A Method for the Quantification of Intracellular Zidovudine Nucleotides
Open Access
- 1 October 1991
- journal article
- Published by Oxford University Press (OUP) in The Journal of Infectious Diseases
- Vol. 164 (4) , 773-776
- https://doi.org/10.1093/infdis/164.4.773
Abstract
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%.Keywords
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