Assessing biomarkers of oxidative stress: analysis of guanosine and oxidized guanosine nucleotide triphosphates by high performance liquid chromatography with electrochemical detection.
Oxidation of the guanosine moiety in DNA has become a hallmark biomarker in assessing oxidative stress. The oxidation of guanosine in the nucleotide triphosphate pool has been overlooked due to the lack of a reliable methodology. This method describes a sample processing and high performance liquid chromatography with electrochemical detection protocol for the analysis of the cellular pool of guanosine triphosphates and oxidized guanosine triphosphates. Validation of this method is demonstrated along with evaluation of these analytes in control and oxidizing conditions in vitro and in HEK 293T cells. Oxidation of this triphosphate pool occurred independently of oxidation to DNA.
Bolin
Celeste
C
Center for Environmental Health Sciences, Department of Pharmaceutical Sciences, University of Montana, Missoula, MT 59812, USA.
Cardozo-Pelaez
Fernando
F
D015415:Biomarkers / Q000032:analysis*; D002138:Calibration; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004563:Electrochemistry / Q000379:methods*; D006151:Guanosine / Q000032:analysis*; D006160:Guanosine Triphosphate / Q000032:analysis*; D006801:Humans; D018384:Oxidative Stress; D010766:Phosphorylation; D012015:Reference Standards
D015415:Biomarkers; D006151:Guanosine; D006160:Guanosine Triphosphate