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Updated: Aug 3, 2026
HPLC-based Assay to Monitor Extracellular Nucleotide/Nucleoside Metabolism in Human Chronic Lymphocytic Leukemia Cells
Published on: July 20, 2016
Simple and sensitive method for quantification of fludarabine triphosphate intracellular concentration in leukemic
Takahiro Yamauchi1, Takanori Ueda
1First Department of Internal Medicine, Fukui Medical University, 23 Shimoaizuki, Matsuoka, Fukui 910-1193, Japan. tyamauch@fmsrsa.fukui-med.ac.jp
Abstract:
A simple, isocratic HPLC method was newly developed for quantitating intracellular fludarabine triphosphate (F-ara-ATP). Samples (500 microl) were injected onto an anion-exchange column and eluted isocratically with phosphate-acetonitrile buffer (flow rate: 0.7 ml/min) at an ambient temperature. F-ara-ATP was quantitated according to its peak area at the absorbance of 261 nm. The standard curve was linear with minimal within-day and inter-day variability. The low and high quantification limits were 50 pmol and 20 nmol, respectively. The method was capable of measuring F-ara-ATP generated in cultured leukemic cells in vitro. Thus, our method will be useful because of its sensitivity and simplicity as well as applicability to biological materials.
