A Rapid Method of Quantitating Steroids Resulting from the Incubation of Gonadal Tissues with Radioactive Precursors

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Abstract
A rapid method was developed for the quantitation of steroid metabolites resulting from the incubation of specific gonadal cell types or gonadal tissue [human testis] with radioactive precursors [pregnenolone]. The method involves the use of high performance liquid chromatography (HPLC) for separating the steroids and a flow-through radioactive detector (Flo-One HP) for quantitating the radioactive 3H precursor and metabolites in the presence or absence of 14C-steroid recovery tracers. A comparison is made between the results obtained directly by the Flo-One HP radioactivity detector and the fraction collection method, (counting aliquots from individual fractions in the liquid scintillation counter). In addition, the results using an electronic stream splitter in the analysis of a percentage of the effluent directly by Flo-One HP are evaluated. The remaining percentage is collected in a fraction collector and is used for further analysis (e.g., recrystallization, radioimmunoassay, further purification and characterization).