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Calcified Tissue Research

, Volume 4, Issue 1, pp 202–209 | Cite as

Determination of45Calcium and14Carbon ascorbic acid in bone using a liquid scintillation system

  • Susanne L. von Schuching
  • Normal N. Willingham
Original Papers
  • 22 Downloads

Abstract

Methods were developed for the determination of45Ca plus L-[1-14C] asorbic acid in bones of animals following administration of both isotopes. Asorbic acid and calcium were extracted quantitatively with 6% trichloracetic acid from fresh bone under such mild conditions that asorbic acid was not destroyed. The chemical determinations of calcium and ascorbic acid according to standard procedures were performed in aliquots of the same extract. The determination of the radioactivity of calcium and ascorbic acid were then carried out in aliquots of the same extract. A Triton-containing liquid scintillation fluid was employed for the radioassay of45Ca and L-[1-14C] ascorbic acid in the liquid scintillation spectrometer. A procedure allowing the determination of both isotopes in the same extract is described. A number of criteria were applied to each step, in order to determine the validity and limitations of the procedure.

Key words

Bone Radiocalcium Ascorbic acid Scintillimetry Radiocarbon 

Résumé

Les auteurs on décrit des méthodes pour la détermination de45Ca plus L-1-14C acide asorbique dans les os d'animaux aprés administration des deux isotopes.

Calcium et acide asorbique ont été extraits totalement par l'acide trichloracétique a 6% des os a l'état frais en évitant de détruire l'acide asorbique. La détermination chimique du calcium et de l'acide asorbique selon les méthodes classique a été faite dans des portions de l'échantillon.

La détermination de la radioactivité du calcium et de l'acide asorbique a été faite ensuite dans des portions du même échantillon.

Du Triton a servi pour le scintillant liquide dans le compteur de scintillation liquide.

Un procédé de détermination des deux isotopes dans le meme echantillon a été décrit.

Différents critéres on été appliques a chacque étape afin d'établir la validité et les limites d'application de la méthode.

Zusammenfassung

Eine Methode ist beschrieben, die die quantitative und radioaktive Bestimmung von45Ca und L-[-L14C] Ascorbinsäure in Knochen von Tieren, denen die beiden Isotopen verabreicht worden sind, erlaubt. Calcium und Ascorbinsäure werden quantitativ mit 6% Trichloressigsäure ausgezogen, wobei die Ascorbinsäure erhalten bleibt.

Quantitative Analysen für Calcium und Ascorbinsäure werden nach bekannten Methoden in einem Teil des Extraktes bestimmt und die Bestimmung der Radioaktivität in einem anderen Anteil mit Hilfe eines Flüssigkeits-Szintillations-Zählers durchgeführt. Dem flüssigen Szintillanten wird Triton zugesetzt. Diese Versuchsanordnung ermöglicht außerdem die gleichzeitige Bestimmung der beiden Isotopen im Knochen-Extrakt.

Die Methode wurde sorgfältig auf ihre Verwendbarkeit ausgewertet.

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References

  1. Bauer, G. C. H.: Rate of bone salt formation in a healing fracture determined in rats by means of radiocalcium. Acta orthop. scand.23, 169–191 (1954).Google Scholar
  2. Benson, R. H.: Limitations of tritium measurements of liquid scintillation counting of emulsions. Anal. Chem.38, 1353–1356 (1966).Google Scholar
  3. Firschein, H. E., Miller, O. P.: Comparison of calcium 48 and carbon 14 proline as indicators of bone metabolism. Nature (Lond.)212, 1252–1253 (1966).Google Scholar
  4. Fox, B. W.: The application of triton-X-100 colloid scintillation counting in biochemistry. Int. J. appl. Radiat. Isotopes19, 717–730 (1968).Google Scholar
  5. Fuwa, K., Valley, B.: The physical basis of analytical atomic absorption spectrometry. Anal. Chem.34, 942–946 (1963).Google Scholar
  6. Glascock, R. F.: Combustion of labeled compounds. In: Isotopic gas analysis for biochemists, p. 87–115. New York: Academic Press 1954.Google Scholar
  7. Hardcastle, J. E., Hannagel, R. J., Fuller, H. H.: A liquid scintillation technique for the radioassay of calcium-45. Int. J. appl. Radiat. Isotopes18, 193–199 (1967).Google Scholar
  8. Hawk, Ph. B., Oser, B. L., Summerson, W. H.: Determination of calcium in practical physiological chemistry, p. 589–593. New York: The Blakiston Co. 1951.Google Scholar
  9. Hendler, R. W.: Procedure for simultaneous assay of two β-emitting isotopes with the liquid scintillation technique. Anal. Biochem.7, 110–120 (1964).Google Scholar
  10. Humphreys, E. R.: An improved method for determining calcium-45 in biological materials. Int. J. appl. Radiat. Isotopes16, 345–348 (1965).Google Scholar
  11. Hutchinson, F.: Assay of45Ca in biological materials by liquid scintillation counting. Int. J. appl. Radiat. Isotopes18, 136–137 (1967).Google Scholar
  12. Laarse, J. D. Van der: Experience with emulsion counting of tritium. Int. J. appl. Radiat. Isotopes18, 485–491 (1967).Google Scholar
  13. McLean, F. C., Urist, M. V.: Structure and chemical composition of bone matrix. In: Bone, An introduction to the physiology of skeletal tissue, p. 24–30. Chicago: Chicago University Press 1955.Google Scholar
  14. McPherson, G. D.: Stable calcium isotopes as tracers in studies of mineral metabolism. Acta orthop. scand., Suppl. 78 (1965).Google Scholar
  15. Passman, J. M., Radin, N. S., Cooper, J.: Liquid scintillation technique for measuring carbon-14-dioxide activity. Anal. Chem.28, 484–486 (1956).Google Scholar
  16. Patterson, M. S., Greene, R. C.: Measurement of low energy β-emitters in aqueous solution by liquid scintillation counting of emulsions. Anal. Chem.37, 854–857 (1965).Google Scholar
  17. Roe, J. H.: Chemical determination of ascorbic, dehydroascorbic and diketogulonic acid. In: D. Glick (ed.), Methods of biochemical analysis, p. 115–149. New York: Interscience Publ. 1954.Google Scholar
  18. Sarnat, M., Jeffay, H.: Determination of radioactive calcium by liquid scintillation counting. Anal. Chem.34, 643–646 (1962).Google Scholar
  19. Schuching, S. Von, Abt, A. F.: Ascorbic acid distribution and localization in animal tissues and cells, autoradiographic studies. Proc. VII. Internat. Congr. on Nutrition. vol. 1, 315–320. Braunschweig: Friedr. Vieweg 1967.Google Scholar
  20. Turner, J. C.: Triton-X-100 Scintillant for carbon-14 labeled material. Int. J. appl. Radiat. isotopes19, 557–563 (1968).Google Scholar
  21. Way, K., Fano, L., Scott, M. R., Thew, K.: Nuclear data. National Bureau of Standards Circular 499, p. 10 and 37, Sept. 1, 1950.Google Scholar
  22. Williams, J. B., Irvine, J. W.: Preparation of the inorganic matrix of bone. Science119, 771–753 (1954).Google Scholar
  23. Woeller, F. H.: Counting14Co2 with phenylethylamine. Anal. Biochem.2, 508–511 (1961).Google Scholar

Copyright information

© Springer-Verlag 1969

Authors and Affiliations

  • Susanne L. von Schuching
    • 1
  • Normal N. Willingham
    • 1
  1. 1.Radioisotope ServiceVeterans Administration CenterMartinsburgUSA

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