Direct detection and quantification of 19-norandrosterone sulfate in human urine by liquid chromatography-linear ion trap mass spectrometry

详细信息    查看全文

• 作者：Emmanuel Strahm ; Christophe Saudan ; Pierre-Edouard Sottas ; Patrice Mangin ; Martial Saugy
• 关键词：Nandrolone ; 19-Norandrosterone sulfate ; Liquid chromatography-mass spectrometry ; Doping control
• 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
• 出版年：2007
• 出版时间：1 June 2007
• 年：2007
• 卷：852
• 期：1-2
• 页码：491-496
• 全文大小：184 K

文摘

19-Norandrosterone sulfate (19-NAS) is the sulfoconjugated form of 19-norandrosterone (19-NA), the major metabolite of the steroid nandrolone. A sensitive and accurate liquid chromatography/tandem mass spectrometry (LC-MS/MS) assay was developed for the direct measurement of 19-NAS in human urine samples. The method involved a quaternary amine SPE protocol and subsequently injection of the extract onto an analytical column (Uptisphere^® ODB, 150 mm × 3.0 mm, 5 μm) for chromatographic separation and mass spectrometry detection in negative electrospray ionisation mode. The sulfoconjugate of 19-NA was identified in urine by comparison of mass spectra and retention time with a reference substance. The limit of detection (LOD) and lowest limit of quantification (LLOQ) of 19-NAS were of 40 pg/mL and 200 pg/mL, respectively. For a nominal concentration of 2 ng/mL, recovery (94 % ), intra-day precision (2.7 % ), intra-assay precision (6.6 % ) and inter-assay precision (14.3 % ) were determined. Finally, this analytical method was applied for quantifying the concentration of 19-NAS in doping samples, using calibration curves (0.2–20 ng/mL) and the standard-addition method. The results show the feasibility of applying this LC-MS/MS assay as a complementary tool to detect misuse of nandrolone or nandrolone precursors.