Feasibility of a liquid-phase microextraction sample clean-up and liquid chromatographic/mass spectrometric screening method for selected anabolic steroid glucuronides in biological samples

Feasibility of a liquid-phase microextraction sample clean-up and liquid chromatographic/mass spectrometric screening method for selected anabolic steroid glucuronides in biological samples / Tiia Kuuranne, Tapio Kotiaho, Stig Pedersen-Bjergaard, Knut Einar Rasmussen, Antti Leinonen, Steven Westwood, Risto Kostiainen. - (Journal of Mass Spectrometry 38 (2003) 1 (January); p. 16-26)

  • PMID: 12526002
  • DOI: 10.1002/jms.393


Abstract

Anabolic androgenic steroids (AAS) are metabolized extensively in the human body, resulting mainly in the formation of glucuronide conjugates. Current detection methods for AAS are based on gas chromatographic/mass spectrometric (GC/MS) analysis of the hydrolyzed steroid aglycones. These analyses require laborious sample preparation steps and are therefore time consuming. Our interest was to develop a rapid and straightforward method for intact steroid glucuronides in biological samples, using liquid-phase microextraction (LPME) sample clean-up and concentration method combined with liquid chromatographic/tandem mass spectrometric (LC/MS/MS) analysis. The applicability of LPME was optimized for 13 steroid glucuronides, and compared with conventional liquid-liquid extraction (LLE) and solid-phase extraction (SPE) procedures. An LC/MS/MS method was developed for the quantitative detection of AAS glucuronides, using a deuterium-labeled steroid glucuronide as the internal standard. LPME, owing to its high specificity, was shown to be better suited than conventional LLE and SPE for the clean-up of urinary AAS glucuronides. The LPME/LC/MS/MS method was fast and reliable, offering acceptable reproducibility and linearity with detection limits in the range 2-20 ng ml(-1) for most of the selected AAS glucuronides. The method was successfully applied to in vitro metabolic studies, and also tested with an authentic forensic urine sample. For a urine matrix the method still has some unsolved problems with specificity, which should be overcome before the method can be reliably used for doping analysis, but still offering additional and complementary data for current GC/MS analyses.

Original document

Parameters

Science
Study
Date
2 January 2003
People
Kostiainen, Risto
Kotiaho, Tapio
Kuuranne, Tiia
Leinonen, Antti
Pedersen-Bjergaard, Stig
Rasmussen, Knut Einar
Westwood, Steven
Country
Australia
Finland
Norway
Language
English
Other organisations
Helsingin yliopisto - University of Helsinki
Universitetet i Oslo - University of Oslo
Laboratories
Helsinki, Finland: Doping Control Laboratory
Analytical aspects
Mass spectrometry analysis
Testing method development
Doping classes
S1. Anabolic Agents
Document category
Scientific article
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Date generated
2 March 2021
Date of last modification
3 March 2021
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