
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  givenname: Leen
  orcidid: 0000-0002-1079-2860
  surname: Antonio
  fullname: Antonio, Leen
  organization: Laboratory of Clinical and Experimental Endocrinology, KULeuven, Herestraat 49, 3000 Leuven, Belgium
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  givenname: Tom
  surname: Fiers
  fullname: Fiers, Tom
  organization: Department of Clinical Chemistry, Ghent University Hospital, Corneel Heymanslaan 10, 9000 Ghent, Belgium
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  orcidid: 0000-0002-1584-4965
  surname: Lapauw
  fullname: Lapauw, Bruno
  organization: Department of Endocrinology, Ghent University Hospital; Department of Internal Medicine & Pediatrics, Ghent University, Corneel Heymanslaan 10, 9000 Ghent, Belgium
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Keywords LC-MS/MS
reference ranges
free testosterone
men
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