SEMIAUTOMATED METHOD FOR THE QUANTITATION OF PLASMA OR SERA ANDROSTENEDIONE, TESTOSTERONE, AND DIHYDROTESTOSTERONE IN POPULATION STUDIES

A method is presented that analyzes quantitatively and reproducibly th e androgens testosterone, androstenedione, and dihydrotestosterone fro m human sera or plasma. The chromatographic separation step generates an unattended throughput of one preparative separation per hour. Contr ols are built into the method to account for changing chromatographic conditions that otherwise result in shifts in retention characteristic s. Separation factors for the three androgens are as follows (mean +/- standard deviation): alpha = 1.23 +/- 0.011 between androstenedione a nd testesterone and alpha = 1.38 +/- 0.025 between testosterone and di hydrotestosterone. Sensitivities of the method are androstenedione 5 p g, testosterone 3 pg, and dihydrotestosterone 14 pg. A study of proced ural losses associated with initial sample processing, a validation, a nd application to two sample sets which demonstrates the methods utili ty for the analysis of hypoandrogenic populations (postmenopausal wome n) and hyperandrogenic groups (prostate cancer patients) is also repor ted. The precision for replicate aliquots of control plasma is androst enedione and testosterone = 5-11% CV and dihydrotestosterone = 10-20% CV. (C) 1996 Academic Press, Inc.