SIMULTANEOUS ANALYSIS OF AZIDOTHYMIDINE AND ITS MONOPHOSPHATE, DIPHOSPHATE AND TRIPHOSPHATE DERIVATIVES IN BIOLOGICAL-FLUIDS, TISSUE AND CULTURED-CELLS BY A RAPID HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC METHOD

G MOLEMA*, RW JANSEN, Jan Visser, DKF MEIJER

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

27 Citations (Scopus)

Abstract

A rapid high-performance liquid chromatographic (HPLC) method for the simultaneous analysis of the antiviral drug azidothymidine (AZT), AZT monophosphate, AZT diphosphate and AZT triphosphate, with ultraviolet detection in the nanomolar range, is described. Determination of these compounds in vitro in the human MT-4 lymphocyte cell line did not require a prior extraction, and AZT and its phosphorylated derivatives could be accurately analysed in one HPLC run. However, plasma, bile, liver homogenate and urine samples could not be injected directly into the chromatograph. Therefore, a solid-phase extraction procedure was developed. using azidodideoxyinosine as internal standard. The extractions of the compounds of interest from all but urine samples were reproducible, with recoveries between 65% (AZT triphosphate from plasma) and 100% (AZT from plasma).

Original languageEnglish
Pages (from-to)107-114
Number of pages8
JournalJournal of chromatography-Biomedical applications
Volume579
Issue number1
DOIs
Publication statusPublished - 7-Aug-1992

Keywords

  • HUMAN-IMMUNODEFICIENCY-VIRUS
  • COLONY-STIMULATING FACTOR
  • INTRACELLULAR METABOLISM
  • 3'-AZIDO-3'-DEOXYTHYMIDINE
  • MACROPHAGE
  • INFECTION
  • INVITRO
  • 3'-AZIDO-2',3'-DIDEOXYTHYMIDINE
  • PHOSPHORYLATION
  • 5'-TRIPHOSPHATE

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