Development of a sensitive and selective LC/MS/MS method for the simultaneous determination of intracellular 1-beta-D-arabinofuranosylcytosine triphosphate (araCTP), cytidine triphosphate (CTP) and deoxycytidine triphosphate (dCTP) in a human follicular lymphoma cell line

J Chromatogr B Analyt Technol Biomed Life Sci. 2009 May 15;877(14-15):1417-25. doi: 10.1016/j.jchromb.2009.02.071. Epub 2009 Mar 17.

Abstract

A method was developed for the quantification of araCTP, CTP and dCTP in a human follicular lymphoma cell line. This method involves solid phase extraction (SPE) using a weak anion-exchanger (WAX) cartridge, a porous graphitic carbon high-performance liquid chromatography (HPLC) column separation, and tandem mass spectrometry (MS/MS) detection. By using a triple quadrupole mass spectrometer operating in negative ion multiple reaction monitoring (MRM) mode, the method was able to achieve a lower limit of quantification (LLOQ) of 0.1 microg mL(-1) for araCTP and of 0.01 microg mL(-1) for both CTP and dCTP. The method was validated and used to determine the amount of araCTP, CTP and dCTP formed after incubation of araC and an araCMP prodrug in the human follicular lymphoma cell line RL.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabinofuranosylcytosine Triphosphate / chemistry*
  • Cell Line, Tumor
  • Chromatography, High Pressure Liquid / methods*
  • Cytidine Triphosphate / chemistry*
  • Deoxycytosine Nucleotides / chemistry*
  • Humans
  • Lymphoma, Follicular / chemistry*
  • Sensitivity and Specificity
  • Tandem Mass Spectrometry / methods*
  • Thionucleotides / chemistry*

Substances

  • Deoxycytosine Nucleotides
  • Thionucleotides
  • Arabinofuranosylcytosine Triphosphate
  • deoxycytidine thiotriphosphate
  • Cytidine Triphosphate