SYNTHESIS OF A DEUTERIUM-LABELED CORTISOL FOR THE STUDY OF ITS RATE OF 11-BETA-HYDROXY DEHYDROGENATION IN MAN

11-beta-Hydroxy dehydrogenation of cortisol to cortisone is specifically impaired in the syndrome of apparent mineralocorticoid excess. This defect bears on the pathogenesis of the disorder by unmasking the potential mineralocorticoid agonism of unmetabolized cortisol at or near mineralocorticoid target tissues. A specific index of this defect is provided by measurement of the formation of tritiated water following the administration of [H-3]11-alpha-cortisol. We have explored the use of a non-radioactive tracer to follow this unidirectional dehydrogenation reaction but because of the relatively lower sensitivity of measurement of H-2(2)O compared to H-3(2)O in body fluids, use of the corresponding [H-2]11-alpha-cortisol was not feasible. We have devised instead a method incorporating additional deuterium atoms into cortisol to measure unidirectional 11-beta-hydroxy dehydrogenation not by the formation of labeled water but by the determination of the dehydrogenated cortisol product from its residual deuterium content. Cortisol-d4 metabolized to cortisone-d3 is conveniently measured by the techniques of organic mass spectrometry. The synthesis of cortisol-9-alpha,11-alpha,12-alpha-12-beta-d4 and the validation of its isotopic distribution by mass spectrometry and nuclear magnetic resonance is described.