A method for the estimation of 6-oxygenated metabolites of progesterone in urine

Abstract

1. A method is described for the estimation of the 6-oxygenated metabolites of progesterone in urine. After hydrolysis the extract of urine is chromatographed on alumina to obtain a fraction containing mainly the 6-oxygenated metabolites. This fraction is oxidized to convert the metabolites into pregnane-3,6,20-triones, which are estimated as the dinitrophenylhydrazones. 2. The reliability criteria of the method are presented. Normal subjects excrete 0.1-0.6mg./day, and at the end of pregnancy values of 3.3-11.6mg./day are obtained.

Metabolism of Progesterone in the Pig-Tail Monkey (Macaca Nemestrina)

“…Fraction 4 was not expected to contain 5/?-pregnane-3a,17a,20a-triol, or such possible metabolites as ll/?-hydroxy or 16-hydroxy compounds, however, because they are not stable under the conditions of hydrolysis used. Certain 6-oxygenated pregnanes would appear in this fraction (see James & Fotherby, 1965). These three fractions together accounted for most of the radioactivity present in the neutral extract (Table 1), although the distribution of radioactivity between them varied from monkey to monkey, and from day to day.…”

EXCRETION OF 6-Oxygenated METABOLITES OF PROGESTERONE AND 5β-Pregnane-3α,17α,20α-Triol DURING PREGNANCY

“…Between the 11th week and the end of pregnancy there was a fivefold increase in excretion of the 6-oxygenated metabolites of progesterone and the values obtained in the 11th week of pregnancy were approximately twice as high as those found in the urine of normal subjects during the luteal phase of the menstrual cycle (James & Fotherby, 1965). During pregnancy, with the possible exception of the last few weeks, there was a close correlation between the excretion of the 6-oxygenated metabolites and that of pregnanediol, suggesting that the 6-oxygenated metabolites probably had the same precursor as pregnanediol.…”

“…Table 3. Excretion of 6-oxygenated metabolites of progesterone (6-OP), pregnanetriol (PT) and pregnanediol (PD) after progesterone administration The method described by James & Fotherby (1965) for the estimation of the 6-oxygenated metabolites of progesterone in urine measures the 3,6-dihydroxypregnan-20-oxosteroids and the pregnane-3,6,20-triols but does not measure any of the less polar 6-oxygenated metabolites that might be present in urine. It should also be emphasized that although the term 6-oxygenated metabolites of progesterone suggests that these metabolites arise solely from progesterone, the possibility exists that these metabolites arise also from 6-oxo or 6-hydroxyprogesterone secreted by the endocrine glands.…”

“…James & Fotherby (1965) recently described a method for the estimation of some of the metabolites of progesterone oxygenated at position 6, and it was of interest to ascertain the excretion of these compounds by a number of Postgraduate Medical School, Ducane Road, London, W. 12, Obstetric Medicine Research Unit, The Medical School, Foresterhill, Aberdeen, Scotland (Received 1 February 1965) women at regular intervals throughout pregnancy. This provided an opportunity to see whether there was a disproportionate increase in the excretion of the 6-oxygenated metabolites as progesterone secretion increased.…”

“…This study pre¬ sented, therefore, the opportunity of measuring the excretion of pregnanetriol throughout pregnancy in the same series of patients, and to examine the specificity of the method of Fotherby & Love (1960) for estimating the level of pregnanetriol in urine from pregnant subjects. METHODS Pregnanediol, pregnanetriol and the 6-oxygenated metabolites of progesterone in urine were estimated by the methods of Klopper, Michie & Brown (1955), Fotherby & Love (1960) and James & Fotherby (1965), respectively.…”