Determination of nonconjugated and conjugated steroid levels in plasma and prostate after separation on C-18 columns.

An accurate method is described for analysis of C-21, C-19, and C-18 steroids as well as steroid conjugates, namely, androstane-3 alpha,17 beta-diol glucuronide, androsterone glucuronide, estradiol glucuronide, and estrone glucuronide as well as dehydroepiandrosterone sulfate and androst-5-ene-3 beta,17 beta-diol sulfate. This technique involves an extraction step, aimed at solubilizing the nonconjugated steroids as well as the steroid sulfates and glucuronides, C-18 column chromatography, permitting the separation of nonconjugated steroids and the conjugated group followed by specific hydrolysis of the glucuronide and, finally, solvolysis of the steroid sulfates. Our data indicated that using 1 ml of plasma or 1 g of prostate, good recovery of the three groups of steroids was obtained. Moreover, an accurate determination of steroids could be achieved. The plasma levels of steroids in normal adult women and men found using our technique were within the range of those previously reported by us and other authors.