Extraction of steroids from urine with C18 solid-phase extraction cartridges results in an extract containing impurities. If, during the extraction of hydrolyzed urine, an amino (NH2) column is placed in series with the C18 column, then one obtains a sample that is sufficiently clean for gas-chromatographic analysis. Analytical recovery of dehydroepiandrosterone from urine is considerably decreased by the use of increasing amounts of Helix pomatia enzyme preparation. Extraction of the steroid conjugates from urine with C18 columns before the hydrolysis stage is essential for hydrolysis with an amount of enzyme preparation that suffices for complete splitting of the polar steroid conjugates but not so much as to cause insufficient analytical recovery of dehydroepiandrosterone.

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