Assays of oxidative DNA damage biomarkers 8-oxo-2'-deoxyguanosine and 8-oxoguanine in nuclear DNA and biological fluids by high-performance liquid chromatography with electrochemical detection.
High-performance liquid chromatography with electrochemical detection is a highly sensitive and selective method for detecting oxo8dG and oxo8Gua, biomarkers of oxidative DNA damage. When employed together with the DNA isolation and monoclonal antibody-based immunoaffinity purification methods described, oxo8dG and oxo8Gua in DNA and urine can be readily detected and quantitated, offering a powerful approach for assessing oxidative DNA damage in vivo. Application of the technique to the detection of oxo8dG from DNA permits quantitation of the steady-state levels of this oxidatively modified deoxynucleoside and overcomes the detection problems associated with the extremely low levels present in DNA. In addition, the selectivity gained by this detection method eliminates the problem of separating the signal for oxo8dG from those of normal deoxynucleosides. The quantitation of oxo8dG and oxo8Gua in biological fluids is noninvasive and complements the measurement of oxo8dG in DNA by estimating the rate of oxidative DNA damage occurring within the body or in a population of cells. This analytical approach may allow one to estimate oxidative DNA damage in an animal or individual exposed to prooxidant conditions associated with lifestyle, genetic predisposition, degenerative diseases, or environmental toxins. Furthermore, these assays may allow one to assess the potentially beneficial effects of intervention strategies that protect DNA from such damage.
Shigenaga
M K
MK
Department of Molecular and Cell Biology, University of California at Berkeley 94720.
Aboujaoude
E N
EN
Chen
Q
Q
Ames
B N
BN
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000818:Animals; D017209:Apoptosis; D015415:Biomarkers / Q000032:analysis; D001826:Body Fluids / Q000737:chemistry*; D002250:Carbon Radioisotopes; D002467:Cell Nucleus / Q000737:chemistry*; D002478:Cells, Cultured; D002846:Chromatography, Affinity / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003600:Cytosol / Q000737:chemistry; D004247:DNA / Q000737:chemistry* / Q000302:isolation & purification; D004249:DNA Damage; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000032:analysis; D004563:Electrochemistry / Q000379:methods; D006147:Guanine / Q000031:analogs & derivatives* / Q000032:analysis / Q000138:chemical synthesis; D006801:Humans; D006868:Hydrolysis; D007553:Isotope Labeling / Q000379:methods; D008322:Mammals; D009336:Necrosis; D010084:Oxidation-Reduction; D011865:Radioisotope Dilution Technique; D014316:Tritium
D015415:Biomarkers; D002250:Carbon Radioisotopes; D014316:Tritium; C024829:8-hydroxyguanine; D006147:Guanine; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; D003849:Deoxyguanosine