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

Abstract:

BACKGROUND/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.

journal_name

Anticancer Res

journal_title

Anticancer research

authors

Yamauchi T,Nishi R,Ueda T

subject

Has Abstract

pub_date

2011-09-01 00:00:00

pages

2863-7

issue

9

eissn

0250-7005

issn

1791-7530

pii

31/9/2863

journal_volume

31

pub_type

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