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Analysis of anabolic androgenic steroids as sulfate conjugates using high performance liquid chromatography coupled to tandem mass spectrometry
Author(s) -
Rzeppa S.,
Heinrich G.,
Hemmersbach P.
Publication year - 2015
Publication title -
drug testing and analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.065
H-Index - 54
eISSN - 1942-7611
pISSN - 1942-7603
DOI - 10.1002/dta.1895
Subject(s) - chromatography , chemistry , glucuronic acid , solid phase extraction , tandem mass spectrometry , mass spectrometry , liquid chromatography–mass spectrometry , sulfation , high performance liquid chromatography , sulfate , biochemistry , polysaccharide , organic chemistry
Improvements in doping analysis can be effected by speeding up analysis time and extending the detection time. Therefore, direct detection of phase II conjugates of doping agents, especially anabolic androgenic steroids (AAS), is proposed. Besides direct detection of conjugates with glucuronic acid, the analysis of sulfate conjugates, which are usually not part of the routine doping control analysis, can be of high interest. Sulfate conjugates of methandienone and methyltestosterone metabolites have already been identified as long‐term metabolites. This study presents the synthesis of sulfate conjugates of six commonly used AAS and their metabolites: trenbolone, nandrolone, boldenone, methenolone, mesterolone, and drostanolone. In the following these sulfate conjugates were used for development of a fast and easy analysis method based on sample preparation using solid phase extraction with a mixed‐mode sorbent and detection by high performance liquid chromatography coupled to tandem mass spectrometry (HPLC‐MS/MS). Validation demonstrated the suitability of the method with regard to the criteria given by the technical documents of the World Anti‐Doping Agency (WADA). In addition, suitability has been proven by successful detection of the synthesized sulfate conjugates in excretion urines and routine doping control samples. Copyright © 2015 John Wiley & Sons, Ltd.