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

Cortol compounds have been not determined as 170HCS 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 17OHCS 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 (17KS and pregnandiol) and 11 fraction (17OHCS, pregnantriol and cortol compounds) by first florisil column chromatography. (4a) Separation of 17OHCS fraction by thin layer chromatography (solvent system: chloroform: methanol: water). (5a) Colorimetry as Porter-Silber chromogen. (5b) Separation of cortol compounds from other HIO(4)-17KS 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 Zimmerman 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) THE 486±94, THE 1153±535, THS 256±57, (secretory phase) THE 457±58, THE 801±189, THS 261±46, normal males: THE 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.