[Radioimmunoassay of 18-hydroxy-11-desoxycorticosterone (18-OH-DOC). Preliminary studies]. / Dosage radioimmunologique de la 18-hydroxy-11-désoxycorticostérone (18-OH-DOC)

An antiserum anti 18-OH-DOC-21-hemisuccinate-BSA was induced in the Rabbit. Its radioimmunological properties were studied. In this preliminary study, the classical reaction antigen-antibody was examined, principaly in the procedure for the separation of free antigen from antigen bound to antibody.

Separation and characterization of the reduced metabolites of the 18-hydroxydeoxycorticosterone hormone by gas-liquid chromatography-mass spectrometry. Occurrence of stereoisomeric forms in rat adrenals and liver.

In order to study the metabolism of 18-OH-11-deoxycorticosterone, the mineralocorticoid hormone responsible for hypertension in rats and humans, we have synthesized the following dihydrogenated and tetrahydrogenated reference derivatives: 18,21-dihydroxy-5alpha-pregnane-3,20-dione, 18,21-dihydroxy-5beta-pregnane-3,20-dione, 3alpha,18,21-trihydroxy-5alpha-pregnan-20-one (I), 3beta,18,21-trihydroxy-5alpha-pregnan-20-one (II), 3alpha,18,21-trihydroxy-5beta-pregnan-20-one (III) and 3beta,18,21-trihydroxy-5beta-pregnan-20-one (IV). A complete separation of these compounds from each other and from tetrahydrocorticosterone isomers has been realized only by the association of thin-layer chromatography and gas-liquid chromatography on high-efficiency glass capillary columns. Characterization by gas-liquid chromatography-mas spectrometry is described. The stereoisomer distribution in rats is: adrenals of adult males and females (compounds I and II in the ratio 4:1); adrenals of 23-day-old males and females (compounds I, II and III, 11:11:3); liver of females (compound I and traces of III); and the liver of adult male (compounds II, III and IV, 4.5:4.5:1).

Production and characterisation of antisera to 18-hydroxy-11-deoxycorticosterone.

The production of highly sensitive and specific antisera to 18-hydroxy-11-deoxycorticosterone (18,21-dihydroxy-4-pregnene-3,20-dione) is reported. The antisera were generated in rabbits and guinea pigs with a 3-carboxymethoxime derivative of the steroid coupled to rabbit serum albumin. Antibody characteristics were determined by a radioimmunoassay procedure. Only minor differences between the two animal species were observed. Antibody titers ranged from 10 to 8000. Association constants were in the order of 10(8) to 10(10) 1/mole. A minimal amount of 40 pg unlabeled steroid was necessary to displace 50% of the tritiated steroid. Cross reaction with cortisol was 0.0002% to 0.031%, with aldosterone 0.0007% to 1.09%, with corticosterone 0.0025% to 1%, with 18-hydroxy-corticosterone 0.05% to 1% and with progesterone 0.0048% to 1.5%.