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

• Published in: Analytical and bioanalytical chemistry Vol. 380; no. 3; pp. 524 - 536
• Main Authors: Pujos, E, Flament-Waton, M M, Goetinck, P, Grenier-Loustalot, M F
• Format: Journal Article
• Language: English
• Published: Germany Springer Verlag 01.10.2004
• Subjects: Adrenal Cortex Hormones - isolation & purification; Adrenal Cortex Hormones - urine; Analytical chemistry; Androgens - isolation & purification; Androgens - urine; Calibration; Chemical Fractionation; Chemical Sciences; Child; Creatine - metabolism; Dehydroepiandrosterone Sulfate - isolation & purification; Dehydroepiandrosterone Sulfate - urine; Female; Gas Chromatography-Mass Spectrometry; Health; Humans; Hydrocortisone - pharmacology; Male; Molecular Structure; Reproducibility of Results; Sensitivity and Specificity; Urinalysis - methods; DHEA; Solid phase extraction; Purification; urine; Metabolite; Liquid liquid extraction; Flame ionization detector; Flame ionization detection; analysis; Sulfates; Chemical enrichment; Gas chromatography; Sample preparation; corticosteroids; steroids; Mass spectrometry
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-004-2763-2

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.