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. 1968 Jan;106(1):77–86. doi: 10.1042/bj1060077

The determination of oestradiol and oestrone in the plasma of the domestic fowl by a method involving the use of labelled derivatives

J E O'Grady 1,*
PMCID: PMC1198471  PMID: 5721473

Abstract

1. A method involving the use of triple-labelled derivatives has been developed for the determination of total oestrone and oestradiol in the plasma of the domestic fowl. The double-labelling technique devised by Svendsen (1960) for the determination of free oestrogens in human plasma was modified to enable the total oestrogen recovery to be determined for each sample. 2. [6,7-3H2]Oestradiol-17β is added to the plasma samples (1–10ml.), which are hydrolysed with acid and the phenolic steroids then extracted and partially purified. The extract is esterified with iodobenzene-p[35S]-sulphonyl chloride of high specific activity. After addition of standard oestrogen [131I]iodobenzene-p-sulphonates the esters are finally purified by paper chromatography. 3. The oestrogens are determined by comparing the 3H/35S and 131I/35S ratios in the purified esters with similar ratios of appropriate standards. 4. With this procedure the recoveries of oestrone and oestradiol after hydrolysis were 70–85% and 72–84% respectively, and after hydrolysis and preliminary purification 38–53% and 39–51% respectively. With this procedure up to 500ng. of oestradiol can be determined. The sensitivity of the technique for oestrone is 3·0ng. and for oestradiol 2·1ng. 5. The ranges of oestradiol and oestrone concentrations found in six plasma samples were 8·3–21·4ng./ml. and 15·2–31·6ng./ml. respectively.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BOJESEN E. Determination of 17- hydroxycorticosterone in peripheral plasma from dogs and humans with radioactive p- iodophenylsulfonylic acid anhydride (pipsan). Scand J Clin Lab Invest. 1956;8(1):55–66. doi: 10.3109/00365515609049244. [DOI] [PubMed] [Google Scholar]
  2. BROWN J. B. A chemical method for the determination of oestriol, oestrone and oestradiol in human urine. Biochem J. 1955 Jun;60(2):185–193. doi: 10.1042/bj0600185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BUSH I. E. Methods of paper chromatography of steroids applicable to the study of steroids in mammalian blood and tissues. Biochem J. 1952 Jan;50(3):370–378. doi: 10.1042/bj0500370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. HEALD P. J., MCLACHLAN P. M. THE SYNTHESIS OF PHOSVITIN IN VITRO BY SLICES OF LIVER FROM THE LAYING HEN. Biochem J. 1965 Jan;94:32–39. doi: 10.1042/bj0940032. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. HEALD P. J., ROOKLEDGE K. A. EFFECT OF GONADAL HORMONES, GONADOTROPHINS AND THYROXINE ON PLASMA FREE FATTY ACIDS IN THE DOMESTIC FOWL. J Endocrinol. 1964 Aug;30:115–130. doi: 10.1677/joe.0.0300115. [DOI] [PubMed] [Google Scholar]
  6. HENDLER R. W. PROCEDURE FOR SIMULTANEOUS ASSAY OF TWO BETA-EMITTING ISOTOPES WITH THE LIQUID SCINTILLATION COUNTING TECHNIQUE. Anal Biochem. 1964 Jan;7:110–120. doi: 10.1016/0003-2697(64)90125-3. [DOI] [PubMed] [Google Scholar]
  7. LAYNE D. S., COMMON R. H., MAW W. A., FRAPS R. M. Presence of oestrone, oestradiol and oestriol in extracts of ovaries of laying hens. Nature. 1958 Feb 1;181(4605):351–352. doi: 10.1038/181351b0. [DOI] [PubMed] [Google Scholar]
  8. O'GRADY J. E., HEALD P. J. IDENTIFICATION OF OESTRADIOL AND OESTRONE IN AVIAN PLASMA. Nature. 1965 Jan 23;205:390–390. doi: 10.1038/205390a0. [DOI] [PubMed] [Google Scholar]
  9. SVENDSEN R., SORENSEN B. THE CONCENTRATION OF UNCONJUGATED OESTRONE AND 17 BETA-OESTRADIOL IN PLASMA DURING PREGNANCY. Acta Endocrinol (Copenh) 1964 Oct;47:237–244. doi: 10.1530/acta.0.0470237. [DOI] [PubMed] [Google Scholar]
  10. SVENDSEN R., SORENSEN B. THE PLASMA CONCENTRATION OF UNCONJUGATED OESTRONE AND 17 BETA-OESTRADIOL DURING THE NORMAL MENSTRUAL CYCLE. Acta Endocrinol (Copenh) 1964 Oct;47:245–254. doi: 10.1530/acta.0.0470245. [DOI] [PubMed] [Google Scholar]

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