Nuklearmedizin 1967; 06(03): 311-320
DOI: 10.1055/s-0037-1621339
Originalarbeiten — Original Articles — Travaux Originaux
Schattauer GmbH

Isotopic Determination of Serum Vitamin B12 Level using Sephadex G-25

Détermination isotopique du taux de Vitamine B12 dans le sérum au moyen de Sephadex G-25Isotopenbestimmung des Vitamin-B12-Serumspiegels mittels Sephadex G-25
J. Mantzos
1   From the Department of Clinical Therapeutics, University of Athens and the Radioisotope Laboratory, Alexandra Hospital, Athens, Greece
,
H. Gyftaki
1   From the Department of Clinical Therapeutics, University of Athens and the Radioisotope Laboratory, Alexandra Hospital, Athens, Greece
,
V. Alevizou
1   From the Department of Clinical Therapeutics, University of Athens and the Radioisotope Laboratory, Alexandra Hospital, Athens, Greece
› Author Affiliations
Further Information

Publication History

Received: 08 March 1967

Publication Date:
05 February 2018 (online)

Summary

A radioisotope method for the determination of the serum vitamin B12 level is described. The method is based on the principle of saturation analysis and on the ability of Sephadex to separate free from bound vitamin.

We have found the method to be sensitive, reproducible and fairly rapid. Because of its simplicity, it can be used in any laboratory equipped for the use of radioisotopes.

The method was applied to determine the level of Vitamin B12 in 71 normal subjects and in 15 patients with different diseases.

Il est décrit une méthode radioisotopique de la détermination du taux de Vitamine B12 dans le sérum. Cette méthode est basée sur le principe de l’analyse de saturation et de la capacité de Sephadex de séparer les vitamines non-liées et liées.

Nous avons trouvé cette méthode sensible, reproductible et assez rapide. Sa simplicité permet son utilisation dans tout laboratoire installé pour Pemploy de radioisotopes.

La méthode a été utilisée pour la détermination du taux de Vitamine B12 chez 71 sujets normaux et chez 15 sujets affectés de différentes maladies.

Eine Radioisotopenmethode zur Bestimmung des Vitamin-B12-Serumspiegels wird beschrieben. Die Methode beruht auf dem Prinzip der Saturationsanalyse und dem Vermögen von Sephadex, freies von gebundenem Vitamin zu trennen.

Wir fanden die Methode empfindlich, reproduzierbar und ziemlich schnell. Ihre Einfachheit erlaubt ihre Anwendung in jedem Laboratorium, das zur Verwendung von Radioisotopen eingerichtet ist.

Die Methode wurde zur Bestimmung des Vitamin-B12-Spiegels bei 71 normalen Personen und bei 15 Patienten mit verschiedenen Krankheiten verwendet.

 
  • References

  • 1 Barakat M. R, Ekins P. R. Assay of Vitamin B10 in blood. Lancet I: 25 1961;
  • 2 Barakat M. R, Ekins P. R. An isotopic method for the determination of Vitamin B12 levels in blood. Blood 21: 1 1963;
  • 3 Bauriedel W. R, Ricken J. C, Underkofler L. A. Reactions of cyanocobalamin and aquocobalamin with proteins. Proc. Soc. Biol. Med 91: 377 1956;
  • 4 Daislay K. N. Gel filtration of sea water: separation of free and bound forms of Vitamin B12 . Nature (Lond.) 191: 868 1961;
  • 5 Ekins P. R, Scherzi M. A. The microssay of Vitamin B12 in human plasma by the saturation assay technique. Symposium on Radiochemical Methods of Analysis. October 1964. Salzburg, Austria.:
  • 6 Grossowicz D, Sulitzeanu D, Merzbach D. Isotopic determination of Vitamin B12 binding capacity and concentration. Proc. Soc. Exp. Biol 109: 6C4 1962;
  • 7 Kakei M, Jerzy Glass B. G. Separation of bound and free Vitamin B12 on Sephadex G-25 column. Proc. Soc. Exp. Biol. Med 3: 270 1962;
  • 8 Lindstrand K, Stahlberg G. K, Ehrenvard G, Norden A. Studies on free and serum protein-bound Vitamin B12 by the use of Sephadex G-25 and high voltage electrophoresis. Acta Med. Scand 173: 5 1963;
  • 9 Lau K. S, Gottlieb C, Wasserman R. L, Herbert V. Measurement of serum Vitamin B12 level using radioisotope dilution and coated charcoal. Blood 26: 2 1965;
  • 10 Miller A. The in vitro binding of cobalt60 labelled Vitamin B12 by normal and leukemic sera. J. Clin. Invest 37: 556 1958;
  • 11 Pitney W. R, Beard M. F, van Loon E. J. Observations on the bound form of Vitamin B12 in human serum. J. Biol. Chem 207: 143 1954;
  • 12 Ross G. I. M. Vitamin B12 assay in body fluids using Euglena gracilis. J. Clin. Path 5: 250 1952;
  • 13 Rothenberg S. P. Assay of serum Vitamin B12 concentration using 57Co-B12 and intrinsic factor. Proc. Soc. Exp. Biol 108: 45 1961;
  • 14 Rothenberg S. P. Radioassay of serum Vitamin B12 by quantitating the competition between 57Co B12 and unlabelled B12 for binding sites of intrinsic factor. J. Clin. Invest 42: 1391 1963;
  • 15 Thompson H. J, Deitrich L. S, Elvehjem C. A. The use of Lactobacillus leismanii in the estimation of Vitamin B12 activity. J. Biol. Chem 184: 175 1950;
  • 16 Weinstein I. B, Weissman S. M, Watkins D. M. The plasma Vitamin B12 binding substance. J. Clin. Invest 38: 1904 1959;