title: Capillary electrophoresis-laser induced fluorescence analysis of endogenous damage in mitochondrial and genomic DNA.
abstract: Reactive oxygen molecules are formed in vivo as by-products of normal aerobic metabolism. All organisms dependent on oxygen are inevitably exposed to these species so that DNA damage can occur in both genomic and mitochondrial DNA (mtDNA). In order to determine endogenous DNA damage we have developed an analytical method that involves the isolation and hydrolysis of genomic DNA or mtDNA, the labeling of modified and unmodified nucleotides and micellar electrokinetic chromatography with laser-induced fluorescence detection. With this method we have found etheno-adenine, thymine glycol, uracil, hypoxanthine, and 5-methylcytosine. These were identified by the addition of internal standards to the genomic or mtDNA. There are a large number of other signals in the electropherograms of mtDNA that we have never found in genomic DNA analysis because they are at lower concentration in the genome. In the DNA of untreated patients with chronic lymphocytic leukemia (CLL), uracil and high levels of etheno-adenine were found, which can be explained by antioxidant enzyme alterations and oxidative stress in the CLL lymphocytes.
authors.affiliation: Department of Analytical Chemistry, University of Wuppertal, Wuppertal, Germany.
authors.forename: Michaela
authors.initials: M
authors.lastname: Wirtz
authors.forename: Claus A
authors.initials: CA
authors.lastname: Schumann
authors.forename: Marc
authors.initials: M
authors.lastname: Schellenträger
authors.forename: Siegmar
authors.initials: S
authors.lastname: Gäb
authors.forename: Jochen
authors.initials: J
authors.lastname: Vom Brocke
authors.forename: Michael A L
authors.initials: MA
authors.lastname: Podeschwa
authors.forename: Hans-J
authors.initials: HJ
authors.lastname: Altenbach
authors.forename: David
authors.initials: D
authors.lastname: Oscier
authors.forename: Oliver J
authors.initials: OJ
authors.lastname: Schmitz
mesh_terms: D044503:5-Methylcytosine / Q000032:analysis / Q000302:isolation & purification; D000249:Adenosine Monophosphate / Q000031:analogs & derivatives / Q000032:analysis / Q000302:isolation & purification; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002417:Cattle; D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods; D018736:DNA Adducts / Q000032:analysis / Q000302:isolation & purification*; D004249:DNA Damage; D019175:DNA Methylation; D004272:DNA, Mitochondrial / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D016678:Genome; D006801:Humans; D007834:Lasers; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000378:metabolism; D008099:Liver / Q000737:chemistry; D018384:Oxidative Stress; D013050:Spectrometry, Fluorescence
chemical_list: C503731:1,N(6)-etheno-2'-deoxyadenosine-3'-monophosphate; D015415:Biomarkers; D018736:DNA Adducts; D004272:DNA, Mitochondrial; D000249:Adenosine Monophosphate; D044503:5-Methylcytosine