Development and validation of a qualitative screening method for the detection of exogenous anabolic steroids in urine by liquid chromatography-tandem mass spectrometry

Anal Bioanal Chem. 2007 Oct;389(4):1209-24. doi: 10.1007/s00216-007-1530-6. Epub 2007 Aug 28.


A screening method for the urinary detection of 34 exogenous anabolic steroids has been developed. The method involves an enzymatic hydrolysis, liquid-liquid extraction and detection by liquid chromatography-tandem mass spectrometry. The use of some adducts such as [M+NH(4)](+), [M+CH(3)COO](-) and [M+H+MeOH](+) was necessary in order to detect some analytes at the required level (lower than 10 ng/ml). Two transitions were selected for each analyte. Different concentration factors have been studied in order to increase the sensitivity. A concentration factor of 50 was selected for the screening method although the high ion suppression observed under these conditions can hamper its application as a quantitative method. The method was validated and limits of detection were obtained by spiking ten different blank urine samples at five different concentration levels. Up to 29 analytes were detected in all spiked urines at the required level. Limits of detection between 1 and 10 ng/ml were obtained for most analytes which fulfil current requirements. The applicability of the method was shown by analysing positive samples.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Acetates / chemistry
  • Anabolic Agents / urine*
  • Chromatography, Liquid
  • Doping in Sports*
  • Humans
  • Molecular Structure
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Steroids / chemistry
  • Steroids / urine*
  • Substance Abuse Detection / methods*
  • Substance-Related Disorders / diagnosis
  • Substance-Related Disorders / urine
  • Tandem Mass Spectrometry / methods*


  • Acetates
  • Anabolic Agents
  • Steroids
  • ammonium acetate