尿中17-HydroxycorticosteroidsおよびCortol compoundsの新分画測定法に関する研究

抄録

Cortol compounds have been not determined as 17 OHCS or Porter-Silber chromogen, although they form one of the main groups of urinary metabolites of glucocorticoids.
In this study, a new method for fractional determination of these three cortol compounds, and 17 OHCS in urine was established. The new method consisted of the following procedure. (1) Hydrolysis with, β-glucuronidase. (2) Extraction with ethyl acetate. (3) Separation of 1 fraction (17 KS and pregnandiol) and 11 fraction (17 OHCS, pregnantriol and cortol compounds) by first florisil column chromatography. (4a) Separation of 17 OHCS fraction by thin layer chromatography (solvent system: chloroform: methanol: water). (5a) Colorimetry as Porter-Silber chromogen. (5b) Separation of cortol compounds from other HIO₄-17 KS steroids by second florisil column chromatography. (6b) Oxidation with periodic acid.
(7a) Fractionation of three converted steroids by 6% water alumina column chromatogrphy. (7b) Colorimetry as Zimmermann chromogen.
This procedures was cosisted from its fundamental experiments.
Total recovery rate of β-cortol 100μg was average 66.2±2.0%.
Identification of cortol compounds messured by this method was carried out on the Rf. values by TLC, RRT values by GLC, RRT values of TMSi-ether deliverts by GLC.
5β-pregnan-3α, 17α, 20, 21-tetrol named “11-deoxy-Cortol” was messured for excretion values in urine. Normal mean excretion values were as follow.
Normal females (proliferative phase) THF 486±94, THE 1153±535, THS 256±57, (secretory phase) THF 457±58, THE 801±189, THS 261±46, normal males: THF 1789±360, THE 2741±568, THS 225±52, ug/day.
Normal females (proliferative phase): Cortol 734.9±198.1, Cortolone 1120.6±241.2, 11-deoxy-Cortol 178.8±98.2, (secretory phase): Cortol 462.8±169.6, Cortolone 975.2±132.4, 11-deoxy-Cortol 207.6±58.9, normal males: Cortol 757.5±181.2, Cortolone 1514.9±227.3, 11-deoxy-Cortol 264.9±49.2 ug/day.