ABSTRACT
[4(-14C)]Estrone was injected intramuscularly into two mature laying Rhode Island Red hens. Radioactive steroids and steroid conjugates recovered from the urine on Amberlite XAD-2 columns were fractionated on columns (100 cm) of DEAE-Sephadex A-25 by NaC1 gradients. The presence of the following were confirmed, the figures in brackets indicating average proportions as per cent of total radioactivity recovered after Sephadex column chromatography:-the 3-beta-glucuronides of estrone (10.9) and of estradiol-17alpha plus estradiol-17beta(9.8): the 17-beta-glucuronides of estradiol-17alpha plus estradiol-17beta (2.1); the 3-sulfates of estrone (14.5) and of estradiol-17alpha plus estradiol-17beta(27.4); and the disulfates of estradiol-17alpha plus estradiol-17beta (2.3). The following additional conjugates were identified:-a beta-glucuronide of 16-epiestriol (0.2) and a beta-glucuronide of 16-keto-estradiol-17beta (0.2); the 3-sulfates of 16-epiestriol (1.4), of 17-epiestriol (0.9), of 16,17-epiestriol (0.7), of 16-keto-estradiol-17beta (1.1), and of 2-methoxyestrone (0.7). Some evidence was obtained for the presence of 16,17-epoxy-estratrienol-3-sulfate (1.9).
Subject(s)
Estrone/urine , Animals , Biotransformation , Chickens , Chromatography, Ion Exchange , Estrone/administration & dosage , Female , Injections, IntramuscularABSTRACT
The phytoestrogen formononetin was injected intramuscularly as [4-14C]formononetin into two adult hens. Radioactive materials in the urine for the succeeding 14 days (hen 1) or 16 days (hen 2) were fractionated on DEAE-Sephadex-25 columns by elution with a gradient of NaCl; the four major fractions thus separated were examined by solvent partition, thin-layer chromatography, and enzymic cleavage. The following seven radioactive components were identified in the urine, the average proportions of each being given in terms of percentage of total 14C recovered from the urine: [14C]formononetin (4.3%); [14C]diaidzein (11.4%); [14C]equol (6.8%); [14C]daidzein monosulfate (30.4%); [14C]equol monosulfate (5.8%); [14C]diadzein disulfate (19.8%); and [14C]equol disulfate (6.5%). Small proportions of sulfates of unidentified radioactive phenols were present. Tests for presence of glucosiduronates of 14C-labelled material gave negative results. Radioactive formononetin sulfate was not detected in the urine of either hen.
Subject(s)
Estrogens/urine , Flavonoids/urine , Isoflavones/urine , Animals , Chickens , Chromans/urine , Chromatography, Thin Layer , Female , Sulfuric Acid Esters/urineABSTRACT
[4-14C]-Estrone in 2 ml. ethylene glycol was injected into the breast muscles of S.C. White Leghorn laying hens and the amounts of plasma [14C] per ml. whole blood were measured at various times thereafter. Maximal values were observed at 30 to 50 minutes after injection. The values then declined logarithmically with time and with a half-life of about 80 minutes between 40 and 270 minutes after injection.
Subject(s)
Chickens/blood , Estrone/blood , Animals , Estrone/administration & dosage , Female , Injections, IntramuscularABSTRACT
Two previously uncharacterized radioactive estrogen conjugates, 17beta-estradiol-17-beta-D-glucuronide (3-hydroxyestra-1,3,5(10)-trien-17beta-D-glucopyranosiduronate) and 17alpha-estradiol-17beta-D-glucuronide (3-hydroxyestra-1,3,5(10)-trien-17alpha-yl-beta-D-glucopyranosiduronate), have been identified in small but significant amounts in avian urine and in a ratio of approximately 2:1 after intramuscular injection of [4-14C]estrone.
Subject(s)
Estradiol/urine , Estrone/pharmacology , Glucuronates/urine , Animals , Chickens , Chromatography, Thin Layer , Estrogens, Conjugated (USP)/urine , FemaleABSTRACT
[4--14C] Estrone was injected intramuscularly into six laying hens. Fifty minutes later the hens were exsanguinated. The plasmas were examined for conjugates of radioactive phenolic steroids by recovery on columns of Amberlite XAD-2 or by extraction with tetrahydrofuran followed by chromatography on a column of DEAE-Sephadex A-25 in a gradient of NaCl. The biggest Sephadex chromatographic fraction (50,4% of total) contained about 42% of its radioactivity as estradiol-17alpha-3-sulfate and 18% as estradiol-17beta-3-sulfate and the remaining 40% was identified tentatively as estradiol-17alpha-17-sulfate plus a small proportion of estradiol-17beta-17-sulfate. The second biggest Sephadex chromatographic fraction (12.7% of total) was a mixture of conjugates not further identified. Minor fractions identified comprised estrone-beta-glucuronide (2.8%), estradiol-17alpha-3-beta-glucuronide (2.8%), estradiol-17beta-3-beta-glucuronide (2.3%) and estrone sulfate (6.0%). Evidence was obtained for the presence of small proportions of estradiol-17alpha disulfate and estradiol-17beta disulfate.
Subject(s)
Estrogens/blood , Estrone/administration & dosage , Animals , Carbon Radioisotopes , Chickens , Chromatography, Ion Exchange , Chromatography, Thin Layer , Estradiol/blood , Estrone/blood , Female , Glucuronates/blood , Injections, Intramuscular , Methods , Sulfuric Acids/blood , TritiumABSTRACT
The phytoestrogen, diadzein, was injected intramuscularly as [4-14-C]daidzein into two laying hens. The radioactive materials in the urine for the succeeding 23 (hen 1) or 14 (hen 2) days were fractionated on a DEAE-Sephadex column by a gradient of NaCl and the fractions thus separated were further analyzed by solvent partition, susceptability to enzymic cleavage and thin-layer chromatography. The sic following components were identified and quantitated: [14-C]diadzein, [14-C]equol, [14-C]diadzein monosulfate, [14-C]equol monosulfate, [14-C]diazein disulfate, and [14-C]equol disulfate. The urine from hen 2 yielded also the sulfate of an unidentified conversion product of [14-C]daidzein. Repeared tests for glucuronides of [14-C]daidzein or its conversion products gave negative results, excluding the possibility that any appreciable proportion of the radioactivity in the urine was in the form of beta-glucuronide. It is concluded that the diadzein and the equol excretion in the urine of the laying hen are present for the most part as monosulfates and disulfates.