High-performance liquid chromatography method for the determination and quantitation of arabinosylguanine triphosphate and fludarabine triphosphate in human cells

Carlos O. Rodriguez, William Plunkett, Melanie T. Paff, Min Du, Billie Nowak, Prameen Ramakrishna, Michael J. Keating, Varsha Gandhi

Research output: Contribution to journalArticle

37 Scopus citations

Abstract

A gradient anion-exchange high-performance liquid chromatographic assay was developed for the simultaneous determination and quantitation of the cytotoxic triphosphates of arabinosylguanine (ara-GTP) and fludarabine (F- ara-ATP). The method was validated with respect to selectivity, recovery, linearity, precision, and accuracy using authentic standards. To test this assay in a more complex biological matrix, perchloric acid extracts of circulating human leukemia cells spiked with known concentrations of ara-GTP and F-ara-ATP were examined. Finally, to assess the clinical utility of our method, perchloric acid extracts of circulating human leukemia cells isolated from patients treated with fludarabine and nelarabine were analyzed. The range of quantitation was 0.0125-10 nmol for the ara- and native NTPs in cellular extracts. This assay should be helpful in establishing the mechanistic rationales for drug scheduling and combinations of nelarabine and fludarabine, and for correlating the therapeutic efficacy and levels of the cytotoxic triphosphates in target cells. (C) 2000 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)421-430
Number of pages10
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Volume745
Issue number2
DOIs
StatePublished - Aug 18 2000
Externally publishedYes

Keywords

  • 506U78
  • Arabinosylguanine
  • Fludarabine
  • Nelarabine

ASJC Scopus subject areas

  • Chemistry(all)

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