Analysis of anabolic steroids by partial filling micellar electrokinetic capillary chromatography and electrospray mass spectrometry

Lotta Kaarina Amundsen, Juha T. Kokkonen, Stella Rovio, Heli Sirén (Corresponding Author)

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A partial filling micellar electrokinetic capillary chromatography (PF-MEKC) separation of six anabolic androgenic steroids (androstenedione, metandienone, fluoxymesterone, methyltestosterone, 17-epimetandienone and testosterone) is introduced. The method utilises a mixed micellar solution consisting of sodium dodecyl sulphate (SDS) and sodium taurocholate. The analytes are detected with a photodiode array detector at 247 nm wavelength. Methyltestosterone is used as internal standard. The detection limits were 39 μg/L for androstenedione, 40 μg/L for testosterone, 45 μg/L for fluoxymesterone, 45–90 μg/L for 17-epimetandienone, 59 μg/L for methyltestosterone and 90 μg/L for metandienone. Linear correlation between concentration (0.1–5.0 mg/L) and detector response was obtained with r2 of 0.994 for fluoxymesterone, 0.998 for 17-epimetandienone and 0.999 for androstenedione, metandienone and testosterone. In addition, ionisation of the investigated compounds in electrospray mass spectrometry (ESI-MS) was studied in positive ion mode. The most intense signal (100%) was the protonated molecular ion [M+H]+, except for 17-epimetandienone, which gave its strongest signal at m/z corresponding to [M−H2O+H]+. Finally, separation and identification of fluoxymesterone, androstenedione and testosterone by PF-MEKC–ESI-MS is described. This is the first use of PF-MEKC and PF-MEKC–ESI-MS assays for anabolic androgenic steroids.
Original languageEnglish
Pages (from-to)123-131
JournalJournal of Chromatography A
Issue number1
Publication statusPublished - 2004
MoE publication typeA1 Journal article-refereed



  • Doping analysis
  • Partial filling micellar electrokinetic chromatography
  • Steroids
  • Hormones

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