Quantitative Extraction and Separation of Conjugated Steroid Metabolites from Human Urine

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Abstract
A procedure has been described for the quantitative extraction of steroid conjugates from human urine. The steroid conjugates so obtained have been further studied by chromatographic separation. The following conclusions have been reached: 1. Conjugated metabolites of many classes of steroids can be studied by this method. 2. Conjugates of cortisol and testosterone metabolites can be separated chromatographically into 3 major groups: “sulfates,” “glucuronides” and more polar conjugates of unknown structure. The proportion of these groups was variable and cannot yet be correlated with clinical factors. 3. The yield of neutral steroid after β-glucuronidase hydrolysis of the chromatographic fraction considered to comprise the glucuronides was essentially identical with the yield of neutral steroid after β-glucuronidase hydrolysis of the raw urine. Similar results were obtained for sulfate, comparing hydrolysis of the pertinent chromatographic fraction to that of raw urine. Therefore, the amount of apparent “glucuronide” and “sulfate” estimated to be present in raw urine by application of appropriate hydrolytic procedures is probably an excellent measure of the respective amounts of glucuronide and sulfate actually present. 4. The present procedure does not introduce any artifacts into the metabolities of cortisol or testosterone.