Quantitation of deoxyribonucleoside 5'-triphosphates by a sequential boronate and anion-exchange high-pressure liquid chromatographic procedure

Anal Biochem. 1992 Oct;206(1):178-82. doi: 10.1016/s0003-2697(05)80030-2.


A rapid method for the quantitative determination of cellular deoxyribonucleoside 5'-triphosphates is described. Cell extracts are first separated by boronate chromatography at pH 8.9, which removes 99% of the ribonucleoside triphosphate (rNTPs) from the deoxyribonucleoside triphosphates (dNTPs). The resulting dNTP fraction is analyzed by gradient high-pressure liquid chromatography utilizing a strong anion-exchange column, which can separate minor rNTP peaks from the corresponding dNTPs. This sequential procedure, which requires less than 1 h per sample for both chromatographic steps, results in the quantitative recovery of greater than 98% of the dNTPs from cell extracts. Nucleotide analogs, such as 1-beta-D-arabinofuranosylcytosine-5'-triphosphate and 5-bromo-2'-deoxyuridine-5'-triphosphate, can also be quantitated efficiently by this method.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Boronic Acids*
  • Buffers
  • Cell Extracts / chemistry
  • Cells, Cultured
  • Chromatography, High Pressure Liquid / methods
  • Chromatography, Ion Exchange / methods
  • Deoxyribonucleotides / analysis*
  • Deoxyribonucleotides / isolation & purification
  • Glioma / chemistry
  • Humans
  • Hydrogen-Ion Concentration
  • Lymphocytes / chemistry
  • Sensitivity and Specificity
  • Tumor Cells, Cultured


  • Boronic Acids
  • Buffers
  • Cell Extracts
  • Deoxyribonucleotides