Analysis of anabolic steroids in human hair using LC–MS/MS

Analysis of anabolic steroids in human hair using LC–MS/MS / Nawed Deshmukh, Iltaf Hussain, James Barker, Andrea Petroczi, Declan P. Naughton. - (Steroids 75 (2010) 10 (October); p. 710-714)

  • PMID: 20435054
  • DOI: 10.1016/j.steroids.2010.04.007


New highly sensitive, specific, reliable, reproducible and robust LC-MS/MS methods were developed to detect the anabolic steroids, nandrolone and stanozolol, in human hair for the first time. Hair samples from 180 participants (108 males, 72 females, 62% athletes) were screened using ELISA which revealed 16 athletes as positive for stanozolol and 3 for nandrolone. Positive samples were confirmed on LC-MS/MS in selective reaction monitoring (SRM) mode. The assays for stanozolol and nandrolone showed good linearity in the range 1-400pg/mg and 5-400pg/mg, respectively. The methods were validated for LLOD, interday precision, intraday precision, specificity, extraction recovery and accuracy. The assays were capable of detecting 0.5pg stanozolol and 3.0pg nandrolone per mg of hair, when approximately 20mg of hair were processed. Analysis using LC-MS/MS confirmed 11 athletes' positive for stanozolol (5.0pg/mg to 86.3pg/mg) and 1 for nandrolone (14.0pg/mg) thus avoiding false results from ELISA screening. The results obtained demonstrate the application of these hair analysis methods to detect both steroids at low concentrations, hence reducing the amount of hair required significantly. The new methods complement urinalysis or blood testing and facilitate improved doping testing regimes. Hair analysis benefits from non-invasiveness, negligible risk of infection and facile sample storage and collection, whilst reducing risks of tampering and cross-contamination. Owing to the wide detection window, this approach may also offer an alternative approach for out-of-competition testing.

Original document


Research / Study
30 April 2010
Barker, James
Deshmukh, Nawed
Hussain, Iltaf
Naughton, Declan P.
Petróczi, Andrea
United Kingdom
Other organisations
Kingston University London
Analytical aspects
Mass spectrometry analysis
Testing method development
Doping classes
S1. Anabolic Agents
Nandrolone (19-nortestosterone)
Document category
Scientific article
Document type
Pdf file
Date generated
28 September 2020
Date of last modification
1 October 2020
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