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Determination of mepitiostane metabolites in human urine by liquid chromatography/tandem mass spectrometry for sports drug testing
Determination of mepitiostane metabolites in human urine by liquid chromatography/tandem mass spectrometry for sports drug testing
- Source :
- Journal of Pharmaceutical and Biomedical Analysis. 115:236-244
- Publication Year :
- 2015
- Publisher :
- Elsevier BV, 2015.
-
Abstract
- Mepitiostane (2α,3α-epithio-17β-(1-methoxycyclopentyloxy)-5α-androstane), which is a prodrug of epitiostanol (2α,3α-epitio-5α-androstane-17β-ol), is an epitiosteroid having anti-estrogenic and weak androgenic anabolic activities. The World Anti-Doping Agency prohibits the misuse of mepitiostane by athletes. Detection of the urinary metabolites epitiostanol sulfoxide and epitiostanol was studied using liquid chromatography/mass spectrometry (LC-MS) for doping control purposes. The use of LC-MS provided advantages over gas chromatography/mass spectrometry for detecting heat labile steroids because epitiostanol and epitiostanol sulfoxide were primarily pyrolized to 5α-androst-2-en-17β-ol. The method consists of enzymatic hydrolysis using β-glucuronidase (Escherichia coli), liquid-liquid extraction, and subsequent ultra-performance liquid chromatography/electrospray-tandem mass spectrometry. Epitiostanol sulfoxide was determined at urinary concentrations of 0.5-50ng/mL, recovery was 76.2-96.9%, and assay precision was calculated as 0.9-1.7% (intra-day) and 2.0-6.6% (inter-day). Epitiostanol was determined at urinary concentrations of 0.5-50ng/mL, recovery was 26.1-35.6% and assay precision was calculated as 4.1-4.6% (intra-day) and 3.3-8.5% (inter-day). The limits of detection for epitiostanol sulfoxide and epitiostanol were 0.05ng/mL and 0.10ng/mL, respectively. Epitiostanol sulfoxide and epitiostanol, as their gluco-conjugates, were identified in human urine after oral administration of 10mg mepitiostane. Epitiostanol sulfoxide and epitiostanol could be detected up to 48h and 24h after administration, respectively. The results showed that the detection window of epitiostanol is much shorter than that of epitiostanol sulfoxide. The LC-MS detection of urinary epitiostanol sulfoxide, a specific metabolite with a sulphur atom in its molecular structure, is likely to be able to identify the abuse of mepitiostane.
- Subjects :
- Spectrometry, Mass, Electrospray Ionization
Metabolite
Clinical Biochemistry
Pharmaceutical Science
Urine
Tandem mass spectrometry
Mass spectrometry
Analytical Chemistry
chemistry.chemical_compound
Limit of Detection
Tandem Mass Spectrometry
Liquid chromatography–mass spectrometry
Drug Discovery
Humans
Androstanols
Chromatography, High Pressure Liquid
Spectroscopy
Doping in Sports
Detection limit
Chromatography
Molecular Structure
Chemistry
Reproducibility of Results
Sulfoxide
Sulfoxides
Gas chromatography
Metabolic Networks and Pathways
Subjects
Details
- ISSN :
- 07317085
- Volume :
- 115
- Database :
- OpenAIRE
- Journal :
- Journal of Pharmaceutical and Biomedical Analysis
- Accession number :
- edsair.doi.dedup.....c64ae218fdec178dfbb3dbccf5d844c7
- Full Text :
- https://doi.org/10.1016/j.jpba.2015.07.016