Determination of steroid hormones in human plasma by GC-triple quadrupole MS

Steroids. 2015 Jul;99(Pt B):151-4. doi: 10.1016/j.steroids.2015.01.016. Epub 2015 Feb 2.


A fast and sensitive GC-MS/MS method is proposed to determine pregnenolone, dehydroepiandrosterone (DHEA), testosterone and dihydrotestosterone from human plasma. Steroids were extracted by liquid/liquid extraction, and derivatized with N-methyl-N-trimethylsilyl-trifluoracetamide. Electron ionization at 30eV was applied to generate high abundant precursor ions. Specific precursor/product transitions were generated a priori for each compound of interest and these assays were then used to selectively detect and quantify these steroids. Using the selected reaction monitoring mode, detection limits in the pg/ml (subnmol/l) range could be achieved for all four steroid compounds. The method was validated for imprecision, and recovery and applied in a pilot study in patients with hypercholesterolemia. Associations of increased cholesterol biosynthesis and low testosterone and dihydrotestosterone concentrations were found.

Keywords: DHEA; Dihydrotestosterone; Pregnenolone; Steroid hormones; Testosterone.

MeSH terms

  • Adult
  • Circadian Rhythm
  • Dehydroepiandrosterone / blood
  • Female
  • Gas Chromatography-Mass Spectrometry / methods*
  • Hormones / blood*
  • Humans
  • Male
  • Middle Aged
  • Reference Standards
  • Steroids / blood*
  • Sterols / blood
  • Time Factors


  • Hormones
  • Steroids
  • Sterols
  • Dehydroepiandrosterone