Evaluation of urinary excretion of androgens conjugated to cysteine in human pregnancy by mass spectrometry

J Steroid Biochem Mol Biol. 2014 Jan;139:192-200. doi: 10.1016/j.jsbmb.2013.01.014. Epub 2013 Feb 11.


Alterations in the maternal excretion of steroids during pregnancy are not restricted to the production of progesterone and estriol by the fetoplacental unit. Although there is a lack of longitudinal data on urinary androgen concentrations during pregnancy, some studies revealed that modifications in the excretions of androgens might be significant. Recently, several testosterone metabolites excreted as cysteine conjugates have been reported in human urine. We conducted a longitudinal study on androgens conjugated with cysteine and major androgens and estrogens excreted as glucuronides in three pregnant women by mass spectrometric techniques. The urinary concentrations obtained in samples weekly collected during each of the three trimesters and samples collected before pregnancy were compared. Results showed a significant increase in urinary estrogens and norandrosterone and a moderate decrease in the urinary concentrations for most of the androgens. The most significant exception to this behavior was the rise observed for epitestosterone glucuronide when comparing basal levels with the first trimester. Cysteinyl conjugates of testosterone metabolites showed a different behavior. Whereas 4,6-androstanedione remained almost constant through the three trimesters, and Δ(6)-testosterone decreased as the majority of androgens, the excretion profile of 1,4-androstanedione notably increased, reaching a maximum at the third trimester. Alterations in the steroid profile are used in doping control analysis for the screening of endogenous anabolic androgenic steroid misuse. In this study, the main parameters proposed for doping control have been determined for basal samples and samples collected in the first trimester and they have been compared. In spite of the limited number of cases, significant variations have been found in all pregnancies studied. These alterations have to be taken into consideration if anabolic steroids are included into the Athlete Biological Passport. This article is part of a Special Issue entitled 'Pregnancy and Steroids'.

Keywords: 5α-androstane-3α,17β-diol; 5α-dihydrotestosterone; 5α-diol; 5β-androstane-3α,17β-diol; 5β-diol; Androgens; DHEA; DHT; Doping; MSTFA; Mass spectrometry; N-methyl-N-trimethylsilyl-trifluoroacetamide; Pregnancy; SRM; T/E; Urine; dehydroepiandrosterone; ratio between testosterone and epitestosterone excreted as glucuronides; selected reaction monitoring.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Androgens / urine*
  • Androstadienes / urine
  • Androstenedione / urine
  • Cysteine / urine*
  • Dehydroepiandrosterone / urine
  • Dihydrotestosterone / urine
  • Doping in Sports
  • Estrogens / urine
  • Female
  • Gas Chromatography-Mass Spectrometry / standards
  • Humans
  • Longitudinal Studies
  • Pregnancy
  • Reference Standards
  • Reference Values
  • Tandem Mass Spectrometry / standards
  • Testosterone / analogs & derivatives
  • Testosterone / urine


  • Androgens
  • Androstadienes
  • Estrogens
  • Dihydrotestosterone
  • testosterone glucuronate
  • 1,4-androstadiene-3,17-dione
  • Testosterone
  • Androstenedione
  • Dehydroepiandrosterone
  • 4,6-androstadiene-3,17-dione
  • Cysteine