Isolation and quantification by high-performance liquid chromatography–ion-trap mass spectrometry of androgen sulfoconjugates in human urine

详细信息    查看全文

• 作者：Emmanuel Strahm ; Isabelle Kohler ; Serge Rudaz ; Sophie Martel ; Pierre-Alain Carrupt ; Jean-Luc Veuthey ; Martial Saugy ; Christophe Saudan
• 关键词：Endogenous steroids ; Phase II metabolism ; Mixed-mode polymeric anion-exchange ; Solid-phase extraction ; Linear ion-trap mass spectrometry
• 刊名：Journal of Chromatography A
• 出版年：2008
• 出版时间：4 July 2008
• 年：2008
• 卷：1196-1197
• 期：Complete
• 页码：153-160
• 全文大小：555 K

文摘

Together with steroid glucuronides, sulfoconjugates may be used as markers of steroid administration as well as endogenous steroid production. A fast and sensitive analytical procedure has been developed for the simultaneous separation, determination and quantification of sulfate and glucuronide derivatives of testosterone (T), epitestosterone (E), androsterone (A), etiocholanolone (Etio) and dehydroepiandrosterone (DHEA) in human urine. First, a weak anion-exchange solid-phase extraction support (SPE Oasis WAX) was used for complete and rapid separation of sulfates and glucuronides in two extracts after loading of urine sample (2 mL). Then sulfates were analyzed directly by high-performance liquid chromatography–ion-trap mass spectrometry (LC–MS/MS) with electrospray ionization in negative mode. Chromatographic separation of the targeted sulfoconjugates was achieved using a Waters XBridge C₁₈ column (150 mm × 4.6 mm I.D., 5 μm) with gradient elution. Assay validation demonstrated good performance for instance for T sulfate (TS) and E sulfate (ES) in terms of trueness (89–107 % ), repeatability (3.4–22 % ) and intermediate precision (5.8–22 % ) over the range of 2–200 ng/mL (corresponding to 1.5–147 ng/mL as free steroids). Results obtained on biological samples demonstrated the suitability of this analytical strategy for direct measurement of androgen sulfoconjugates and glucuroconjugates in human urine.