Quantitative Determination of Metanephrine and Normetanephrine in Urine

  • Kazumi Taniguchi
  • Yasuo Kakimoto
  • Marvin D. Armstrong
Part of the Recent Results in Cancer Research book series (RECENTCANCER, volume 2)


Details of a fluorometric method for the separate determination of free and conjugated metanephrine and normetanephrine in urine are presented. The procedure consists of: (1) a separation and concentration of free amines by absorption on a column of Dowex-50 resin followed by elution with a small volume of aqueous ammonia; (2) acid hydrolysis of the initial effluent from the Dowex-50 column to release conjugated amines, which are then separated in the same manner as for free amines; (3) separation of the metanephrine and normetanephrine in each fraction by chromatography on a column of Amberlite CG-50; and (4) a fluorometric estimation of the amines by a variation of the trihydroxyindole method. Metanephrine may be estimated in samples which contain between 0.25 and 50 μg. and normetanephrine between 0.35 and 120 μg. No substances have been encountered in urine which interfere with fluorescence development or which respond to the test. The average daily excretion of the amines by 6 male subjects was: metanephrine, free, 29.5 μg., conjugated, 134.1 μg.; total, 163.6 μg.; and normetanephrine, free, 21.2 μg.; conjugated, 223.2 μg.; and total 244.4 μg. The amounts of the different fractions excreted by an individual appear to be relatively constant, and marked differences in the pattern of excretion was observed in different persons. There were marked diurnal variations in the rate of excretion of the amines but there was no consistent pattern to the variation.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. [1]
    Euler, U., S. Von, and I. Floding: Diagnosis of pheochromocytoma by fluorometric estimation of adrenaline and noradrenaline in urine. Scand. J. Clin. Lab. Invest. 8, 288 (1956).CrossRefGoogle Scholar
  2. [2]
    Axelrod, J., S. Senoh, and B. Witkop: O-Methylation of catechol amines in vivo. J. Biol. Chem. 233, 697 (1958).PubMedGoogle Scholar
  3. [3]
    Armstrong, M. D., A. Mcmillan, and K. N. F. Shaw: 3-Methoxy-4-hydroxy-D-man-delic acid, a urinary metabolite of norepinephrine. Biochim. Biophys. Acta 25, 422 (1957).PubMedCrossRefGoogle Scholar
  4. [4]
    Yoshinaga, K., C. Itoh, N. Ishida, T. Sato, and Y. Wada: Quantitative determination of metadrenaline and normetadrenaline in normal human urine. Nature 191, 599 (1961).PubMedCrossRefGoogle Scholar
  5. [5]
    Kraupp, O., H. Bernheimer, and P. Papistas: Isolierung und Quantitative Bestimmung von 3-O-Methyladrenalin und 3-O-Methylnoradrenalin im Harn. Clin. Chim. Acta 6, 851 (1961).PubMedCrossRefGoogle Scholar
  6. [6]
    Smith, E. B., and H. Weil-Malherbe: Metanephrine and normetanephrine in human urine: Method and results. J. Lab. Clin. Med. 60, 212 (1962).PubMedGoogle Scholar
  7. [7]
    Brunjes, S., D. Wybenga, and J. J. Varner Jr.: Fluorometric determination of urinary metanephrine and normetanephrine. Clin. Chem. 10, 1 (1964).PubMedCrossRefGoogle Scholar
  8. [8]
    Kakimoto, Y., and M. D. Armstrong: The phenolic amines of human urine. J. Biol. Chem. 237, 208 (1962).PubMedGoogle Scholar
  9. [9]
    Bertler, A., A. Carlsson, and E. Rosengren: Fluorimetric method for differential estimation of the 3-O-methylated derivatives of adrenalin and Noradrenalin (Metanephrine and Normetanephrine). Clin. Chim. Acta 4, 456 (1959).PubMedCrossRefGoogle Scholar
  10. [10]
    Randrup, A.,: On the differential fluorometric determination of metadrenaline and normetadrenaline. Clin. Chim. Acta 6, 584 (1961).PubMedCrossRefGoogle Scholar
  11. [11]
    Adler, E., and R. Magnusson: Periodate oxidation of phenols. I. Monoethers of pyro-catechol and hydroquinone. Acta Chem. Scand. 13, 505 (1959).CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin · Heidelberg 1966

Authors and Affiliations

  • Kazumi Taniguchi
    • 1
  • Yasuo Kakimoto
    • 1
  • Marvin D. Armstrong
    • 1
  1. 1.The Fels Research InstituteYellow SpringsUSA

Personalised recommendations