Androgen profiling by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in healthy normal-weight ovulatory and anovulatory late adolescent and young women

J Clin Endocrinol Metab. 2013 Jul;98(7):3058-67. doi: 10.1210/jc.2013-1381. Epub 2013 Jun 18.


Context: Physiological transient imbalance typical of adolescence needs to be distinguished from hyperandrogenism-related dysfunction. The accurate determination of circulating androgens is the best indicator of hyperandrogenism. However, reliable reference intervals for adolescent and young women are not available.

Objective: The aim of the study was to define androgen reference intervals in young women and to analyze the impact of the menstrual phase and ovulation efficiency over the androgen profile as assessed by reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) technique.

Participants: Female high school students aged 16-19 years were included in the study.

Main outcome measures: The study was performed on reference subjects properly selected among an unbiased population. Normal-weight, drug and disease free, eumenorrheic females with no signs of hyperandrogenism were included. The steroid hormone profile was determined by a validated in-house LC-MS/MS method. A statistical estimation of overall and menstrual phase-specific reference intervals was performed. A subgroup of anovulatory females was identified based on progesterone circulating levels. The impact of ovulation efficiency over hormonal profile was analyzed.

Results: A total of 159 females satisfied healthy criteria. Androgen levels did not vary according to menstrual phase, but a significantly higher upper reference limit was found for T in the luteal phase compared to the follicular phase. Higher T and androstenedione levels were observed in anovulatory compared to ovulatory females, paralleled by higher LH and FSH and lower 17-hydroxyprogesterone and 17β-estradiol levels.

Conclusions: This is the first study providing LC-MS/MS-based, menstrual phase-specific reference intervals for the circulating androgen profile in young females. We identified a subgroup of anovulatory healthy females characterized by androgen imbalance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 17-alpha-Hydroxyprogesterone / blood
  • 17-alpha-Hydroxyprogesterone / chemistry
  • Adolescent
  • Algorithms
  • Androgens / blood*
  • Androgens / chemistry
  • Anovulation / blood*
  • Chromatography, High Pressure Liquid
  • Cohort Studies
  • Cross-Sectional Studies
  • Estradiol / blood
  • Estradiol / chemistry
  • Female
  • Follicle Stimulating Hormone / blood
  • Humans
  • Italy
  • Luteinizing Hormone / blood
  • Menstrual Cycle / blood
  • Ovulation / blood*
  • Progesterone / blood
  • Progesterone / chemistry
  • Reference Values
  • Tandem Mass Spectrometry
  • Testosterone Congeners / blood
  • Testosterone Congeners / chemistry


  • Androgens
  • Testosterone Congeners
  • Progesterone
  • Estradiol
  • 17-alpha-Hydroxyprogesterone
  • Luteinizing Hormone
  • Follicle Stimulating Hormone