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Analysis of 8-hydroxy-2′-deoxyguanosine in human urine using hydrophilic interaction chromatography with tandem mass spectrometry

Journal of Chromatography B
Publication Date
DOI: 10.1016/j.jchromb.2012.02.043
  • Hydrophilic Interaction Chromatography
  • Tandem Mass Spectrometry
  • 8-Hydroxy-2′-Deoxyguanosine
  • Urine
  • Oxidative Stress


Abstract Urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) is a widely used noninvasive biomarker of oxidative stress. A selective, sensitive and rapid method for determining 8-OHdG in human urine was developed using hydrophilic interaction chromatography–tandem mass spectrometry (HILIC–MS/MS) with electrospray ionization. 8-OHdG and isotopically labeled 8-OHdG (internal standard) were separated on a HILIC column with a mobile phase of 10mM ammonium acetate: acetonitrile (1:9, v/v) within 10min and detected by using a positive electrospray ionization interface under the selected reaction monitoring mode. The detection limits of 8-OHdG (corresponding to a signal-to-noise ratio of 3) for the HILIC-MS/MS system and the conventional method using a reversed-phase column with MS/MS were 1.0 and 26.0fmol/injection, respectively. The proposed method makes it possible to monitor the basal level of urinary 8-OHdG from non-exposed healthy subjects and can be used for large-scale human studies.

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