Development and validation of a high-performance liquid chromatography assay for the quantitative determination of 7-oxo-dehydroepiandrosterone-3beta-sulfate in human plasma.
A new, simple, reproducible and reliable high-performance liquid chromatography method with ultraviolet absorbance detection at 240 nm was developed and validated for the determination of 7-oxo-dehydroepiandrosterone-3beta-sulfate in human plasma. The method was based upon solid-phase (C18) extraction of plasma after addition of 17beta-hydroxy-3beta-methoxyandrost-5-en-7-one as internal standard. Using 1 ml of plasma for extraction, the detection limit of the assay was 3 ng/ml. The standard curve was linear over the concentration range 10-1000 ng/ml. Stored at -20 degrees C for about 4 months at various concentrations in plasma, 7-oxo-dehydroepiandrosterone-3beta-sulfate did not reveal any appreciable degradation. Also included herein is a method for the simultaneous detection and determination of 7-oxo-dehydroepiandrosterone and 7-oxo-dehydroepiandrosterone-3beta-acetate in plasma.
Institute for Enzyme Research, University of Wisconsin at Madison, 53705, USA.
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Marwah
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Marwah
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Lardy
D000738:Androsterone / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D003687:Dehydroepiandrosterone / Q000031:analogs & derivatives / Q000097:blood; D006801:Humans; D012015:Reference Standards; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
C119255:7-oxo-dehydroepiandrosterone-sulfate; C002914:7-oxodehydroepiandrosterone; D003687:Dehydroepiandrosterone; C117281:3-acetoxyandrost-5-ene-7,17-dione; D000738:Androsterone