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A Method for the Quantification of Intracellular Zidovudine Nucleotides

Kuster, Herbert ; Vogt, Markus ; Joos, Beda ; Nadai, Vera ; Lüthy, Ruedi

In: Journal of Infectious Diseases, 1991, vol. 164, no. 4, p. 773-776

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    Titre
    • A Method for the Quantification of Intracellular Zidovudine Nucleotides
    Auteur
    • Kuster, Herbert. Division of Infectious Diseases, Department of Medicine, University Hospital, Zurich, Switzerland
    • Vogt, Markus. Division of Infectious Diseases, Department of Medicine, University Hospital, Zurich, Switzerland
    • Joos, Beda. Division of Infectious Diseases, Department of Medicine, University Hospital, Zurich, Switzerland
    • Nadai, Vera. Division of Infectious Diseases, Department of Medicine, University Hospital, Zurich, Switzerland
    • Lüthy, Ruedi. Division of Infectious Diseases, Department of Medicine, University Hospital, Zurich, Switzerland
    Type de document
    • Postprint
    Langue
    • Anglais
    Publié dans
    • Journal of Infectious Diseases, 1991, vol. 164, no. 4, p. 773-776. The University of Chicago Press
    Autre version électronique
    Classification
    • Santé
    Identifiant OAI-PMH
    • oai:doc.rero.ch:294562
    Summary
    An assay to quantify the phosphorylation products of zidovudine (AZT) in peripheral blood mononuclear cells (PBMC) was developed. Extracts of PBMC were separated by high-performance liquid chromatography. Eluted AZT mono- (MP), di- (DP), and triphosphate (TP) were collected in separate portions. Treatment with alkaline phosphatase yielded equimolar amounts ofAZT, which after solid-phase enrichment were assayed by radioimmunoassay. Detection limit was 0.1 pmol/106 PBMC for each nucleotide. Recoveries of 102%-118% were observed. AZT nucleotides were measured in samples from three patients receiving 250 mg ofAZT every 12 h. Intracellular concentrations of AZT-MP after 1-2 h ranged from 0.9 to 1.4 pmol/106 PBMC and then declined to 0.3-1.1 pmol/106 PBMC after 4 h. AZT-DP and AZT-TP reached concentrations of 0.3-0.5 pmol/106 PBMC after 1-2 h and could not be detected after 4 h in any of the three patients. Duplicate determinations deviated by <20%