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SUMMARY
[4-14C]Progesterone, [4-14C]20α- and 20β-hydroxypregn-4-en-3-one were injected intravenously into deeply anaesthetized rabbits and the bile and urine collected for 6 h. Neutral metabolites recovered after Ketodase hydrolysis of bile accounted for a mean of 82·8% (range 68·0–98·8%), 68·4% (61·0–80·0%) and 59·0% (57·0–61·0%) of the metabolites of progesterone, 20α-and20β-hydroxypregn-4-en-3-one, respectively. Acidic metabolites accounted for no more than 14·7% of the biliary radioactivity. Similar recoveries of neutral metabolites were obtained after hydrolysis of bile with 15% HCl, but only 41·0% (22·2–62·0%), 45·3% (36·4–55·4%) and 13·7% (10·6–16·9%) of the urinary metabolites were extractable as neutral metabolites. Acidic metabolites represented a further 33·9% (28·9–49·5%), 34·9% (27·5–40·4%) and 38·6% (29·4–47·8%) of the urinary radioactivity, respectively.
Neutral ether soluble metabolites more polar than 5β-pregnane-3α,20α-diol were the most abundant metabolites. 3α-Hydroxy-5β-pregnan-20-one was identified in the bile as a metabolite of [4-14C]progesterone whereas 20α-hydroxypregn-4-en-3-one was present in the urine. 5β-Pregnane-3α, 20α-diol was identified as a biliary metabolite of [4-14C]20β-hydroxypregn-4-en-3-one.
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SUMMARY
Two rabbits were injected intravenously with [4-14C]progesterone and the bile and urine collected for 6 h. The crude bile was administered through an intraduodenal cannula to four rabbits as a single injection, and to three rabbits at a constant rate of infusion for 1·5 h. Bile and urine were collected hourly for up to 7 h. In the single injection experiments 42·3–64·4% of the radioactivity was excreted in the bile and 3·8–13·5% in the urine in 5–6 h. After constant rate infusion 35·4–64·8% and 4·7–9·5% of the radioactivity was excreted in the bile and urine respectively during 6–7 h. The hourly recovery of radioactivity in the neutral fractions of bile and urine decreased with time, whereas radioactivity not accounted for increased. Acidic metabolites represented an insignificant proportion of both biliary and urinary metabolites after enterohepatic recirculation.
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MUN Research Unit, Medical School, Memorial University of Newfoundland, St John's, Newfoundland, AIA 1E5, Canada
(Received 22 July 1975)
Acidic steroids are the major progesterone metabolites excreted in rabbit urine (Cooke, Rogers & Thomas, 1963) and are C-21-carboxy compounds (Allen, Cooke & Thomas, 1968; Senciall & Dey, 1975). Resolution from steroid conjugates was obtained by alumina column chromatography (Senciall, 1973), and a major derivative, 3,6,20-trioxo-5α-pregnane-21-methyl ester, was identified by g.l.c. after oxidation and methylation (Senciall & Dey, 1975). Pregnancy and non-pregnancy urine samples of several species have now been screened for acidic metabolites of progesterone.
[4-14C] Progesterone (52·8 mCi/mmol) was administered to female non-pregnant rabbits (i.v.; 5 μCi); pigs (i.m.; 10 μCi) and women (oral; 5 μCi);and [21-14C] progesterone (55·6 mCi/mmol) i.m. to guinea-pigs (5 μCi) and rats (5 μCi). Urine collected for 24 h contained means of 24·9; 25·9; 30·5; 15·9 and 11·8% of the doses respectively. Table 1
