Radio gas chromatography for evaluation of sub-cellular hormone synthesis in the androgen insensitivity syndrome

Abstract

This paper gives a description of a radio gas-liquid chromatographic method for the evaluation of androgen hormone synthesis patterns in the testicular tissue of a patient suffering from the androgen insensitivity syndrome (AIS). A modified dual-column gas chromatography system, equiped with column switching facilities and a radioactivity monitor run parallel to a flame ionisation detector, enables the monitoring of radioactive intermediates of testosterone anabolism and catabolism, generated from a labelled precursor. Tissue preparations were incubated with tritiated pregnenolone for 45 min at 37°C. Steroid hormones were stripped from the aqueous phase by solvent extraction and analysed by gas chromatography as methoxitrimethylsilyl (MO-TMS) derivatives on a 15 m × 0.32 mm I.D. fused-silica capillary column coated with DB-5. The results reveal abnormal enzyme kinetics due to accelerated precursor utilisation. The findings reflect the pathophysiology of AIS at the sub-cellular level of the androgen hormone target organ.