Feasibility of a liquid-phase microextraction sample clean-up and liquid chromatographic/mass spectrometric screening method for selected anabolic steroid glucuronides in biological samples
Version of Record online: 2 JAN 2003
Copyright © 2003 John Wiley & Sons, Ltd.
Journal of Mass Spectrometry
Volume 38, Issue 1, pages 16–26, January 2003
How to Cite
Kuuranne, T., Kotiaho, T., Pedersen-Bjergaard, S., Einar Rasmussen, K., Leinonen, A., Westwood, S. and Kostiainen, R. (2003), Feasibility of a liquid-phase microextraction sample clean-up and liquid chromatographic/mass spectrometric screening method for selected anabolic steroid glucuronides in biological samples. J. Mass Spectrom., 38: 16–26. doi: 10.1002/jms.393
- Issue online: 8 JAN 2003
- Version of Record online: 2 JAN 2003
- Manuscript Accepted: 1 OCT 2002
- Manuscript Received: 18 JUL 2002
- National Technology Agency (TEKES).
- anabolic steroids;
- glucuronide conjugates;
- liquid-phase microextraction;
- liquid chromatography;
- tandem mass spectrometry
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. Copyright © 2003 John Wiley & Sons, Ltd.