New methodology in analysis of steroids using solid phase extraction and micellar electrokinetic chromatography (poster)

Lotta Amundsen, Tarja Nevanen, Kristiina Takkinen, Heli Sirén

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

Abstract

Steroid specific methodologies for monitoring body fluids are essential goals in clinical and doping analyses. The conventional GC-MS methods are laborious and time-consuming, since deconjugation, concentration and derivatisation of the steroid analytes are required. More straightforward analysis methods are thus desired. Separation of androgenic steroids with micellar electrokinetic chromatography has been studied very little. The separation of endogenous alpha-epimers testosterone and epitestosterone has not been reported with any electroseparation techniques before. Here, a novel partial filling micellar electrokinetic chromatography (PF-MEKC) method for determination of three endogenous steroid hormones (androstenedione, testosterone, epitestosterone) and two synthetic anabolic steroids (fluoxymesterone, methyltestosterone) is introduced. Discontinuous solvent pair made of single-isomer sulphated gamma-cyclodextrins and surfactants sodium dodecyl sulphate and sodium taurocholate is exploited by using two different pseudostationary phases in the capillary. The fast steroid separation is completed in 10 minutes. The PF-MEKC method was used for pretreated human urine samples. The one of the new sample preparation methods allows determination of testosterone in male urine after enzymatic hydrolysis with beta-glucuronidase from E. Coli. It is based on a highly testosterone specific microscale immunoextraction prior to PF-MEKC. The other method allows simultaneous determination of testosterone and epitestosterone in male urine after the enzymatic hydrolysis followed by their quantification in urine samples.
Original languageEnglish
Title of host publication30th International Symposium on High Performance Liquid Phase Separations and Related Techniques, HPLC 2006
Subtitle of host publicationBook of Abstracts
Publication statusPublished - 2006
Event30th International Symposium on High Performance Liquid Phase Separations and Related Techniques, HPLC 2006 - San Francisco, United States
Duration: 17 Jun 200622 Jun 2006

Conference

Conference30th International Symposium on High Performance Liquid Phase Separations and Related Techniques, HPLC 2006
Abbreviated titleHPLC 2006
CountryUnited States
CitySan Francisco
Period17/06/0622/06/06

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Keywords

  • capillary electrophoresis
  • micellar electrokinetic chromatography
  • steroid
  • antibody fragments

Cite this

Amundsen, L., Nevanen, T., Takkinen, K., & Sirén, H. (2006). New methodology in analysis of steroids using solid phase extraction and micellar electrokinetic chromatography (poster). In 30th International Symposium on High Performance Liquid Phase Separations and Related Techniques, HPLC 2006: Book of Abstracts [67]