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Optimizing the extraction and analysis of DHEA sulfate, corticosteroids and androgens in urine: application to a study of the influence of corticosteroid intake on urinary steroid profiles

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Abstract

A method of detecting and quantifying dehydroepiandrosterone (DHEA) sulfate, corticosteroids, and androgens has been developed. All of the compounds were first extracted from urine using solid phase extraction (SPE), enzymatically hydrolyzed, and separated into three samples using a second SPE. A DHEA sulfate sample was acetylated and re-extracted using SPE for purification before analysis. Corticosteroid samples were oxidized and re-extracted using liquid/liquid extraction for analysis. Androgen samples were acetylated and re-extracted using SPE prior to analysis. The extraction and analysis methods were investigated and optimized. Analyses were performed with gas chromatography/mass spectrometry (GC/MS) and gas chromatography/flame ionization detection (GC/FID). The entire procedure was then applied to the study of urine profiles of healthy volunteers and patients treated with corticosteroids. The results showed that the quantities of androgens found in patient urines were lower than in those of healthy volunteers. In addition, other metabolites were detected in patient urines.

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Acknowledgements

We thank Dr. F. Wright of the Medical Biochemistry Department of the Pitié Salpêtrière Hospital in Paris for providing urine from patients treated with hydrocortisone. This project was financed by the European Community (ISOTRACE project [9]).

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  1. Service Central d’Analysis, USR 059, CNRS, BP 22, 69390, Vernaison, France

    E. Pujos, M. M. Flament-Waton, P. Goetinck & M. F. Grenier-Loustalot

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  1. E. Pujos
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  2. M. M. Flament-Waton
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  3. P. Goetinck
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  4. M. F. Grenier-Loustalot
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Correspondence to M. F. Grenier-Loustalot.

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Pujos, E., Flament-Waton, M.M., Goetinck, P. et al. Optimizing the extraction and analysis of DHEA sulfate, corticosteroids and androgens in urine: application to a study of the influence of corticosteroid intake on urinary steroid profiles. Anal Bioanal Chem 380, 524–536 (2004). https://doi.org/10.1007/s00216-004-2763-2

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  • DOI: https://doi.org/10.1007/s00216-004-2763-2

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