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Fluoroenzymatic cycling assay (FECA) for the determination of catechol estrogen monomethyl ethers in human urine

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Abstract

In the present study a method is described for the quantitative determination of the methylated metabolites of catechol estrogens in human urine. Following initial enzymatic hydrolysis the urine samples are extracted with ethyl acetate. The monomethyl ethers of catechol estrogens are then selectively fractionated with straight phase chromatography on Lipidex-5000 gel. Finally, samples are quantitated using enzymatic cycling with 17β-estradiol dehydrogenase combined with fluorometry. The method is sensitive, reproducible and reasonably rapid for routine analysis and avoids the hazards of radioisotopes. Preliminary values of normal males and non-pregnant females are presented.

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References

  1. Ball, P., and Knuppen, R., 1980. Catechol estrogens. Acta Endocrinol. 93: Suppl. 232:1–127.

    Google Scholar 

  2. Ball, P., Knuppen, R., Haupt, M., and Breuer, H. 1972. Interactions between estrogens and catecholamines. III. Studies on the methylation of catechol estrogens, catecholamines and other catechols by the catechol-O-methyltransferase of human liver. J. Clin. Endocrinol. Metab. 34:736–746.

    Google Scholar 

  3. Breuer, H., and Koster, G. 1974. Interaction between oestrogens and neurotransmitters at the hypophysialhypothalamic level. J. Steroid Biochem. 5:961–967.

    Google Scholar 

  4. Lloyd, I., and Weisz, J. 1978. Direct inhibition of tyrosine hydroxylase activity by catechol estrogens. J. Biol. Chem. 253:4841–4844.

    Google Scholar 

  5. Ball, P., Haupt, M., and Knuppen, R. 1978. Comparative studies on the metabolism of estradiol in the brain, the pituitary and liver of the rat. Acta Endocrinol. 87:1–11.

    Google Scholar 

  6. Fishman, J., and Norton, B. 1975. Catechol estrogen formation in the central nervous system of the rat. Endocrinology 109:1799–1801.

    Google Scholar 

  7. Paul, S. M., and Axelrod, J. 1977. Catechol estrogens: Presence in brain and endocrine tissues. Science 197:657–659.

    Google Scholar 

  8. Jellink, P. H., and Shin, S. H. 1983. Catechol estrogen action on gonadotropin and prolactin secretion in experimental animals. Pages 225–236, in G. R. Merriam, and M. B. Lipsett (eds.). Catechol Estrogens, Raven Press, New York.

    Google Scholar 

  9. Merriam, G. R., Loriaux, L., and Lipsett, M. B. 1983. Effects of catechol estrogens on gonadotropins and prolactin in man. Pages 237–248, in G. R. Merriam, and M. B. Lipsett (eds.). Catechol Estrogens, Raven Press, New York.

    Google Scholar 

  10. Bates, G. W., Edman, C. D., Porter, J. C., and Mac Donald, P. C. 1977. Metabolism of catechol estrogens by human erytrocytes. J. Clin. Endocrinol. Metab. 45:1120.

    Google Scholar 

  11. Kono, S., Merriam, G. R., Brandon, D. D., Loriaux, D. L., and Lipsett, M. B. 1982. Radioimmunoassay and metabolism of the catechol estrogen 2-hydroxyestradiol. J. Clin. Endocrinol. Metab. 54:150–154.

    Google Scholar 

  12. Adlercreutz, H., and Fotsis, T. 1983. Chemical assays of catechol estrogens and their monomethyl ethers. Pages 57–70, in G. R. Merriam, and M. B. Lipsett (eds.). Catechol Estrogens, Raven Press, New York.

    Google Scholar 

  13. Emons, G., Ball, P., and Knuppen, R. 1983. Radioimmunoassays of catechol estrogens. Pages 71–81, in G. R. Merriam, and M. B. Lipsett (eds.), Catechol Estrogens, Raven Press, New York.

    Google Scholar 

  14. London, J., Crookall, J., and Mc Gregor, R. 1975. Enzymelabeled immunoassays for steroids. Pages 183–188, in E. H. D. Cammeron, S. G. Hillier, and K. Griffiths (eds.). Steroid Immunoassay, Alpha Omega Publishing, Cardiff, Wales.

    Google Scholar 

  15. Wisdom, B. 1976. Enzyme-immunoassay, Clin. Chem. 22:1243–1255.

    Google Scholar 

  16. Härkönen, M., Adlercreutz, H., and Groman, E. V. 1974. Enzymatic techniques in steroid assay. J. Steroid Biochem. 5:717–725.

    Google Scholar 

  17. Nicolas, J. C., Boussioux, A. M., Descomps, B., and Crastes De Paulet, A. 1979. Enzymatic determination of estradiol and estrone in plasma and urine. Clin. Chim. Acta 92:1–9.

    Google Scholar 

  18. Nicolas, J. C., Boussioux, A. M., Boularan, A. M., Descomps, B., and Crastes De Paulet, A. 1983. Bioluminescent assay of femtomole levels of estrone and estradiol. Anal. Biochem. 135:141–145.

    Google Scholar 

  19. Axelrod, J., and Tomchick, R. 1958. EnzymaticO-methylation of epinephrine and other catechols. J. Biol. Chem. 233:702–705.

    Google Scholar 

  20. Endert, E. 1979. Determination of noradrenaline and adrenaline in plasma by a radioenzymatic assay using high pressure liquid chromatography for the separation of the radiochemical products. Clin. Chim. Acta 96:233–239.

    Google Scholar 

  21. Fotsis, T., and Heikkinen, R. 1983. Selective chromatographic fractionation of catechol estrogens on anion exchangers in borate form. J. Steroid Biochem. 18:357–363.

    Google Scholar 

  22. Fotsis, T., Adlercreutz, H., Järvenpää, P., Setchell, K. D. R., Axelson, M., and Sjövall, J. 1981. Group separation of steroid conjugates by DEAE-Sephadex anion exchange chromatography. J. Steroid Biochem. 14:457–463.

    Google Scholar 

  23. Kushinsky, S., and Anderson, H. 1974. Creepage of estrogens vs loss by sorption on glassware. Clin. Chem. 20:1528–1534.

    Google Scholar 

  24. Adams, J. A., Jarabak, J., and Talalay, A. 1962. The steroid specificity of the 17β-hydroxysteroid dehydrogenase of human placenta. J. Biol. Chem. 237:3069–3073.

    Google Scholar 

  25. Langer, L. S., Alaxander, J. A., and Engel, L. L. 1959. Human placental estradiol-17β dehydrogenase II. Kinetics and substrate specificities. J. Biol. Chem. 234:2609–2614.

    Google Scholar 

  26. Fotsis, T., Adlercreutz, H., and Bannwart, C. 1985. Determination of the estrogen profile in non-pregnancy urine utilizing ion exchange chromatography and selected ion monitoring: preliminary results. Pages 389–398, in S. Görög (ed.). Steroid Analysis, Vol. 2, Akadémiai Kiadó, Budapest.

    Google Scholar 

  27. Ball, P., Reu, G., Schwab, J., and Knuppen, R. 1979. Radioimmunoassay of 2-hydroxyestrone and 2-methoxyestrone in human urine. Steroids 33:563–576.

    Google Scholar 

  28. Berg, D., Sonsalla, R., and Kuss, E. 1983. Concentrations of 2-methoxyoestrogens in human serum measured by a heterologous immunoassay with an125I-labeled ligand. Acta Endocrinol. 103:282–288.

    Google Scholar 

  29. Adlercreutz, H., Fotsis, T., Bannwart, C., Hämäläinen, E., Bloigu, S., and Ollus, A. 1986. Urinary estrogen profile determination in young Finnish vegetarian and omnivorous women. J. Steroid Biochem. 24:289–296.

    Google Scholar 

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Authors and Affiliations

  1. Department of Clinical Chemistry, University of Helsinki, Meilahti Hospital, SF-00290, Helsinki, Finland

    Theodore Fotsis, Haresh P. Shah, Matti Härkönen & Herman Adlercreutz

Authors
  1. Theodore Fotsis
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  2. Haresh P. Shah
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  3. Matti Härkönen
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  4. Herman Adlercreutz
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Special Issue dedicated to Dr. O. H. Lowry.

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Fotsis, T., Shah, H.P., Härkönen, M. et al. Fluoroenzymatic cycling assay (FECA) for the determination of catechol estrogen monomethyl ethers in human urine. Neurochem Res 12, 507–513 (1987). https://doi.org/10.1007/BF01000234

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