Should testosterone and Sex Hormone-Binding Globulin (SHBG) levels be measured in the morning in female patients?

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Morning Testosterone and SHBG Testing in Women

Obtaining testosterone levels in women in nonresearch settings is not recommended, and therefore timing of collection (morning vs. other times) is generally not clinically relevant for routine practice. 1

Guideline-Based Recommendation

The HIV Medicine Association of the Infectious Diseases Society of America explicitly states that obtaining testosterone levels in women in nonresearch settings is not recommended (strong recommendation, low quality evidence). 1 This guideline specifically recommends morning serum testosterone levels only for adult men with symptoms such as decreased libido, erectile dysfunction, reduced bone mass, or low trauma fractures. 1

When Testing May Be Considered

If testosterone testing is pursued in women despite guideline recommendations against routine testing, the following considerations apply:

Timing of Collection

  • Morning collection (8:30 AM) yields slightly higher testosterone levels than afternoon collection (1:30 PM) in women, with mean levels of 23.4 ± 12.4 ng/dL vs. 21.7 ± 12.9 ng/dL respectively (p=0.03). 2
  • The diurnal variation in women is relatively small compared to overall variability, so reference ranges can be applied irrespective of the day in the menstrual cycle or time of day the sample is taken. 3
  • Morning fasting samples are preferred when accurate measurement is needed, consistent with general principles for hormone testing. 4, 5

SHBG Measurement

  • SHBG should be measured alongside testosterone to calculate the free testosterone index (ratio of total testosterone/SHBG), as this provides better assessment of androgen status than total testosterone alone. 6
  • SHBG levels remain relatively stable across age ranges in premenopausal women (reference range 18-86 nmol/L for a 30-year-old woman). 3
  • A free testosterone index <0.3 indicates hypogonadism. 6

Measurement Methodology

If testing is performed:

  • Use liquid chromatography-tandem mass spectrometry in a laboratory certified by the CDC's Hormone Standardization Program, as immunoassays lack sufficient sensitivity for accurately measuring the low testosterone concentrations found in women. 4, 7
  • Free testosterone should ideally be measured by equilibrium dialysis method if needed. 4, 5
  • For premenopausal women, reference ranges for a 30-year-old are: total testosterone 15-46 ng/dL, free testosterone 1.2-6.4 pg/mL, and bioavailable testosterone 1.12-7.62 ng/dL. 3

Critical Caveats

  • Most immunoassay-based methods used in clinical laboratories are inadequate for measuring testosterone in women due to insufficient low-end sensitivity, leading to inaccurate results. 7
  • The clinical utility of testosterone measurement in women remains controversial and limited due to lack of well-established diagnostic criteria for female androgen deficiency. 7
  • Clinical diagnosis must correlate laboratory values with symptoms, not rely on laboratory values alone. 6

References

Guideline

Guideline Directed Topic Overview

Dr.Oracle Medical Advisory Board & Editors, 2025

Research

Accurate Measurement and Harmonized Reference Ranges for Total and Free Testosterone Levels.

Endocrinology and metabolism clinics of North America, 2022

Research

Accurate measurement of total and free testosterone levels for the diagnosis of androgen disorders.

Best practice & research. Clinical endocrinology & metabolism, 2022

Guideline

Diagnosing Hypogonadism with Testosterone Labs

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2025

Professional Medical Disclaimer

This information is intended for healthcare professionals. Any medical decision-making should rely on clinical judgment and independently verified information. The content provided herein does not replace professional discretion and should be considered supplementary to established clinical guidelines. Healthcare providers should verify all information against primary literature and current practice standards before application in patient care. Dr.Oracle assumes no liability for clinical decisions based on this content.

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