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2 protocols using 7 oxo dhea

1

Steroid Extraction and Quantification

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Cortisol, cortisone and dehydroepiandrosterone (DHEA) were purchased form Koch-Light Laboratories LTD (Colnbrook, Great Britain), 7α-OH-DHEA, 7β-OH-DHEA, 7-oxo-DHEA, 16α-OH-DHEA and D3-DHEA were from Steraloids (Newport, USA). D4-Cortisol was from CDN isotopes (Ponte-Claire, Canada).
2-hydrazinopyridine, ammonium formate, methyl tertbutyl ether and trifluoroacetic acid were from Sigma-Aldrich (St. Louis, USA). LC-MS grade methanol, water and diethyl ether were from Merck AG (Darmstadt, Germany). The physiological solution (0.9 % sodium chloride) was from B-Braun (Melsungen AG, Germany). [1,2,6,7-3H]Cortisol, specific radioactivity 3.04 TBq/mmol was from Amesrham Biosciences, Inc.
(Amersham, UK).
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2

Steroid Quantification Methodology

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Cortisol, cortisone and dehydroepiandrosterone (DHEA) were purchased from Koch-Light Laboratories LTD (Colnbrook, Great Britain); 7α-hydroxy-DHEA, 7β-hydroxy-DHEA, 7-oxo-DHEA, 16α-hydroxy-DHEA and D3-DHEA were from Steraloids (Newport, USA). D4-Cortisol was from CDN isotopes (Ponte-Claire, Canada). 2-hydrazinopyridine, ammonium formate, methyl tert-butyl ether, trifluoroacetic acid, homocystine, dithiothreitol, pyridine, p-fluoro-DL-phenylalanine, ethyl chloroformate, isooctane, ethanol, chloroform, sodium chloride, hydrochloric acid and potassium oxalate monohydrate were from Sigma-Aldrich (St. Louis, USA). LC-MS grade methanol and water as well as diethyl ether were from Merck AG (Darmstadt, Germany). The physiological solution was from B-Braun (Melsungen AG, Germany).
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