Aldosteroneを中心とした尿中Corticosteroids分離定量法に関する研究

抄録

Metabolism of body fluids and electrolytes which related not only to aldosterone alone but to other corticosteroids, for example, glucocorticoids had mineral action. And the simultaneous administration of aldosterone and corticosterone consistently resulted in a greater potassium diuresis than did the aldosterone alone.
Therefore, body fluids and electrolytes are aummation of many corticosteroids which had biological action, and can't know by the determination of simple corticosteroids.
The author report on the systemic determination of aldosterone, Comp. B, Comp. E, Comp. F, and the newly separated contaminants.
Method :
The 24 hours urine specimen is pooled at 5°C and acidified to pH 1.5 with conc. HCL, then standing 24 hours at 15°C.
Urine was extracted by chloroform and dried. Extract was treated with Florisil column chromatographies and 4 consecutered and different paper chromatographies with Chloroform-Formamide System, Bush-C System, E₂B System, and Bush-B5 System, to separated corticosteroid partitions.
Results :
1. Recoveries of 5 mg/100 ml H₂O standard samples after 5 times continued extraction by 0.1 volume chloroform, for Comp. B, Comp. E, and DOC is 95%, and Comp. F is 83%.
2. Additional chloroform re-extraction, Comp. B, Comp. E, and DOG do not appear in the 7th chloroform extraction, but Comp. F appear in the amount of 60μg in 8th chloroform and 9th extraction is nothing, thus this determination necessitate at least 8 consecutered extractions.
3. Comp. B, Comp. E, and Comp. F appear in 25% methanol in chloroform fraction by the use of the Florisil column chromatography is made, and recovery is about 100% in condition of 60 drops/minutes.
4. Separate Comp. III, Comp. V, Comp. VI, and Comp. VII from aldosterone partition in Neher-Wettstein's method.
5. New contaminants were discover from Comp. E and Comp. F partition after Bush-C System paper chromatography, called Comp. E₂-E4₂, Comp. F₂ ; and, other conatminants named M₁-M₈ Z₁-Z₈, HC₁-HC₃, AC₁ and AC₂ in Chloroform-Formamide and Bush-C System paper chromatography.
6. Examination of the purities of Comp. B, Comp. E, Comp. F, and aldosterone partitions is made use of the ultra violet absorption spectrum, and necessitates 4 (C-F→Bush-C→E₂B→Bush-B₅), or 3 (C-F→E₂B→Bush-B₅), paper chromatographies to satisfactorily separate these compounds.
7. The H₂SO₄ chromogen spectrum of these partition correspond with the standard compounds.