Simultaneous quantitative determination of alpha-ketoglutaric acid and 5-hydroxymethylfurfural in human plasma by gas chromatography-mass spectrometry

Anal Bioanal Chem. 2010 Apr;396(7):2629-37. doi: 10.1007/s00216-010-3479-0. Epub 2010 Feb 14.


Alpha-ketoglutaric acid (alpha-KG) and 5-hydroxymethylfurfural (5-HMF) are currently under investigation as promising cancer cell damaging agents. A method for the simultaneous quantitative determination of alpha-KG and 5-HMF in human plasma was established for screening these compounds in human plasma. Plasma samples were directly treated with O-(2,3,4,5,6-pentafluorobenzyl) hydroxylamine hydrochloride to form the corresponding oximes, thus facilitating subsequent liquid-liquid extraction. After formation of the trimethylsilyl ethers, samples were analyzed by gas chromatography with electron ionization mass spectrometry. Stable isotope labeled standards were used, the preparation of (13)C(6)-5-HMF is described. Limits of quantitation were set to 0.938 microg/mL for alpha-KG and 0.156 microg/mL for 5-HMF. Inter-day accuracy was < or = 93.7% (alpha-KG) and < or = 92.8% (5-HMF). Inter-day precision was < or = 6.0% (alpha-KG) and < or = 4.6% (5-HMF). The method has been successfully applied to pharmacokinetic profiling of the compounds after intravenous application.

Publication types

  • Evaluation Study

MeSH terms

  • Antineoplastic Agents / blood
  • Blood Chemical Analysis / methods*
  • Complex Mixtures / analysis
  • Furaldehyde / analogs & derivatives*
  • Furaldehyde / blood
  • Gas Chromatography-Mass Spectrometry / methods*
  • Humans
  • Ketoglutaric Acids / blood*
  • Reproducibility of Results
  • Sensitivity and Specificity


  • Antineoplastic Agents
  • Complex Mixtures
  • Ketoglutaric Acids
  • 5-hydroxymethylfurfural
  • Furaldehyde