Determination of 3H-labelled cytosine arabinoside and eight metabolites in cell extracts by high-performance liquid chromatography and scintillation spectrometry

J Chromatogr. 1989 Jul 21;491(2):331-40. doi: 10.1016/s0378-4347(00)82851-0.

Abstract

We have developed a method for the separation and quantitation of radiolabeled cytosine arabinoside and its eight metabolites in cell extracts by anion-exchange gradient high-performance liquid chromatography. Baseline separation of cytosine arabinoside and uracil arabinoside and their respective 5'-mono-, di- and triphosphates, as well as cytosine arabinoside diphosphocholine was obtained with the shortest interval between peaks being 3 min. This degree of separation was found to be essential for quantitation of 3H-labeled metabolites by scintillation counting of 1-min fractions. Application of this procedure to the quantitation of [3H] cytosine arabinoside and its metabolites from HL60 human leukemia cells is demonstrated.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Arabinofuranosyluracil / analysis
  • Cell Line
  • Chromatography, High Pressure Liquid / methods
  • Cytarabine / analysis*
  • Cytarabine / metabolism
  • Humans
  • Hydrogen-Ion Concentration
  • Leukemia / metabolism
  • Scintillation Counting
  • Spectrophotometry, Ultraviolet
  • Tumor Cells, Cultured

Substances

  • Cytarabine
  • Arabinofuranosyluracil