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Evaluation of various approaches to the isolation of steroid hormones from urine samples prior to FASS‐MEKC analysis
Author(s) -
Olędzka Ilona,
Kowalski Piotr,
Plenis Alina,
Bączek Tomasz
Publication year - 2017
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201600509
Subject(s) - chromatography , chemistry , epitestosterone , analyte , methyltestosterone , urine , steroid , hormone , medicine , biochemistry
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 ( R 2 above 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.