RADIOIMMUNOASSAYS OF TETRAHYDROALDOSTERONE (TH-ALDO) IN HUMAN URINE

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Abstract
Specific antisera against tetrahydroaldosterone (TH-Aldo) were raised in 2 white New Zealand rabbits. 3.alpha.,5.beta.-TH-aldo-20-oxime-bovine-serum albumin complex was used as antigen. The resulting titers were 1:18,000 and 1:16,000. Except tetrahydrocortisol (THF) (0.23%) and tetrahydro-18-hydroxy-11-dehydrocorticosterone (18-OH-THA) (3.2%), all steroids and steroid metabolites gave negligible cross-reactions. Immunograms of the paper chromatograms made from the n-butanol-extract of the urines, as well as after .beta.-glucuronidase treatment and dichlormethane extraction, were studied to further define the specificity of the antiserum. Antibody H1 reacted with aldosterone-18-gluc. [glucuronide], a TH-aldosterone-glucuronide (probably the 21-glucuronide) and an unidentified less polar material. Two methods were developed: (a) TH-Aldo-glucuronide(s) estimation after ethylacetate pre-extraction as a rapid screening test of endogenous aldosterone production; (b) estimation of TH-aldosterone using 1 chromatographic system. The results of method a showed a significant correlation with the values obtained by technique b. Normal values (method b) were 25.88 .+-. 16.50 .mu.g/24 h (range 9.5-64.8 .mu.g/24 h). A significant correlation was also shown between the TH-aldo (technique b and 18-gluc. values.

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