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

J. Mantzos; H. Gyftaki; V. Alevizou

doi:10.1055/s-0037-1621339

Nuklearmedizin - NuclearMedicine, Table of Contents

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

¹From the Department of Clinical Therapeutics, University of Athens and the Radioisotope Laboratory, Alexandra Hospital, Athens, Greece

,

H. Gyftaki

¹From the Department of Clinical Therapeutics, University of Athens and the Radioisotope Laboratory, Alexandra Hospital, Athens, Greece

,

V. Alevizou

¹From the Department of Clinical Therapeutics, University of Athens and the Radioisotope Laboratory, Alexandra Hospital, Athens, Greece

› Author Affiliations

Abstract

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.

Full Text

References

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;