Direct quantitative digital autoradiography-thin-layer chromatography of 3 alpha,3 beta- and 5 alpha-reduced and 17 beta-dehydrogenated androgens derived from testosterone metabolism.

A digital autoradiographic-thin-layer chromatographic method involving simple steps is described for thorough separation of eight major androgens (testosterone, androstenedione, dihydrotestosterone, androsterone, epiandrosterone, androstanedione, 3 alpha-androstanediol and 3 beta-androstanediol) derived from testosterone metabolism. Their direct quantification is performed by radioscanning, which avoids tedious plate-scraping and liquid scintillation counting. Satisfactory accuracy is obtained by both external standardization of plates by calibrated amounts of radiolabelled standard steroids and internal standardization of bioassays by radiolabelled squalane. Coefficients of variation are below 8.5% in the range 100-500 dpm. Some analytical criteria related to chromatographic conditions and quantification parameters depending on position-sensitive proportional counter are discussed.