title: Evaluation of various approaches to the isolation of steroid hormones from urine samples prior to FASS-MEKC analysis.
abstract: The goal of this study was to assess various analytical approaches for the simultaneous and efficient extraction of steroid hormones (cortisone, cortisol, prednisolone, corticosterone, testosterone, 17α-methyltestosterone, epitestosterone, progesterone) from urine samples prior to separation based on field-amplified sample stacking MEKC (FASS-MEKC). FASS-MEKC successfully allowed the compounds to be separated within 12 min using a BGE composed of 5 mM sodium tetraborate, 150 mM boric acid, 50 mM SDS, and 15% methanol. Therefore, many procedures such as solid-phase microextraction, SPE, and dispersive liquid-liquid microextraction (DLLME) were tested and compared using a multivariate tool, namely, cluster analysis. Finally, DLLME-FASS-MEKC was validated and proved a good linearity of calibration curves (R2above 0.9948) in a concentration range from 50 to 1000 ng/mL for all analytes. The LOD was established at 15 ng/mL, whereas the LOQ was 50 ng/mL. The intra- and interday precision, expressed as RSD%, did not exceed 9.97%. The DLLME-FASS-MEKC method was successfully applied to the analysis of urine samples from healthy volunteers and sportsmen. This methodology could prove to be useful in clinical studies and/or doping control depending on the steroid concentrations required in biomedical applications.
authors.affiliation: Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Medical University of Gdańsk, Gdańsk, Poland.
authors.forename: Ilona
authors.initials: I
authors.lastname: Olędzka
authors.affiliation: Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Medical University of Gdańsk, Gdańsk, Poland.
authors.forename: Piotr
authors.initials: P
authors.lastname: Kowalski
authors.affiliation: Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Medical University of Gdańsk, Gdańsk, Poland.
authors.forename: Alina
authors.initials: A
authors.lastname: Plenis
authors.affiliation: Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Medical University of Gdańsk, Gdańsk, Poland.
authors.forename: Tomasz
authors.initials: T
authors.lastname: Bączek
mesh_terms: D020374:Chromatography, Micellar Electrokinetic Capillary; D004300:Doping in Sports; D019075:Electrophoresis, Capillary; D012739:Gonadal Steroid Hormones / Q000302:isolation & purification / Q000652:urine*; D057230:Limit of Detection; D059627:Liquid Phase Microextraction; D012680:Sensitivity and Specificity; D045165:Testosterone Congeners / Q000652:urine
chemical_list: D012739:Gonadal Steroid Hormones; D045165:Testosterone Congeners
keywords: Cluster analysis; Dispersive liquid-liquid microextraction; Field-amplified sample stacking-micellar electrokinetic chromatography; Solid-phase extraction; Solid-phase microextraction
  