Detection, quantification and confirmation of anabolic steroids in equine plasma by liquid chromatography and tandem mass spectrometry

Fuyu Guan, Cornelius E. Uboh, Lawrence R. Soma, Yi Luo, Jeffery Rudy, Thomas Tobin

Research output: Contribution to journalArticlepeer-review

85 Scopus citations


Anabolic androgenic steroids are related to the male sex hormones and are abused in equine sports. In an effort to deter the abuse of anabolic steroids, a sensitive LC-MS/MS method was developed for detection, quantification and confirmation of eight major anabolic steroids (testosterone, normethandrolone, nandrolone, boldenone, methandrostenolone, tetrahydrogestrinone (THG), trenbolone, and stanozolol) in equine plasma. Formation of solvent adduct ions of the analytes was observed under electrospray ionization (ESI) conditions, and desolvation of the solvent adduct ions by source collision-induced decomposition (CID) increased the abundance of the [M + H]+ ions as well as the multiple-reaction monitoring (MRM) signals. ESI (+) and APCI (+) were compared with respect to sensitivity for the analytes and the former provided better sensitivity. The matrix effect on ion suppression or enhancement was evaluated, and was negligible. Confirmation of the analytes was performed using criteria of three ion transitions and LC retention time of each analyte. The limit of detection (LOD) and quantification (LOQ) was 25 pg/mL. The limit of confirmation (LOC) was 25 pg/mL for boldenone; 50 pg/mL for normethandrolone, nandrolone, and methandrostenolone; and 100 pg/mL for testosterone, THG, trenbolone, and stanozolol. The analytes were evaluated for stability and found to be stable in plasma for 24 h at room temperature, 13 days at 4°C, and 34 days at -20 and -70°C. The method was successfully applied to analyses of equine plasma samples for pharmacokinetics study. This method is sensitive and useful for detection, quantification and confirmation of these anabolic steroids in equine plasma.

Original languageEnglish
Pages (from-to)56-68
Number of pages13
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Issue number1-2
StatePublished - Dec 27 2005

Bibliographical note

Funding Information:
This study was funded by the Pennsylvania Horse and Harness Racing Commissions to which the authors are very grateful. Financial contribution was also made by the Pocono Downs Horsemen Association. The authors are very thankful to Dr. Donald Catlin of the Olympic Analytical Laboratory at the University of California, Los Angeles, CA, USA for information on dose of THG for administration. The authors acknowledge Dr. Susan McDonalds of the University of Pennsylvania, School of Veterinary Medicine at New Bolton Center Campus, Kennett Square, PA, USA for providing the stallion plasma samples, and for the excellent technical support of Fengyu Hao, Anne Hess, Deborah Tsang, and Donna Telies.


  • Anabolic steroid
  • Boldenone
  • Doping control
  • Horse
  • LC-MS
  • Plasma
  • THG
  • Testosterone

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology


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