1967 Volume 58 Issue 12 Pages 1191-1201
A gas chromatographic method was used for the estimation of urinary pregnanetriol (5β-pregnane-3α, 17α, 20α-triol), pregnanetriolone (5β-pregnane-3α, 17α, 20α-triol-11-one) and pregnanetetrol (5β-pregnane-3α, 11β, 17α, 20α-tetrol) in 28 normal subjects and 12 patients with congenital adrenal hyperplasia of simple virilizing type.
After alumina column chromatography of the extract from the urine, hydrolysed by β-glucuronidase, a fraction containing pregnanetriol, pregnanetriolone and pregnanetetrol was made trimethylsilyl-ether derivatives in pyridine, as the solvent. The reaction mixture was dried under nitrogen stream and the residue was dissolved in a known amount of tetrahydrofuran, an aliquot of which was subjected to gas chromatography on SE30 column. This method was proved to be sensitive enough to measure as little as 10μg/day of each of the steroids using 1/10 aliquot of the 24-hour urine.
In normal subjects the mean urinary excretion of pregnanetriol was 0.42±0.20 (S. D.) mg/day and neither pregnanetriolone nor pregnanetetrol was detected. These three steroids were found in all 12 patients with congenital adrenal hyperplasia.
The ratio (pregnanetriolone+pregnanetetrol)/pregnanetriol in patients of 16-month-old infant and 2-year-old boy was 2.33 and 0.56 respectively, while in older patients it was in a range of 0.19 to 0.39. After administration of corticosteroids an increase of the ratio was observed and this may be indicative of the existence of an abnormal 11β-hydroxylase which acts specifically on 21-deoxysteroid, as suggested by Finkelstein. This ratio decreased markedly following the administration of metopirone.
It was also observed that the ratio pregnanetetrol/pregnanetriolone decreased following the metopirone administration and increased with the dexamethasone treatment. The mechanism of these changes of the ratio is obscure, however it might be useful for the index of transformation of 11-hydoxysteroids to 11-ketosteroids in steroid metabolism.
