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Determination of clofarabine triphosphate concentrations in leukemia cells using sensitive, isocratic high-performance liquid chromatography.

AbstractBACKGROUND/AIM:
An active metabolite of the anti-leukemia agent clofarabine (Cl-F-ara-A) is an intracellular triphosphate form, Cl-F-ara-ATP. Monitoring this active form could provide crucial information for optimizing treatment regimens based on Cl-F-ara-A. A simple, isocratic HPLC method was developed.
MATERIALS AND METHODS:
Samples (500 μl) from leukemic cells were loaded onto an anion-exchange column and eluted with a phosphate-acetonitrile buffer (flow rate: 0.7 ml/min) at ambient temperature. The Cl-F-ara-ATP concentration was determined by measuring absorbance at 254 nm.
RESULTS:
The standard curve was linear, with minimal within-day and inter-day variability. Recovery was excellent; low and high quantitation limits were 10 pmol and 5,000 pmol, respectively. Cl-F-ara-ATP eluted independently of ATP, GTP, UTP, and CTP. Production of Cl-F-ara-ATP was successfully measured in cultured leukemia HL-60 cells treated in vitro with Cl-F-ara-A.
CONCLUSION:
This method is simple, sensitive and applicable for determination of the Cl-F-ara-ATP content of biological materials.
AuthorsTakahiro Yamauchi, Rie Nishi, Takanori Ueda
JournalAnticancer research (Anticancer Res) Vol. 31 Issue 9 Pg. 2863-7 (Sep 2011) ISSN: 1791-7530 [Electronic] Greece
PMID21868530 (Publication Type: Journal Article, Validation Study)
Chemical References
  • Adenine Nucleotides
  • Antineoplastic Agents
  • Arabinonucleosides
  • Clofarabine
Topics
  • Adenine Nucleotides (analysis)
  • Antineoplastic Agents (analysis)
  • Arabinonucleosides (analysis)
  • Chromatography, High Pressure Liquid (methods)
  • Clofarabine
  • HL-60 Cells
  • Humans
  • Leukemia (pathology)
  • Reproducibility of Results

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